Oronasal or Intramuscular Immunization with a Thermo-Attenuated ASFV Strain Provides Full Clinical Protection against Georgia 2007/1 Challenge

Figures

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  Figure 1. Representation of the genes and the DIVA PCR system in the deleted region of ASFV-989 strain. (A) Comparison of the Georgia 2007/1 and ASFV-989 genomes at the site of the deletion. (B) Position of the primers and probes for PCR 505 (L1, P1, and R1) to detect the Georgia 2007/1 strain and PCR 989 (Del_Fow, Del_Probe, and Del_Rev) to detect ASFV-989.

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  Figure 2. Viral replication of Georgia 2007/1 and ASFV-989 strains in porcine alveolar macrophages (titer in Log10 HAD50/mL, time in days post-inoculation).

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  Figure 3. Evolution of rectal temperature and growth performances after Georgia or ASFV-989 strain inoculation. (A) Evolution of rectal temperature (°C) after IM inoculation with Georgia or ASFV-989 strains (time in days post-inoculation). Control—data from group A1; 989 IM—data from groups C and D; Geo IM—data from group E that were euthanized/died at 6 dpi. (B) Evolution of growth performances: average daily weight gain (Kg/D) after IM inoculation with Georgia or ASFV-989 strains (time in days post-inoculation). Control—data from groups A1 and A2; 989 IM—data from groups C and D; Geo IM—data from group E. (C) Evolution of rectal temperature (°C) after ON inoculation with Georgia or ASFV-989 strains (time in days post-inoculation). Control—data from group A1; 989 ON—data from groups B, H, and I; Geo ON—data from group F that were euthanized/died at 6 dpi. (D) Evolution of growth performances: average daily weight gain (Kg/D) after ON inoculation with Georgia or ASFV-989 strains (time in days post-inoculation). Control—data from group A1 and A2; 989 ON—data from groups B, H, and I; Geo ON—data from group F.

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  Figure 4. Evolution of ASFV viremia after Georgia or ASFV-989 strain inoculation. (A) Evolution of ASFV viremia (Log10 eq HAD50/mL) after inoculation with Georgia or ASFV-989 strains using a qPCR test [41] that detect both ASFV-989 and Georgia strains (time in days post-inoculation). Geo IM—data from groups E and F; Geo ON—data from groups F and J; 989 IM—data from groups C and D; 989 ON—data from groups B, H, and I. (B) Evolution of ASFV viremia (PCR 989, Log10 eq HAD50/mL) after IM or ON inoculation with ASFV-989 strain (time in days post-inoculation). 989 IM—data from groups C and D; 989 ON—data from groups B, H, and I.

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  Figure 5. Development of the ASFV-specific immune response after ASFV-989 or Georgia strain inoculation. (A) Detection of ASFV-specific antibodies (competition ELISA) after Georgia or ASFV-989 strain inoculation (time in days post-inoculation). The dotted line indicates the positivity threshold of ELISA test. “–“ indicates seronegative samples, “+” indicates seropositive ones. 989 IM—group C; 989 ON—group B; Geo IM—group E; Geo ON—group F. (B) Detection of the cell-mediated immune response (number of IFNγ-secreting cells/106 PBMC) after inoculation with ASFV-989 strain (time in days post-inoculation). 989 IM—data from groups C and D; 989 ON—data from groups B, H, and I.

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  Figure 6. Evolution of rectal temperature and growth performance after Georgia challenge occurring 28 days after immunization. (A) Evolution of rectal temperature (°C) after IM challenge with Georgia ASFV strain (time in days post-challenge). Control—data from group A2; 989 IM/Geo IM—data from group D; 989 ON/Geo IM—data from group I; Geo IM—data from group G. (B) Evolution of rectal temperature (°C) after ON challenge with Georgia ASFV strain (time in days post-challenge). Control—data from group A2; 989 ON/Geo ON—data from group H; Geo ON—data from group J. (C) Evolution of growth performance: average daily weight gain (Kg/D) after IM or ON challenge with Georgia ASFV strain (time in days post-challenge). Control—data from group A1 and A2; 989 IM/Geo IM—data from group D; 989 ON/Geo ON—data from group H; 989 ON/Geo IM—data from group I; Geo ON—data from group J.

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  Figure 7. Evolution of rectal temperature, growth performance, and ASFV viremia after ASFV-989 inoculation and Georgia challenge occurring 14 days after inoculation. (A) Evolution of rectal temperature (°C) (time in days post-inoculation). (B) Evolution of growth performance: average daily weight gain (Kg/D) (time in days post-inoculation). (C) Evolution of ASFV viremia (PCR 989, Log10 eq HAD50/mL) (time in days post-inoculation). Control—data from group A3; 989 IM/Geo IM-14—data from group K; 989 ON/Geo ON-14—data from group L.

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  Figure 8. Survival curves for pigs inoculated with Georgia or ASFV-989 strains at D0 and then challenged or not with the Georgia strain. (A) Survival curves for pigs inoculated with ASFV-989 or Georgia by intramuscular route and challenged (or not) at D14 or D28 with Georgia strain via the same route (time in days post-inoculation). Control—data from group A1, A2, and A3; 989 IM D0—data from groups C, D, and K; Geo IM D0 or D28—data from groups E and G. (B) Survival curves for pigs inoculated with ASFV-989 or Georgia by oronasal route and challenged (or not) at D14 or D28 with Georgia strain via IM or ON route (time in days post-inoculation). Control—data from group A1, A2, and A3; 989 ON D0—data from groups B, H, I and L; Geo ON D0 or D28—data from groups F and J.