Repeated and Delayed Challenge Study Evaluating the Durability of Protection Induced by the Live Attenuated ASF Vaccine ASFV-G-ΔI177L/ΔLVR

Background/Objectives: African swine fever (ASF) is a highly lethal disease of domestic pigs and wild suids that continues to cause substantial economic losses worldwide. Despite recent progress in live-attenuated ASF vaccine development, evidence supporting durable protection under repeated exposure conditions representative of endemic settings remains limited. Here, we assessed the long-term safety and protective efficacy of a live-attenuated ASFV-G-ΔI177L/ΔLVR vaccine using a repeated-challenge experimental design intended to model re-exposure in ASF-endemic regions. Methods: Vaccinated pigs were subjected to homologous virulent ASF virus challenges at multiple intervals, including repeated challenges (three sequential inoculations) and single challenges administered at 8 and 12 weeks post-vaccination. Results: Across all challenge regimens, vaccinated animals survived and remained clinically healthy, including those receiving three challenges, supporting sustained protection under repeated exposure pressure. Animals challenged at 8 or 12 weeks post-vaccination likewise exhibited complete survival, indicating maintained efficacy through at least 12 weeks. No vaccine-associated adverse clinical outcomes were detected over the study period, and post-challenge viral shedding was minimal. Conclusions: Overall, these data demonstrate that the candidate live-attenuated ASF vaccine provides excellent protective efficacy and confers sustained protection against homologous ASF virus infection. This result is expected to be equally applicable under repeated exposure conditions in regions with unstable ASF biosecurity, making it a sufficiently promising model experiment for field application in ASF epidemic areas.