Previously, abobotulinumtoxinA (aboBoNT-A) injected intraoperatively resulted in effective, but delayed post-surgical analgesia in pigs. Here, we explore the efficacy of preemptively administered aboBoNT-A in intact animals on pain and associated behaviors following a full-skin-muscle incision and retraction surgery on the lower back. AboBoNT-A (200 U/animal) or saline, distributed across ten points, were injected around anticipated incision 15, 5, or 1 day before surgery via ID route (part A) or 15 days before surgery via ID, intramuscular (IM) or subcutaneous (SC) routes (part B). We assessed mechanical sensitivity (withdrawal force
WF), distress behavior score (DBS), and latency to approach the investigator before and after surgery for 7 days.AboBoNT-A, injected ID 15 days before surgery, didn't alter any baseline behaviors, but resulted in 5-fold increases in WF, 75% reduction in DBS and 70% reduction in approach latencies (all p <
0.01). Injections 5 days before surgery led to similar effects, albeit with a fewer animals reaching thresholds, while those made 1 day before surgery were less effective. SC and IM injections were ineffective. Thus, aboBoNT-A administered ID 15 days before surgery represents the most optimal condition for postoperative analgesia. These findings warrant for clinical investigation of preemptively administered aboBoNT-A in postsurgical pain.