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To evaluate the effects of lactylate and Bacillus subtilis on growth performance, complete blood cell count, and microbial changes, 264 weaning pigs were assigned to four treatments (1) control (Con) basal diets that met the nutrient requirement for each phase, (2) 0.2% lactylate (LA), (3) 0.05% Bacillus subtilis strains mixtures (BM), or (4) the combination of LA and BM (LA+BM) added to the control basal diet at their respective inclusion rates in each of the three phases. Dietary lactylate tended to increase weight gain, significantly increased feed intake, and reduced fecal total E. coli and enterotoxigenic E. coli counts during Phase 1. Pigs fed Bacillus subtilis had a greater gain to feed ratio (G:F) during Phases 1 and 2. Pigs fed lactylate had an increased peripheral absolute neutrophil count on D14 but a decreased eosinophil percentage. Pigs fed Bacillus subtilis had an elevated peripheral total white blood cell count at study completion. The addition of lactylate increased microbiota richness, reduced E. coli, and increased Prevotella, Christensenellaceae, and Succinivibrio. Bacillus subtilis supplementation-enriched f_Ruminococcaceae_unclassified and S24-7_ unclassified had positive relationships with feed efficiency. Collectively, these findings suggested that lactylate can be added to diets to balance gut microbiota and improve growth performance during the early postweaning period. The combination of lactylate and Bacillus subtilis strains exerted a synergic effect on the growth performance of nursery pigs.