Neonatal meconium represents a relatively underexplored microbial source that may harbour microorganisms with probiotic potential, uniquely adapted for intestinal colonization and metabolic health. From 120 meconium samples, 167 spore-forming isolates were screened to identify strains exhibiting desirable probiotic functional and safety attributes. Three strains (ZBOE6, ZBRV8, and ZBTM2) were selected and identified as Bacillus subtilis through whole-genome sequencing. These strains showed non-h