An experiment was carried out to investigate the effect of Bacillus subtilis RX7 and B2A supplementation on egg production, egg quality, blood profile, and excreta Salmonella population in laying hens. A total of 192 Hy-Line Brown laying hens (40-wk-old) were randomly assigned into four dietary treatments, each of which was replicated eight times with six hens per replicate in a 5 wk trial. The dietary treatments were NC, which is a basal diet without antibiotics or Bacillus subtilis; PC, which is an NC + 0.5 g kg^-1 antibiotic (virginiamycin) diet; BSR, which is an NC + 0.5 g kg^-1 Bacillus subtilis RX7 (1.0 × 10⁹ colony forming units g^-1); and BSB, which is an NC + 0.5 g kg^-1Bacillus subtilis B2A (1.0 × 10⁹ colony forming units g^-1). The results showed that hens receiving BSB diets had increased (P < 0.05) eggshell thickness over hens fed the NC, PC, and BSR diets (week 3). The excreta Salmonella counts in the PC, BSR, and BSB groups were lower than the NC group (P < 0.05). These results indicate that dietary inclusion of Bacillus subtilis RX7 and B2A strains effectively decreased excreta Salmonella counts without any detrimental effects on the performance of laying hens.