This study evaluated the effect of the replacement of AGPs by a probiotic in diets for piglets in the nursery stage. The dietary treatments were as follows: CON-basal diet (BD); ANT-BD+antibiotic; and PRO-BD+probiotic. From d 35–42 of age, the piglets that received ANT-BD showed a higher average daily weight gain (ADWG) (p = 0.0296), followed by those that received PRO and the control. The average daily feed intake (ADFI) was higher (p = 0.0224) for PRO- and ANT-fed piglets when compared to CON. From d 43–56, the ADWG was the highest (p = 0.0207) in piglets fed ANT. The ADFI was also higher (p = 0.0258) in ANT and PRO. Final body weight (BW) was also influenced (p = 0.0291), whereas ANT-fed animals, followed by PRO, showed a higher BW compared to CON. For overall nursery performance, PRO showed the highest (p = 0.046) ADFI compared to all other treatments. Piglets fed PRO and ANT also showed the highest (p = 0.05) end weights. There was no significant difference in the fecal concentration of Escherichia coli. Concentrations for the sugar absorption test were higher on days 34 and 49 (p < 0.05). The inclusion of B. subtilis DSM 32315 does not replace the use of antibiotics with the same level of results but can provide benefits compared to diets without the use of antibiotics.