This experiment was carried out to investigate the potential probiotics properties of Bacillus subtilis in protecting of juvenile freshwater prawn, Macrobrachium rosenbergii against Aeromonas hydrophila infection. A B. subtilis bacterium isolated from gut of juvenile prawns was added to the prawns feed at 10 8 cells g -1 feed. There were significant differences (P \ 0.05) between B. subtilis-treated and control groups in growth and survival enhancement of juvenile prawns after 60 days of feeding trial. Sixty days after B. subtilis feeding trial, the prawns were challenged by bath exposure to A. hydrophila (10 7 cells mL -1 ) for 28 days. Four weeks after challenge, there was significant difference in the survival of prawns between B. subtilis-treated groups (88.33 %) and control groups (20.81 %; P \ 0.05). In addition, the control groups had an unhealthy external appearance, while the treated groups, appeared healthy and normal. From this, it was concluded that the selected B. subtilis may be a promising probiotics for protection of prawns from A. hydrophila infection.