A good level of biosecurity practice is important for efficient porcine reproductive and respiratory syndrome (PRRS) control. In the current study, disinfecting ability of ozone against PRRS virus (PRRSV) was evaluated in comparison with ultraviolet (UV) and an organic acid-based disinfectant to assess the possible use of ozone for disinfecting farm vehicles, equipments, and materials to reduce the risk of new virus introduction. For in vitro evaluation, the levels of infectious virus and viral RNA were determined on the swabs collected from the floor surface of each room treated with either ozone, UV or the disinfectant up to 30 min after contamination with 100 mL of VR2332 ($10^5;TCID_{50}/mL$). For in vivo evaluation, 3, 4-week old, PRRS-free pigs were housed into those rooms right after the last swab collection. Then the pigs in each room were injected intramuscularly with the corresponding swab samples collected at the last time point and pooled per each room. Although ozone, UV, and the disinfectant significantly reduced the levels of PRRSV RNA contamination, ozone was most effective in removing the viral RNA. In addition, the virus collected after at least 10 min exposure to ozone failed to replicate in pigs while the virus collected after treatment with UV and the disinfectant for 30 min still replicated in pigs. Based on the results, it was concluded that ozone is more effective in inactivating PRRSV as compared with UV and the organic acid-based disinfectant.