Existence of bioaerosol contaminants in farms and outbreaks of some infectious organisms with the ability of transmission by air increase the need for enhancement of biosecurity, especially for the application of aerosol disinfectants. Here we selected slightly acidic hypochlorous acid water (SAHW) as a candidate and evaluated its virucidal efficacy toward a virus in the air. Three-day-old conventional chicks were challenged with 25 doses of Newcastle disease live vaccine (B1 strain) by spray with nebulizer (particle size <3 μm in diameter), while at the same time reverse osmosis water as the control and SAHW containing 50 or 100 parts per million (ppm) free available chlorine in pH 6 were sprayed on the treated chicks with other nebulizers. Exposed chicks were kept in separated cages in an isolator and observed for clinical signs. Oropharyngeal swab samples were collected from 2 to 5 days postexposure from each chick, and then the samples were titrated with primary chicken kidney cells to detect the virus. Cytopathic effects were observed, and a hemagglutination test was performed to confirm the result at 5 days postinoculation. Clinical signs (sneezing) were recorded, and the virus was isolated from the control and 50 ppm treatment groups, while no clinical signs were observed in and no virus was isolated from the 100 ppm treatment group. The virulent Newcastle disease virus (NDV) strain Sato, too, was immediately inactivated by SAHW containing 50 ppm chlorine in the aqueous phase. These data suggest that SAHW containing 100 ppm chlorine can be used for aerosol disinfection of NDV in farms.