Conditions for the disinfectant efficacy test under subzero temperatures

To establish appropriate conditions for a disinfectant efficacy test at subzero temperatures, this study examined mixtures of frozen foot-and-mouth disease virus or avian influenza virus solutions and disinfectant diluents at $-5^{\circ}C$ and monitored temperature and freezing status of an anti-freezing diluent (AFD, 15% ethanol + 30% propylene glycol + 55% distilled water) over time at various subzero temperatures. Viral solutions and disinfectant diluents froze before the mixtures reached $-5^{\circ}C$, whereas the AFD was not frozen at $-30^{\circ}C$. The times taken for the AFD to reach -10, -20, -30, and $-40^{\circ}C$ from room temperature were 36, 39, 45, and 48 min, respectively.