Survivability, Partitioning, and Recovery of Enveloped Viruses in Untreated Municipal Wastewater

Many of the devastating pandemics and outbreaks of the 20th and 21st centuries have involved enveloped viruses, including influenza, HIV, SARS, MERS, and Ebola. However, little is known about the presence and fate of enveloped viruses in municipal wastewater. Here, we compared the survival and partitioning behavior of two model enveloped viruses (MHV and phi 6) and two nonenveloped bacteriophages (MS2 and T3) in raw wastewater samples. We showed that MHV and phi 6 remained infective on the time scale of days: Up to 26% of the two enveloped viruses adsorbed to the solid fraction of wastewater compared to 6% of the two non enveloped viruses. Based on this partitioning behavior, we assessed and optimized methods for recovering enveloped viruses from wastewater. Our optimized ultrafiltration method resulted in mean recoveries (+/- SD) of 25.1% (+/- 3.6%) and 18.2% (+/- 9.5%) for the enveloped MHV and phi 6, respectively, and mean recoveries of 55.6% (+/- 16.7%) and 85.5% (+/- 24.5%) for the nonenveloped MS2 and T3, respectively. A maximum of 3.7% of MHV and 2% of MS2 could be recovered from the solids. These results shed light on the environmental fate of an important group of viruses and the presented methods will enable future research on enveloped viruses in water environments.