Development of a Ultra-Filtration Method for the Concentration of Male-Specific Coliphage as a Surrogate for Enteric Viruses in Seawater

Human enteric viruses are frequently discharged from municipal wastewater and certain non-point wastewater streams to estuarine and marine environments.  These discharges pose a significant public health risk if they are adjacent to recreational waters and shellfish harvesting areas.  The public health impact of viruses in these waste streams is difficult to assess since traditional indicator microorganisms are unreliable for indexing the possible presence of pathogenic enteric viruses.  The objective of this study was to characterize the recovery percentage of male-specific coliphage MS2, a surrogate of pathogenic enteric viruses, by tangential flow filtration (TFF) using seawater collected near Dauphin Island, AL.  Polysulfone membrane filters with a 30 KDa cutoff were pre-treated with 400 ml of a 1 % polyethylene glycol (PEG 8000) solution.  Ten liters of seawater (pH ~ 7) was seeded with bacteriophage MS2 to a final concentration of either 10² PFU/liter and 10⁵ PFU/liter. Seeded water was passed through the TFF using a peristaltic pump at a flow of 2 l/min.  Elution of MS2 phage was achieved by passing the retentate, sodium polyphosphate (NaPP) or NaPP/Tween-80 based buffers in the opposite direction of the original flow.  Using this approach bacteriophage MS2 recovery average range was 87-93%, 64-65% and 53-54% from seawater with low, medium and high turbidities, respectively.  These findings suggest the use of TFF as a promising, cost-effective, and reliable approach for the detection of enteric pathogens in seawater.