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Simultaneous Concentration of Enterococci and Coliphage from Marine Waters using a Dual Layer Filtration System

^a Univ. of Miami, Oceans and Human Health Center, Key Biscayne, FL 33149
^b Dep. of Civil, Architectural, and Environmental Engineering, Univ. of Miami, Coral Gables, FL 33146-0630
^c Dep. of Infectious Disease and Pathology, Univ. of Florida, Gainesville, FL 32608
^d BCS Labs., Miami, FL 33155

^* Corresponding author (hmsolo{at}miami.edu).

Received for publication November 21, 2008. Concentrating diverse microbes in a time and cost effective manner is an essential component in water quality monitoring of recreational beaches. Historically, detection of bacteria and viruses requires two different capture methods to detect both types of organisms in a given water sample. The purpose of this present study was to evaluate a newly devised dual layered filtration system, which was developed to simultaneously concentrate both viruses and bacteria in one step from marine waters. An apparatus was designed to accommodate two 90-mm diam., 0.45µm pore size membranes in series, one on top of the other. The top polyvinylidene fluoride (PVDF) membrane was used to filter bacteria by physical straining while the bottom HA membrane retained viruses through adsorption. Results indicated that the dual layer filtration system recovered 83 ± 14% of the test bacteria (Enterococcus fecalis) and 81 ± 28% of the test virus (MS2 coliphage) on the top and bottom membranes, respectively. This research demonstrates the potential of using a dual layered filtration system for the simultaneous concentration of both bacteria and viruses on separate filters from recreational beach waters. This system is relatively simple to use, inexpensive, and has the potential to be suitable for routine monitoring. This study serves as a proof of concept for the technique. Additional experiments are needed to evaluate the system on a variety of different bacteria and viruses as well as on water with different physical and chemical parameters.

Abbreviations: CFU, colony forming units • PFU, plaque forming units • PVDF, polyvinylidene fluoride • qPCR, quantitative polymerase chain reaction