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Appl. Environ. Microbiol. doi:10.1128/AEM.02713-06
Copyright (c) 2007, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

Multi-State Evaluation of an Ultrafiltration-Based Procedure for Simultaneous Recovery of Enteric Microbes in 100-L Tap Water Samples

Centers for Disease Control and Prevention, National Center for Zoonotic, Vector-Borne, and Enteric Diseases, Division of Parasitic Diseases, Atlanta, GA, USA; Atlanta Research and Education Foundation

^* To whom correspondence should be addressed. Email: vhill{at}cdc.gov.

	Abstract

Ultrafiltration is increasingly being recognized as a potentially effective procedure for concentrating and recovering microbes from large volumes of water and treated wastewater. Because of their very small pore sizes, ultrafilter (UF) membranes are capable of simultaneously concentrating viruses, bacteria and parasites based on size exclusion. In this study, an ultrafiltration-based water sampling procedure was used to simultaneously recover representatives of these three microbial classes seeded into 100-L samples of tap water collected from eight cities covering six hydrologic areas of the United States. The UF-based procedure included hollow-fiber ultrafiltration as the primary step for concentrating microbes, then used membrane filtration for bacterial culture assays, immunomagnetic separation (IMS) for parasite recovery and quantification, and centrifugal UFs for secondary concentration of viruses. Water samples were tested for nine water quality parameters to investigate whether water quality data correlated with measured recovery efficiencies and molecular detection levels. Average total method recovery efficiencies were 71, 97, 120, 110, and 91% for phi X174 bacteriophage, MS2 bacteriophage, Enterococcus faecalis, Clostridium perfringens spores, and Cryptosporidium parvum oocysts, respectively. Real-time PCR and reverse transcription-PCR (RT-PCR) for seeded microbes and controls indicated that tap water quality could affect the analytical performance of molecular amplification assays, although no specific water quality parameter was found to correlate with reduced PCR or RT-PCR performance.

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