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Applied and Environmental Microbiology, July 2007, p. 4218-4225, Vol. 73, No. 13
0099-2240/07/$08.00+0     doi:10.1128/AEM.02713-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Multistate Evaluation of an Ultrafiltration-Based Procedure for Simultaneous Recovery of Enteric Microbes in 100-Liter Tap Water Samples{triangledown}

Vincent R. Hill,1* Amy M. Kahler,1,2 Narayanan Jothikumar,1 Trisha B. Johnson,1,2 Donghyun Hahn,1,2 and Theresa L. Cromeans1,2

Centers for Disease Control and Prevention, National Center for Zoonotic, Vector-Borne, and Enteric Diseases, Division of Parasitic Diseases, Atlanta, Georgia,1 Atlanta Research and Education Foundation, Atlanta, Georgia2

Received 20 November 2006/ Accepted 27 April 2007

Ultrafiltration (UF) 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, UF membranes are capable of simultaneously concentrating viruses, bacteria, and parasites based on size exclusion. In this study, a UF-based water sampling procedure was used to simultaneously recover representatives of these three microbial classes seeded into 100-liter samples of tap water collected from eight cities covering six hydrologic areas of the United States. The UF-based procedure included hollow-fiber UF as the primary step for concentrating microbes and then used membrane filtration for bacterial culture assays, immunomagnetic separation for parasite recovery and quantification, and centrifugal UF 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.

* Corresponding author. Mailing address: Centers for Disease Control and Prevention, National Center for Infectious Diseases, Division of Parasitic Diseases, 4770 Buford Highway, Mail Stop F-36, Atlanta, GA 30341-3724. Phone: (770) 488-4432. Fax: (770) 488-4253. E-mail: vhill{at}cdc.gov

{triangledown} Published ahead of print on 4 May 2007.

Applied and Environmental Microbiology, July 2007, p. 4218-4225, Vol. 73, No. 13
0099-2240/07/$08.00+0     doi:10.1128/AEM.02713-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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