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Applied and Environmental Microbiology, November 2005, p. 6878-6884, Vol. 71, No. 11
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.11.6878-6884.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Development of a Rapid Method for Simultaneous Recovery of Diverse Microbes in Drinking Water by Ultrafiltration with Sodium Polyphosphate and Surfactants

Vincent R. Hill,^1* Amy L. Polaczyk,^1,2 Donghyun Hahn,^1,2 Jothikumar Narayanan,^1,2 Theresa L. Cromeans,^1,2 Jacquelin M. Roberts,¹ and James E. Amburgey^1,3

Centers for Disease Control and Prevention, National Center for Infectious Diseases, Division of Parasitic Diseases,¹ Atlanta Research and Education Foundation,² CDC/APHL Emerging Infectious Diseases Laboratory Fellowship Program, Atlanta, Georgia³

Received 2 December 2004/ Accepted 13 June 2005

The ability to simultaneously concentrate diverse microbes is an important consideration for sample collection methods that are used for emergency response and environmental monitoring when drinking water may be contaminated with an array of unknown microbes. This study focused on developing a concentration method using ultrafilters and different combinations of a chemical dispersant (sodium polyphosphate [NaPP]) and surfactants. Tap water samples were seeded with bacteriophage MS2, Escherichia coli, Enterococcus faecalis, Cryptosporidium parvum, 4.5-µm microspheres, Salmonella enterica serovar Typhimurium, Bacillus globigii endospores, and echovirus 1. Ten-liter tap water samples were concentrated to 250 ml in 12 to 42 min, depending on the experimental condition. Initial experiments indicated that pretreating filters with fetal bovine serum or NaPP resulted in an increase in microbe recovery. The addition of NaPP to the tap water samples resulted in significantly higher microbe and microsphere recovery efficiencies. Backflushing of the ultrafilter was found to significantly improve recovery efficiencies. The effectiveness of backflushing was improved further with the addition of Tween 80 to the backflush solution. The ultrafiltration method developed in this study, incorporating the use of NaPP pretreatment and surfactant solution backflushing, was found to recover MS2, C. parvum, microspheres, and several bacterial species with mean recovery efficiencies of 70 to 93%. The mean recovery efficiency for echovirus 1 (49%) was the lowest of the microbes studied for this method. This research demonstrates that ultrafiltration can be effective for recovering diverse microbes simultaneously in tap water and that chemical dispersants and surfactants can be beneficial for improving microbial recovery using this technique.

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* 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.

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Applied and Environmental Microbiology, November 2005, p. 6878-6884, Vol. 71, No. 11
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.11.6878-6884.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

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