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Applied and Environmental Microbiology, July 2003, p. 4098-4102, Vol. 69, No. 7
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.7.4098-4102.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Optimization of a Reusable Hollow-Fiber Ultrafilter for Simultaneous Concentration of Enteric Bacteria, Protozoa, and Viruses from Water

Hugo A. Morales-Morales, Guadalupe Vidal, John Olszewski, Channah M. Rock, Debanjana Dasgupta, Kevin H. Oshima, and Geoffrey B. Smith*

Biology Department, New Mexico State University, Las Cruces, New Mexico 88003

Received 25 September 2002/ Accepted 25 March 2003

The detection and identification of pathogens from water samples remain challenging due to variations in recovery rates and the cost of procedures. Ultrafiltration offers the possibility to concentrate viral, bacterial, and protozoan organisms in a single process by using size-exclusion-based filtration. In this study, two hollow-fiber ultrafilters with 50,000-molecular-weight cutoffs were evaluated to concentrate microorganisms from 2- and 10-liter water samples. When known quantities (105 to 106 CFU/liter) of two species of enteric bacteria were introduced and concentrated from 2 liters of sterile water, the addition of 0.1% Tween 80 increased Escherichia coli strain K-12 recoveries from 70 to 84% and Salmonella enterica serovar Enteritidis recoveries from 36 to 72%. An E. coli antibiotic-resistant strain, XL1-Blue, was recovered at a level (87%) similar to that for strain K-12 (96%) from 10 liters of sterile water. When E. coli XL1-Blue was introduced into 10 liters of nonsterile Rio Grande water with higher turbidity levels (23 to 29 nephelometric turbidity units) at two inoculum levels (9 x 105 and 2.4 x 103 per liter), the recovery efficiencies were 89 and 92%, respectively. The simultaneous addition of E. coli XL1-Blue (9 x 105 CFU/liter), Cryptosporidium parvum oocysts (10 oocysts/liter), phage T1 (105 PFU/liter), and phage PP7 (105 PFU/liter) to 10 liters of Rio Grande surface water resulted in mean recoveries of 96, 54, 59, and 46%, respectively. Using a variety of surface waters from around the United States, we obtained recovery efficiencies for bacteria and viruses that were similar to those observed with the Rio Grande samples, but recovery of Cryptosporidium oocysts was decreased, averaging 32% (the site of collection of these samples had previously been identified as problematic for oocyst recovery). Results indicate that the use of ultrafiltration for simultaneous recovery of bacterial, viral, and protozoan pathogens from variable surface waters is ready for field deployment.

* Corresponding author. Mailing address: Biology Department, New Mexico State University, Las Cruces, NM 88003. Phone: (505) 646-6080. Fax: (505) 646-5665. E-mail: gsmith{at}nmsu.edu.

Applied and Environmental Microbiology, July 2003, p. 4098-4102, Vol. 69, No. 7
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.7.4098-4102.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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