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Int. J. Environ. Res. Public Health 2015, 12(7), 7118-7132;

Quantification of Protozoa and Viruses from Small Water Volumes

Oceans and Human Health Center, University of Miami, Key Biscayne, FL 33149, USA
Department of Biology, University of Wisconsin-River Falls, River Fall, WI 54022, USA
Corporate Research Materials Laboratory, St. Paul, MN 55144, USA
Department of Civil, Arch., and Environmental Engineering, University of Miami, Coral Gables, FL 33126, USA
Hydros Coastal Solutions, Inc.-Miami, FL 33126, USA
BCS Laboratories, Inc., Gainesville FL 32609, USA
BioStar Consulting, Inc., Greenbrier, TN 37073, USA
Author to whom correspondence should be addressed.
Academic Editor: Samuel Dorevitch
Received: 8 April 2015 / Revised: 16 June 2015 / Accepted: 18 June 2015 / Published: 24 June 2015
Full-Text   |   PDF [270 KB, uploaded 26 June 2015]


Large sample volumes are traditionally required for the analysis of waterborne pathogens. The need for large volumes greatly limits the number of samples that can be processed. The aims of this study were to compare extraction and detection procedures for quantifying protozoan parasites and viruses from small volumes of marine water. The intent was to evaluate a logistically simpler method of sample collection and processing that would facilitate direct pathogen measures as part of routine monitoring programs. Samples were collected simultaneously using a bilayer device with protozoa capture by size (top filter) and viruses capture by charge (bottom filter). Protozoan detection technologies utilized for recovery of Cryptosporidium spp. and Giardia spp. were qPCR and the more traditional immunomagnetic separation—IFA-microscopy, while virus (poliovirus) detection was based upon qPCR versus plaque assay. Filters were eluted using reagents consistent with the downstream detection technologies. Results showed higher mean recoveries using traditional detection methods over qPCR for Cryptosporidium (91% vs. 45%) and poliovirus (67% vs. 55%) whereas for Giardia the qPCR-based methods were characterized by higher mean recoveries (41% vs. 28%). Overall mean recoveries are considered high for all detection technologies. Results suggest that simultaneous filtration may be suitable for isolating different classes of pathogens from small marine water volumes. More research is needed to evaluate the suitability of this method for detecting pathogens at low ambient concentration levels. View Full-Text
Keywords: Cryptosporidium; Giardia; enterovirus; quantitative PCR; water quality Cryptosporidium; Giardia; enterovirus; quantitative PCR; water quality
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited (CC BY 4.0).

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Bonilla, J.A.; Bonilla, T.D.; Abdelzaher, A.M.; Scott, T.M.; Lukasik, J.; Solo-Gabriele, H.M.; Palmer, C.J. Quantification of Protozoa and Viruses from Small Water Volumes. Int. J. Environ. Res. Public Health 2015, 12, 7118-7132.

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