title: Quantification of Protozoa and Viruses from Small Water Volumes.
abstract: 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.
authors.affiliation: Oceans and Human Health Center, University of Miami, Key Biscayne, FL 33149, USA. j.alfred.bonilla@uwrf.edu.|Department of Biology, University of Wisconsin-River Falls, River Fall, WI 54022, USA. j.alfred.bonilla@uwrf.edu.
authors.forename: J Alfredo
authors.initials: JA
authors.lastname: Bonilla
authors.affiliation: Oceans and Human Health Center, University of Miami, Key Biscayne, FL 33149, USA. tdbonilla@mmm.com.|Corporate Research Materials Laboratory, St. Paul, MN 55144, USA. tdbonilla@mmm.com.
authors.forename: Tonya D
authors.initials: TD
authors.lastname: Bonilla
authors.affiliation: Oceans and Human Health Center, University of Miami, Key Biscayne, FL 33149, USA. amabdelzaher@gmail.com.|Department of Civil, Arch., and Environmental Engineering, University of Miami, Coral Gables, FL 33126, USA. amabdelzaher@gmail.com.
authors.forename: Amir M
authors.initials: AM
authors.lastname: Abdelzaher
authors.affiliation: Oceans and Human Health Center, University of Miami, Key Biscayne, FL 33149, USA. drtmscott@gmail.com.|Hydros Coastal Solutions, Inc.-Miami, FL 33126, USA. drtmscott@gmail.com.
authors.forename: Troy M
authors.initials: TM
authors.lastname: Scott
authors.affiliation: BCS Laboratories, Inc., Gainesville FL 32609, USA. Lukasik@microbioservices.com.
authors.forename: Jerzy
authors.initials: J
authors.lastname: Lukasik
authors.affiliation: Oceans and Human Health Center, University of Miami, Key Biscayne, FL 33149, USA. hmsolo@miami.edu.|Department of Civil, Arch., and Environmental Engineering, University of Miami, Coral Gables, FL 33126, USA. hmsolo@miami.edu.
authors.forename: Helena M
authors.initials: HM
authors.lastname: Solo-Gabriele
authors.affiliation: Oceans and Human Health Center, University of Miami, Key Biscayne, FL 33149, USA. carolpalmer@biostarconsulting.com.|BioStar Consulting, Inc., Greenbrier, TN 37073, USA. carolpalmer@biostarconsulting.com.
authors.forename: Carol J
authors.initials: CJ
authors.lastname: Palmer
mesh_terms: D003458:Cryptosporidium / Q000302:isolation & purification*; D005374:Filtration / Q000379:methods; D005872:Giardia / Q000302:isolation & purification*; D018189:Immunomagnetic Separation; D017955:Poliovirus / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction; D012623:Seawater / Q000382:microbiology* / Q000469:parasitology; D014871:Water Microbiology; D060753:Water Quality
keywords: Cryptosporidium; Giardia; enterovirus; quantitative PCR; water quality