Please use this identifier to cite or link to this item: http://hdl.handle.net/2440/116001
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Type: Journal article
Title: Integrated Cryptosporidium assay to determine oocyst density, infectivity, and genotype for risk assessment of source and reuse water
Author: King, B.
Fanok, S.
Phillips, R.
Swaffer, B.
Monis, P.
Citation: Applied and Environmental Microbiology, 2015; 81(10):3471-3481
Publisher: American Society for Microbiology
Issue Date: 2015
ISSN: 0099-2240
1098-5336
Statement of
Responsibility: 
Brendon King, Stella Fanok, Renae Phillips, Brooke Swaffer, Paul Monis
Abstract: Cryptosporidium continues to be problematic for the water industry, with risk assessments often indicating that treatment barriers may fail under extreme conditions. However, risk analyses have historically used oocyst densities and not considered either oocyst infectivity or species/genotype, which can result in an overestimation of risk if the oocysts are not human infective. We describe an integrated assay for determining oocyst density, infectivity, and genotype from a single-sample concentrate, an important advance that overcomes the need for processing multiple-grab samples or splitting sample concentrates for separate analyses. The assay incorporates an oocyst recovery control and is compatible with standard primary concentration techniques. Oocysts were purified from primary concentrates using immunomagnetic separation prior to processing by an infectivity assay. Plate-based cell culture was used to detect infectious foci, with a monolayer washing protocol developed to allow recovery and enumeration of oocysts. A simple DNA extraction protocol was developed to allow typing of any wells containing infectious Cryptosporidium. Water samples from a variety of source water and wastewater matrices, including a semirural catchment, wastewater, an aquifer recharge site, and storm water, were analyzed using the assay. Results demonstrate that the assay can reliably determine oocyst densities, infectivity, and genotype from single-grab samples for a variety of water matrices and emphasize the varying nature of Cryptosporidium risk extant throughout source waters and wastewaters. This assay should therefore enable a more comprehensive understanding of Cryptosporidium risk for different water sources, assisting in the selection of appropriate risk mitigation measures.
Keywords: Cryptosporidium
Rights: © 2015, American Society for Microbiology
RMID: 0030045984
DOI: 10.1128/AEM.00163-15
Appears in Collections:Ecology, Evolution and Landscape Science publications

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