Integrated cryptosporidium assay to determine oocyst density, infectivity, and genotype for risk assessment of source and reuse water

Applied and Environmental Microbiology
B J KingP T 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 catch...Continue Reading

References

Jul 1, 1993·The Journal of Applied Bacteriology·G VeseyC R Fricker
Aug 23, 2002·Microbes and Infection·Rebecca A DillinghamRichard L Guerrant
Jul 8, 2005·Applied and Environmental Microbiology·Brendon J KingChristopher P Saint
Dec 1, 2009·The Science of the Total Environment·E CumminsM Cormican
Dec 24, 2010·Journal of Clinical Microbiology·Stephen J HadfieldRachel M Chalmers
Mar 19, 2011·Parasitology·B J KingP T Monis

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Citations

Sep 26, 2015·International Journal of Environmental Research and Public Health·Jean-Baptiste BurnetHenry-Michel Cauchie
Jan 1, 2017·Applied and Environmental Microbiology·Brendon KingPaul Monis
Jun 6, 2021·The Science of the Total Environment·S PettersonP Byleveld

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