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Effects of Seeding Procedures and Water Quality on Recovery of Cryptosporidium Oocysts from Stream Water By Using U.S. Environmental Protection Agency Method 1623

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Abstract:U.S.EPA Methods 1622 and 1623 are used to detect and quantify Cryptosporidium oocysts in water. The protocol consists of filtration, immunomagnetic separation (IMS), staining with a fluorescent antibody, and microscopic analysis. Microscopic analysis includes detection by fluorescent antibody and confirmation by the demonstration of 1-4 sporozoites or nuclei after staining with 4',6-diamidino-2-phenyl indole dihydrochloride (DAPI). The purpose of this study was to evaluate a new IMS dissociation, a 10 minute incubation at 80 ° C. Heat dissociation improved the average oocyst recovery from 41% to 71% in seeded reagent water, and from 10% to 51% in seeded river samples. The average DAPI confirmation rate improved from 49 % to 93% in reagent water, and from 48% to 73% in river samples. This modification improved both oocyst recovery and confirmation.
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Citation:Francy, D. S., O. D. Simmons, M. W. Ware, E. J. Granger, M. D. Sobsey, and F. W. Schaefer III. Effects of Seeding Procedures and Water Quality on Recovery of Cryptosporidium Oocysts from Stream Water By Using U.S. Environmental Protection Agency Method 1623. APPLIED AND ENVIRONMENTAL MICROBIOLOGY 70(7):4118-4128, (2004).
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Contact: Mary P. O'Bryant - (919)-541-4871 or obriant.mary@epa.gov
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Division: Microbiological & Chemical Exposure Assessment Division
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Branch: Biohazard Assessment Research Branch
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Product Type: Journal
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Published: 07/01/2004
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Downloads:
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Bullet Item Effects of Spiking Procedures and Water Quality on Recovery of Cryptosporidium in Stream Water Using USEPA Method 1623
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