NEMI Method Summary

Method Summary Information

Method Number: ICR Media: WATER
Revision: Jun-95
Method Source: U.S. EPA National Exposure Research Laboratory (NERL), Microbiological and Chemical Exposure Assessm
Analytes in this method
Subcategory: MICROBIOLOGICAL
Official Name: Information Collection Rule Protozoan Method for Detecting Giardia Cysts and Cryptosporidium Oocysts in Water by A Fluorescent Antibody Procedure
Descriptive Name ICR method for cryptosporidium and giardia by filtration, flotation, and immunofluorescence microscopy
Source Info: U.S.EPA National Exposure Research Laboratory (NERL)
Microbiological and Chemical Exposure Assessment Research Division (MCEARD)
[formerly the Environmental Monitoring Systems Laboratory (EMSL), Cincinnati, OH]
26 West Martin Luther King Drive
Cincinnati, Ohio 45268-0001
Fax: 513-569-7757
email: DWmethods.help@epa.gov
Citation: USEPA. 1995 (June). Information Collection Rule Protozoan Method for Detecting Giardia Cysts and Cryptosporidium Oocysts in Water by A Fluorescent Antibody Procedure. EPA/814-B-95-003. US EPA: Wash
2790916
Brief Method
Summary:
 Method is superceded by USEPA Methods 1622 and 1623. Protozoa are concentrated from a large volume of water (typically 100 L) by retention on a yarn-wound filter on site. Retained particulates are eluted from the filter with an eluting solution and are concentrated by centrifugation. Cysts and oocysts are separated from other particulate debris by flotation on a Percoll-sucrose solution. A monolayer of the water/Percoll-sucrose interface is placed on a membrane filter, indirectly stained with fluorescent antibody, and examined under a microscope. Specialized equipment and supplies are required and include, but are not limited to the following: sampling apparatus; a microscope capable of epifluorescence and differential interference contrast; reagent-grade chemicals and water; centrifuge; vortex mixer; membrane filtration manifold. Cost of analysis (USEPA Fed. Reg. Aug 2001): $400-500
Scope And
Application:
 ambient, compliance monitoring (unregulated contaminants): all water. USEPA. 1996 (May 14). National primary drinking water regulations: monitoring requirements for public drinking water supplies - Cryptosporidium, Giardia, viruses, disinfection byproducts, water treatment plant data and other information requirements. Fed. Reg. 61(94):24354-24388.
Applicable
Conc Range:
 40 - 10,000 oocysts/100 L and 40 - 10,000 cysts/100 L (without correction for incomplete recovery)
Interferences: Turbidity due to inorganic and organic debris, clays and algae, iron and alum coagulants may interfere with concentration, purification and examination of the sample. Organisms that autofluoresce may contribute to false positives. Chlorine compounds and other disinfection chemicals may interfere with visualization of the internal structures of cysts and oocysts. Freezing filtered samples is not recommended
QC Requirements: Requirements defined in: Fout, G. S.; Schaefer, F. W.; Messer, J.W.; Dahling, D. R.; Stetler, R. E. 1996. (April). ICR Microbial Laboratory Manual. EPA/600/R-95/178. USEPA: Cincinnati, OH. 233 pp. http://www.epa.gov/microbes/icrmicro.pdf
Sample Handling: Sample preservation: filters are chilled at 2 to 5oC. Techniques for collection: A 100-L sample is filtered in the field (USEPA, 1996). Method is superceded by USEPA Methods 1622 and 1623.
See USEPA. 2001. Implementation and Results of the Information Collection Rule Supplemental Surveys. EPA-815-R-01-003. Office of Water, Office of Ground Water and Drinking Water, Standards and Risk Management Division, Washington, DC.
Max Holding Time:
 
Relative Cost: Greater than $400
URL to obtain ordering information for this method:
http://www.epa.gov/nerlcwww/pvid.pdf
5614
Page 1