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Microbiological and Chemical Exposure Assessment

Microbiological Methods/Online Publications

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Bacteria

  • Method 1605: Aeromonas in Finished Water by Membrane Filtration Using Ampicillin-Dextrin Agar with Vancomycin (ADA-V) (October 2001) (PDF) (26 pp, 443K) — This method (EPA 821-R-01-034) describes a performance-based membrane filtration technique for the detection and enumeration of Aeromonas species. This method uses a selective medium that partially inhibits the growth of non-target bacterial species while allowing Aeromonas to grow and be detected by the production of acid from dextrin fermentation (causing a pH change producing yellow colonies). The method will be proposed for use in EPA's data gathering and monitoring programs under the Unregulated Contaminant Monitoring Rule.
  • Method 1604: Total Coliforms and Escherichia coli in Water by Membrane Filtration Using a Simultaneous Detection Technique (MI Medium) (September 2002) (PDF) (18 pp, 486K) — This document (EPA 821-R-02-024) is identical to the February 2000 version of the MI Agar Method (Membrane Filter Method for the Simultaneous Detection of Total Coliforms and Escherichia coli in Drinking Water, EPA-600-R-00-13), with one exception, the addition of MI Broth. The MI Agar Method was approved for use in compliance monitoring of drinking water and source water in support of the National Primary and Secondary Drinking Water Regulations (Federal Register, Vol. 64, No. 230, 1 December 1999, p.67450-67467) becoming effective 3 January 2000. On 6 November 2001, MI Broth was approved as a minor modification. This method, now designated as Method 1604, has been approved for use in monitoring ambient water (Federal Register, Vol. 68, No. 139, 21 July 2003, p.43272-43283) becoming effective 4 August 2003. Four color photos of coliform and E. coli colonies on MI agar are also included in Method 1604. The procedure is in the format of the Environmental Monitoring Management Council (EMMC). The method may also be viewed as it appeared upon 1993 publication in the journal Applied and Environmental Microbiology (Vol 59, p.3534-3544) PDF (11 pp, 2.2MB), (EPA 600-J-99-225) but it does not include photos.
  • Method 1603: Escherichia coli (E.coli) in Water by Membrane Filtration Using Modified membrane-Thermotolerant Escherichia coli Agar (Modified mTEC) (September 2002) (PDF) (13 pp, 129K) — This method (EPA 821-R-02-023) combines information from a 1985 publication (Test methods for Escherichia coli and Enterococci in Water by the Membrane Filter Procedure, EPA 600-4-85-076) and a subsequent March 2000 manual (Improved Enumeration Methods for the Recreational Water Quality Indicators: Enterococci and Escherichia coli, EPA/821/R-97/004). Method 1603 is a revised membrane filter (MF) procedure, a single-step method that uses one medium, modified mTEC Agar, and does not require the transfer of the membrane filter to another medium or other substrate.
  • Method 1600: Enterococci in Water by Membrane Filtration Using membrane-Enterococcus Indoxyl-ß-D-Glucoside Agar (mEI) (September 2002) (PDF) (14 pp, 125K) — This method (EPA 821-R-02-022) combines information previously published in the May 1997 version of Method 1600 (Membrane Filter Test Method for Enterococci in Water, EPA-821-R-97-004a) and a subsequent March 2000 manual (Improved Enumeration Methods for the Recreational Water Quality Indicators: Enterococci and Escherichia coli, EPA/821/R-97/004). Method 1600 is a revision of EPA's previous enterococci method, used since 1985 in ambient water quality monitoring. It reduces analysis time to 24 hours and improves analytical quality. The method has been validated in single- and multi-laboratory studies and has undergone peer review.
  • Method 1106.1: Enterococci in Water by Membrane Filtration Using membrane-Enterococcus-Esculin Iron Agar (mE-EIA) (September 2002) (PDF) (16 pp, 806K) — This method (EPA 821-R-02-021) combines information from a 1985 publication (Test methods for Escherichia coli and Enterococci in Water by the Membrane Filter Procedure, EPA 600-4-85-076) and a subsequent March 2000 manual (Improved Enumeration Methods for the Recreational Water Quality Indicators: Enterococci and Escherichia coli, EPA/821/R-97/004). Method 1106.1 is a membrane filter (MF) procedure for the detection and enumeration of enterococci in water. The method incorporates the sequential use of a selective medium, mE agar, and a differential medium, EIA agar.
  • Method 1103.1: Escherichia coli (E. coli) in Water by Membrane Filtration Using membrane-Thermotolerant Escherichia coli Agar (mTEC) (September 2002) (PDF) (17 pp, 471K) — This method (EPA 821-R-02-020) combines information from a 1985 publication (Test methods for Escherichia coli and Enterococci in Water by the Membrane Filter Procedure, EPA 600-4-85-076) and a subsequent March 2000 manual (Improved Enumeration Methods for the Recreational Water Quality Indicators: Enterococci and Escherichia coli, EPA/821/R-97/004). Method 1106.1 is a membrane filter (MF) procedure for the detection and enumeration of E. coli in water. The method incorporates the use of a selective and differential medium, mTEC agar followed by incubation with urea substrate medium.
  • Improved Enumeration Methods for the Recreational Water Quality Indicators: Enterococci and Escherichia coli (March 2000) (PDF) (49 pp, 350K) — This manual (EPA 821-R-97-004) describes four test methods for measuring bacteriological densities in ambient waters: the original and a revised method for detecting enterococci, and the original and a revised method for detecting E. coli. All four methods use a membrane filter procedure. The manual complements a training video on this topic but may be used independently. (To obtain the video, see pg 49 of the manual.)

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Protozoans

  • Method 1623.1: Cryptosporidium and Giardia in Water by Filtration/IMS/FA (PDF) (83 pp, 1.1MB) — June 2012 Update —On 06/28/2012 the Environmental Protection Agency expedited Approval of EPA Method 1623.1 as a revision of EPA Method 1623 (USEPA 2005) for the detection of Cryptosporidium in water. The primary change in EPA Method 1623.1 relative to the Method 1623 is the addition of sodium hexametaphosphate (NaHMP) after filtration of the water sample. More significant improvement in Cryptosporidium recovery with EPA Method 1623.1 was associated with samples that had low recovery using EPA Method 1623 during two side-by-side studies. The Cryptosporidium recoveries during the validation of EPA Method 1623.1 were at least 20 percentage points higher than the recoveries cited in the validation study for EPA Method 1623. In addition, the precision for Cryptosporidium measurements was improved in the validation of EPA Method 1623.1 compared with the validation of Method 1623. Laboratories performing Method 1623.1 are expected to have better detection rates for Cryptosporidium oocysts than those laboratories following Method 1623. Method 1623.1 includes more stringent quality control measures.
  • Method 1622: Cryptosporidium in Water by Filtration/IMS/FA (PDF) (75 pp, 1.3MB) — December 2005 Update — This method (EPA 815-R-05-001) is a performance-based method applicable to the determination of Cryptosporidium in aqueous matrices. Method 1622 requires filtration, immunomagnetic separation (IMS) of the oocysts from the material captured, and immunofluorescent assay (FA) for determination of the oocyst concentration. There are also forms associated with Method 1622. A Training module is also available.
  • Method 1622 and 1623 Forms — These forms are in PDF format and include text-only PDF versions for vision-assisted software:
  • Criteria for Evaluation of Proposed Protozoan Detection Methods (PDF) (14 pp, 79K) — This document (EPA 815-K-99-02) provides a framework for evaluating methods for detecting Giardia, Cryptosporidium, and other protozoa.

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Viruses and Coliphage

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Recreational Water Publications:

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ICR-Related Materials:

Note: Sample collection for the Information Collection Rule ended in December, 1998. The following ICR publications are maintained online as informational resources.

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