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ASTM D4454

85th Edition, January 25, 1985

Complete Document

Standard Test Method for Simultaneous Enumeration of Total and Respiring Bacteria in Aquatic Systems by Microscopy

Includes all amendments and changes through Reapproval Notice , 2009


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Product Details:

  • Revision: 85th Edition, January 25, 1985
  • Published Date: January 2009
  • Status: Not Active, See comments below
  • Document Language: English
  • Published By: ASTM International (ASTM)
  • Page Count: 3
  • ANSI Approved: No
  • DoD Adopted: No

Description / Abstract:

This test method covers the detection and enumeration of aquatic bacteria by the use of an acridine-orange epifluorescence direct-microscopic counting procedure. This test method is applicable to environmental waters and potable waters.

Certain types of debris and other microorganisms may fluoresce in acridine-orange stained smears.

The procedure described requires a trained microbiologist or technician who is capable of distinguishing bacteria from other fluorescing bodies on the basis of morphology when viewed at higher magnifications.2

Use of bright light permits differentiation of single bacteria where reduced formazan is deposited at the polar ends.

Approximately 104 cells/mL are required for detection by this test method.2

Minimal cell size which allows the detection of formazan deposits is represented by bacteria of 0.4 μm.2

The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.

This standard does not purport to address the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety and health practices and determine the applicability of regulatory limitations prior to use.

2 DIFCO Technical Information—Bacto Acridine Orange Stain, is available from Difco Laboratories, P.O. Box 1058, Detroit, MI 48201.