ASTM D4454 85th Edition, January 25, 1985
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
Additional Comments: W/D NO S/S
Published By:ASTM International (ASTM)
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.