ASTM D4201-96

Standard Test Method for Coliphages in Water (Withdrawn 2005)

SCOPE
1.1 This test method covers the determination of coliphages infective for E. coli C in water. The test method is simple, inexpensive, and yields rapid water quality data. Its sensitivity is limited to 5 coliphages per 100 mL of water sample. This test method is applicable to natural fresh water samples.
1.2 This standard does not purport to address all of 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.
WITHDRAWN RATIONALE
This test method covers the determination of coliphages infective for E. coli C in water. The test method is simple, inexpensive, and yields rapid water quality data. Its sensitivity is limited to 5 coliphages per 100 mL of water sample. This test method is applicable to natural fresh water samples.
Formerly under the jurisdiction of Committee D19 on Water, this test method was withdrawn in August 2005 in accordance with section 10.5.3.1 of the Regulations Governing ASTM Technical Committees, which requires that standards shall be updated by the end of the eighth year since the last approval date.

General Information

• Status: Withdrawn
• Publication Date: 09-May-1996
• Withdrawal Date: 02-Aug-2005
• ICS: 13.060.70 - Examination of biological properties of water
• Technical Committee: D19 - Water
• Drafting Committee: D19.24 - Water Microbiology

Standards Content (Sample)

NOTICE: This standard has either been superseded and replaced by a new version or withdrawn.
Contact ASTM International (www.astm.org) for the latest information
An American National Standard
Designation:D 4201–96
Standard Test Method for
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Coliphages in Water
This standard is issued under the fixed designation D 4201; the number immediately following the designation indicates the year of
original adoption or, in the case of revision, the year of last revision.Anumber in parentheses indicates the year of last reapproval.A
superscript epsilon (e) indicates an editorial change since the last revision or reapproval.
1. Scope 5. Significance and Use
1.1 This test method covers the determination of coliphages 5.1 Coliphage organisms may serve as indicators of fecal
infective for E. coli C in water. The test method is simple, pollution.Thepresenceofcoliphagesinwaterintheabsenceof
inexpensive, and yields rapid water quality data. Its sensitivity adisinfectantindicatestheprobablepresenceoffecalcontami-
islimitedto5coliphagesper100mLofwatersample.Thistest nation, but the absolute relationship between the number of
method is applicable to natural fresh water samples. coliforms and coliphages in natural waters has not been
3, 4, 5
1.2 This standard does not purport to address all of the demonstrated conclusively.
safety concerns, if any, associated with its use. It is the 5.2 The detection of coliphages in a water sample depends
responsibility of the user of this standard to establish appro- upon the use of a sensitive-host strain in the coliphage assay.
priate safety and health practices and determine the applica- Coliphages may be detected in 4 to6hto provide important
bility of regulatory limitations prior to use. same-day information on the sanitary quality of a water. The
lowerdetectionlimitis5coliphagesper100mLoffreshwater
2. Referenced Documents
sample.
2.1 ASTM Standards:
2
6. Interferences
D 1129 Terminology Relating to Water
2
D 1193 Specification for Reagent Water 6.1 Highsaltconcentrations,suchasthosefoundinsalineor
D 3370 Practices for Sampling Water from Closed Con- brackish water, interfere in this test method by inhibiting
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duits plaque formation.
6.2 Analysisforcoliphagescanbeperformedondisinfected
3. Terminology
waters. However, the quantitative relationship between coliph-
3.1 Definitions—For definitions of terms used in this test ages and coliform bacteria is different from that observed in
method, refer to Terminology D1129. natural fresh waters. This difference is due to variations in the
3.2 Definitions of Terms Specific to This Standard:
survival rates of coliphages and coliform bacteria exposed to
3.2.1 bacterial lawn—confluent growth of bacteria. disinfectants. For example, coliphages may have been shown
3.2.2 coliphage—as used in this procedure, bacterial virus
to be more resistant to chlorine disinfection than fecal or total
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capable of replication using a specific strain of coliform coliforms.
bacteria (E. coli C) as a host.
7. Apparatus
4. Summary of Test Method
7.1 Water Bath, 44.56 0.2°C.
4.1 A measured water sample is added to a tube of melted
7.2 Incubator,35 6 0.5°C.
modifiednutrientagar.An E. coliChostcultureisaddedtothe 7.3 Balance.
tube, and the contents of the tube are mixed and poured into a
7.4 Petri Dishes, sterile, 100 by 15-mm.
petri dish. The dish is incubated at 35°C. The coliphages 7.5 Pipets, plugged, sterile, 1-mL and 5-mL.
presentinthewatersamplelysethebacteriaandformplaques;
the total number of plaques represents the number of coliph-
3
Kenard, R.P., and Valentine, R.S., “Rapid Determination of the Presence of
ages in the volume of water sample tested.
Enteric Bacteria in Water,” Applied Microbiology, Vol 27, 1974, p. 484.
4
Scarpino, P.V., “Bacteriophage Indicators,” Berg, G., editor, Indicators of
Viruses in Water and Food, Ann Arbor Science, Ann Arbor, Mich., 1978, p. 201.
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This method is under the jurisdiction ofASTM Committee D-19 on Water and Kott,Y.,Ari,B.,andBuras,N.,“TheFateofVirusesinaMarineEnvironment,”
is the direct responsibility of Subcommittee D19.24 on Water Microbiology. Proceedings 4th International Conference on Water Pollution Research, Jenkins,
Current edition approved May 10, 1996. Published July 1996. Originally S.H., editor, Pergamon Press, Oxford, 1969, p. 823.
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published as D4201–82. Last previous edition D4201–82(1989). Kott, Y., Roze, N., Sperber, S., and Betzer, N., “Bacteriophages as Viral
2
Annual Book of ASTM Standards, Vol 11.01. Pollution Indicators,” Water Research, Vol 8, p. 165.
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D 4201–96
7.6 Test Tubes with close fitting or airtight caps, 16 by nm with a spectrophotometer that has been previously cali-
125-mm and 25 by 150-mm. brated with sterile TSB (containing 10% v/v glycerin).
7.7 Platinum Transfer Loop, sterilized by flaming. 10.1.3 PlacetheErlenmeyerflaskfr
...