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ASTM D5246-92(2004)

(Test Method)

Standard Test Method for Isolation and Enumeration of Pseudomonas aeruginosa from Water

Standard Test Method for Isolation and Enumeration of <i>Pseudomonas aeruginosa</i> from Water

SIGNIFICANCE AND USE
Pseudomonas aeruginosa is an opportunistic pathogen, and has been linked as the causative agent of numerous infections that may be transmitted through a contaminated water supply to a susceptible host. In addition to its direct pathogenicity, the association of P. aeruginosa with human fecal waste indicates that where elevated levels of P. aeruginosa are found, a serious health hazard may exist due to the presence of other pathogens.
The membrane filtration procedure described is a rapid and reliable test method of detecting P. aeruginosa in water.
SCOPE
1.1 This test method covers the isolation and enumeration of Pseudomonas aeruginosa (P. aeruginosa) from surface waters; recreational waters; ground water, water supplies; especially rural nonchlorinated sources; waste water; and saline waters. The detection limit of this test method is one microorganism per 100 mL.
1.2 This test method was used successfully with reagent water and it is the user's responsibility to ensure the validity of this test method for waters of untested matrices.
1.3 The values stated in SI units are to be regarded as the standard.
1.4 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. Specific hazard statements are given in Section 10.

General Information

Status
Historical
Publication Date
31-May-2004
ICS
07.100.20 - Microbiology of water
Technical Committee
D19 - Water
Drafting Committee
D19.24 - Water Microbiology
Current Stage
Ref Project

Relations

Replaces
ASTM D5246-92(1998) - Standard Test Method for Isolation and Enumeration of <I>Pseudomonas aeruginosa</I> from Water
Effective Date
01-Jun-2004
Replaced By
ASTM D5246-13 - Standard Test Method for Isolation and Enumeration of <emph type="ital">Pseudomonas aeruginosa </emph> from Water
Effective Date
01-May-2013

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ASTM D5246-92(2004) - Standard Test Method for Isolation and Enumeration of <i>Pseudomonas aeruginosa</i> from WaterASTM D5246-92(2004) - Standard Test Method for Isolation and Enumeration of <i>Pseudomonas aeruginosa</i> from Water
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ASTM D5246-92(2004) - Standard Test Method for Isolation and Enumeration of <i>Pseudomonas aeruginosa</i> from Water
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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
Designation: D5246 − 92 (Reapproved2004)
Standard Test Method for
Isolation and Enumeration of Pseudomonas aeruginosa from
1
Water
This standard is issued under the fixed designation D5246; 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 (´) indicates an editorial change since the last revision or reapproval.
1. Scope 3.2.1 Pseudomonas aeruginosa—an aerobic, motile, gram
negative rod that produces fluorescent pigments and pyocya-
1.1 Thistestmethodcoverstheisolationandenumerationof
nin. It is oxidase and caseinase positive, is able to grow at
Pseudomonas aeruginosa(P. aeruginosa)fromsurfacewaters;
42°C,isrelativelyresistanttomanyantibiotics,andmayutilize
recreational waters; ground water, water supplies; especially
acetamide.
rural nonchlorinated sources; waste water; and saline waters.
3.2.2 refrigeration—storage at 2 to 8°C.
The detection limit of this test method is one microorganism
per 100 mL.
4. Summary of Test Method
1.2 This test method was used successfully with reagent
4.1 A water sample is passed through a 0.45 mm or
wateranditistheuser’sresponsibilitytoensurethevalidityof
equivalent membrane filter. The filter carrying the retained
this test method for waters of untested matrices.
3
organisms is placed on a selective medium (M-PA-C) and is
1.3 The values stated in SI units are to be regarded as the
incubated at 41.5 6 0.5°C for 48 to 72 h. The resulting
standard.
pink-brown to black colonies of Pseudomonas aeruginosa are
1.4 This standard does not purport to address all of the
counted and reported per 100 mLof the sample. Colonies may
safety concerns, if any, associated with its use. It is the
be verified on skim milk agar.
responsibility of the user of this standard to establish appro-
priate safety and health practices and determine the applica-
5. Significance and Use
bility of regulatory limitations prior to use. Specific hazard
5.1 Pseudomonas aeruginosa is an opportunistic pathogen,
statements are given in Section 10.
and has been linked as the causative agent of numerous
infections that may be transmitted through a contaminated
2. Referenced Documents
water supply to a susceptible host. In addition to its direct
2
2.1 ASTM Standards:
pathogenicity, the association of P. aeruginosa with human
D1129Terminology Relating to Water
fecal waste indicates that where elevated levels of P. aerugi-
D1193Specification for Reagent Water
nosa are found, a serious health hazard may exist due to the
D2777Practice for Determination of Precision and Bias of
presence of other pathogens.
Applicable Test Methods of Committee D19 on Water
5.2 The membrane filtration procedure described is a rapid
D3370Practices for Sampling Water from Closed Conduits
and reliable test method of detecting P. aeruginosa in water.
3. Terminology
6. Interferences
3.1 Definitions:
6.1 For certain samples, bacterial cells may have been
3.1.1 For definitions of terms used in this test method, refer
exposed to adverse environmental factors that lower their
to Terminology D1129.
probability for survival and growth on a membrane filter
3.2 Definitions of Terms Specific to This Standard:
medium.Thiseffectmaybepronouncedinthistestmethoddue
to the presence of antibiotics and the elevated incubation
1
temperature.
This test method is under the jurisdiction of ASTM Committee D19 on Water
and is the direct responsibility of Subcommittee D19.24 on Water Microbiology.
6.2 The selection of an appropriate dilution volume is
Current edition approved June 1, 2004. Published June 2004. Originally
essential.Too small a dilution volume may fail to detect any P.
approved in 1992. Last previous edition approved in 1998 as D5246–92 (1998).
DOI: 10.1520/D5246-92R04.
2
For referenced ASTM standards, visit the ASTM website, www.astm.org, or
contact ASTM Customer Service at service@astm.org. For Annual Book of ASTM
3
Standards volume information, refer to the standard’s Document Summary page on Available from BBL Microbiological Systems, Division of Becton Dickinson
the ASTM website. and Co., Cockeysville, MD 21030. Other suppliers may be utilized if equivalent.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
1

---------------------- Page: 1 ----------------------
D5246 − 92 (2004)
aeruginosa organisms, while too large a volume may cause an 8.5 Magnesium Chloride Solution (81.1 g/L)—Dissolve
overabundance of colonies that would interfere with an accu- 81.1 g magnesium chloride (MgCl 6H O) in water and dilute
2 2
rate count. to 1 L with water.
6.3 Chemicals or a combination of chemicals in certain
8.6 Potassium Hydroxide Solution(5.6g/L)—Dissolve5.6g
samples can have
...

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5.1 Sulfate reducing archaea and bacteria are known to contribute to microbiologically influenced corrosion.  
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