ASTM D932-85(2009)

Standard Test Method for Iron Bacteria in Water and Water-Formed Deposits

SIGNIFICANCE AND USE
Iron bacteria is a general classification for microorganisms that utilize ferrous iron as a source of energy and are characterized by the deposition of ferric hydroxide in their mucilaginous sheaths. The process is continuous with these growths, and over a period of time large accumulations of slimey brown deposits can occur. Iron bacteria may clog water lines, reduce heat transfer, and cause staining; objectionable odors may arise following death of the bacteria. The organic matter in the water is consequently increased, and this in turn favors the multiplication of other bacteria.
SCOPE
1.1 This test method covers the determination of iron bacteria by examination under the microscope. The method provides for the identification of the following genera of bacteria found in water and water-formed deposits: Siderocapsa, Gallionella (Dioymohelix), Sphaerotilus, Crenothrix, Leptothrix, and  Clonothrix.
1.2 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.
1.3 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.

General Information

• Status: Historical
• Publication Date: 30-Apr-2009
• ICS: 13.060.50 - Examination of water for chemical substances
• Technical Committee: D19 - Water
• Drafting Committee: D19.24 - Water Microbiology

Relations

• Replaces: ASTM D932-85(2002) - Standard Test Method for Iron Bacteria in Water and Water-Formed Deposits
• Effective Date: 01-May-2009
• Replaced By: ASTM D932-15 - Standard Practice for Filamentous Iron Bacteria in Water and Water-Formed Deposits
• Effective Date: 01-Feb-2015
• Referred By: ASTM D2331-08(2022) - Standard Practices for Preparation and Preliminary Testing of Water-Formed Deposits
• Effective Date: 01-May-2009
• Referred By: ASTM C1840/C1840M-22 - Standard Practice for Inspection and Acceptance of Installed Reinforced Concrete Culvert, Storm Drain, and Storm Sewer Pipe
• Effective Date: 01-May-2009
• Referred By: ASTM D4025-18 - Standard Practice for Reporting Results of Examination and Analysis of Deposits Formed from Water for Subsurface Injection
• Effective Date: 01-May-2009

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:D932 −85(Reapproved2009)
StandardTest Method for
Iron Bacteria in Water and Water-Formed Deposits
This standard is issued under the fixed designation D932; 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 accomplished by microscopical examination of sediment from
the sample. Table 1 and Figs. 1-10 (3) may be used to
1.1 This test method covers the determination of iron
differentiate the various types. This test method provides an
bacteria by examination under the microscope. The method
indicationofthedensityoftheironbacteriaandtheseverityof
provides for the identification of the following genera of
the clogging problem in pipes caused by these bacteria.
bacteria found in water and water-formed deposits:
Siderocapsa, Gallionella (Dioymohelix), Sphaerotilus ,
5. Significance and Use
Crenothrix, Leptothrix , and Clonothrix.
5.1 Iron bacteria is a general classification for microorgan-
1.2 The values stated in SI units are to be regarded as
isms that utilize ferrous iron as a source of energy and are
standard. No other units of measurement are included in this
characterized by the deposition of ferric hydroxide in their
standard.
mucilaginous sheaths. The process is continuous with these
1.3 This standard does not purport to address the safety
growths, and over a period of time large accumulations of
concerns, if any, associated with its use. It is the responsibility
slimey brown deposits can occur. Iron bacteria may clog water
of the user of this standard to establish appropriate safety and
lines, reduce heat transfer, and cause staining; objectionable
health practices and determine the applicability of regulatory
odors may arise following death of the bacteria. The organic
limitations prior to use.
matter in the water is consequently increased, and this in turn
favors the multiplication of other bacteria.
2. Referenced Documents
6. Apparatus
2.1 ASTM Standards:
D887Practices for Sampling Water-Formed Deposits
6.1 Centrifuge, complete with conical tubes.
D1129Terminology Relating to Water
6.2 Microscope that provides a magnification of 400 to
D1193Specification for Reagent Water
1000×andiscompletewithasuitablelightsource.Adark-field
D3370Practices for Sampling Water from Closed Conduits
condenser is desirable.
3. Terminology
6.3 Pipets, Mohr-type, 10-mL, with an opening 3 to 4 mm
in diameter, for thick samples, and 1-mLMohr-type pipets for
3.1 Definitions—For definitions of terms used in this test
thin samples.
method, refer to Terminology D1129.
6.4 Spatula, small and narrow, for handling thick samples.
4. Summary of Test Method
6.5 Membrane Filter, with appropriate filter-holding assem-
4.1 The iron bacteria are generally filamentous, typically
bly (see 9.2).
found in fresh water, and frequently surrounded by a sheath
which is usually encrusted with iron or manganese, or both (1,
7. Reagents
2). However, Starkey (3) reports another type which is classi-
7.1 Purity of Reagents—Reagent grade chemicals shall be
fied among the true bacteria. Detection and identification is
used in all tests. Unless otherwise indicated, it is intended that
all reagents shall conform to the specifications of the Commit-
This test method is under the jurisdiction ofASTM Committee D19 on Water
tee onAnalytical Reagents of theAmerican Chemical Society,
and is the direct responsibility of Subcommittee D19.24 on Water Microbiology.
where such specifications are available. Other grades may be
Current edition approved May 1, 2009. Published June 2009. Originally
approved in 1947. Last previous edition approved in 2002 as D932–85 (2002).
DOI: 10.1520/D0932-85R09.
2 4
For referenced ASTM standards, visit the ASTM website, www.astm.org, or Reagent Chemicals, American Chemical Society Specifications, American
contact ASTM Customer Service at [e-mail]. For Annual Book of ASTM Chemical Society, Washington, DC. For Suggestions on the testing of reagents not
Standards volume information, refer to the standard’s Document Summary page on listed by the American Chemical Society, see Annual Standards for Laboratory
the ASTM website. Chemicals, BDH Ltd., Poole, Dorset, U.K., and the United States Pharmacopeia
Theboldfacenumbersinparenthesesrefertothelistofreferencesattheendof and National Formulary, U.S. Pharmacopeial Convention, Inc. (USPC), Rockville,
this test method. MD.
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D932−85(Reapproved2009)
TABLE 1 Key for Identification of Bacteria
FIG. 1 Siderocapsa treubii. Multiple colonies surrounded by ferric
FIG. 2 Gallionella major. Cells at the ends of excretion bands un-
hydrate. Magnification about 500×. Fig. 4 of Ref (5)
dergoing division. Magnification about 1180×. Fig. 3 of Ref (6)
D932−85(Reapproved2009)
FIG. 3 Gallionella major. Curved cells at the ends of excretion
bands. Magnification about 1120×. Fig. 6 of Ref (6)
FIG. 5 Crenothrix polyspora. Sketch showing details of false
branching of cells within sheath. Magnification about 380×.
Plate 1, Fig. A of Ref (8)
sufficiently high purity to permit its use without lessening the
accuracy of the determination.
7.2 Purity of Water—Unless otherwise indicated, references
towatershallbeunderstoodtomeanreagentwaterconforming
to Specification D1193, Type II.
7.3 Ammonium Oxalate-Crystal Violet Solution—Prepare
Hucker’s modification of the Gram stain (4) by mixing a
solution of 2.0 g of crystal violet (90% dye content) in 20 mL
of ethyl alcohol (95% with a solution of 0.8 g of ammonium
oxalate monohydrate (NH ) C O ·H O) in 80 mL of water.
4 2 2 4 2
7.4 HydrochloricAcid(1+4)—Mix1volumeofhydrochlo-
ric acid (HCl, sp gr 1.19) with 4 volumes of water.
7.5 Iodine Solution—Prepare Gram’s modification of
Lugol’s solution (4) by dissolving1gof iodine in a solute
containing2gof potassium iodide (KI) in 10 mLof water and
diluting the resulting solution to 300 mL with water.
FIG. 4 Sphaerotilus dichotoma. Sketch showing false branching.
7.6 Filter Paper or Blotter.
Magnification about 230×. Fig. 3b of Ref (7)
7.7 Slides, standard type, 25 by 76-mm (1 by 3 in.) with
either plain or frosted end.
7.8 Cover Glasses, round or square type, 19 mm ( ⁄4 in.) in
used, provided it is first ascertained that the reagent is of diameter.
D932−85(Reapproved2009)
FIG. 6 Crenothrix polyspora. Cells enclosed within a sheath of ferric hydrate and showing false branching. Magnification about 390×.
Plate3,Fig.BofRef(8)
8. Sampling 8.7 As soon as possible after collection of the solids,
microscopically examine them
...