Standard Practice for Filamentous Iron Bacteria in Water and Water-Formed Deposits

SIGNIFICANCE AND USE
5.1 Filamentous 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 slimy 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 practice covers the determination of filamentous iron bacteria (FIB) by examination under the microscope. The practice 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.

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NOTICE: This standard has either been superseded and replaced by a new version or withdrawn.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 − 15
Standard Practice for
Filamentous Iron Bacteria in Water and Water-Formed
1
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.
3
1. Scope 2). However, Starkey (3) reports another type which is
classified among the true bacteria. Detection and identification
1.1 This practice covers the determination of filamentous
isaccomplishedbymicroscopicexaminationofsedimentfrom
iron bacteria (FIB) by examination under the microscope. The
the sample.
practice provides for the identification of the following genera
of bacteria found in water and water-formed deposits:
4.2 This practice provides a qualitative indication of the
Siderocapsa, Gallionella (Dioymohelix), Sphaerotilus,
density of the filamentous iron bacteria and the severity of the
Crenothrix, Leptothrix, and Clonothrix.
clogging problem in pipes caused by these bacteria.
1.2 The values stated in SI units are to be regarded as
5. Significance and Use
standard. No other units of measurement are included in this
standard.
5.1 Filamentous iron bacteria is a general classification for
microorganisms that utilize ferrous iron as a source of energy
1.3 This standard does not purport to address the safety
concerns, if any, associated with its use. It is the responsibility and are characterized by the deposition of ferric hydroxide in
their mucilaginous sheaths. The process is continuous with
of the user of this standard to establish appropriate safety and
health practices and determine the applicability of regulatory thesegrowths,andoveraperiodoftimelargeaccumulationsof
slimy brown deposits can occur. Iron bacteria may clog water
limitations prior to use.
lines, reduce heat transfer, and cause staining; objectionable
2. Referenced Documents
odors may arise following death of the bacteria. The organic
2
matter in the water is consequently increased, and this in turn
2.1 ASTM Standards:
favors the multiplication of other bacteria.
D887Practices for Sampling Water-Formed Deposits
D1129Terminology Relating to Water
6. Apparatus
D1193Specification for Reagent Water
D3370Practices for Sampling Water from Closed Conduits
6.1 Centrifuge, complete with 250 mL conical bottles.
D5465Practice for Determining Microbial Colony Counts
3
6.2 Cover Glasses, round or square type, 19 mm ( ⁄4 in.) in
from Waters Analyzed by Plating Methods
diameter.
3. Terminology
6.3 Filter Paper or Blotter.
3.1 Definitions—For definitions of terms used in this
6.3.1 For 8.3.2.1 – Grade 5 (nominal 2.5 µm particle-size
practice, refer to Terminology D1129.
retention).
6.3.2 For 9.3 – any absorbent paper medium will suffice.
4. Summary of Test Method
6.4 Containers, sterile 1 L glass or plastic (can be autocla-
4.1 The iron bacteria are generally filamentous, typically
vable).
found in fresh water, and frequently surrounded by a sheath
which is usually encrusted with iron or manganese, or both (1,
6.5 Membrane Filter, 0.45 µ nominal pore size, with appro-
priate filter-holding and vacuum assembly (see 9.2).
1
This practice is under the jurisdiction ofASTM Committee D19 on Water and
6.6 Microscope that provides a magnification of 400 to
is the direct responsibility of Subcommittee D19.24 on Water Microbiology.
1000×andiscompletewithasuitablelightsource.Adark-field
Current edition approved Feb. 1, 2015. Published March 2015. Originally
condenser is desirable.
approved in 1947. Last previous edition approved in 2009 as D932– 85 (2009).
DOI: 10.1520/D0932-15.
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 The boldface numbers in parentheses refer to a list of references at the end of
the ASTM website. this standard.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
1

---------------------- Page: 1 ----------------------
D932 − 15
6.7 Pipets, Mohr-type, 10-mL, with an opening 3 to 4 mm 8.3.2 Filtration—Useasmallsidestreamfiltertocollectthe
in diameter, for thick samples, and 1-mLMohr-type pipets for sample to be examined.
thin samples or equivalent disposable plastic pipettes. 8.3.2.1 Filterthewatersuspectedofcontainingironbacteria
through a Grade 5 (nominal 2.5 µm particle-size retention)
6.8 Slides, glass, standard type, 25 by 76-mm (1 by 3 in.)
filter paper (6.3.1 or some other comparable media) for 24 h.
with either plain or frosted end.
8.3.2.2 Adjust the side-stream filter flow rat
...

This document is not an ASTM standard and is intended only to provide the user of an ASTM standard an indication of what changes have been made to the previous version. Because
it may not be technically possible to adequately depict all changes accurately, ASTM recommends that users consult prior editions as appropriate. In all cases only the current version
of the standard as published by ASTM is to be considered the official document.
Designation: D932 − 85 (Reapproved 2009) D932 − 15
Standard Test Method Practice for
Filamentous Iron Bacteria in Water and Water-Formed
1
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. A number in parentheses indicates the year of last reapproval. A
superscript epsilon (´) indicates an editorial change since the last revision or reapproval.
1. Scope
1.1 This test method practice covers the determination of filamentous iron bacteria (FIB) by examination under the microscope.
The methodpractice provides for the identification of the following genera of bacteria found in water and water-formed deposits:
Siderocapsa,Gallionella (Dioymohelix), Sphaerotilus ,Sphaerotilus,Crenothrix, Leptothrix ,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.
2. Referenced Documents
2
2.1 ASTM Standards:
D887 Practices for Sampling Water-Formed Deposits
D1129 Terminology Relating to Water
D1193 Specification for Reagent Water
D3370 Practices for Sampling Water from Closed Conduits
D5465 Practice for Determining Microbial Colony Counts from Waters Analyzed by Plating Methods
3. Terminology
3.1 Definitions—For definitions of terms used in this test method, refer to Terminology D1129.
3.1 Definitions—For definitions of terms used in this practice, refer to Terminology D1129.
4. Summary of Test Method
4.1 The iron bacteria are generally filamentous, typically found in fresh water, and frequently surrounded by a sheath which is
3
usually encrusted with iron or manganese, or both ((1, 2).). However, Starkey ((3)) reports another type which is classified among
the true bacteria. Detection and identification is accomplished by microscopicalmicroscopic examination of sediment from the
sample.Table 1 and Figs. 1-10 (3) may be used to differentiate the various types. This test method provides an indication of the
density of the iron bacteria and the severity of the clogging problem in pipes caused by these bacteria.
4.2 This practice provides a qualitative indication of the density of the filamentous iron bacteria and the severity of the clogging
problem in pipes caused by these bacteria.
5. Significance and Use
5.1 Iron Filamentous 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 slimeyslimy 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.
1
This test method practice is under the jurisdiction of ASTM Committee D19 on Water and is the direct responsibility of Subcommittee D19.24 on Water Microbiology.
Current edition approved May 1, 2009Feb. 1, 2015. Published June 2009March 2015. Originally approved in 1947. Last previous edition approved in 20022009 as
D932 – 85 (2002).D932 DOI: 10.1520/D0932-85R09. – 85 (2009). DOI: 10.1520/D0932-15.
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 Standards
volume information, refer to the standard’s Document Summary page on the ASTM website.
3
The boldface numbers in parentheses refer to thea list of references at the end of this test method.standard.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
1

---------------------- Page: 1 ----------------------
D932 − 15
TABLE 1 Key for Identification of Bacteria
TABLE 1 Key for Identification of Bacteria
6. Apparatus
6.1 Centrifuge, complete with 250 mL conical tubes.bottles.
3
6.2 Cover Glasses, round or square type, 19 mm ( ⁄4 in.) in diameter.
6.3 Filter Paper or Blotter.
6.3.1 For 8.3.2.1 – Grade 5 (nominal 2.5 μm particle-size retention).
6.3.2 For 9.3
...

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