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ASTM D6530-00

(Test Method)

Standard Test Method for Total Active Biomass in Cooling Tower Waters (Kool Kount Assay; KKA)

Standard Test Method for Total Active Biomass in Cooling Tower Waters (Kool Kount Assay; KKA)

SCOPE
1.1 This test method covers the determination of viable active biomass in cooling tower water in the range from 102 to 108 cfu/mL (1). It is a semiquantitative test method.
1.2 This test method was used successfully with reagent water, physiologic saline, and cooling tower waters. It is the user's responsibility to ensure the validity of this test method for waters of untested matrices.
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. For specific hazard statements, see Section .

General Information

Status
Historical
Publication Date
09-Mar-2000
ICS
27.200 - Refrigerating technology
Technical Committee
D19 - Water
Drafting Committee
D19.24 - Water Microbiology
Current Stage
Ref Project

Relations

Replaced By
ASTM D6530-00(2006) - Standard Test Method for Total Active Biomass in Cooling Tower Waters (Kool Kount Assay; KKA)
Effective Date
01-Jul-2006

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ASTM D6530-00 - Standard Test Method for Total Active Biomass in Cooling Tower Waters (Kool Kount Assay; KKA)ASTM D6530-00 - Standard Test Method for Total Active Biomass in Cooling Tower Waters (Kool Kount Assay; KKA)
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ASTM D6530-00 - Standard Test Method for Total Active Biomass in Cooling Tower Waters (Kool Kount Assay; KKA)
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NOTICE: This standard has either been superseded and replaced by a new version or withdrawn.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:D6530–00
Standard Test Method for
Total Active Biomass in Cooling Tower Waters (Kool Kount
Assay; KKA)
This standard is issued under the fixed designation D6530; 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 4. Summary of Test Method
1.1 This test method covers the determination of viable 4.1 Thistestmethodconsistsofaddingaspecificvolumeof
active biomass in cooling tower water in the range from 10 to water to nutrients and a color indicator contained in a glass
10 cfu/mL (1). It is a semiquantitative test method. vial. The contents of the vial are then mixed and incubated at
1.2 This test method was used successfully with reagent 95°F (35 6 3°C; that is, in a shirt pocket, incubator, or heat
water, physiologic saline, and cooling tower waters. It is the block). The color of the sample after addition into the vial
user’s responsibility to ensure the validity of this test method containing the nutrients and color indicator is yellow. Viable
for waters of untested matrices. active biomass in the sample replicates using the nutrients
1.3 This standard does not purport to address all of the provided and reduces the color indicator.At a critical biomass
safety concerns, if any, associated with its use. It is the concentration, sufficient quantities of the color indicator are
responsibility of the user of this standard to establish appro- reduced resulting in a visible change in the indicator from the
priate safety and health practices and determine the applica- original yellow sample color to orange. The time required for
bility of regulatory limitations prior to use. For specific hazard conversion of the oxidized indicator to the reduced indicator
statements, see Section 9. resulting in an orange color as directly correlated with the
concentration of viable active biomass in the water sample
2. Referenced Documents
tested. High concentrations of active biomass in the sample
2.1 ASTM Standards:
produce the positive orange color more rapidly than low
D1129 Terminology Relating to Water concentrations of viable biomass.
D1193 Specification for Reagent Water
5. Significance and Use
D1192 Specification for Equipment for Sampling Water
and Steam in Closed Conduits 5.1 This test method is useful for rapid determination of
D3370 PracticesforSamplingWaterfromClosedConduits viable active biomass concentrations in cooling tower waters.
The efficiency of cooling towers is directly affected by the
3. Terminology
concentration of biomass in the cooling tower waters. As
3.1 Definitions—For definitions of terms used in this test
biomass concentrations increase, biofilm formation occurs
method, refer to Terminology D1129. resulting in a decrease in the efficiency of heat exchange in the
3.2 Definitions of Terms Specific to This Standard:
tower. Current tests for monitoring the biomass concentration
3.2.1 snapping cup—container provided for holding the in cooling towers require at least 36 h for growth of the
sample and snapping tip of the vial.
microorganisms on a solid agar surface for counting. Replica-
3.2.2 vial—sealed glass ampoule under vacuum containing tion of microorganisms over the 36-h period before results are
reagents for the Kool Kount Test.
available creates an aqueous environment which is no longer
3.3 Symbol: representedbythedatagenerated.Timelytestresultscanassist
cfu/mL—colony forming units per millilitre
in minimizing biocide addition to control biomass concentra-
tions.KoolKountprovidesdatawithinhourstoallowformore
precise control of active biomass concentrations in the waters.
This test method is under the jurisdiction of ASTM Committee D19 on Water
6. Interferences
and is the direct responsibility of Subcommittee D19.24 on Water Microbiology.
Current edition approved March 10, 2000. Published June 2000.
6.1 Halogens interfere with this test method by inhibiting
For referenced ASTM standards, visit the ASTM website, www.astm.org, or
microbialgrowthresultinginlengthyincubationperiodsbefore
contact ASTM Customer Service at service@astm.org. For Annual Book of ASTM
a positive orange color is produced suggesting better water
Standards volume information, refer to the standard’s Document Summary page on
the ASTM website. quality.Addition of thiosulfate eliminates this interference and
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959, United States.
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.
D6530–00
allows for testing of waters previously treated with halogens snapping cup. Add two drops of the thiosulfate solution
(not immediately prior to testing). provided. Mix and allow sample to sit quiescently in the
6.2 Reducing agents (that is, beta mercaptoethanol) may snapping cup for 15 min.
interfere in this test method by reducing the color indicator 11.2 SubmergethetipofglassVialAcontainingreagentsin
chemically. Rapid color development upon filling of the vials the sample to be tested (in the snapping cup). Place the tip in
suggests a chemical rather than a biological reaction. Waters oneofthegroovesinthebottomofthesnappingcup.Carefully
containingreducingagentswhichreactwiththecolorindicator press the vial toward the opposite wall of the cup to snap the
are not suitable for testing with Kool Kount. tipallowingthevialtofillasaresultofthevacuuminthevial.
6.3 Avoidprolongedexposure(greaterthan30min)offilled 11.3 Submerge the tip of control glass Vial B (no glass rod)
or unopened KKA vials to sunlight to avoid false positive in the same sample. Place the tip in one of the grooves in the
reactions. bottomofthesnappingcup.Carefullypressthevialtowardthe
6.4 Testing must not take place within 24 h of biocide wall of the cup to snap the tip allowing the vial to fill.
addition. 11.4 Place a protective sleeve on the neck of each vial to
cover the sharp edges. Carefully invert vials several times to
7. Apparatus
completely mix the reagent powders with the water sample.
7.1 TheschematicarrangementoftheKKAtestkitisshown
11.5 Prepare the label with the sample designation, sample
in Fig. 1.
pH, sample temperature, and the time at which test was
7.2 (Parts) of the KKA Test K—VialA(test vial), vial under
initiated. Place the sample label on the appropriate vial and
vacuum containing nutrient and reagent on glass rod; Vial B
label the control vial. Incubate vials at approximately 95°F (35
(control vial), vial under vacuum containing nutrient only
6 3°C; heat block, shirt pocket, incubator).
(does not contain a glass rod); snapping cup; and plastic safety
11.6 Examine Vials A and B after 10 to 15 min for
sleeve.
development of pink to red color indicative of chemical
reaction, not biological activity.
8. Reagents and Materials
11.7 Examine sample Vial A for color change (yellow
8.1 Purity of Reagents—Reagent grade chemicals shall be
[negative] to orange [positive]) after 30 min of incubation by
used in all tests. Unless otherwise indicated, it is intended that
looking through the flat base of the vials comparing the test
all reagents shall conform to the specifications of the Commit-
sample vial (A) with the control vial (B). Examine Vial A for
tee onAnalytical Reagents of theAmerican Chemical Society
color change at hourly intervals by looking t
...

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SIGNIFICANCE AND USE
5.1 This test method is useful for rapid determination of viable active biomass concentrations in cooling tower waters. The efficiency of cooling towers is directly affected by the concentration of biomass in the cooling tower waters. As biomass concentrations increase, biofilm formation occurs resulting in a decrease in the efficiency of heat exchange in the tower. Current tests for monitoring the biomass concentration in cooling towers require at least 36 h for growth of the microorganisms on a solid agar surface for counting. Replication of microorganisms over the 36-h period before results are available creates an aqueous environment which is no longer represented by the data generated. Timely test results can assist in minimizing biocide addition to control biomass concentrations. Kool Kount provides data within hours to allow for more precise control of active biomass concentrations in the waters.
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1.1 This test method covers the determination of viable active biomass in cooling tower water in the range from 102 to 108 cfu/mL. It is a semiquantitative test method.  
1.2 This test method was used successfully with reagent water, physiologic saline, and cooling tower waters. It is the user’s responsibility to ensure the validity of this test method for waters of untested matrices.  
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1.5 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.

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5.1 This test method is useful for rapid determination of viable active biomass concentrations in cooling tower waters. The efficiency of cooling towers is directly affected by the concentration of biomass in the cooling tower waters. As biomass concentrations increase, biofilm formation occurs resulting in a decrease in the efficiency of heat exchange in the tower. Current tests for monitoring the biomass concentration in cooling towers require at least 36 h for growth of the microorganisms on a solid agar surface for counting. Replication of microorganisms over the 36-h period before results are available creates an aqueous environment which is no longer represented by the data generated. Timely test results can assist in minimizing biocide addition to control biomass concentrations. Kool Kount provides data within hours to allow for more precise control of active biomass concentrations in the waters.
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5.4 Research O.N. is important in relation to the specifications for spark-ignition engine fuels used in stationary and other nonautomotive engine applications.
SCOPE
1.1 This laboratory test method covers the quantitative determination of the knock rating of liquid spark-ignition engine fuel in terms of Research O.N., including fuels that contain up to 25 % v/v of ethanol. However, this test method may not be applicable to fuel and fuel components that are primarily oxygenates.2 The sample fuel is tested using a standardized single cylinder, four-stroke cycle, variable compression ratio, carbureted, CFR engine run in accordance with a defined set of operating conditions. The O.N. scale is defined by the volumetric composition of PRF blends. The sample fuel knock intensity is compared to that of one or more PRF blends. The O.N. of the PRF blend that matches the K.I. of the sample fuel establishes the Research O.N.  
1.2 The O.N. scale covers the range from 0 to 120 octane number but this test method has a working range from 40 to 120 Research O.N. Typical commercial fuels produced for spark-ignition engines rate in the 88 to 101 Research O.N. range. Testing of gasoline blend stocks or other process stream materials can produce ratings at various levels throughout the Research O.N. range.  
1.3 The values of operating conditions are stated in SI units and are considered standard. The values in parentheses are the historical inch-pound units. The standardized CFR engine measurements continue to be in inch-pound units only because of the extensive and expensive tooling that has been created for this equipment.  
1.4 For purposes of determining conformance with all specified limits in this standard, an observed value or a calculated value shall be rounded “to the nearest unit” in the last right-hand digit used in expressing the specified limit, in accordance with the rounding method of Practice E29.  
1.5 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, health, and environmental practices and determine the applicability of regulatory limitations prior to use. For specific warning statements, see Section 8, 14.4.1, 15.5.1, 16.6.1, Annex A1, A2.2.3.1, A2.2.3.3 (6) and (9), A2.3.5, X3.3.7, X4.2.3.1, X4.3.4.1, X4.3.9.3, X4.3.11.4, and X4.5.1.8.  
1.6 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Gu...

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