ASTM E1839-96

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Designation: E 1839 – 96
Standard Test Method for
Efficacy of Slimicides for the Paper Industry—Bacterial and
Fungal Slime
This standard is issued under the fixed designation E 1839; 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 (e) indicates an editorial change since the last revision or reapproval.
1. Scope Hardwood pulp is made from trees, such as maples or oaks, and
softwood pulp is produced from trees, such as pines.
1.1 This test method presents a procedure to evaluate the
3.1.3 pulp slurry, n—an aqueous combination of cellulosic
efficacy of slimicides for the control of bacterial and fungal
fibers, fillers, and other additives used for specific grades of
slimes in paper mill systems and their counterparts.
paper.
1.2 It is the responsibility of the investigator to determine
3.1.4 slimicides, n—chemicals added during pulp and paper
whether Good Laboratory Practices (GLP) are required and to
processing to control the growth of slime-forming microorgan-
follow them where appropriate (40 CFR 160).
isms.
1.3 This standard does not purport to address all of the
safety concerns, if any, associated with its use. It is the
4. Summary of Test Method
responsibility of the user of this standard to establish appro-
4.1 Bacterial cells or fungal spores are added to acid or
priate safety and health practices and determine the applica-
alkaline pulp slurries, or both, treated with slimicides to
bility of regulatory limitations prior to use.
6 7
achieve final concentrations of 2 3 10 to 1 3 10 bacteria/mL
5 6
2. Referenced Documents or 10 to 10 fungal spores/mL, and incubated at appropriate
temperature for determined time periods. Aliquots of the test
2.1 ASTM Standards:
suspension are then neutralized, plated onto bacterial or fungal
D 1193 Specification for Reagent Water
medium, and observed for growth. Results with biocide are
E 1054 Practices for Evaluating Inactivators of Antimicro-
compared to results without biocide (control).
bial Agents Used in Disinfection, Sanitizer, Antiseptic, or
3 4.2 As a performance standard, an effective slimicide is one
Preserved Products
that shows a continued reduction in bacterial and fungal counts
2.2 TAPPI Standard:
4 relative to the control over the duration of the test.
T 205 Forming Handsheets for Physical Tests of Pulp
2.3 CFR Standard:
5. Significance and Use
Title 40, Code of Federal Regulations (CFR), Part 160,
5 5.1 This test method is to be used to determine if a slime
Good Laboratory Practice Standards
control agent has application in the paper industry for control
3. Terminology of bacterial or fungal slime.
5.2 This test method is run in acid, alkaline, or acid and
3.1 Definitions:
alkaline conditions to determine the efficacy of the slime
3.1.1 furnish, n—pulp slurry fed to a paper machine. The
control agent.
type of pulp (sulfite, Kraft, mechanical), the source of fiber
5.3 The test conditions may be modified to reflect intended
(virgin, recycled including pre- or post-consumer waste paper),
use patterns in typical paper mill systems, including use of
and the pH are used to designate a specific type of furnish.
actual paper mill furnish.
3.1.2 pulp, n—wood separated by chemical or mechanical
means into their fibrous components. The pulp is used to make
6. Apparatus
paper, paper board, or pulp sheets after specific treatments.
6.1 Balance:
6.1.1 Plant Balance, sensitive to 0.1 g and used to weight
This test method is under the jurisdiction of ASTM Committee E-35 on
furnish.
Pesticides and is the direct responsibility of Subcommittee E35.15 on Antibacterial
6.1.2 Analytical Balance, sensitive to 0.1 mg and employed
Agents.
Current edition approved Oct. 10, 1996. Published December 1996. Replaces
to weigh the candidate slime control agent to be used in the
Test Methods E 599 and E 600.
preparation of the stock solutions.
Annual Book of ASTM Standards, Vol 11.01.
3 6.2 Sample Containers (Sterile), 120-mL plastic specimen
Annual Book of ASTM Standards, Vol 11.05.
containers with screw-cap lids are ideal for holding test
Forming Handsheets for Physical Tests of Pulp, TAPPI Test Method T 205
on-88, 1994–1995, TAPPI, Atlanta, GA, 11994.
materials. Other suitable containers include 150/160-mL milk
Available from U.S. Government Printing Office, Superintendent of Docu-
dilution bottles or WHIRL-PAKS.
ments, Mail Stop: SSOL, Washington, DC 20402-9328.
Copyright © ASTM, 100 Barr Harbor Drive, West Conshohocken, PA 19428-2959, United States.
E 1839
6.3 Culture Containers, Petri plates, tissue culture bottles or accuracy of the determination.
glass tubes (15 3 125 mm or 18 3 150 mm without lip, 7.2 Purity of Water—Unless otherwise indicated, references
preferably of borosilicate glass).
to water shall be understood to mean distilled water or water of
6.4 Closures, for tubes and containers. equal purity (see Specification D 1193, Type III).
6.5 Disintegrators
7.3 Buffer for Suspending Spores and for Dilutions, sample
6.6 Flaming Equipment—Depending upon circumstances, containers having 100-mL phosphate buffer dilution water,
either an alcohol lamp, a bunsen burner, or electric incinerator
sterile, for spore suspension have solid, sterile glass beads in
may be used to flame inoculating needles and other equipment. container.
6.7 Reliable incubators that control at the temperature re-
7.3.1 0.25 M Phosphate Buffer Stock Solution—Dissolve 34
quired, 6 2°C. Temperatures used should be consistent with g of reagent grace KH PO in 500 mL of distilled water and
2 4
the temperatures of the systems.
mix. Adjust to pH 7.2 with 1 N NaOH and dilute to 1 l.
6.8 pH Meter—Any reliable pH meter is suitable to stan- 7.3.2 Phosphate buffer dilution water. Add 1.25 mL of 0.25
dardize the pH of the culture.
M phosphate buffer stock solution to 1 L of distilled water and
6.9 Pipets—1.1-mL milk dilution type, 1.0 mL graduated in mix. Dispense to sample container and sterilize.
0.01 mL, and 10 mL graduated in 0.1 mL. Pipetters may be
7.4 Aluminum Sulfate (Alum) [Al (SO ) 18H O]—Prepare
2 4 3 2
used, but not for highly viscous materials. a 0.4 % solution of the hydrated aluminum in distilled water
6.9.1 Pipetting Aid—Rubber bulb or other device to accom-
and sterilize in an autoclave. Any loss of water during
plish the transfer of liquid. sterilization is made up by adding sterile distilled water.
6.10 Sterilizers, steam sterilizer (121°C) or hot-air oven
Alternately, the solution may be filter sterilized.
(180 6 2°C for 2 h), or both.
7.5 Acid and Base for pH Adjustment to Make Acid and
6.11 Filter Apparatus for Filter Sterilizing, Disposable filter
Alkaline Furnish:
units, appropriate volume, 0.22-μm pore size.
7.5.1 Prepare a 2 N solution of sulfuric acid in water.
6.12 Sterile Funnel, with sterile glass wool or sterile cotton
Sterilize by filtration.
gauze for filtration of spores.
7.5.2 Prepare a 2.0 N solution of sodium hydroxide in water.
6.13 Colony counter, manual, such as the Quebec, Buck,
Sterilize by filtration.
Wolffhuegel, or equivalent; or a colony image analyzer
7.6 Pulp—A two-third hardwood and one-third softwood
(electronic/scanner type) are suitable for counting plates after
pulp, typical of current production techniques, and that has
incubation. A hand tally for recording of bacteria count is
been produced without slimicide is needed. Disintegrate the
recommended.
sheet in distilled water until free of fiber clots and undispersed
6.14 Swabs, sterile, for aiding in removal of fungal spores
fiber bundles. Avoid methods which involve extensive cutting
from agar surface.
of fibers. The concentration of the pulp in water should be 1 %.
6.15 Hemacytometer, for counting spore suspension.
7.7 Bacterial and Fungal Culture Medium:
6.16 Microscope, that provides a magnification of 400 to
7.7.1 Bacteria—Standard dehydrated tryptone glucose ex-
10003 and is complete with a suitable light source. Phase
tract agar or equivalent is recommended. Adjust pH of culture
contrast or dark field capability is desirable.
medium to pH of the test system.
6.17 Constant Temperature Shaker—A reliable constant-
7.7.2 Fungi—Sabouraud Dextrose Agar or Potato Dextrose
temperature shaker (water bath or incubator type), shall be
Agar are recommended for enumeration. Adjust pH of culture
used to provide mixing and aeration and to maintain a selected
medium to pH of the test system.
temperature (6 2°C) during the contact period.
7.8 Stock Slimicide Solution—Weigh appropriate amount of
6.18 Mechanical Stirrer—Magnetic or propeller-type stir-
slimicide and add to distilled water so that when 1 mL is added
rers or any other suitable device.
to the furnish it gives the desired final concentration.
7. Reagents and Materials
8. Test Organisms
7.1 Purity of Reagents—Reagent chemicals shall be used in
8.1 Bacteria: Enterobacter aerogenes (ATCC 13048) and
all tests. Unless otherwise indicated, it is intended that all
Pseudomonas aeruginosa (ATCC 15442)—Maintain these or-
reagents shall conform to the specifications of the Committee
ganisms on tryptone glucose extract agar at 32 6 2°C.
on Analytical Reagents of the American Chemical Society,
8.2 Fungi: Aspergillus niger (ATCC 6275)—Maintain and
where such specifications are available. Other grades may be
grow for sporulation on Sabouraud Dextrose Agar, grow at 25
used, provided it is first ascertained that the reagent is of
6 2°C. Chaetomium globosum (ATCC 6205)—Maintain and
sufficiently high purity to permit its use without lessening the
grow for sporulation on Mineral Slats Agar with sterile filter
paper at 25 6 2°C.
Forming Handsheets for Physical Tests of Pulp. Appendix A: Specifications and
Care of Apparatus (Disintegrator), TAPPI Test Method T 205 on-88. 1994–1995.
TAPPI, Atlanta, GA 11994.
7 8
Reagent Chemicals, American Chemical Society Specifications, American The sole source of supply of the apparatus known to the committee at this time
Chemical Society, Washington, DC. For suggestions o
...