E24-06 AMERICAN WOOD-PRESERVERS’ ASSOCIATION STANDARD
© 2007 All Rights Reserved
STANDARD METHOD OF EVALUATING THE RESISTANCE
OF WOOD PRODUCT SURFACES TO MOLD GROWTH
Jurisdiction: AWPA Subcommittee P-6
This Standard was developed by AWPA’s Technical Committees in an open, consensus based process. The intent of this Standard is to provide
meaningful data to AWPA’s Technical Committees for the sole purpose of developing other AWPA Standards. Modifications, deviations, or exceptions
to this Standard invalidate any references to this Standard unless documented and attached to a valid standardization proposal form submitted to
AWPA in accordance with its Technical Committee Regulations. It consists of 5 pages.
1. Scope
1.1 This Standard provides an evaluation method for
assessing the resistance of surfaces of wood products,
treated or untreated, coated or uncoated, to the growth of
molds. The main objective of the method is to determine
the relative resistance of wood products to surface growth
of molds in an environment favorable for growth of fungi.
Resistance to mold growth is evaluated relative to reference
products and unprotected sapwood of a species of pine.
Pine sapwood is included as an untreated control substrate
to indicate viability of molds in the test chamber and
severity of growth conditions. This test method may also be
used to evaluate compounds or formulations that may
inhibit fungal growth and the aggregate levels for their use.
Products evaluated with this method would be intended for
use in interior environments where the products may be
unintentionally subjected to conditions favorable to the
growth of molds. This Standard is based on the ASTM
Standard Test Method D 3273-00, Standard Test Method
for Resistance to Growth of Mold on the Surface of Interior
Coatings in an Environmental Chamber.
1.2 The method is applicable to the testing of
commercial or experimental wood products designed to be
resistant to the growth of molds. Performance at a certain
rating does not imply any specific period of time for a
fungal free surface in service, however products with a
better rating would likely perform better in actual end use.
1.3 The requirements for preparation of the material for
testing and the test procedure appear in the following
sections:
Outline of Method .......................................2
Apparatus.....................................................3
Reagents and Materials................................4
Test Specimens ............................................5
Treatment Procedure....................................6
Preparation of Inoculum ..............................7
Exposure in Environment Chambers ...........8
Assessment of Mold Growth .......................9
Interpretation of Results ............................10
2. Outline of Method
Samples of wood products are exposed in an
environment chamber where temperature and relative
humidity are controlled to provide ideal conditions for the
growth of molds. The chamber and samples are inoculated
with specified molds, and circulating air within the chamber
continually subjects samples to spores for the duration of
the test. The method is non-sterile, and therefore molds
from the air or soil may be present and compete with the
inoculated molds for colonization on surfaces of test
products. Samples are removed from the chamber and
evaluated for growth of molds on the sample surfaces every
two weeks for eight weeks. Each sample is assigned a
rating for extent and intensity of mold growth. Average
results for replicate samples are calculated and compared to
results obtained for reference products and untreated pine
sapwood samples to evaluate the relative resistance of
products to mold growth.
3. Apparatus
3.1 Wood working equipment capable of producing
samples of wood products and pine sapwood samples of
required dimensions, and for drilling appropriate size holes
in the samples to accommodate an eyehook screw.
3.2 Latex or vinyl gloves for handling samples.
3.3 A balance capable of weighing to within 0.1 g.
3.4 Appropriate laboratory facilities for preparation and
disposal of mold inoculum, including air handling systems,
a laboratory blender, distilled water, flasks, pipettes, and a
trigger sprayer.
3.5 A temperature controlled room to house the
environment chamber capable of being maintained
approximately 5°C cooler than the selected test temperature
within the chamber so that there is sufficient heat loss from
the cabinet that a minimum of 95% relative humidity is
maintained at the test temperature, and to prevent excessive
condensation occurring inside the environment chamber.
The room should be maintained at negative pressure with
respect to all other rooms in the building and should be
separately vented to the outside. All persons entering the
room should wear a NIOSH N95 mask1.
3.6 A self-contained environment chamber containing
water, soil, and racks for suspending test samples, with a
pitched roof to prevent deposition of condensation onto
samples. The environment chamber shall be temperature
controlled to within ±1°C of the desired test temperature.
The chamber shall be constructed of materials that do not
support mold growth, and shall consist of a small cabinet,
suitable for holding about 28 approximately 75 by 100-mm
1
This standard does not purport to address all safety concerns, if any,
associated with its use. It is the responsibility of the user of the standard
to establish appropriate safety and health practices and determine the
applicability of regulatory limitation prior to use.
 STANDARD METHOD FOR EVALUATING THE RESISTANCE
E24-06 OF WOOD PRODUCT SURFACES TO MOLD GROWTH
(3 by 4 in.) samples under test conditions. It can be
constructed as follows (Fig 1):
Figure 1: Components of Controlled Environment Mold
Growth Chamber
Tank, polypropylene or polyethylene, with an offset
shoulder at the top rim is used as the chamber.2 A pitched
(90 degree angle) top with straight sides should be
constructed out of acrylic plastic so moisture condensation
will run down the sides and be re-circulated within the
chamber instead of dripping onto the test panels. Gasket
material may be required to achieve a seal where the top
rests on the lip of the tank.
An immersion heater3 is installed in the bottom of the
chamber by connections through the end wall. It is so
placed that it is immersed when there are approximately 80
mm (3 in) of water in the bottom of the chamber. The
heater is regulated by a solid-state electronic temperature
controller4 to maintain a temperature of 25.0°C ± 1.0°C in
proximity to the samples, as measured by a thermocouple
located amongst the samples. Other operating temperatures
may be selected between 15°C and 32.5°C to simulate
2
Nalgene #14100-0045 (approximately 24 x 18 x 18 inches), or
equivalent.
3 6
Mighty Watt immersion heater, part # mwej06a0119-N, or equivalent
4 7
Ogden model ETR-9200-1321 universal controller, or equivalent
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© 2007
specific exposure situations. Generally, lower temperatures
favor growth of staining fungi and higher temperatures
favor growth of molds. To aid in even heat distribution,
water within the tank is constantly stirred by an aquarium
circulating pump5set at maximum flow. An acrylic tray
approximately 25 mm (1 in.) smaller than the inside
dimensions of the chamber and approximately 76 mm (3
in.) deep with a non-corrodible metal6 mesh bottom shall be
supported approximately 25 mm (1 in.) above the water
level and centered in the chamber. One layer of fine mesh
plastic or fiberglass screen should be placed over the metal
mesh for holding soil.
A small fan,7 mounted perpendicular to and just above
the surface of the soil bed in the tray to provide dispersion
and circulation of the fungal spores to achieve continuous
inoculation of the surfaces of the samples.
A series of sample holding supports, suspended across
the width of the chamber at a height and spacing that allows
the use of samples approximately 75 by 100 mm (3 by 4
in.), situated vertically, with 75-mm (approximately 3 in.)
clearance above the inoculated soil with a suitable method
of fastening. Fasteners and supports shall be of materials
that do not support mold growth to avoid contamination of
samples or the chamber environment. Stainless steel
eyehook screws may be attached to the samples for
suspension from the sample holding supports.
3.7 A good quality greenhouse-grade potting soil,
suitable for plant propagation, containing about 25% peat
moss, with a pH of between 5.5 and 7.6, should be used.
The moisture content of the soil should be at 100% of water
holding capacity at the beginning of the test. Place the soil
in the tray to a depth of about 8 cm (3 in.). Soil should not
be compacted.
3.8 Proper relative humidity is indicated by the
constant formation of condensate on the pitched-roof cover.
A data logger for measuring temperature and relative
humidity may be used in the test panel area to confirm test
conditions.
4. Reagents and Materials
4.1 Tween 80 dispersion agent for preparation of mold
inoculum.
4.2 Two-part epoxy for sealing unprotected edges of
samples to simulate a mid-panel section, if appropriate.
5. Test Specimens
5.1 Wood Species
Sapwood of a pine species, free of defects and fungal
growth, or another softwood species equally susceptible to
fungal growth, shall be used as an untreated negative
control and as a substrate for testing of surface- applied
treatments. The pine species used shall be reported. All test
samples should be handled with latex or vinyl gloves to
avoid soiling test surfaces.
5
Aqua-pump-1, variable flow, or equivalent
Stainless steel is well suited for this purpose
A 4-inch Muffin fan, model MU2A1, or equivalent
 STANDARD METHOD FOR EVALUATING THE RESISTANCE
E24-06 OF WOOD PRODUCT SURFACES TO MOLD GROWTH
5.2 Reference Products
At least one reference wood treatment or in-process
treated wood product shall be included in the test for
comparative purposes. The product chosen should be
commercially available and in current common use in the
marketplace. Source material from which reference samples
are prepared should be obtained from a wholesale or retail
outlet. If obtained directly from a manufacturer, the
material shall be from a production run. Unique labels
should be applied to samples to identify replicates and
source material from which the samples were cut.
5.3 Preparation of test pieces
The material should be cut into a series of 75 x 100mm
samples, 12.5mm thick or normal product thickness.
Should cutting of a treated panel into samples
compromise the integrity of a treatment or coating, a two-
part epoxy sealant may be used to seal exposed edges. A
minimum of two coats should be applied and allowed to
thoroughly dry before entry into the environment chamber.
5.4 Replication
A minimum of six replicates shall be used in each test,
with at least one replicate included in each environment
chamber if multiple chambers are used simultaneously for
one test. Samples of each control, reference, and test
product shall be retained unexposed for future comparison
with exposed samples.
6. Treatment Procedure
6.1 Surface-Applied Treatments
The test preservative supplied shall be accompanied by
an analytical certificate. The material supplied shall be a
representative sample of the product to be tested. Samples
shall be stored and handled in accordance with any written
requirements such as provided in a material safety data
sheet.
Apply the test preservative to the samples using the
process specified by the manufacturer. Examples include
immersion, spraying or flow coating. For products at an
early stage of development it is desirable to test at least
three retentions of the active ingredients ranging around the
anticipated effective retention.
Weigh all samples before and after treatment to
determine uptake. Retentions should be expressed as
micrograms per square centimeter of surface. Treat
sufficient samples to provide the necessary replication plus
additional samples to allow for elimination of samples with
preservative retention deviating more than 15% from the
mean. Substitute an appropriate alternative that falls within
this range.
Samples should be labeled to identify the preservative
and retention.
Dry treated samples in a well ventilated protected area.
If samples are stacked for drying, use supporting rods of a
material that does not react with the preservative and does
not support growth of molds.
6.2 In-Process-Treated Wood Products
Wood products should be prepared with care to ensure
uniformity of substrate, treatment, or coating. Preservative
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retentions should be expressed as kilograms per cubic meter
of wood product. If possible, reference or analytical
samples should be obtained immediately adjacent to test
sample locations on the same source material. Unique
labels should be applied to samples to identify replicates
and source material from which samples were cut.
Should cutting of source material into samples
compromise the integrity of a treatment or coating, a two-
part epoxy sealant may be used to seal exposed edges. A
minimum of two coats should be applied and allowed to
thoroughly dry before entry into the environment chamber.
7. Preparation of Inoculum
7.1 Preparation of mold cultures
Pure cultures of the following molds should be used to
prepare inoculum for soil in the environment chamber and
for application to sample surfaces:
Aureobasidium pullulans (d. By.) Arnaud, ATCC 9348
Aspergillus niger v. Tiegh, ATCC 6275
Penicillium citrinum Thom, ATCC 9849
Alternaria tenuissima group (Kunze) Wiltshire Ftk 691B
The ATCC numbers refer to strains maintained by the
American Type Culture Collection (ATCC), PO Box 1549,
Manassas, VA 20108, USA (www.atcc.org).
The cultures are introduced onto 1.5% malt extract, 2%
agar or potato dextrose agar (Difco or equivalent) 100 mm
Petri plates and incubated until spores form at about 25-
30°C. At least three plates should be prepared for
inoculation of the soil and an additional three plates should
be prepared for inoculation of the sample surfaces two
weeks later.
Other mold species may be added to, or substituted for,
the above if it is necessary to demonstrate resistance to
specific molds. For certain mold species a Type II
Biohazard safety cabinet may be required1.
7.2 Inoculation of Soil in the Environment Chamber
Place the soil in the tray in the chamber and add water
to the tank chamber to the appropriate depth. Allow the
cabinet to equilibrate at test temperature for at least 24
hours before inoculating the soil with the specified mold
suspensions.
Use 14 to 20 day old pure mold cultures to prepare mold
suspensions using the following procedure.
An inoculation suspension is prepared by adding
approximately 10 ml of distilled water containing 2 drops
of Tween 80 and rubbing the surface with a glass rod to
dislodge spores. Decant the spore suspension and rinse the
plate surface with an additional 10 ml of distilled water.
Combine the spore suspension and rinsing from all the
plates and filter through a coarse filtration medium like
glass wool or cheese cloth. Make up to about one liter with
distilled water.
Distribute the inoculum evenly over the soil surface in
the chamber. Allow two weeks of continuous operation for
the mold to sporulate and equilibrate within the
environment chamber before introducing test samples.
 STANDARD METHOD FOR EVALUATING THE RESISTANCE
E24-06 OF WOOD PRODUCT SURFACES TO MOLD GROWTH
Additional inoculum should be prepared to apply to sample
surfaces immediately before placement of samples in the
chamber. Inoculate all sample surfaces with a trigger or
chromatography sprayer. Apply evenly and lightly to
minimize surface wetting; do not apply to the point of
runoff. Application should be done in an appropriate air
exhaustion hood with samples arranged to allow for
consistency of application.
8. Exposure in Environment Chambers
Use gloves while handling samples. Samples shall be
conditioned at ambient laboratory temperature and relative
humidity before installed in the environment chamber.
Hang the inoculated samples vertically with the bottom
approximately 75mm. (3 in) above the surface of the
inoculated soil and with sufficient spacing to allow free
circulation of air. The samples are suspended with the long
dimension vertical and parallel to each other so that the
faces are perpendicular to the fan airflow. Sample spacing
should be a minimum of 2 cm between faces. A minimum
of 5 cm clearance should be allowed between the outermost
samples and the sides of the chamber. Sample locations in
the chamber should be randomized. If more than one
chamber is utilized in a test, samples should be equally
distributed between chambers, ensuring at least one
replicate of each test group is included in each chamber. A
sample location map should be recorded for each chamber
in the event mold growth was to obscure a sample label.
Viability of the mold growth in the cabinet can be
checked by placing several malt agar or potato dextrose
agar plates,8 open and face up, at several locations on the
sample supports. After 1 hour, cover plates and place in an
incubator at 32.5 ± 1°C (86 ± 2°F) for 3 days. Mold growth
should be medium-heavy to heavy and cover the complete
surface of the agar plate. If an incubator is not available,
leave the covered plates in the cabinet. Mold growth may
be less extensive at cooler chamber temperatures.
The chamber and soil are not sterile; therefore
organisms other than those in the inoculum may be present
and become established on test samples.
The environment chamber is operated at 25.0°C (or
desired temperature) for eight weeks.
9. Assessment of Mold Growth
All persons evaluating samples should, as a minimum,
wear a NIOSH N95 mask. Following 2, 4, 6 and 8 weeks
exposure within the environment chamber, samples are
removed, weighed (optional), and visually rated (unaided)
for the extent and intensity of mold growth using the scale
in Table 1. Observations of mold growth characteristics
shall be recorded. It may be appropriate for some products
to be assigned separate ratings for each face or for surfaces
of sapwood and heartwood. Observations comparing
8
Prepared agar plates can be obtained form Difco Laboratories, Inc.,
Detroit. Mich. 48232; Baltimore Biological Laboratories. Baltimore,
Md. 21218, or from equivalent sources.
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conditions of unexposed and exposed samples shall be
recorded.
Table 1
Rating Description
0 No visible growth
Mold covering up to 10% of surfaces providing
growth is not so intense or colored as to
1
obscure the sample color over more than 5% of
surfaces
Mold covering between 10% and 30% of
surfaces providing growth is not so intense or
2
colored as to obscure the sample color on more
than 10% of surfaces
Mold covering between 30% and 70% of
surfaces providing growth is not so intense or
3
colored as to obscure the sample color on more
than 30% of surfaces
Mold on greater than 70% of surfaces
providing growth is not so intense or colored as
4
to obscure the sample color over more than
70% of surfaces
Mold on 100% of surfaces or with less than
100% coverage and with intense or colored
5
growth obscuring greater than 70% of the
sample color
If the cabinet is operating properly, untreated pine
sapwood control samples should develop a minimum
average mold growth rating of at least 3 within four weeks
(when chamber is operated at 25°C). If this growth is not
obtained, the cabinet conditions are not satisfactory, there is
some interfering substance in the chamber, or inoculation
was unsuccessful. The test shall be abandoned.
Upon completion of the 8-week test, samples may be
weighed to determine moisture uptake during exposure in
the chamber. Samples may be oven-dried at 105°C to
determine pre-exposure and 8-week exposure moisture
contents.
10. Interpretation of Results
Results should be reported giving the mean, standard
deviation and range of the six (or more) replicates. Relative
resistance to mold growth is determined from comparing
average ratings of replicates of each panel product. Report
the results at the end of the 8-week exposure giving the
mean rating, standard deviation and range of the six (or
more) replicate samples. Compare results to the
performance of control and reference samples in the same
test and any dose/efficacy effects determined. The test
report shall include:
• The number of this AWPA Standard and the date of
publication.
• Species of sapwood used as untreated negative control.
• Reference wood product used and pertinent
information such as species, source, etc.
 STANDARD METHOD FOR EVALUATING THE RESISTANCE
E24-06 OF WOOD PRODUCT SURFACES TO MOLD GROWTH
• The number of replicates.
• Operating temperature of the environment chamber.
• Source of soil.
• Species and source of mold cultures.
• Description of wood products tested, including
manufacturing details, information on treatment
chemicals and loading or coatings, and any pre-test
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conditioning or exposure (leaching, weathering,
sterilization), if appropriate.
• Summary of observations of mold growth
characteristics for each wood product tested.
• Summary of observations of unexposed and exposed
sample conditions.
• Any deviations from the Standard and any special
factors that may have influenced the results.