ASTM D5583-00

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Designation: D 5583 – 00
Standard Test Method for
Detection and Estimation of Retention of Wood
Preservatives by Aspergillus Bioassaying
This standard is issued under the fixed designation D 5583; 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 its typical black spores around samples containing an inhibi-
tory amount of preservative; this indicates the presence of
1.1 This test method covers a non-chemical, biological
preservative, and the size of the white area (zone of effect) is
procedure for assaying wood for approximate preservative
the index of preservative retention (see Fig. 1 and Fig. 2). The
content (Note 1). It requires minimal equipment and technical
relation of retention to zone of effect is determined for each
proficiency. No costly equipment is needed other than a small
preservative,fromassaysofspecimenscontainingagradientof
autoclave for preparing fungal media. It does not require
retentions of the preservative. (See example for pentachlo-
composite samples, such as called for in Specification D 1760
rophenol in Fig. 3.) Hereafter, this is called the reference
to ascertain preservation retentions, therefore it is used to
relations.
assess retention at individual locations.
NOTE 1—With appropriate, simple adaptation, the method can be used 4. Significance and Use
withotherproductsbesideswood,suchasfungicidalpaintsandglues.The
4.1 This test method is intended for use with any preserva-
assaying is done wherever it is convenient, since aseptic precautions are
tive that is of uniform composition and not so tightly bound to
not required except for maintaining a pure stock culture of the assay
the wood or so water insoluble that it cannot diffuse in the
fungus.
culture medium on which the assay fungus is grown (1, 2, 3).
1.2 The values stated in SI units are to be regarded as the
It is especially applicable to monitoring non-pressure treated
standard. The inch-pound units given in parentheses are for
millwork and other wood items (4), but it can be used to
information only.
determine approximate preservative retentions at prescribed
1.3 This standard does not purport to address all of the
locations in pressure-treated wood (5). When measuring lon-
safety concerns, if any, associated with its use. It is the
gitudinal preservative penetration, as is especially useful with
responsibility of the user of this standard to establish appro-
millwork,thewoodcanbesampledontheendgrain.Theassay
priate safety and health practices and determine the applica-
also provides a reliable means of observing losses of preser-
bility of regulatory limitations prior to use.
vative retention in wood during service (or experimentally
subjected to particular environmental conditions). This test
2. Referenced Documents
method is not intended for comparing preservatives nor for
2.1 ASTM Standards:
estimating retentions of a preservative of variable composition,
D 1760 Specification for Pressure Treatment of Timber
such as creosote. It can be used, however, for comparing
Products
relative potencies of such preservative.
3. Summary of Test Method
5. Apparatus
3.1 Small wood specimens of prescribed size are placed on
5.1 Conventional Equipment, for culturing and aseptic han-
nutrient agar in a Petri dish freshly seeded with spores of
dling of fungi, such as autoclave, refrigerator, transfer needle,
Aspergillus niger, a black mold, and the culture is allowed to
and gas or alcohol burner.
incubateforthreetofourdays.Thefungusisunabletoproduce
5.2 Glassware, 100 by 15 mm (approximately) Petri dishes,
glass or plastic, 250 mL Erlenmeyer flasks, and a few 150 by
22 mm (approximately) test tubes.
This test method is under the jurisdiction of ASTM Committee D07 on Wood
and is the direct responsibility of Subcommittee D07.06 on Treatments for Wood 5.3 RoomorChamber,inwhichtemperatureiscontrolledat
Products.
22 to 27°C (72 to 82°F).
Current edition approved Oct. 10, 2000. Published December 2000. Originally
published as D 5583 – 94. Last previous edition D 5583 – 94.
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 boldface numbers in parentheses refer to the list of references at the end of
the ASTM website. this test method.
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D5583–00
NOTE 1—The black area shows dense coverage by black spores of the fungus j 6.
FIG. 1 Zone of Effect (d) in an Aspergillus Bioassay of a Pentachlorophenol-Treated Block
NOTE 1—Penta retentions (PCF) here = solution
FIG. 2 An Aspergillus niger Bioassay of Small-Block Specimens
strength 3 approximately 0.369.
Treated to Selected Retentions With Tributyltin Oxides
FIG. 3 Example of Reference Relation Between Strength of
Pentachlorophenol Solution and Zone of Effect in an Aspergillus
5.4 Atomizer, delivering a fine spray (optional).
niger Bioassay Using Small-Treated Blocks
5.5 Tools, such as increment borer, plug cutter, saw, and
knife for extracting samples and cutting them to desired
specimen size.
7. Culture Medium
7.1 The nutrient medium for carrying the stock culture, and
6. Assay Fungus
4 for the bioassaying, shall be potato-dextrose agar (PDA) (Note
6.1 The assay fungus shall be Aspergillus niger Van Tiegh
2). Aspergillus niger tends to grow more rapidly and sporulate
(for example, culture ATCC 64045 from the American Type
more profusely on potato-dextrose than on malt-agar. A com-
Culture Collection).
mercial dry-mix preparation, made up according to directions,
is satisfactory.
Cultures are available from ATCC, 12301 Parklawn Drive, Rockville, MD
20852. NOTE 2—ThePDAmaybepreparedinthelaboratorybycombininghot
D5583–00
water extract of 200 g of potatoes, 20 g of dextrose (glucose), and 15 g of
delivers fine droplets. This operation is facilitated by holding
agar per litre.
the dishes slightly off the vertical. One or two bursts from the
atomizer is sufficient to provide suitably dense and uniform
8. Assay Samples
spore coverage.To prevent spores escaping into the laboratory,
8.1 For standard use, acquire samples of the wood being
this operation should be done under a fume hood (Note 3).
assayed with a conventional increment borer or plug cutter.
NOTE 3—Remaining spore suspension can be stored up to six to eight
8.2 When monitoring treated products for quality control of
weeks for further use if kept near 5°C.
the treating, always take samples at the same location on the
10.5.2 Seeding by Flooding—Keep the spores uniformly
product.
suspended by shaking the flask and pour about 4 mL of the
8.3 The number of samples required may vary with product
spore suspension onto the hardened culture medium in each
and precision needed (see 11.1).
dish.Gentlyrockthedishafewsecondstodistributethespores
9. Assay Specimens
uniformlyoverthemediumandpourofftheexcesssuspension.
10.6 Insert Assay Specimens:
9.1 Reducethesamplestosmallspecimensfortheassaying.
10.6.1 Immediately after seeding, place three or four assay
For standard use, the specimens shall be increment core or
specimens on the agar medium (Note 4). Space the specimens
circular wafers cut from plugs. The core segments shall be at
1 uniformly around the dish, approximately 1 cm from the edge.
least 6 mm ( ⁄4 in.) long. Wafers shall be complete or cut in
Replace the dish covers and mark them to identify the
half, and 3 cm ( ⁄8 in.) thick. If preservative retentions are to be
individual specimens.
estimated,thespecimensshouldbeoftheformsizeandkindof
wood as those used to derive the reference relation.
NOTE 4—More specimens can be used per plate if the preservative
9.2 To assay the preservatives at various depths in the
diffusion is moderate enough to preclude interference between zones of
sampled wood, take the increment core segments and identify effect.
serially.
10.6.2 Once the dishes have been seeded, no precautions
againstcontaminationareneededotherthantokeepthelidson.
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