1、Designation: D 4783 01 (Reapproved 2008)Standard Test Methods forResistance of Adhesive Preparations in Container to Attackby Bacteria, Yeast, and Fungi1This standard is issued under the fixed designation D 4783; the number immediately following the designation indicates the year oforiginal adoption
2、 or, in the case of revision, the year of last revision. A number in parentheses indicates the year of last reapproval. Asuperscript epsilon (e) indicates an editorial change since the last revision or reapproval.This standard has been approved for use by agencies of the Department of Defense.1. Sco
3、pe*1.1 These test methods cover the determination of theresistance of liquid adhesive preparations to microbial attack inthe container by challenging adhesive specimens with culturesof bacteria, yeast, or fungi, and checking for their ability toreturn to sterility. These test methods return qualitat
4、ive results.1.2 The values stated in SI units are to be regarded as thestandard. The values in parentheses are for information only.1.3 This standard does not purport to address all of thesafety concerns, if any, associated with its use. It is theresponsibility of the user of this standard to establ
5、ish appro-priate safety and health practices and determine the applica-bility of regulatory limitations prior to use. These test methodsare designed to be used by persons trained in correct micro-biological technique. Specific precautionary statements aregiven in Section 8.2. Referenced Documents2.1
6、 ASTM Standards:2D 907 Terminology of AdhesivesD 4299 Test Method for Effect of Bacterial Contaminationon Performance of Adhesive Preparations and AdhesiveFilms3D 4300 Test Methods for Ability of Adhesive Films toSupport or Resist the Growth of FungiE 640 Test Method for Preservatives in Water-Conta
7、iningCosmeticsNOTE 1Test Method E 640 is under the jurisdiction of ASTMCommittee E35 on Pesticides. The procedure in this method outlines aserial dilution method of determining plate count using a pour platetechnique.2.2 TAPPI Method:T 487 Fungus Resistance of Paper and Paperboard42.3 CSMA:Cosmetics
8、 Preservation, Method 3853. Terminology3.1 DefinitionsMany terms in these test methods aredefined in Terminology D 907.3.2 Definitions of Terms Specific to This Standard:3.2.1 adhesive preparation, nthe adhesive as packagedfor distribution, storage, and use.3.3 Abbreviations:3.3.1 PBSphosphate buffe
9、red saline.3.3.2 PDApotato dextrose agar.3.3.3 YMPGyeast malt peptone glucose (agar).4. Summary of Test Methods4.1 The adhesive specimen is challenged by inoculationwith a culture of bacteria, yeast, or fungi, which may be asingle species or a mixed culture of several species, followingthe guideline
10、s given in Note 6. The inoculated adhesivespecimen is stored at 21 to 27C (70 to 80F) for 7 days, duringwhich time cultures (streak plates) are made at preset intervals.See Note 2. At any point in the series of challenges, if theinoculated specimen shows microbial growth on the streakplates made dur
11、ing the week following the challenge (indicat-ing that it has not returned to sterility), the test is discontinued,and the sample is reported as not resistant to attack in thecontainer by the species or combination of species used as theinoculum. If the cultures show no growth, the test is repeatedw
12、ith up to four challenges. If the specimen tests out as sterilefollowing the fourth challenge, it is reported to be resistant toattack in the container by the species or combination of speciesof bacteria, fungi, or yeast used as the inoculum. At thediscretion of the biological laboratory, the test m
13、ay be discon-tinued after the second or third challenge. See Section 16 forfurther interpretation.4.2 The time necessary to kill is determined by noting theearliest streak plate to read sterile. If the 4-h plate is positive1These test methods are under the jurisdiction of ASTM Committee D14 onAdhesi
14、ves and are the direct responsibility of Subcommittee D14.30 on WoodAdhesives.Current edition approved April 1, 2008. Published April 2008. Originallyapproved in 1988. Last previous edition approved in 2001 as D 4783 01e1.2For referenced ASTM standards, visit the ASTM website, www.astm.org, orcontac
15、t ASTM Customer Service at serviceastm.org. For Annual Book of ASTMStandards volume information, refer to the standards Document Summary page onthe ASTM website.3Withdrawn.4Available from Technical Association of the Pulp and Paper Industry (TAPPI),15 Technology Parkway South, Norcross, GA 30092, ht
16、tp:/www.tappi.org.5This method is the same as Test Method E 640.1*A Summary of Changes section appears at the end of this standard.Copyright ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959, United States.and the 24-h plate is negative, the kill time could be
17、narroweddown further by repeating the challenge and making streakplates at intervals of 4, 8, 12, and 24 h following the challenge.4.3 The testing laboratory has the option of changing thetiming of the challenges, the sterility checks, and the incuba-tion period.NOTE 2Two proposed schedules for the
18、challenging and sterilitychecks are shown in Table 1 and Table 2, Schedule A for bacteria andyeast, and Schedule B for fungi. The exact format to be followed will vary,according to the convenience of the schedule to the testing laboratory andspecial circumstances relating to the problem being addres
19、sed.NOTE 3A serial-dilution plate-count method of checking for sterilitymay be used when numerical information is needed on the population ofviable organisms or the reduction in population with increasing levels ofbiocide. Letheen broth is recommended for the diluent and Letheen agarfor the pour pla
20、te. See Note 1.5. Significance and Use5.1 These test methods are used to demonstrate whether anadhesive preparation is sufficiently protected with biocide toresist attack by bacteria, yeast, and fungi during its storage life.They are patterned after methods used by biological laborato-ries serving t
21、he adhesive industry.5.2 These test methods may also be used to determine theefficacy of different biocide systems against specific microor-ganisms.5.3 These test methods are especially useful when testedagainst wild-type microorganisms which have been isolatedfrom contaminated adhesives as an aid i
22、n determining theamount and type of biocide necessary to kill or inhibit thegrowth of the contaminants. If an isolated microorganism notgenerally used as a challenge organism, is chosen as theinoculum, it is important to identify the organism and deter-mine on which medium and under what conditions
23、it will grow,in order to demonstrate the efficacy of the biocide.5.4 The results obtained when using the procedures given inthese methods apply only to the species which are used for thetesting. The test species listed in Section 9 are frequently usedby laboratories to test for antimicrobial propert
24、ies, but they arenot the only ones which could be used. Selection of the speciesto use for these test methods requires informed judgment by theTABLE 1 Schedule AProposed Bacteria and Yeast Testing, Covering 4-h, 24-h, 48-h, 72-h, and 7-Day TestsDay of Week Day no. First Challenge Second Challenge Th
25、ird Challenge Fourth ChallengeMonday (1) inoculate fresh bacterialor yeast culture. . .Tuesday 0 prepare suspension . . .Tuesday 0 inoculate specimens . . .Tuesday (0 + 4 h) streak 4-h plate . . .Wednesday 1 streak 24-h plate . . .Thursday 2 streak 48-h plate . . .Friday 3 streak 72-h plate . . .Sat
26、./Sun. 45 . . . .Monday 6 . inoculate fresh bacterialor yeast culture. .Tuesday 7 . prepare suspension . .Tuesday 7 streak 7-day plate inoculate specimens . .Tuesday (7 + 4 h) read 4-h plate streak 4-h plate . .Wednesday 8 read 24-h plate streak 24-h plate . .Thursday 9 read 48-h plate streak 48-h p
27、late . .Friday 10 read 72-h plate streak 72-h plate . .Sat./Sun. 1112 . . . .Monday 13 . . inoculate fresh bacterialor yeast culture.Tuesday 14 . . prepare suspension .Tuesday 14 read 7-day plate streak 7-day plate inoculate specimens .Tuesday (14 + 4 h) . read 4-h plate streak 4-h plate .Wednesday
28、15 . read 24-h plate streak 24-h plate .Thursday 16 . read 48-h plate streak 48-h plate .Friday 17 . read 72-h plate streak 72-h plate .Sat./Sun. 1819 . . . .Monday 20 . . . inoculate freshbacterial or yeastcultureTuesday 21 . . . prepare suspensionTuesday 21 . read 7-day plate streak 7-day plate in
29、oculate specimensTuesday (21 + 4 h) . . read 4-h plate streak 4-h plateWednesday 22 . . read 24-h plate streak 24-h plateThursday 23 . . read 48-h plate streak 48-h plateFriday 24 . . read 72-h plate streak 72-h plateSat./Sun. 2526 . . . .Monday 27 . . . .Tuesday 28 . . read 7-day plate streak 7-day
30、 plateTuesday (28 + 4 h) . . . read 4-h plateWednesday 29 . . . read 24-h plateThursday 30 . . . read 48-h plateFriday 31 . . . read 72-h plateSat./Sun. 3233 . . . .Monday 34 . . . .Tuesday 35 . . . read 7-day plateD 4783 01 (2008)2testing laboratory or by the party requesting the tests. It is alsoi
31、mportant that species which commonly attack adhesives beused. See 9.4.5.5 The presence of an active biocide carried over from theadhesive specimen to the agar could have an inhibiting effecton the growth of microorganisms, resulting in no growthduring the span of a normal incubation period, when in
32、fact,viable microorganisms are present, but their growth has beenslowed down or held in stasis. The use of Letheen agar andbroth is recommended to neutralize the effect of this carry-over.NOTE 4Letheen agar may be used for the streak plates, or if anotheragar is chosen for testing, a Letheen agar pl
33、ate could be streaked as acontrol to test against the neutralizing effect. Even more effective wouldbe diluting the challenged adhesive specimen with Letheen broth andrunning Letheen agar pour plates. See Note 1 and Note 3. Extending theincubation period of negative plates would be another safeguard
34、. Toneutralize thiazoline-based preservatives, 10 to 50 ppm of sodiumthioglycolate can be added to the medium.5.6 These test methods are dependent upon the physiologi-cal action of living microorganisms under a reported set ofconditions. Conclusions about the resistance of the test adhe-sive to micr
35、obiological attack can be drawn by comparing theresults to simultaneously run controls of known resistance. SeeX5.2 for statements regarding test repeatability.6. Apparatus6.1 In addition to the standard equipment found in any fullyequipped microbiological laboratory, the following items aresometime
36、s needed:6.1.1 Autoclave, capable of producing 103 kPa of steampressure at 121C (250F) and maintaining it for a minimum of15 min.6.1.2 Cell Counting Chamber, Petroff-Hausser, cell depth0.02 mm (or equivalent).66.1.3 Bottles, Screwcap, approximately 375 mL, BostonRounds of flint glass. Mold-A-7232-D,
37、 Finish 28-400, and6Available from most laboratory supply houses.TABLE 2 Schedule BProposed Fungi Testing, Covering 4-h, 24-h, 48-h, 72-h, and 7-Day TestsDay of Week Day no. First Challenge Second Challenge Third Challenge Fourth ChallengeFriday (10) inoculate fresh fungalculture. . .Friday (3) . in
38、oculate freshfungal culture. .Monday 0 prepare sporesuspension. . .Monday 0 inoculate specimens . . .Monday (0 + 4 h) streak 4-h plate . . .Tuesday 1 streak 24-h plate . . .Wednesday 2 streak 48-h plate . . .Thursday 3 streak 72-h plate . . .Friday 4 . . inoculate fresh fungalculture.Sat./Sun. 5, 6
39、. . . .Monday 7 . prepare sporesuspension. .Monday 7 streak 7-day plate inoculate specimens . .Monday (7 + 4 h) read 4-h plate streak 4-h plate . .Tuesday 8 read 24-h plate streak 24-h plate . .Wednesday 9 read 48-h plate streak 48-h plate . .Thursday 10 read 72-h plate streak 72-h plate . inoculate
40、 freshfungal cultureFri./Sat./Sun. 11, 12, 13 . . . .Monday 14 . . prepare sporesuspension.Monday 14 read 7-day plate streak 7-day plate inoculate specimens .Monday (14 + 4 h) . read 4-h plate streak 4-h plate .Tuesday 15 . read 24-h plate streak 24-h plate .Wednesday 16 . read 48-h plate streak 48-
41、h plate .Thursday 17 . read 72-h plate streak 72-h plate .Fri./Sat./Sun. 18, 19, 20 . . . .Monday 21 . . . prepare sporesuspensionMonday 21 . read 7-day plate streak 7-day plate inoculate specimensMonday (21 + 4 h) . . read 4-h plate streak 4-h plateTuesday 22 . . read 24-h plate streak 24-h plateWe
42、dnesday 23 . . read 48-h plate streak 48-h plateThursday 24 . . read 72-h plate streak 72-h plateFri./Sat./Sun. 25, 26, 27 . . . .Monday 28 . . read 7-day plate streak 7-day plateMonday (28 + 4 h) . . . read 4-h plateTuesday 29 . . . read 24-h plateWednesday 30 . . . read 48-h plateThursday 31 . . .
43、 read 72-h plateFri./Sat./Sun. 32, 33, 34 . . . .Monday 35 . . . read 7-day plateD 4783 01 (2008)3Black Artmold Caps BM-8041, Size 28-400, with rubber ringliners fastened to caps with steamproof adhesive.76.1.4 Constant Temperature Chamber, capable of beingmaintained at 35 6 0.5C (95 6 1F) for bacte
44、ria, or 30 60.5C (86 6 1F) for fungi, or two chambers if neededsimultaneously.6.1.5 Glass Rods, 305 mm in length having a diameter of6.3 mm.6.1.6 Hemacytometer, Levy Counting Chamber, celldepth0.1 mm, Newbauer rulings.66.1.7 Hood, laminar flow type.86.1.8 Jar, Screw Cap, round, approximately 1 L (1-
45、qt masontype) for samples.6.1.9 Pasteur Pipets.66.1.10 Pipet, 1 mL, disposable, sterile.6.1.11 Pipettes, automatic Oxford or Eppindorf, sterile, withsterile tips.66.1.12 Refrigerator, capable of maintaining temperature of4 6 1C (39 6 2F).6.1.13 Spectrophotometer, capable of measuring cell countat a
46、wavelength of 425 nm.67. Materials7.1 Beef Extract.7.2 Deionized or Distilled Water, sterile.7.3 Disinfectant Solution, Amphyll, Alcide, or equivalent.67.4 Glucose.7.5 Letheen Agar, (Difco or equivalent).67.6 Letheen Broth, (Difco or equivalent).67.7 Mycophil Agar, pH 4.7 (BBL or equivalent).67.8 Nu
47、trient Agar, (BBL or equivalent).67.9 Potato Dextrose Agar (PDA), Dehydrated, (Difco orequivalent).67.10 Phosphate buffered saline (PBS), sterile.7.11 Physiological Saline Solution, 0.85 % NaCl, sterile.7.12 Sorbitan mono-oleate polyoxyethylene.97.13 Tryptone.7.14 Tryptone Glucose Extract Agar, dehy
48、drated (Difco orequivalent).7.15 Yeast Malt Peptone Glucose Agar (YMPGA), dehy-drated (Difco or equivalent).8. Precautions8.1 These test methods employ live cultures of bacteria,fungi, and yeast, some of which are capable of causing disease,and others allergic reaction in some humans. In addition to
49、other precautions, assign laboratory personnel trained in cor-rect microbiological techniques to run these tests. Use propermicrobiological procedures in order to prevent contaminationof the cultures or of the work area. Clean and sterilize in anapproved manner all spills and all equipment coming intocontact with the cultures and the inoculated adhesive speci-mens. Also sterilize in an approved manner all cultures andcontaminated disposable equipment before discarding. See 1.3.9. Test Organisms109.1 Cultures of one or more of the following bacterials
