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166TM 100-2012职称英语考试报名时间 2012
AATCC Technical Manual/2013
Developed in 1961 by AAT CC Commit-tee RA31; revid 1965, 1981, 1988(with title change), 1993, 1999, 2012;editorially revid 1969, 1971, 1974,1985, 2009, 2010; reaffirmed 1977,1981, 1989, 1998, 2008; editorially re-vid and reaffirmed 1986, 2004.
beetle1.Purpo and Scope韩国 旅游
1.1 This test method provides a quanti-tative procedure for the evaluation of the degree of antibacterial activity. Asss-ment of antibacterial finishes on textile materials is determined by the degree of antibacterial activity intended in the u of such materials. If only bacteriostatic activity (inhibition of multiplication) is intended, a qualitative procedure which clearly demonstrates antibacterial activity as contrasted with lack of such activity by an untreated specimen may be accept-
able. However, if bactericidal activity is intended or implied, quantitative evalua-tion is necessary. Quantitative evaluation also provides a clearer picture for possi-ble us of such treated textile materials.initiative
2.Principle
2.1 This test method provides a quanti-tative procedure for the comparison and evaluation of the degree of antibacterial activity after a 24 h exposure to the test bacteria on the test fabric. After incuba-tion, the bacterial challenge is eluted from the swatches and enumerated and a per-cent reduction by the fabric specimen is calculated.
3.Terminology
3.1 activity, n.—of an antibacterial agent , a measure of effectiveness of the agent.
3.2 antibacterial agent, n.—in tex-tiles , any chemical which kills bacteria (bactericide) or interferes with the multi-plication, growth or activity of bacteria (bacteriostat).
4. Safety Precautions
NOTE: The safety precautions are for information purpos only. The pre-cautions are ancillary to the testing proce-dures and are not intended to be all inclu-sive. It is the ur’s responsibility to u safe and proper techniques in handling materials in this test method. Manufac-turers MUST be consulted for specific details such as material safety data sheets
and other manufacturer’s recommenda-tions. All OSHA standards and rules must also be consulted and followed.4.1 This test should be performed only by appropriately trained personnel. The U.S. Department of Health and Human Services publication, Biosafety in Micro-biological and Biomedical Laboratories ,should be consulted (e 12.1).
4.2 CAUTION: Some of the microor-ganisms ud in the tests are allergenic and pathogenic; i.e., capable of infecting humans and producing dia. There-fore, every necessary and reasonable pre-caution must be taken to eliminate this risk to the laboratory personnel and to personnel in the associated environment.Wear protective clothing, respiratory pro-tection, and impervious gloves when working with the organisms.
4.3 Good laboratory practices should be followed. Wear safety glass in all laboratory areas.
4.4 All chemicals should be handled with care.
4.5 An eyewash/safety shower should be located nearby for emergency u.4.6 Sterilize all contaminated samples and test materials prior to disposal.
4.7 Exposure to chemicals ud in this procedure must be controlled at or below levels t by government authorities (e.g.,Occupational Safety and Health Administration’s [OSHA] permissible ex-posure limits [PEL] as found in 29 CFR 1910.1000; e web site: v for latest version). In addition, the Ameri-can Conference of Governmental Indus-trial Hygienists (ACGIH) Threshold Limit V alues (TLVs) comprid of time weighted averages (TLV-TWA), short term exposure limits (TLV-STEL) and ceiling limits (TLV-C) are recommended as a general guide for air contaminant ex-posure which should be met (e 12.2).
5.Limitations
5.1 For a qualitative, relatively quick and easily executed method to determine antibacterial activity of textile materials,refer to AA TCC Method 147, Antibacte-rial Activity Asssment of Textile Mate-rials: Parallel Streak Method.
6.Test Organisms
6.1 Test bacteria:
6.1.1 Staphylococcus aureus , A TCC No. 6538, Gram positive organism, CIP 4.83, DSM 799, NBRC 13276, NCIMB 9518 or equivalent strain (e 12.3 and 12.4).
6.1.2 Klebsiella pneumoniae , ATCC No. 4352, Gram negative organism, CIP 104216, DSM 789, NBRC 13277,NCIMB 10341 or equivalent strain (e 12.3 and 12.4).
6.1.3 Other suitable species can also be ud depending on the intended end-u of the test sample.
7.Materials, Media and Reagents
7.1 Media and reagents. Suitable broth/agar media are:
7.1.1 Nutrient broth/agar.
7.1.2 Tryptica Soy broth/agar.
7.1.3 Brain-Heart Infusion broth/agar.7.1.4 Müller-Hinton broth/agar.
7.1.5 Slurry Inoculum Carrier (for hy-drophobic fabrics):
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Sodium Chloride 8.5 g Agar 3.0 g Sterile Distilled Water 1000 mL.7.2 Materials.
7.2.1 Incubator maintained at 37 ± 2°C (99 ± 4°F).
7.2.2 Inoculating loop.
7.2.3 Bunn Burner or equivalent.7.2.4 Water bath maintained at 45-50°C (113 -122°F).
7.2.5 Pipettes, 1 mL, sterile or equiva-lent.
7.2.6 Culture Tubes with caps; mini-mum 10 mL capacity.
7.2.7 Petri dishes, 100 mm diam. × 15mm deep, sterile.
7.2.8 Forceps, sterile.
7.2.9Stereomicroscope, minimum 40X magnification.7.2.10 Ruler.
8.Test Specimens
8.1 Preparation. The following descrip-tion will be in terms of fabric swatches.Textile materials not in fabric form can likewi be tested with the appropriate modification.
8.1.1 Size and shape of treated swatches: Cut circular swatches 4.8 ±0.1 cm (1.9 ± 0.03 in.) in diameter, from the test fabric (preferably with a steel die). Stack the swatches in a 250 mL wide-mouth glass jar with screw cap or other appropriate clod container. The number of swatches to be ud is depen-dent on the fiber type and fabric con-struction. U that amount of fabric which will absorb the 1.0 ± 0.1 mL of inoculum, and leave no free liquid in the jar. For example, 4 swatches of cotton print cloth will absorb 1 mL. The num-ber of swatches ud per jar should be reported.
AAT CC Test Method 100-2012
Antibacterial Finishes on Textile Materials: Asssment of
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AATCC Technical Manual/2013
TM 100-2012167
8.1.2 Controls. Swatches of the same fiber type and fabric construction as test sample but containing no antibacterial finish (negative control).
8.1.3 Sterilization of samples. This is optional. The method to be ud depends on the type of fiber and finish. Cotton, ac-etate and many manmade fibers can be sterilized in the autoclave. Wool can be sterilized by ethylene oxide or by inter-mittent (fractional) sterilization in flow-ing steam. The latter is also least damag-ing to certain finishes. Report method of sterilization, if ud.
9. Procedure
9.1 Size of inoculum per sample. Ap-ply 1.0 ± 0.1 mL of an appropriate dilu-tion of a 24 h broth culture of the test or-ganism so that recovery from (1)untreated control fabric swatches or (2)treated test fabric swatches at “0” contact time (plated as soon as possible after in-oculation) will show counts of 1-2 × 105organisms. The dilution of the test organ-ism should be made in nutrient (or appro-priate) broth (e 7.1.1, 7.1.5 and 12.5).9.1.1 Inoculation of fabrics. When using Staphylococcus aureus , shake a 24h culture and let stand for 15-20 min be-fore preparing the inoculum.* Place the swatches parately in sterile petri dishes and u a microliter pipette to inocul影视动漫学校
ate them making sure that there is even dis-tribution of the inoculum (e 12.6).Transfer the swatches aptically to the jar. Screw the jar tops on tightly to pre-vent evaporation.
9.1.2 As soon as possible after inocula-tion (“0” contact time), add 100 ± 1 mL of neutralizing solution to each of the jars containing the inoculated untreated con-trol swatches, the inoculated treated test swatches and the uninoculated treated test swatches.
9.1.3 The neutralizing solution should include ingredients to neutralize the spe-cific antibacterial fabric treatment and to take care of any pH requirements of the fabrics (from finishes, antibacterial agents, etc.). The neutralizing solution employed should be reported (e 12.7).9.1.4 Shake the jars vigorously for one minute. Make rial dilutions with sterile distilled water and plate (in duplicate) on nutrient (or appropriate) agar. Dilutions of 100, 101, 102 are usually suitable.
9.1.5 Incubation over contact periods.Incubate additional jars containing inocu-lated untreated control swatches and jars containing inoculated treated test
swatches at 37 ± 2°C (99 ± 3°F) for 18-24 h. Similar jars may be incubated over other periods (e.g., 1 or 6 h) to provide information about the bactericidal activ-ity of the treatment over such periods.9.1.6 Sampling of inoculated and incubated swatches. After incubation,add 100 ± 1 mL of ne
utralizing solution to jars containing untreated control swatches and to jars containing treated test swatches. Shake the jars vigorously for one minute. Make rial dilutions and plate (in duplicate) on nutrient (or appro-priate) agar. Dilutions of 100, 101, 102 are usually suitable for treated test fabrics.Several different dilutions may be re-quired for untreated control fabrics de-pending on the incubation period.
9.1.7 Incubate all plates for 48 h at 37± 2°C (99 ± 3°F) or other optimal temperature.
10. Evaluation
10.1 Report bacterial counts as the number of bacteria per sample (swatches in jar) not as the number of bacteria per mL of neutralizing solution. Report “0”counts at 100 dilution as “less than 100.”10.2 Calculate percent reduction of bacteria by the specimen treatments by one of the following formulas:1)
100(B – A )/B = R
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R =% reduction
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A =the number of bacteria recovered
from the inoculated treated test specimen swatches in the jar in-cubated over the desired contact period
B =the number of bacteria recovered
from the inoculated treated test specimen swatches in the jar im-mediately after inoculation (at “0” contact time)2)
100(C – A )/C = R
where:
C =the number of bacteria recovered
from the inoculated untreated control specimen swatches in the jar immediately after inoculation (at “0” contact time)
If “B and C ” are not similar, the larger number should be ud. If “B ” and “C ”are not significantly different, (B + C )/2should be ud as follows:3)
100(D – A )/D = R
where:
D =(B + C )/2
10.3 If an untreated control is not available, u the following calculation which allows for any background organ-isms that might interfere with the test.
Bg =100 [(B – E ) – (A – F )/B – E ]where:
A ,
B =(e 10.2)
马尔克斯去世E =the number of bacteria initially
recovered from the uninocu-lated treated test sample (exist-ing background organisms)F =The number of bacteria recov-ered from the uninoculated,pre-wet treated test sample after incubation in the jar over the desired contact period (existing background organisms after contact period)
Bg =background organisms 10.4 For a valid test there should be:(1) “0” colonies of test organism recov-ered from the uninoculated treated test specimen swatches and (2) a significant increa in the numbers of bacteria recov-ered from the inoculated untreated con-trol specimen swatches incubated for the specified contact time over the numbers of bacteria recovered from the inoculated untreated specimen swatches at “0” con-tact time (immediately after inoculation).This applies only if dilution was made in broth (e 9.1 and 12.5).
10.5 Report percent reduction of bac-teria by the specimen treatment against each test organism.
10.6 The criterion for passing the test must be determined by the interested parties.
10.7 Report the dilution medium ud.
11. Precision and Bias
rai和ri11.1 Studies (e 12.8) indicate the fol-lowing within-laboratory precision of the Standard Plate Count (SPC) Test: (a)among-analyst variation of 18% and (b)within-analyst variation of 8%.
12. Notes and References
12.1 Publication available from U.S. De-partment of Health and Human Services CDC/NIH-HHS Publication No. (CDC) 84-8395;web site: v.
12.2 Booklet available from Publications Office, ACGIH, Kemper Woods Center, 1330Kemper Meadow Dr., Cincinnati OH 45240;tel: 513/742-2020; web site: www.acgih.12.3 ATCC is the American Type Culture Collection (USA), P.O. Box 1549, Manassas V A 20108; tel: 703/365-2700; fax: 703/365-2701; web site: www.atcc. CIP is the Pas-teur Institute Collection (France), DSM is the German Collection of Microorganisms and Cell Cultures (Germany), NBRC is the NITE Biological Resource Center (Japan), NRRL is the Northern Regional Rearch Lab (USA),NCIMB is the National Collection of Indus-trial Bacteria (UK), and CUG is the Culture Collection University of Göteborg (Sweden).Equivalent bacteria strains obtained from agencies of the World Federation of Culture Collection (WFCC) may be ud by agree-ment between the interested parties. The strains ud in the test shall be documented
*Using a 1 mL pipette, pad the inoculum carefully onto the fabric. If a strain of Pudomonas that forms a pelli-cle is ud, avoid including fragments of the pellicle in the inoculum.
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168TM 100-2012
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with their supply source.
12.4 Consistent and accurate testing re-quires maintenance of a pure, uncontaminated,nonmutant test culture. Avoid contamination by u of good sterile technique in plating and transferring. Avoid mutation by strict adher-ence to monthly stock transfers. Check culture purity by making streak plates periodically and obrving for single species-characteristic type of colonies.12.5 The dilution of the test organism may be prepared in sterile 0.85% saline solution or suitable buffer if a steady-state culture is de-sired during the contact period with a fabric or in the slurry inoculum carrier when hydropho-bic fabrics are being tested.
12.6 A surfactant may be added to the dilu-tion medium to enhance wetting of hydropho-bic fabrics. The surfactant must be shown not to cau a reduction in bacterial numbers, by prior testing at the intended u concentration. Report the u and concentration of surfactant ud.12.7 If sterile distilled water is ud in the place of a neutralizing solution, there will al-ways be the possibility that some of the bio-cide will be carried over.
12.8 Peeler, J. T.; Leslie, J. W.; Mesr,J.W. Replicate counting errors by analysts and bacterial colony counters. J. Food Protec-tion , V ol. 45, 1982, pp238-240.
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