1、BSI Standards PublicationBS ISO 13629-2:2014T e x t i l e s D e t e r m i n a t i o n o f antifungal activity of textile productsPart 2: Plate count methodBS ISO 13629-2:2014 BRITISH STANDARDNational forewordThis British Standard is the UK implementation of ISO 13629-2:2014. The UK participation in
2、its preparation was entrusted to TechnicalCommittee TCI/80, Chemical testing of textiles.A list of organizations represented on this committee can be obtained on request to its secretary.This publication does not purport to include all the necessary provisions of a contract. Users are responsible fo
3、r its correct application. The British Standards Institution 2014.Published by BSI Standards Limited 2014ISBN 978 0 580 80749 7ICS 59.080.01Compliance with a British Standard cannot confer immunity from legal obligations.This British Standard was published under the authority of the Standards Policy
4、 and Strategy Committee on 31 July 2014.Amendments/corrigenda issued since publicationDate Text affectedBS ISO 13629-2:2014 ISO 2014Textiles Determination of antifungal activity of textile products Part 2: Plate count methodTextiles Dtermination de lactivit antifongique des produits textiles Partie
5、2: Mthode par dnombrement sur plaque de gloseINTERNATIONAL STANDARDISO13629-2First edition2014-06-15Reference numberISO 13629-2:2014(E)BS ISO 13629-2:2014ISO 13629-2:2014(E)ii ISO 2014 All rights reservedCOPYRIGHT PROTECTED DOCUMENT ISO 2014All rights reserved. Unless otherwise specified, no part of
6、 this publication may be reproduced or utilized otherwise in any form or by any means, electronic or mechanical, including photocopying, or posting on the internet or an intranet, without prior written permission. Permission can be requested from either ISO at the address below or ISOs member body i
7、n the country of the requester.ISO copyright officeCase postale 56 CH-1211 Geneva 20Tel. + 41 22 749 01 11Fax + 41 22 749 09 47E-mail copyrightiso.orgWeb www.iso.orgPublished in SwitzerlandBS ISO 13629-2:2014ISO 13629-2:2014(E) ISO 2014 All rights reserved iiiContents PageForeword ivIntroduction v1
8、Scope . 12 Normative reference . 13 Terms and definitions . 14 Principle 25 Safety precaution . 26 Reference fungi 27 Apparatus . 28 Reagents and culture media . 48.1 Pure water . 48.2 Anionic surfactant 48.3 Culture medium . 49 Fungi preservation and use . 610 Spore suspension . 610.1 General . 610
9、.2 Suspending spores in culture media 710.3 Collection and dispersion of spore suspension from a culture medium . 710.4 Filtering to remove hyphae and spore thread 710.5 Using centrifuge and re-suspension to remove supernatant 710.6 Confirming the concentration of spore suspension 810.7 Adjusting sp
10、ore suspension for testing . 810.8 Enumeration of inoculum 811 Testing procedure . 811.1 Inoculation and preparation of specimens . 811.2 Plate count method procedure . 1112 Test results .1312.1 Judgment of test effectiveness 1312.2 Calculation of antifungal activity value . 1313 Test report 14Annex
11、 A (normative) Fungi used in this part of ISO 13629 15Annex B (informative) Antifungal efficacy .16Bibliography .17BS ISO 13629-2:2014ISO 13629-2:2014(E)ForewordISO (the International Organization for Standardization) is a worldwide federation of national standards bodies (ISO member bodies). The wo
12、rk of preparing International Standards is normally carried out through ISO technical committees. Each member body interested in a subject for which a technical committee has been established has the right to be represented on that committee. International organizations, governmental and non-governm
13、ental, in liaison with ISO, also take part in the work. ISO collaborates closely with the International Electrotechnical Commission (IEC) on all matters of electrotechnical standardization.The procedures used to develop this document and those intended for its further maintenance are described in th
14、e ISO/IEC Directives, Part 1. In particular the different approval criteria needed for the different types of ISO documents should be noted. This document was drafted in accordance with the editorial rules of the ISO/IEC Directives, Part 2 (see www.iso.org/directives).Attention is drawn to the possi
15、bility that some of the elements of this document may be the subject of patent rights. ISO shall not be held responsible for identifying any or all such patent rights. Details of any patent rights identified during the development of the document will be in the Introduction and/or on the ISO list of
16、 patent declarations received (see www.iso.org/patents).Any trade name used in this document is information given for the convenience of users and does not constitute an endorsement.For an explanation on the meaning of ISO specific terms and expressions related to conformity assessment, as well as i
17、nformation about ISOs adherence to the WTO principles in the Technical Barriers to Trade (TBT) see the following URL: Foreword - Supplementary informationThe committee responsible for this document is ISO/TC 38, Textiles.ISO 13629 consists of the following parts, under the general title Textiles Det
18、ermination of antifungal activity of textile products: Part 1: Luminescence method Part 2: Plate count methodiv ISO 2014 All rights reservedBS ISO 13629-2:2014ISO 13629-2:2014(E)IntroductionThis part of ISO 13629 adopts the plate count method as a basis of quantitative determination of antifungal ac
19、tivity.The following are characteristics of the plate count method: conventional method which is easy to operate in bacteriological laboratories; no need to use special apparatus such as a lumino photometer; long history and common procedure. ISO 2014 All rights reserved vBS ISO 13629-2:2014BS ISO 1
20、3629-2:2014Textiles Determination of antifungal activity of textile products Part 2: Plate count method1 ScopeThis part of ISO 13629 specifies a test method for quantitative determination of antifungal activity by plate count method.This part of ISO 13629 is applicable to various kinds of textile pr
21、oducts such as fibres, yarns, fabrics, clothing, bedclothes, home furnishings, and other miscellaneous goods.2 Normative referenceThe following documents, in whole or in part, are normatively referenced in this document and are indispensable for its application. For dated references, only the editio
22、n cited applies. For undated references, the latest edition of the referenced document (including any amendments) applies.ISO 105-F02, Textiles Tests for colour fastness Part F02: Specification for cotton and viscose adjacent fabricsISO 7218, Microbiology of food and animal feeding stuffs General re
23、quirements and guidance for microbiological examinations3 Terms and definitionsFor the purposes of this document, the following terms and definitions apply.3.1control fabricfabric used to validate the growth condition of test fungiNote 1 to entry: Control specimens are sampled from the control fabri
24、c.Note 2 to entry: The control fabric may be the same fabric as the fabric to be tested but without antifungal treatment. If this is not available, a 100 % cotton fabric without fluorescent brighteners or other finish, complying with the requirements of ISO 105-F02, is used as control fabric, after
25、washing at the temperature of 60 C without detergents or any brighteners, with mechanical agitation and rinsing.3.2antifungal agentchemical agent to prevent or mitigate the growth of fungi or to reduce the number of fungi3.3antifungal treatmenttreatment to prevent or mitigate the growth of fungi or
26、to reduce the number of fungi3.4spore suspensionliquid with evenly dispersed fungal spores in sterilized water containing an anionic surfactantINTERNATIONAL STANDARD ISO 13629-2:2014(E) ISO 2014 All rights reserved 1BS ISO 13629-2:2014ISO 13629-2:2014(E)3.5plate count methodmethod in which the numbe
27、r of fungi present after incubation is calculated by counting the number of colonies according to a ten-time dilution methodNote 1 to entry: The results are expressed in CFU (Colony Forming Unit).3.6neutralizerchemical agent used to inactivate, neutralize, or quench the antifungal properties of anti
28、fungal agents4 PrincipleA test specimen and a control specimen are inoculated with spore suspension of reference fungi and incubated at 30 C for 48 h.In this part of ISO 13629, fungal growth is quantitatively determined by the visual counting of colonies on the agar plate as CFU and the fungal activ
29、ity is calculated by CFU.In case the test specimen absorbs water, the absorption method is recommended. In case the test specimen does not absorb water, the transfer method is recommended.5 Safety precautionThe test method specified herein requires use of fungi.According to ISO 7218, this test shall
30、 be performed only by personnel with training and experience in microbiological techniques.All regulations, rules, and recommendations regarding appropriate safety precautions in the country concerned may be consulted and followed.6 Reference fungiThe fungi to be used shall be selected from Annex A,
31、 Table A.1.The equivalent fungi types obtained from other agencies of the World Federation for Culture Collection (WFCC) shall be used as agreed upon between interested parties.The strain number and supply source of the fungi used shall be stated in the test report.7 ApparatusUsual laboratory appara
32、tuses and, in particular, the following apparatuses are used. When relevant, the items have to be sterilized before using.7.1 Gauze, sterilized.7.2 Petri dish, made of glass or plastic, with a diameter of about 60 mm or 90 mm.7.3 Autoclave, capable of maintaining the temperature of (121 2) C (equiva
33、lent to 103 kPa).7.4 Platinum loop, with a loop of 2 mm to 4 mm in diameter (or plastic equivalent).7.5 L-shaped platinum colony hook (or plastic equivalent).2 ISO 2014 All rights reservedBS ISO 13629-2:2014ISO 13629-2:2014(E)7.6 Incubator, capable of maintaining a temperature in a range from 25 C t
34、o 37 C with a tolerance of 2 C.7.7 Vial, capacity of 30 ml screw-top glass vial with polytetrafluoroethylene or silicone gasket and polypropylene cap. It shall be carefully washed in alkaline or neutral detergent, rinsed, and dried.7.8 Glass funnel.7.9 Pipettes, capacity of 0,2 ml, 1 ml, 5 ml, and 1
35、0 ml with a tolerance of 0,5 % or less and with a tip made of glass or plastic.7.10 Pasteur pipette, for microbiological testing (or plastic equivalent).7.11 Conical flask, capacity of 100 ml to 500 ml.7.12 Tweezers, made of material which can be sterilized.7.13 Centrifuge, with centrifugal accelera
36、tion of approximate 2 000 g.7.14 Centrifuge tube, used for centrifuge.7.15 Hemacytometer, capable of measuring 1 106cells/ml to 3 106cells/ml.7.16 Microscope, capable of 200x magnification.7.17 Ultrasonic cleaner, compact for experiment tools, with frequency of approximately 30 kHz to 50 kHz.7.18 pH
37、 meter, with glass electrodes for biochemical testing or equivalent pH paper.7.19 Erlenmeyer flask, capacity of 100 ml.7.20 Cutting template, made of stainless steel with a diameter of (3,8 0,1) cm.7.21 Stainless steel cylinder, with a weight of (200 10) g and a diameter of (3,5 0,1) cm.7.22 Shaker,
38、 capable of producing a Vortex shaking action.7.23 Paddle blender Stomacher-type, capable of speed 6 blows/s to 8 blows/s with the corresponding disposable containers.7.24 Humidity chamber, a tropical chamber or other container capable of maintaining a high humidity atmospheric condition.7.25 Refrig
39、erator, capable of maintaining a temperature of between 2 C and 8 C with a tolerance of 2 C.7.26 Freezers, adjustable to a temperature below 70 C and below 20 C with a tolerance of 2 C.7.27 Balance, capable of measuring 0,01 g as the readability. ISO 2014 All rights reserved 3BS ISO 13629-2:2014ISO
40、13629-2:2014(E)7.28 Disposable plastic bags, suitable for containing food products to be used for the shake-out of sample.7.29 Microbiological safety cabinet (MSC Type II), designed for microbiological tests use, or other system with equivalent performances.7.30 Water baths, one capable of maintaini
41、ng a constant temperature of (46 2) C and another capable of maintaining a temperature of 70 C to 90 C.8 Reagents and culture mediaReagents used in tests shall be of analytical grade and/or suited for microbiological purposes.Dehydrated products available on the commercial market are recommended for
42、 use in preparing the culture media strictly in accordance with the manufacturers instructions.8.1 Pure waterAnalytical-grade water for microbiological media preparation which is freshly distilled and/or ion-exchanged and/or ultra-filtered and/or filtered with RO (reverse osmosis).It shall be free f
43、rom all toxic or fungi inhibitory substances.8.2 Anionic surfactantDioctyl sodium sulfosuccinate to prepare spore suspension. The concentration of the anionic surfactant in pure water (8.1) is 50 mg/l. Sterilize this solution by an autoclave (7.3) at 121 C for 20 min.8.3 Culture mediumUse a culture
44、medium prepared as described below. Commercially prepared items may be used after appropriate validation.Culture media which will not be used immediately after preparation shall be stored at 5 C to 10 C and discarded after one month.8.3.1 Sabouraud dextrose broth (SDB)Peptone 10 gDextrose 20 gPure w
45、ater 1 000 mlpH after sterilization 5,6 0,28.3.2 Potato dextrose agar (PDA)Potato infusion from 200 gDextrose 20 gAgar 15 gPure water 1 000 mlpH after sterilization 5,6 0,24 ISO 2014 All rights reservedBS ISO 13629-2:2014ISO 13629-2:2014(E)This medium encourages mould sporulation.8.3.3 Sabouraud dex
46、trose agar (SDA)Pepsic meat peptone 10 gDextrose 40 gAgar 15 gPure water 1 000 ml (final volume)Follow the indications of the supplier.pH after sterilization 5,6 0,2NOTE This medium will be used for the transfer method.8.3.4 Slant culture8.3.4.1 Pour approximately 10 ml of pre-heated and fully disso
47、lved PDA (described in 8.3.2) into a sterilized test tube.8.3.4.2 Put a cotton plug on and sterilize it with steam after sterilization.8.3.4.3 Place the test tube at an approximately 15 angle against a level surface on a clean laboratory table, and leave the contents to solidify.8.3.4.4 When there i
48、s no bleed water on the solidified agar, dissolve, and solidify it again for use.8.3.5 Neutralizing solution, SCDLP mediumPolysorbate 80 30 gEgg yolk lecithin 3 gHistidine hydrochloride 1 gMeat or casein peptone 1 gSodium chloride (NaCl) 4,3 gMonopotassium phosphate 3,6 gDisodium phosphate dehydrate
49、 7,2 gWater 1 000 ml (final volume)pH after sterilization 7,2 0,2When sufficient neutralizing power cannot be achieved, the content of polysorbate 80 or lecithin may be adjusted or another neutralizing agent may be added. Commercial solutions of neutralizer can be used after having tested their efficacy (see Annex B). The use of any unspecified neutralizer shall be recorded along with the name and concentration. ISO 2014 All rights reserved 5BS ISO 13629-2:2014ISO 13629-2:201
