1、BSI Standards PublicationBS ISO 16000-21:2013Indoor airPart 21: Detection and enumeration ofmoulds Sampling from materialsBS ISO 16000-21:2013 BRITISH STANDARDNational forewordThis British Standard is the UK implementation of ISO16000-21:2013.The UK participation in its preparation was entrusted to
2、TechnicalCommittee EH/2/5, Emissions to internal environments.A list of organizations represented on this committee can beobtained on request to its secretary.This publication does not purport to include all the necessaryprovisions of a contract. Users are responsible for its correctapplication. The
3、 British Standards Institution 2014. Published by BSI StandardsLimited 2014ISBN 978 0 580 78715 7ICS 13.040.20Compliance with a British Standard cannot confer immunity fromlegal obligations.This British Standard was published under the authority of theStandards Policy and Strategy Committee on 28 Fe
4、bruary 2014.Amendments issued since publicationDate Text affectedBS ISO 16000-21:2013 ISO 2013Indoor air Part 21: Detection and enumeration of moulds Sampling from materialsAir intrieur Partie 21: Dtection et dnombrement des moisissures chantillonnage partir de matriauxINTERNATIONAL STANDARDISO16000
5、-21First edition2013-12-15Reference numberISO 16000-21:2013(E)BS ISO 16000-21:2013ISO 16000-21:2013(E)ii ISO 2013 All rights reservedCOPYRIGHT PROTECTED DOCUMENT ISO 2013All rights reserved. Unless otherwise specified, no part of this publication may be reproduced or utilized otherwise in any form o
6、r 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 in the country of the requester.ISO copyright officeCase postale 56 CH-1
7、211 Geneva 20Tel. + 41 22 749 01 11Fax + 41 22 749 09 47E-mail copyrightiso.orgWeb www.iso.orgPublished in SwitzerlandBS ISO 16000-21:2013ISO 16000-21:2013(E) ISO 2013 All rights reserved iiiContents PageForeword ivIntroduction vi1 Scope . 12 Normative references 13 Terms and definitions . 14 Princi
8、ple of method . 25 Apparatus and materials 25.1 Equipment for sampling . 25.2 Equipment for preparing the agar plates . 35.3 Equipment for processing the bulk samples . 36 Culture media and reagents 36.1 General . 36.2 Dichlorane 18 % glycerol agar (DG-18). 36.3 Malt extract agar 46.4 Potato dextros
9、e agar 46.5 Dilution buffer . 56.6 Staining solution 57 Measurement procedure 67.1 Sampling from surfaces . 67.2 Bulk sampling 77.3 Transport and storage 77.4 Direct microscopy 77.5 Suspension of material and swab samples . 78 Quality assurance 89 Sampling protocol . 810 Performance characteristics
10、8Annex A (informative) Sample exchange for method validation . 9Bibliography .11BS ISO 16000-21:2013ISO 16000-21:2013(E)ForewordISO (the International Organization for Standardization) is a worldwide federation of national standards bodies (ISO member bodies). The work of preparing International Sta
11、ndards 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-governmental, in liaison with ISO, also
12、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 the ISO/IEC Directives, Part 1. In
13、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 possibility that some of the elements
14、 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 patent declarations received (s
15、ee 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 information about ISOs adherence
16、 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 146, Air quality, Subcommittee SC 6, Indoor air.ISO 16000 consists of the following parts, under the general title Indoor a
17、ir: Part 1: General aspects of sampling strategy Part 2: Sampling strategy for formaldehyde Part 3: Determination of formaldehyde and other carbonyl compounds in indoor air and test chamber air Active sampling method Part 4: Determination of formaldehyde Diffusive sampling method Part 5: Sampling st
18、rategy for volatile organic compounds (VOCs) Part 6: Determination of volatile organic compounds in indoor and test chamber air by active sampling on Tenax TAsorbent, thermal desorption and gas chromatography using MS or MS-FID Part 7: Sampling strategy for determination of airborne asbestos fibre c
19、oncentrations Part 8: Determination of local mean ages of air in buildings for characterizing ventilation conditions Part 9: Determination of the emission of volatile organic compounds from building products and furnishing Emission test chamber method Part 10: Determination of the emission of volati
20、le organic compounds from building products and furnishing Emission test cell method Part 11: Determination of the emission of volatile organic compounds from building products and furnishing Sampling, storage of samples and preparation of test specimens Part 12: Sampling strategy for polychlorinate
21、d biphenyls (PCBs), polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs) and polycyclic aromatic hydrocarbons (PAHs)iv ISO 2013 All rights reservedBS ISO 16000-21:2013ISO 16000-21:2013(E) Part 13: Determination of total (gas and particle-phase) polychlorinated dioxin-like
22、 biphenyls (PCBs) and polychlorinated dibenzo-p-dioxins/dibenzofurans (PCDDs/PCDFs) Collection on sorbent-backed filters Part 14: Determination of total (gas and particle-phase) polychlorinated dioxin-like biphenyls (PCBs) and polychlorinated dibenzo-p-dioxins/dibenzofurans (PCDDs/PCDFs) Extraction,
23、 clean-up and analysis by high-resolution gas chromatography and mass spectrometry Part 15: Sampling strategy for nitrogen dioxide (NO2) Part 16: Detection and enumeration of moulds Sampling by filtration Part 17: Detection and enumeration of moulds Culture-based method Part 18: Detection and enumer
24、ation of moulds Sampling by impaction Part 19: Sampling strategy for moulds Part 20: Detection and enumeration of moulds Determination of total spore count Part 21: Detection and enumeration of moulds Sampling from materials Part 23: Performance test for evaluating the reduction of formaldehyde conc
25、entrations by sorptive building materials Part 24: Performance test for evaluating the reduction of volatile organic compound (except formaldehyde) concentrations by sorptive building materials Part 25: Determination of the emission of semi-volatile organic compounds by building products Micro-chamb
26、er method Part 26: Sampling strategy for carbon dioxide (CO2) Part 27: Determination of settled fibrous dust on surfaces by SEM (scanning electron microscopy) (direct method) Part 28: Determination of odour emissions from building products using test chambers Part 29: Test methods for VOC detectors
27、Part 30: Sensory testing of indoor air Part 31: Measurement of flame retardants and plasticizers based on organophosphorus compounds Phosphoric acid ester Part 32: Investigation of buildings for pollutants and other injurious factors InspectionsThe following parts are under preparation: Part 33: Det
28、ermination of phthalates with gas chromatography/mass spectrometry (GC/MS) Part 34: Strategies for the measurement of airborne particles (PM 2,5 fraction) Part 35: Measurement of polybrominated diphenylether, hexabromocyclododecane and hexabromobenzeneA test method for the reduction rate of airborne
29、 bacteria by air purifiers using a test chamber will form a future part 36. ISO 2013 All rights reserved vBS ISO 16000-21:2013ISO 16000-21:2013(E)IntroductionMould is a common name for filamentous fungi from different taxonomic groups (ascomycetes, zygomycetes, and their anamorphic states formerly k
30、nown as deuteromycetes or fungi imperfecti). They form a mycelium and spores by which they become visible macroscopically. Most spores are in the size range of 2 m to 10 m, some up to 30 m, and only few up to 100 m. Spores of some mould genera are small and become airborne very easily (e.g. Aspergil
31、lus, Penicillium) while others are bigger and/or embedded in a slime matrix (e.g. Stachybotrys, Fusarium) and less mobile.Mould spores are widely distributed in the outdoor environment and, therefore, occur in varying concentrations also indoors. Growth of moulds in indoor environments, however, sho
32、uld be considered as a hygienic problem because epidemiological studies have revealed that dampness and/or mould growth in homes and health problems affecting the occupants are closely related.Harmonized methods for sampling, detection, and enumeration of moulds including standards for sampling stra
33、tegies are important for comparative assessment of mould problems indoors. Before doing any measurements, a plan for the measurement strategy should be made.This part of ISO 16000 describes methods for sampling of moulds from building materials.This part of ISO 16000 is based on parts of VDI 4300 Pa
34、rt 10.vi ISO 2013 All rights reservedBS ISO 16000-21:2013INTERNATIONAL STANDARD ISO 16000-21:2013(E)Indoor air Part 21: Detection and enumeration of moulds Sampling from materials1 ScopeThis part of ISO 16000 specifies requirements for sampling of moulds from building materials. Following the instru
35、ctions given, samples are obtained for microscopy or for subsequent detection of moulds by cultivation according to ISO 16000-17.2 Normative referencesThe following documents, in whole or in part, are normatively referenced in this document and are indispensable for its application. For dated refere
36、nces, only the edition cited applies. For undated references, the latest edition of the referenced document (including any amendments) applies.ISO 16000-17, Indoor air Part 17: Detection and enumeration of moulds Culture-based method3 Terms and definitionsFor the purposes of this document, the follo
37、wing terms and definitions apply.3.1colony forming unitcfuunit by which the culturable number of microorganisms is expressedSOURCE: EN 13098:2000Note 1 to entry: One colony can originate from one single microorganism, from aggregates of many microorganisms as well as from one or many microorganisms
38、attached to a particle.Note 2 to entry: The number of colonies can depend on the cultivation conditions.3.2cultivationgrowing of microorganisms on culture mediaSOURCE: ISO 16000-16:2008, 3.63.3filamentous fungusfungus growing in the form of filaments of cells known as hyphaeNote 1 to entry: The term
39、 filamentous fungi differentiates fungi with hyphal growth from yeasts.SOURCE: ISO 16000-16:2008, 3.3 ISO 2013 All rights reserved 1BS ISO 16000-21:2013ISO 16000-21:2013(E)3.4microorganismany microbial entity, cellular or non-cellular, capable of replication or of transferring of genetic material, o
40、r entities that have lost these propertiesSOURCE: EN 13098:20003.5mouldfilamentous fungi from several taxonomic groups, namely ascomycetes, zygomycetes, and their anamorphic states formerly known as deuteromycetes or fungi imperfectiNote 1 to entry: Moulds form different type of spores depending on
41、the taxonomic group they belong to, namely conidiospores (conidia), sporangiospores, or ascospores.3.6myceliumbranched hyphae networkSOURCE: ISO/TS 10832:2009, 3.54 Principle of methodMould-infested materials are examined either by surface sampling (see 7.1) or bulk sampling (see 7.2), i.e. examinat
42、ion of the complete material or defined deeper material layers. The methods used depend on the investigation objective as described in ISO 16000-19. Surfaces are sampled using the contact plate (see 7.1.2), tape-lift (see 7.1.3), or swab method (see 7.1.4). After sampling, the mould spores can be an
43、alysed by direct microscopy (see 7.4) or processed and cultured using the suspension method (see 7.5). The cultivation procedure is described in ISO 16000-17.5 Apparatus and materialsUsual microbiological laboratory equipment, and in particular:5.1 Equipment for sampling5.1.1 Agar plates or flexible
44、 plastic stripes, containing DG-18 agar and malt extract or potato dextrose agar (see Clause 6) with the culture medium slightly projecting over the edge.5.1.2 Cotton swabs, sterile, to take swab samples.5.1.3 Containers to protect the agar plates and material samples during transport, e.g. plastic
45、bags.5.1.4 Disinfectant, e.g. iso-propanol or ethanol (70 % volume fraction) to disinfect sampling tools.5.1.5 Drill, disinfected, with a diameter of at least 3 cm, preferably 5 cm, to take defined cores from the material.5.1.6 Insulated/refrigerated container, for transport of agar plates and mater
46、ial samples below 25 C.5.1.7 Sampling tools, sterile, to take bulk samples of materials in different depths, e.g. spatula, spoons, knives, drilling equipment.2 ISO 2013 All rights reservedBS ISO 16000-21:2013ISO 16000-21:2013(E)5.2 Equipment for preparing the agar plates5.2.1 Autoclave, at (121 3) C
47、 and (115 3) C.5.2.2 Petri dishes, vented, sterile, diameter approximately 9 cm.5.2.3 pH meter, with an accuracy of 0,1.5.3 Equipment for processing the bulk samples5.3.1 Aluminium container, to weigh material samples.5.3.2 Analytical balance, with an accuracy of 0,01 g.5.3.3 Glass flask, baffled fl
48、ask, sterile, 250 ml.5.3.4 Shaking dish, horizontal, 200 rpm.5.3.5 Test tube shaker, e.g. Vortex shaker.6 Culture media and reagents6.1 GeneralAll reagents and chemicals shall be of recognized quality “for microbiology” or better. Water used shall be distilled or of equivalent quality.Use of commerc
49、ially available, dehydrated substrates is encouraged, provided they comply with the descriptions given. These dehydrated substrates shall be prepared according to the instructions from the manufacturer. For surface sampling, agar plates or flexible plastic stripes containing agar medium are also commercially available.6.2 Dichlorane 18 % glycerol agar (DG-18)The components are listed in Table 1.Table 1 Composition of dichlorane 18 % glycerol agar (DG-
