1、BSI Standards PublicationBS ISO 16000-20:2014Indoor airPart 20: Detection and enumeration ofmoulds Determination of total sporecountBS ISO 16000-20:2014 BRITISH STANDARDNational forewordThis British Standard is the UK implementation of ISO16000-20:2014.The UK participation in its preparation was ent
2、rusted to 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 correctapplic
3、ation. The British Standards Institution 2014. Published by BSI StandardsLimited 2014ISBN 978 0 580 81072 5ICS 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 Committe
4、e on 30 November 2014.Amendments issued since publicationDate Text affectedBS ISO 16000-20:2014 ISO 2014Indoor air Part 20: Detection and enumeration of moulds Determination of total spore countAir intrieur Partie 20: Dtection et dnombrement des moisissures Dtermination du nombre total de sporesINTE
5、RNATIONAL STANDARDISO16000-20First edition2014-12-01Reference numberISO 16000-20:2014(E)BS ISO 16000-20:2014ISO 16000-20:2014(E)ii ISO 2014 All rights reservedCOPYRIGHT PROTECTED DOCUMENT ISO 2014All rights reserved. Unless otherwise specified, no part of this publication may be reproduced or utiliz
6、ed 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 in the country of the requester.ISO copyright
7、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 16000-20:2014ISO 16000-20:2014(E)Contents PageForeword ivIntroduction vi1 Scope . 12 Terms and definitions . 13 Principle of method . 24 Apparatus an
8、d materials 25 Reagents 35.1 General . 35.2 Lactophenol blue solution . 36 Measurement procedure 36.1 Sampling . 36.2 Direct microscopy 46.3 Calculation and expression of results 56.4 Transport and storage 67 Quality assurance 68 Calibration of flow rate, function control, and maintenance of the sam
9、pling system 69 Sampling protocol . 610 Performance characteristics 7Annex A (informative) Examples of impactors .11Annex B (normative) Sample exchange for method validation .12Bibliography .13 ISO 2014 All rights reserved iiiBS ISO 16000-20:2014ISO 16000-20:2014(E)ForewordISO (the International Org
10、anization for Standardization) is a worldwide federation of national standards bodies (ISO member bodies). The work 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 establi
11、shed has the right to be represented on that committee. International organizations, governmental and non-governmental, in liaison with ISO, also take part in the work. ISO collaborates closely with the International Electrotechnical Commission (IEC) on all matters of electrotechnical standardizatio
12、n.The procedures used to develop this document and those intended for its further maintenance are described in the 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 t
13、he 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 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 pat
14、ent rights identified during the development of the document will be in the Introduction and/or on the ISO list of 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
15、 an explanation on the meaning of ISO specific terms and expressions related to conformity assessment, as well as information about ISOs adherence to the WTO principles in the Technical Barriers to Trade (TBT) see the following URL: Foreword - Supplementary informationThe committee responsible for t
16、his document is ISO/TC 146, Air quality, Subcommittee SC 6, Indoor air.ISO 16000 consists of the following parts, under the general title Indoor air: Part 1: General aspects of sampling strategy Part 2: Sampling strategy for formaldehyde Part 3: Determination of formaldehyde and other carbonyl compo
17、unds in indoor air and test chamber air Active sampling method Part 4: Determination of formaldehyde Diffusive sampling method Part 5: Sampling strategy for volatile organic compounds (VOCs) Part 6: Determination of volatile organic compounds in indoor and test chamber air by active sampling on Tena
18、x TA sorbent, thermal desorption and gas chromatography using MS or MSFID Part 7: Sampling strategy for determination of airborne asbestos fibre concentrations Part 8: Determination of local mean ages of air in buildings for characterizing ventilation conditions Part 9: Determination of the emission
19、 of volatile organic compounds from building products and furnishing Emission test chamber method Part 10: Determination of the emission of volatile organic compounds from building products and furnishing Emission test cell method Part 11: Determination of the emission of volatile organic compounds
20、from building products and furnishing Sampling, storage of samples and preparation of test specimens Part 12: Sampling strategy for polychlorinated biphenyls (PCBs), polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs) and polycyclic aromatic hydrocarbons (PAHs)iv ISO 201
21、4 All rights reservedBS ISO 16000-20:2014ISO 16000-20:2014(E) Part 13: Determination of total (gas and particle-phase) polychlorinated dioxin-like biphenyls (PCBs) and polychlorinated dibenzo-p-dioxins/dibenzofurans (PCDDs/PCDFs) Collection on sorbent-backed filters Part 14: Determination of total (
22、gas and particle-phase) polychlorinated dioxin-like biphenyls (PCBs) and polychlorinated dibenzo-p-dioxins/dibenzofurans (PCDDs/PCDFs) Extraction, clean-up and analysis by high-resolution gas chromatography and mass spectrometry Part 15: Sampling strategy for nitrogen dioxide (NO2) Part 16: Detectio
23、n and enumeration of moulds Sampling by filtration Part 17: Detection and enumeration of moulds Culture-based method Part 18: Detection and enumeration of moulds Sampling by impaction Part 19: Sampling strategy for moulds Part 20: Detection and enumeration of moulds Determination of total spore coun
24、t Part 21: Detection and enumeration of moulds Sampling from materials Part 23: Performance test for evaluating the reduction of formaldehyde concentrations by sorptive building materials Part 24: Performance test for evaluating the reduction of volatile organic compound (except formaldehyde) concen
25、trations by sorptive building materials Part 25: Determination of the emission of semi-volatile organic compounds by building products Micro-chamber method Part 26: Sampling strategy for carbon dioxide (CO2) Part 27: Determination of settled fibrous dust on surfaces by SEM (scanning electron microsc
26、opy) (direct method) Part 28: Determination of odour emissions from building products using test chambers Part 29: Test methods for VOC detectors Part 30: Sensory testing of indoor air Part 31: Measurement of flame retardants and plasticizers based on organophosphorus compounds Phosphoric acid ester
27、s Part 32: Investigation of constructions on pollutants and other injurious factors InspectionThe following parts are under preparation: Part 33: Determination of phthalates with GC-MS Part 34: Strategies for the measurement of airborne particles (PM 2,5 fraction) Part 35: Measurement of polybromina
28、ted diphenylether, hexabromocyclododecane and hexabromobenzene Part 36: Test method for the reduction rate of airborne bacteria by air purifiers using a test chamber ISO 2014 All rights reserved vBS ISO 16000-20:2014ISO 16000-20:2014(E)IntroductionMould is a common name for filamentous fungi from di
29、fferent taxonomic groups (Ascomycota, Zygomycota, and their anamorphic states former known as Deuteromycota 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.
30、Spores of some mould genera are small and become airborne very easily (e.g. Aspergillus, 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 varyin
31、g concentrations also indoors. Growth of moulds in indoor environments, however, has to be considered 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 sam
32、pling, detection and enumeration of moulds including standards for sampling strategies 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 air sampling of mou
33、ld spores for subsequent microscopic analysis.This part of ISO 16000 is based on parts of VDI 4300 Part 10.6vi ISO 2014 All rights reservedBS ISO 16000-20:2014Indoor air Part 20: Detection and enumeration of moulds Determination of total spore count1 ScopeThis part of ISO 16000 specifies requirement
34、s for sampling of moulds from air. Following the instructions given, samples are obtained for microscopy to determine the total concentration of spores.2 Terms and definitionsFor the purpose of this document, the following terms and definitions apply.2.1cultivationgrowing of microorganisms on cultur
35、e mediaSOURCE: ISO 16000-16:2008, 3.62.2cut-off valueparticle size (aerodynamic diameter) for which the sampling efficiency is 50 %2.3filamentous fungusfungus growing in the form of filaments of cells known as hyphaeSOURCE: ISO 16000-16:2008, 3.3Note 1 to entry: The term “filamentous fungi” differen
36、tiates fungi with hyphal growth from yeasts.2.4impactionsampling of particles suspended in air by inertial separation on a solid surface2.5microorganismany microbial entity, cellular or non-cellular, capable of replication or of transferring of genetic material or entities that have lost these prope
37、rtiesSOURCE: EN 13098:20002.6mouldfilamentous fungi from several taxonomic groups; namely Ascomycota, Zygomycota, and their anamorphic states former known as Deuteromycota or fungi imperfectiSOURCE: ISO 16000-16:2008, 3.9Note 1 to entry: Moulds form different types of spores depending on the taxonom
38、ic group they belong to, namely conidiospores (conidia), sporangiospores, or ascospores.INTERNATIONAL STANDARD ISO 16000-20:2014(E) ISO 2014 All rights reserved 1BS ISO 16000-20:2014ISO 16000-20:2014(E)2.7myceliumbranched hyphae networkSOURCE: ISO/TS 10832:2009, 3.52.8physical sampling efficiencycap
39、acity of the sampler to collect particles with specific sizes suspended in airNote 1 to entry: See Reference 7.3 Principle of methodA defined air quantity is drawn through an impactor containing a sticky solid surface which can subsequently be used for microscopy. The particles in the air stream imp
40、act on the surface, due to their inertia, when the air flows bend to bypass the solid surface.Airborne moulds are thereby collected directly on the sticky surface.Physical sampling efficiency is influenced by the geometry of the slit, air velocity, and the adhesion capability of the surface.The samp
41、ling device is constructed for the detection of particles in the size of mould spores (1 m to ca. 30 m). To achieve this, the cut-off value of the sampling device should preferably be 1 m or less and shall not be more than 2,6 m.NOTE Three main types of impactors are widely used and available commer
42、cially: samplers with replaceable slides and air velocity of ca. 30 L/min, e.g. PS 30 and MBASS30, samplers1)with replaceable slides and air velocity of ca. 15 L/min, e.g. Allergenco MK32), and samplers with disposable cassettes and air velocity of ca. 15 L/min (see Annex A).After sampling, the moul
43、d spores are counted under a microscope. No cultivation is performed. Therefore, the total spore concentration, including culturable and non-culturable spores can be determined.4 Apparatus and materialsUsual microbiological laboratory equipment, and in particular:5.1 Stand, for positioning the impac
44、tor at the sampling height needed.5.2 Impactor, with disposable slides or cassettes.5.3 Vacuum pump, for ensuring a constant flow rate during continuous operation.5.4 Gas volume meter, for determining the gas volume sucked at the sampling head, in operating cubic meters.5.5 Timer, for presetting the
45、 time and duration of sampling.5.6 Protective housing, for protecting the impactor from harmful environmental conditions (optional, mainly for outdoor use).1) PS 30 and MBASS30 are examples of suitable products available commercially. This information is given for the convenience of users of this do
46、cument and does not constitute an endorsement by ISO of this product.2) Allergenco MK3 is an example of a suitable product available commercially. This information is given for the convenience of users of this document and does not constitute an endorsement by ISO of this product.2 ISO 2014 All righ
47、ts reservedBS ISO 16000-20:2014ISO 16000-20:2014(E)5.7 Microscope, equipped with 40 and 100 objectives for ca. 400 and 1000 magnification.5 Reagents5.1 GeneralAll reagents and chemicals shall be of recognized quality “for microbiology” or better. Water used shall be distilled or of equivalent qualit
48、y.5.2 Lactophenol blue solutionThe components of the staining solution are listed in Table 1.WARNING Lactophenol blue solution is toxic and can lead to adverse health reactions. Exposure through direct contact or inhalation has to be avoided.Table 1 Composition of staining solutionComponent Quantity
49、Cotton blue 0,5 gLactic acid (% by mass of 80 % to 85 %) 4,0 gPhenol 4,0 gGlycerol 8,0 gDistilled water 100 mlAdd ingredients in 100 ml water and dissolve.6 Measurement procedure6.1 SamplingSampling is usually conducted at a height of 0,75 m to 1,5 m above ground. For special questions, other heights might be applicable. Take care when sampling at low heights that no settled house dust is sucked in the sampling device.Prepare the required number of imp
