SIST EN ISO 16000-19:2014

SLOVENSKI STANDARD
01-december-2014
1RWUDQML]UDNGHO6WUDWHJLMDY]RUþHQMDJOLY,62
Indoor air - Part 19: Sampling strategy for moulds (ISO 16000-19:2012)
Innenraumluftverunreinigungen - Teil 19: Probenahmestrategie für Schimmelpilze (ISO
16000-19:2012)
Air intérieur - Partie 19: Stratégie d'échantillonnage des moisissures (ISO 16000-
19:2012)
Ta slovenski standard je istoveten z: EN ISO 16000-19:2014
ICS:
13.040.20 Kakovost okoljskega zraka Ambient atmospheres
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

EUROPEAN STANDARD
EN ISO 16000-19
NORME EUROPÉENNE
EUROPÄISCHE NORM
October 2014
ICS 13.040.20
English Version
Indoor air - Part 19: Sampling strategy for moulds (ISO 16000-
19:2012)
Air intérieur - Partie 19: Stratégie d'échantillonnage des Innenraumluftverunreinigungen - Teil 19:
moisissures (ISO 16000-19:2012) Probenahmestrategie für Schimmelpilze (ISO 16000-
19:2012)
This European Standard was approved by CEN on 9 February 2014.

CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulate the conditions for giving this European
Standard the status of a national standard without any alteration. Up-to-date lists and bibliographical references concerning such national
standards may be obtained on application to the CEN-CENELEC Management Centre or to any CEN member.

This European Standard exists in three official versions (English, French, German). A version in any other language made by translation
under the responsibility of a CEN member into its own language and notified to the CEN-CENELEC Management Centre has the same
status as the official versions.

CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia,
Finland, Former Yugoslav Republic of Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania,
Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and United
Kingdom.
EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION

EUROPÄISCHES KOMITEE FÜR NORMUNG

CEN-CENELEC Management Centre: Avenue Marnix 17, B-1000 Brussels
© 2014 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN ISO 16000-19:2014 E
worldwide for CEN national Members.

Contents
Page
Foreword .3

Foreword
The text of ISO 16000-19:2012 has been prepared by Technical Committee ISO/TC 146 “Air quality” of the
International Organization for Standardization (ISO) and has been taken over as EN ISO 16000-19:2014 by
Technical Committee CEN/TC 264 “Air quality” the secretariat of which is held by DIN.
This European Standard shall be given the status of a national standard, either by publication of an identical
text or by endorsement, at the latest by April 2015, and conflicting national standards shall be withdrawn at the
latest by April 2015.
Attention is drawn to the possibility that some of the elements of this document may be the subject of patent
rights. CEN [and/or CENELEC] shall not be held responsible for identifying any or all such patent rights.
According to the CEN-CENELEC Internal Regulations, the national standards organizations of the following
countries are bound to implement this European Standard: Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech
Republic, Denmark, Estonia, Finland, Former Yugoslav Republic of Macedonia, France, Germany, Greece,
Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal,
Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and the United Kingdom.
Endorsement notice
The text of ISO 16000-19:2012 has been approved by CEN as EN ISO 16000-19:2014 without any
modification.
INTERNATIONAL ISO
STANDARD 16000-19
First edition
2012-06-01
Indoor air —
Part 19:
Sampling strategy for moulds
Air intérieur —
Partie 19: Stratégie d'échantillonnage des moisissures

Reference number
ISO 16000-19:2012(E)
©
ISO 2012
ISO 16000-19:2012(E)
©  ISO 2012
All rights reserved. Unless otherwise specified, no part of this publication may be reproduced or utilized in any form or by any means,
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Published in Switzerland
ii © ISO 2012 – All rights reserved

ISO 16000-19:2012(E)
Contents Page
Foreword . iv
Introduction . vi
1  Scope . 1
2  Normative references . 1
3  Terms and definitions . 1
4  Properties, origin and occurrence of moulds in indoor environments . 4
5  Sampling and detection methods . 5
6  Measurement strategy . 6
6.1  General aspects . 6
6.2  Selection of appropriate procedure . 9
7  Quality requirements and uncertainty considerations . 17
Annex A (informative) Moisture damage indicators . 18
Annex B (informative) Devices for total spore count and detection of culturable fungi . 19
Annex C (informative) Field inspection report to describe sampling procedure and to document
potential mould damage . 21
Bibliography . 27

ISO 16000-19:2012(E)
Foreword
ISO (the International Organization 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
established 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 standardization.
International Standards are drafted in accordance with the rules given in the ISO/IEC Directives, Part 2.
The main task of technical committees is to prepare International Standards. Draft International Standards
adopted by the technical committees are circulated to the member bodies for voting. Publication as an
International Standard requires approval by at least 75 % of the member bodies casting a vote.
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.
ISO 16000-19 was prepared by Technical Committee 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 compounds 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 ®
Tenax TA sorbent, thermal desorption and gas chromatography using MS or MS–FID
 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 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 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)
 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
iv © ISO 2012 – All rights reserved

ISO 16000-19:2012(E)
 Part 14: Determination of total (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 (NO )
 Part 16: Detection 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 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) concentrations 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 (CO )
 Part 28: Determination of odour emissions from building products using test chambers
The following parts are under preparation:
 Part 21: Detection and enumeration of moulds — Sampling from materials
 Part 27: Determination of settled fibrous dust on surfaces by SEM (scanning electron microscopy) (direct
method)
 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
 Part 32: Investigation of constructions on pollutants and other injurious factors — Inspections

ISO 16000-19:2012(E)
Introduction
Mould spores and metabolites can be inhaled via the air and cause allergic and irritating reactions and/or
complex symptoms in humans. Moreover, mould growth can be associated with severe odour nuisances. In
[14][18][19]
rare cases, some mould species can cause infections (so-called mycoses) in certain risk groups.
There is sufficient epidemiological evidence that damp and mouldy buildings increase the risk of respiratory
[8]
symptoms, respiratory infections and enhances asthma symptoms of the occupants. In addition, there is
some evidence for increased risk of development of allergic rhinitis and asthma. Furthermore, there is clinical
evidence for rare symptoms like allergic alveolitis, chronic rhinosinusitis and allergic sinusitis. Toxicological
studies in vivo and in vitro show irritating and toxic reactions of microorganisms (including spores, cell
[8]
components and metabolites) from damp buildings.
Growth of microorganisms in damp buildings can lead to increased concentrations of spores, cell fragments,
allergens, mycotoxins, endotoxins, -glucanes and MVOC (microbial volatile organic compounds). From the
studies conducted so far it is not clear which compounds are the causative agents of the health effects
observed. Nevertheless, increased concentrations of each of these compounds are considered a potential
[8][18]
health risk and growth of mould in buildings should, therefore, be avoided.
The prime objective of this part of ISO 16000 is to provide assistance in identifying mould sources in indoor
environments.
vi © ISO 2012 – All rights reserved

INTERNATIONAL STANDARD ISO 16000-19:2012(E)

Indoor air —
Part 19:
Sampling strategy for moulds
1 Scope
This part of ISO 16000 describes the measurement strategy for the detection of fungi in indoor environments.
It describes suitable sampling and analysis methods together with a description of the applicability and the
interpretation of the measurement results to maximize the comparability of the measured data obtained for a
given measurement objective. It does not include details on recording building characteristics or field
inspections by qualified professionals which have to take place prior to any microbiological measurement.
This part of ISO 16000 is not applicable to a detailed description of the building physics- and building-
engineering-related procedures applicable to field inspections. The methods and procedures presented do not
allow quantitative exposure assessment with regard to the room occupants.
The application of this part of ISO 16000 presupposes the knowledge of ISO 16000-1.
2 Normative references
The following referenced documents are indispensable for the application of this document. For dated
references, only the edition cited applies. For undated references, the latest edition of the referenced
document (including any amendments) applies.
ISO 16000-16, Indoor air — Part 16: Detection and enumeration of moulds — Sampling by filtration
ISO 16000-18, Indoor air — Part 18: Detection and enumeration of moulds — Sampling by impaction
3 Terms and definitions
For the purposes of this document, the following terms and definitions apply.
3.1
pre-existing mouldy condition
desiccated “old” mould growth, where additional biomass growth no longer occurs and the indoor air mould
spore concentration gradually decreases with time
3.2
biological preservation efficiency
capacity of the sampler to maintain the viability of the airborne microorganisms during collection and also to
keep the microbial products intact
[6]
[SOURCE: EN 13098:2000 ]
ISO 16000-19:2012(E)
NOTE The biological collection efficiency considers the sampling stress occurring during sampling and analysis in
addition to the physical collection efficiency.
3.3
identification of moulds
assignment of moulds to spore types or groups on the basis of defined properties (e.g. morphological,
biochemical, molecular-biological properties)
NOTE The term “differentiation” is frequently used instead of identification. The term “differentiation” is, however,
misleading because the intention is not to merely differentiate the moulds but to identify them, i.e. to assign them, e.g. to
genera or species.
3.4
filamentous fungus
fungus growing in the form of filaments of cells known as hyphae
NOTE 1 Hyphae aggregated in bundles are called mycelia.
NOTE 2 The term “filamentous fungi” differentiates fungi with hyphal growth from yeasts.
3.5
filtration
collection of particles suspended in a gas or liquid by flow through a porous medium
[6]
[SOURCE: EN 13098:2000 ]
NOTE In this part of ISO 16000, filtration is understood as the separation of microorganisms or moulds from a
defined volume of air by means of filters.
3.6
total spore count
number of (culturable and non-culturable) spores that are collected and enumerated under the microscope
NOTE For the term “spores”, see 3.19, Note 2.
3.7
yeast
unicellular fungus that does not normally produce a mycelium and reproduce by budding (budding fungi) as
against moulds, which reproduce by sporulation
3.8
impaction
sampling of particles suspended in air by inertial separation on a solid surface (culture medium or adhesive-
coated slides)
NOTE 1 See 16000-18.
NOTE 2 Sampling is carried out using either round-hole or slit impactors, for instance. As the air passes through the
orifices, it is accelerated and the particles are impacted on the medium located directly behind the nozzles as a result of
their inertia, while the air flows around the culture medium and exits the sampler. Impaction samples are only suitable for
direct analysis without further resuspension of the sample.
3.9
colony forming unit
cfu
air quality unit by which the culturable number of microorganisms is expressed
[6]
[SOURCE: EN 13098:2000 ]
NOTE 1 One colony can originate from one single microorganism, from aggregates of many microorganisms as well as
from one or many microorganisms attached to a particle.
NOTE 2 The number of colonies can depend on the cultivation conditions.
2 © ISO 2012 – All rights reserved

ISO 16000-19:2012(E)
3.10
colony morphology type
group of colonies which due to their morphological appearance seem to belong to a specific species
3.11
colony count
air quality number of all microorganism colonies visible on a culture medium after incubation under the
selected cultivation conditions
3.12
culturable mould
mould that can be cultured under the selected cultivation conditions
NOTE Parameters governing the culturability are, for instance, the type of culture medium and the incubation
temperature.
3.13
cultivation
growing of microorganisms on culture media
3.14
mycotoxin
secondary metabolites of moulds which are toxic to humans and animals
3.15
mycelium
total of fungal hyphae
3.16
non-culturable mould
mould that cannot be cultured under the selected cultivation conditions
3.17
physical sampling efficiency
capacity of the sampler to collect particles with specific aerodynamic diameters suspended in air
[6]
[SOURCE: EN 13098:2000, modified — "aerodynamic diameters" has replaced "sizes".]
3.18
sampling stress
damage suffered by the microorganisms during sampling (e.g. through mechanical and chemical effects or
through water deprivation)
3.19
mould
filamentous fungi from several taxonomic groups, namely Ascomycetes, Zygomycetes, and their anamorphic
states formerly known as Deuteromycetes or fungi imperfecti
NOTE 1 Taxonomically, moulds do not represent a uniform group.
NOTE 2 Moulds form different types of spores depending on the taxonomic group they belong to, namely
conidiospores (conidia), sporangiospores or ascospores. In practice, all these reproductive stages are summarized under
the term “spores”.
3.20
mould damage
damage caused to building materials and surfaces by mould growth
NOTE Mould damage can result in loss in value, health risks and restrict the occupancy of the affected sites.
ISO 16000-19:2012(E)
3.21
secondary colony
colony that does not originate from the “primary” sampling of airborne spores but from a spore released from a
colony growing on the agar plates
3.22
secondary contamination
mould contamination of surfaces not caused by mould growth but originating from a (contaminated) primary
source after aerial dispersion
3.23
cut-off value
particle size (aerodynamic diameter) for which the sampling efficiency is 50 %
3.24
total sampling efficiency
product of the physical sampling efficiency and the biological preservation efficiency
[6]
[SOURCE: EN 13098:2000 ]
4 Properties, origin and occurrence of moulds in indoor environments
Moulds are ubiquitous on our planet. They are involved in the decomposition of organic material and,
therefore, play an important role in the natural carbon cycle. Their concentration in the ambient air depends,
inter alia, on location, climate, time of the day and season. Airborne mould concentrations are subject to great
[9][10][11]
variability. This is due to the following reasons.
The mould concentration in local ambient air is mainly determined by the location relative to the respective
mould sources, wind direction and wind force. Mould spores are frequently released by specific sources such
as decaying material. Both natural processes and production processes, such as composting, recycling,
animal production facilities, grain and food processing plants as well as horticulture facilities, can be sources
of mould dispersion.
Sporulation, i.e. the production of mould spores occurs discontinuously. It is governed, inter alia, by the mould
lifecycle phase, the environmental conditions, stress factors, humidity as well as substrate composition and
availability.
Factors governing the dispersion of spores, most of which have aerodynamic diameters in the range of 2 µm
to 40 µm, are mechanically or thermally induced air movements, drying phases (leading e.g. to de-
[12][13][14]
agglomeration of deposited dust) and the capability of air dispersal of the mould spores.
Due to the ubiquitous nature of moulds, it can be assumed that they are always present in indoor air. The
presence of moulds in indoor air can be due to spores originating from ambient air on the one hand and to
recent active mould growth, pre-existing mouldy conditions or mould deposits (settled spores) on the other. To
distinguish between sources, it is, therefore, important to perform ambient air measurements for reference
[14][15]
whenever conducting indoor air measurements for moulds. In addition, the collection of a control sample
from a suitable reference room may be helpful.
Possible causes of indoor mould sources are surface moisture on building materials or moisture in the building
structure, but also rotting food, potted plants, biowaste collection, source separation of waste, deposited dust
due to poor cleaning as well as the keeping of animals in residential settings. Moisture damage can be
attributable to building defects, inappropriate ventilation and heating or unfavourable arrangement of furniture
as well as water damage (e.g. plumbing leaks or flooding events). Elevated mould levels in indoor
environments and the occurrence of certain mould species (see Annex A) are indicative of excessive moisture.
When residential environments or occupational settings are infested with moulds, the mould source shall be
located to be able to plan remedial measures.
4 © ISO 2012 – All rights reserved

ISO 16000-19:2012(E)
Main factors affecting the intensity of mould growth and the mould species developing are moisture,
temperature, nutrient supply and the pH. If environmental conditions are favourable, a great variety of moulds
can develop. Once environmental conditions become less favourable, the species best adapted to the given
[16]
conditions will predominate.
Mould sources can release spores, mycelial fragments, but also cell components and metabolic products such
as -glucans (polysaccharides contained in the cell wall of fungi), ergosterol (steroid compound contained in
the cell membrane of fungi), toxins and MVOCs (microbial volatile organic compounds such as certain
aldehydes, alcohols, esters, ketones). On cultivation, colonies can grow not only from spores, but also from
mycelial fragments.
The number and airborne dissemination of spores released vary with the type of mould damage. For an
assessment of indoor mould sources, it is, therefore, important to differentiate the individual mould species by
their type of spore dispersal. Experience has shown that even minor mould contamination of materials can
result in elevated indoor air mould levels if the species involved have dry spores with good air dispersal
capabilities (e.g. Penicillium and Aspergillus). By contrast, airborne spore concentrations are much lower
when materials are colonized, for instance, by moulds of the genera Acremonium, Fusarium or the species
Stachybotrys chartarum that have relatively large spores embedded in slimy substances and, therefore, have
poor air dispersal capabilities.
Furthermore, it should be taken into account that mould spores are not necessarily present as individual
spores in the air or settled dust, but also occur in the form of spore aggregates or are particle-borne.
Depending on the analysis method, they are determined individually or as spore aggregate. Materials, indoor
air and house dust contain not only culturable but also non-culturable mould spores, some of which can have
the same allergenic and toxic effects as culturable spores. For this reason, techniques have been developed
that allow the microscopic determination of both culturable and non-culturable moulds.
Mould detection and identification are performed either after cultivation based on morphological criteria,
biochemical reactions and/or molecular techniques or by direct microscopic examination. Identification based
on the morphological structure (macroscopic examination, stereo-microscopy and microscopy) either after
prior cultivation or by direct microscopy is still the most prevalent approach for the detection of moulds.
Besides, there are other analytical methods based on the determination of cell components and metabolites of
[17]
moulds such as -glucans, ergosterol, toxins and MVOCs. The determination of these compounds serves,
however, only as supplementary information.
The sampling methods employed for detection of moulds are determined by the objective of the investigation.
Depending on the sampling method, the moulds suffer a sampling stress during sample collection and
preparation, which can lead to their drying-out or dying. Factors affecting the culturability of mould spores are
their physiological state as well as the culture medium employed. Some mould species cannot be cultured at
all under laboratory conditions.
NOTE The genera Stachybotrys and Chaetomium hardly grow and sporulate only poorly, if at all, on DG18 agar. The
use of this culture medium for culture-based analysis of these genera is therefore not recommended (see ISO 16000-17).
For a literature summary see References [8]–[10], [12], and [14]–[18].
5 Sampling and detection methods
Depending on the objective of the investigation, materials (see ISO 16000-21, in preparation), air (see
ISO 16000-16 and ISO 16000-18) and house dust may be sampled and analysed for culturable moulds (see
ISO 16000-17). Moulds can also be quantified and, to some extent, differentiated without prior cultivation. For
this purpose, airborne mould spores are collected on filters or directly on an adhesive-coated microscope slide,
followed by staining and subsequent direct microscopy.
Annex B gives an overview on the most common devices for total spore count measurements as well as for
sampling devices for filtration and impaction and the respective analysis methods.
ISO 16000-19:2012(E)
6 Measurement strategy
6.1 General aspects
There is no standard procedure for measurement and assessment of mould damage. The type and amount of
measurements as well as the analytical methods employed are determined by the circumstances triggering
the investigation and the investigation objectives. A visual field inspection (walk through) by technically
qualified professionals prior to sampling is a key prerequisite for detecting and assessing mould sources in
indoor environments. Besides a good knowledge of building engineering and building physics, the
professionals conducting the inspection should have a sufficient background in indoor air hygiene and
microbiology.
Investigations are conducted with the objective of locating mould sources in indoor environments. To support
findings from visual observations and confirm suspected mould growth, professionals can draw on a variety of
sampling and analysis methods. These include methods for determining mould concentrations in or on
materials, procedures for the measurement of mould concentrations in indoor air as well as procedures for
determining mould concentrations in house dust. An example for a report accompanying sampling is attached
as Annex C.
Circumstances triggering a microbiological investigation of indoor environments may include the following (see
also Table 1):
 visible mould damage;
 material dampness without presence of visible mould growth;
 structural or non-structural building anomalies without presence of visible mould growth;
 health problems without presence of visible mould growth;
 odour problems without presence of visible mould growth;
 verification measurements during and after remediation.
In the case of visible mould damage with known source, the remediation and elimination of the underlying
causes should be addressed as a priority. In many cases, microbiological investigations is not necessary.
If mould damage is suspected without the presence of visible sources, the indoor environment can be
examined for the presence of elevated mould concentrations. Depending on the circumstances triggering the
investigation, the following media may be sampled and analysed:
a) materials and their surfaces (see 6.1.1);
b) indoor air in comparison with ambient air (see 6.1.2);
c) house dust (see 6.1.3).
The results of the measurements described in the following sections provide only indications on the damage
stage. Assessing the actual age of the mould growth is not feasible as the state of mould growth can change
drastically within very short time intervals.
The inspection of HVAC systems for lack of hygiene is not the subject of this part of ISO 16000.
In planning and performing measurements, the specific field conditions and influencing factors having a major
impact on the investigation results shall be taken into account and documented.
6 © ISO 2012 – All rights reserved

ISO 16000-19:2012(E)
6.1.1 Analysis of surfaces or materials
For the selection of a suitable sampling method and the definition of the sampling locations, the following
questions have to be clarified.
 Is mould growth or secondary contamination expected on the surface or the material?
 Is a surface colonization or a colonization of deeper layers expected?
 Are the moulds expected or under study culturable?
 Are criteria available for an assessment of the analysis result?
Criteria for differentiating between active mould growth and mould deposits on material surfaces or in wall
cavities originating from natural sedimentation are the mould concentration and evidence of mould structures,
e.g. mycelium or spore carriers, in the material or on its surface. The mould concentration in the material or on
the material surface varies with the type of material, especially the density of the material, and the mould
species. Different mould species grow on or in a material depending on moisture, temperature and nutrient
source. Suspected mould contamination of surfaces may be confirmed by surface sampling using the tape-lift
and contact plate methods. The contact plate method presupposes that the moulds are culturable. If the
surface has already been disinfected or if contamination with Stachybotrys chartarum is suspected, the
contact plate method is not applicable. In such cases, it is necessary that tape-lift samples of the surfaces be
examined by direct microscopy.
Surface sampling (contact plate and tape-lift methods) has limited suitability for materials with rough surfaces
(e.g. plaster, insulation materials). Usually, the samples are suspended in a buffer followed by determination
[25]
of the mould concentration by cultivation or direct microscopy.
Where no empirical threshold levels exist for classifying a material as “contaminated” or “not contaminated”,
materials displaying no visible mould growth are sampled as controls for comparison.
6.1.2 Analysis of indoor air
The objective of indoor air sampling and analysis is to determine the concentration of moulds in a
representative air sample in order to assess the likelihood of mould sources in the indoor environment.
Depending on the investigation objective, this requires a more or less complete identification of the moulds
(see 6.2). In analysing air samples, special attention is given to differences in the species spectrum present in
the indoor compared to ambient air. Moreover, the presence of moisture-indicator species (see Table A.1)
should be taken into account. At high mould spore concentrations in the ambient air due to the specific
weather conditions, the concentration of ambient air species in the indoor air can be many times higher than
the concentrations of the moisture-indicator species of interest. If the concentration of typical ambient air
species exceeds that of indoor-environment-specific moisture-indicators by a factor of more than 10, indoor air
sampling allows no conclusions as to the presence of a potential mould source because moisture indicators
may be overgrown by fungi from ambient air.
To avoid interferences, rooms in which the lowest airborne concentrations are expected should be sampled
first.
The sampling methods in accordance with ISO 16000-16 (filtration) and ISO 16000-18 (impaction) are based
on different measurement principles and do not produce the same results for all measurement tasks. For the
selection of the sampling procedure and the determination of the required number of sampling locations and
the sampling duration, the influencing parameters and conditions prevailing in the specific situation shall be
established by a prior field inspection.
For this purpose, the following questions shall be clarified.
 Is a largely constant mould concentration expected in the room?
 Are air movements present that reflect a normal activity in the room?
ISO 16000-19:2012(E)
 Are major fluctuations in the mould concentration expected as a result of short-term influences (e.g.
occupant influences, convection or downward air flows)?
When no major occupancy-related air movements are expected and no influences leading regularly to major
variations in the airborne mould levels are evident in the rooms being assessed (e.g. residential rooms), both
short-term sampling (sampling period 1 min to 10 min) and long-term sampling (sampling period > 30 min) are
appropriate methods. In practice, sampling by impaction is the preferred procedure for short-term sampling.
This sampling method requires a prior estimate of the expected mould concentration. Different air volumes are
sampled at each sampling location to be able to cover a broader concentration range. This is accomplished by
collecting impaction samples over different sampling durations. The detection of indoor-relevant moulds
presupposes that particles with a diameter greater than 2 µm can be quantitatively collected on the culture
medium or adhesive-coated slide (for the determination of the total spore count). This presupposes that the
impactors are designed for a cut-off d < 2 µm (see Table B.2). All short-term measurements should be
conducted over a minimum period of 1 min. The sample volume should not be less than 50 l. In unoccupied
rooms, short-time measurements may be performed without occupancy simulation, since experience has
shown that especially the installation of the sampling equipments as well as their operation usually results in
air movements at the sampling location that are comparable to those during normal conditions of use.
In rooms with major “old” dust deposits, sampling can cause unintentional disturbance of settled dust, which
can lead to false positive results.
If a sampling device generates major exit air flows resulting in the disturbance of deposited dust, the exit air
stream should be conducted of the room being investigated and/or care should be taken to ensure that it is not
directed at potential mould sources, such as the floor or dusty or mouldy materials.
Filtration methods are the sole applicable option for long-term measurements. Filtration sampling is the
method of choice when sampling is carried out during normal activities in the room and when major air
movements and fluctuating mould concentrations are expected. Filtration sampling is also the preferred
method for culturable sampling when airborne mould concentrations are expected to exceed 2 000 cfu/m . At
sampling durations of 1 h and longer in unoccupied rooms, additional occupancy simulations are needed
during the sampling period. The occupancy simulation should reflect the usual occupancy of the room.
Regardless of the sampling method, the windows and doors of the room shall be closed approx. 8 h before
commencing sampling and kept closed during the sampling process. Samples should preferably be collected
in the centre of the room with a minimum distance of 1 m from enclosing walls and at a height of approx.
0,75 m to 1,5 m. In all cases, an ambient air sample shall be collected for reference. Moreover, the collection
of air samples in an appropriate reference room can be useful. Indoor and ambient air samples shall be
collected on the same day with as short a time interval in-between as possible.
NOTE In buildings with air-conditioning, shorter time intervals (2 h) between closing the windows and sampling can
be sufficient. Measurement of fungi in ambient air might not be necessary in buildings with filtered incoming air and no
windows that can be opened by the occupants.
The specific conditions at the sampling location and the climatic conditions during sampling are documented
in a sampling report (see Annex C).
6.1.3 Analysis of house dust
House dust analyses are normally only conducted to complement the results from indoor air measurements.
As there are currently no suitable procedures for the determination of non-culturable moulds in house dust,
the analysis is limited to the detection of culturable moulds. House dust analyses are a useful tool to check the
results from indoor air measurements for plausibility. Before commencing sampling, it should be clarified
[26][27]
whether suitable sampling locations with sufficient quantities of settled dust exist.
Reference data used for the assessment of the analysis results shall have been obtained by the same
sampling, sample preparation and analysis methods. Results differ greatly if total house dust or fine dust of a
certain size fraction is being used.
8 © ISO 2012 – All rights reserved

ISO 16000-19:2012(E)
6.2 Selection of appropriate procedure
6.2.1 Field inspection
The first step of a mould assessment in indoor environments is a field inspection in order to take an inventory.
On this occasion, the circumstances triggering the investigation and details of the condition of the building,
room furnishings, etc. are assembled.
Table 1 — Recommendations to help decision-making for sampling after a field inspection
Finding and objective Matter being examined Further
procedure
Material Indoor air House dust
a b b
Identify and, if
A B B
Visible mould
1 applicable, eliminate the
damage
moisture source
A B B Identify and, if
2.1 Material dampness applicable, eliminate the
moisture source
— A B Check anomalies,
Suspected
Non-structural /
2.2 identify source, if
mould
structural anomalies
applicable, and remedy
damage
— A B Identify and remedy
2.3 Health problems
source
2.4 Odour problems — A B Identify odour source
3 Remediation monitoring A A B —
A Suitable examination to answer the questions of interest
B Supplementary examination to answer the questions of interest (optional)
a
Material sampling can be useful to answer specific questions concerning major mould damage (see 6.2.2.1).
b
If it is necessary to analyse a dispersion of the contamination.

The building structure, especially the surfaces of critical building components, is visually examined during the
inspection. Moreover, information on potential causes of mould growth should be collected. For this purpose,
relevant physical parameters such as temperature and humidity in the room and on materials (e.g.
condensate) are recorded. If the findings give no clear picture, further non-structural building investigations
(e.g. moisture measurements, thermography, Blower-Door test) may be performed. The detailed description of
the procedures used during inspection is not covered by this part of ISO 16000.
Qualified professionals normally recognize visible mould growth without the need for any elaborate sampling
and analysis methods. If sampling is required, information on the sampling location can be gathered using the
sampling report in Annex C. Table 1 lists possible options for further analysis depending on the findings of the
field inspection. The specific procedures are described in detail in 6.2.2 to 6.2.6.
When performing air and house-dust sampling after field inspection, it is necessary to take into account that
any intrusive inspections into the building structure carried out during the field inspection can have released
additional moulds to the indoor environment that are not attributable to the presence of mould growth.
ISO 16000-19:2012(E)
6.2.2 Investigations prompted by questions related to visible mould damage
6.2.2.1 General aspects
For major visible mould damage, materials may be investigated to answer the following questions:
 confirmation of mould contamination or growth (see 6.2.2.2);
 establishing the damage extent and potential secondary contamination (see 6.2.2.3);
 contamination assessment (see 6.2.2.4);
 establishing prerequisites for remediation monitoring (see 6.2
...