CEN/TS 16115-1:2011
(Main)Ambient air quality - Measurement of bioaerosols - Part 1: Determination of moulds using filter sampling systems and culture-based analyses
Ambient air quality - Measurement of bioaerosols - Part 1: Determination of moulds using filter sampling systems and culture-based analyses
This Technical Specification describes the measurement of moulds in ambient air in order to identify, quantify and characterize bioaerosol pollution in ambient air resulting from emissions from different sources.
The method described specifies the sampling of moulds as part of the suspended particulate matter (SPM, here particles with aerodynamic diameter up to ca. 30 µm) using a filter sampling system with gelatine/poly-carbonate filter combination followed by the culture-based analyses on DG18 agar. The sampling duration can be varied between 10 min to 24 h. The health effect of bioaerosols is not limited to any particle fraction, therefore, this document describes the sampling of moulds as part of the suspended particulate matter as a convention method.
NOTE The sampling method described in this document in principle is likely to be appropriate for the sampling of actinomycetes and other spore-forming bacteria (resistant to desiccation). For these species a special analytical procedure using different culture media should be applied, but this is not within the scope of this document.
The standard method set out in this Technical Specification is accepted by convention as reference method. The measured quantity, here the number of colony forming units per cubic meter (CFU/m3), is determined by the inlet design of the sampling head, the associated operational parameters and the analytical procedure.
Standardized methods for sampling, detection and enumeration of moulds including standards for sampling strategies are important for comparative assessment of moulds in ambient air. Before doing any measurements a plan for the measurement strategy is necessary (see CEN/TS 16115-2 [5]).
WARNING - The use of this Technical Specification may involve hazardous materials, operations and equipment. This Technical Specification does not purport to address all the safety problems associated with its use. (...)
Luftbeschaffenheit - Messen von Bioaerosolen - Teil 1: Bestimmung von Schimmelpilzen mittels Probenahme auf Filtern und kulturellem Nachweis
Diese Technische Spezifikation beschreibt die Messung von Schimmelpilzen in der Außenluft, um die
Verunreinigung der Außenluft durch Bioaerosole, die von unterschiedlichen Quellen emittiert werden, zu
identifizieren, zu quantifizieren und zu charakterisieren.
Das beschriebene Verfahren legt die Probenahme von Schimmelpilzen als Anteil des Schwebstaubes (SPM,
hier Partikel mit dem aerodynamischen Durchmesser von bis zu etwa 30 µm) mit einem
Filterprobenahmesystem bestehend aus einer Gelatine/Polycarbonat-Filterkombination, gefolgt von einem
kulturellen Nachweis auf DG-18-Agar, fest. Die Probenahmedauer kann zwischen 10 min min bis 24 h h
variiert werden. Die gesundheitliche Wirkung von Bioaerosolen ist nicht auf eine Partikelfraktion begrenzt,
deshalb legt dieses Dokument die Probenahme von Schimmelpilzen als Anteil des Schwebstaubes mittels
eines Konventionsverfahrens fest.
ANMERKUNG Das in diesem Dokument festgelegte Probenahmeverfahren ist wahrscheinlich prinzipiell auch für die
Probenahme von Actinomyceten und anderen sporenbildenden Bakterien (widerstandsfähig gegen Austrocknen)
geeignet. Bei diesen Arten sollte ein besonderes Analysenverfahren mit anderen Kultivierungsmedien angewendet
werden, dieses liegt aber nicht im Anwendungsbereich des vorliegenden Dokumentes.
Das in dieser Technischen Spezifikation festgelegte Standardverfahren wird per Konvention als Referenzverfahren
anerkannt. Die gemessene Größe, hier die Anzahl der koloniebildenden Einheiten pro Kubikmeter
(KBE/m3), wird durch die Auslegung des Einlasses des Probenahmekopfes, die dazugehörigen
betriebsbedingten Parameter und das Analysenverfahren bestimmt.
Standardisierte Verfahren für Probenahme, Nachweis und Zählung von Schimmelpilzen sowie für die Probenahmestrategien
sind wichtig für eine vergleichende Bewertung von Schimmelpilzen in der Außenluft. Vor
Beginn der Messungen ist ein Plan für die Messstrategie erforderlich (siehe CEN/TS 16115-2 [5]).
Qualité de l'air ambiant - Mesurage de bioaérosols - Partie 1: Dosage des moisissures à l'aide de systèmes de prélèvement sur filtres et d'analyses de cultures
La présente Spécification technique décrit le mesurage des moisissures dans l'air ambiant afin d'identifier, de
quantifier et de caractériser la pollution par les bioaérosols dans l'air ambiant résultant des émissions de
différentes sources.
La méthode décrite spécifie l'échantillonnage de moisissures incluses dans la matière particulaire en
suspension (MPS, ici les particules ayant un diamètre aérodynamique pouvant atteindre environ 30 µm) en
utilisant un système d'échantillonnage sur filtres avec une combinaison de filtre en polycarbonate/gélatine
puis des analyses de cultures sur gélose DG18. La durée d’échantillonnage peut varier de 10 min à 24 h.
L'effet des bioaérosols sur la santé ne se limite pas à une fraction particulaire. Par conséquent, le présent
document décrit l'échantillonnage de moisissures faisant partie de la matière particulaire en suspension
comme étant une méthode conventionnelle.
NOTE La méthode d'échantillonnage décrite dans le présent document est en principe susceptible de convenir à
l'échantillonnage d'actinomycètes et d'autres bactéries sporulées (résistantes au desséchement). Pour ces espèces, il
convient d'appliquer un mode opératoire d'analyse spécial utilisant différents milieux de culture mais ne faisant par partie
du domaine d'application du présent document.
La méthode normalisée décrite dans la présente Spécification technique est acceptée par convention comme
méthode de référence. La quantité mesurée, ici le nombre d’unités formant colonie par mètre cube (UFC/m3),
est déterminée par la conception de l’entrée de la tête de prélèvement, les paramètres de fonctionnement
associés et le mode opératoire d'analyse.
L'existence de méthodes normalisées pour l'échantillonnage, la détection et le dénombrement des
moisissures, y compris des normes relatives à des stratégies d’échantillonnage, est importante pour
l'évaluation comparative des moisissures dans l’air ambiant.
Kakovost zunanjega zraka - Meritve bioaerosolov - 1. del: Določevanje gliv z uporabo sistemov vzorčenja s filtri in analizatorji, temelječimi na kulturi
Ta tehnična specifikacija opisuje meritve gliv v zunanjem zraku za identifikacijo, kvantifikacijo in karakterizacijo onesnaženja z bioaerosoli, ki je posledica emisij iz različnih virov (npr. kompostiranja odpadkov, hladilnih stolpov, kmetijstva).
Opisana metoda določa dolgotrajno vzorčenje gliv (od 10 minut do 24 ur) kot del suspendiranih delcev (SPM, tukaj so to delci z aerodinamičnim premerom do ca. 30 µm) z uporabo filtrskega sistema vzorčenja s kombinacijo želatinskega/polikarbonatnega filtra, ki ji sledijo analize na osnovi gojenja na agarju DG18. Učinek bioaerosolov na zdravje ni omejen na konkretno frakcijo delcev, zato ta dokument opisuje vzorčenje gliv kot del suspendiranih delcev kot dogovorno metodo.
OPOMBA: Metoda vzorčenja, opisana v tem dokumentu, je načelno primerna za vzorčenje aktinomicet in drugih sporogenih bakterij (odpornih proti izsušitvi). Za te vrste se mora uporabiti poseben analitski postopek z različnimi gojišči, vendar to ne spada v obseg tega dokumenta.
OPOZORILO - Uporaba tega standarda lahko vključuje nevarne materiale, delovne postopke in opremo. Ta dokument ne obravnava vseh varnostnih problemov, povezanih z njegovo uporabo. Odgovornost uporabnika tega standarda je, da vzpostavi primerne varnostne in zdravstvene prakse in pred uporabo določi veljavnost regulativnih omejitev.
General Information
Standards Content (Sample)
SLOVENSKI STANDARD
01-december-2011
.DNRYRVW]XQDQMHJD]UDND0HULWYHELRDHURVRORYGHO 'RORþHYDQMHJOLY]
XSRUDERVLVWHPRYY]RUþHQMDVILOWULLQDQDOL]DWRUMLWHPHOMHþLPLQDNXOWXUL
Ambient air quality - Measurement of bioaerosols - Part 1: Determination of moulds using
filter sampling systems and cultivation based analyses
Luftbeschaffenheit - Messen von Bioaerosolen - Teil 1: Bestimmung von Schimmelpilzen
mittels Probenahme auf Filtern und kulturellem Nachweis
Qualité de l'air ambiant - Mesurage de bioaérosols - Dosage des moisissures à l'aide de
systèmes de prélèvement sur filtres et d'analyses de cultures
Ta slovenski standard je istoveten z: CEN/TS 16115-1:2011
ICS:
13.040.20 Kakovost okoljskega zraka Ambient atmospheres
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.
TECHNICAL SPECIFICATION
CEN/TS 16115-1
SPÉCIFICATION TECHNIQUE
TECHNISCHE SPEZIFIKATION
April 2011
ICS 13.040.20
English Version
Ambient air quality - Measurement of bioaerosols - Part 1:
Determination of moulds using filter sampling systems and
culture-based analyses
Qualité de l'air ambiant - Mesurage de bioaérosols - Partie Luftbeschaffenheit - Messen von Bioaerosolen - Teil 1:
1: Dosage des moisissures à l'aide de systèmes de Bestimmung von Schimmelpilzen mittels Probenahme auf
prélèvement sur filtres et d'analyses de cultures Filtern und kulturellem Nachweis
This Technical Specification (CEN/TS) was approved by CEN on 4 October 2010 for provisional application.
The period of validity of this CEN/TS is limited initially to three years. After two years the members of CEN will be requested to submit their
comments, particularly on the question whether the CEN/TS can be converted into a European Standard.
CEN members are required to announce the existence of this CEN/TS in the same way as for an EN and to make the CEN/TS available
promptly at national level in an appropriate form. It is permissible to keep conflicting national standards in force (in parallel to the CEN/TS)
until the final decision about the possible conversion of the CEN/TS into an EN is reached.
CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia,
Finland, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland,
Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland and United Kingdom.
EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION
EUROPÄISCHES KOMITEE FÜR NORMUNG
Management Centre: Avenue Marnix 17, B-1000 Brussels
© 2011 CEN All rights of exploitation in any form and by any means reserved Ref. No. CEN/TS 16115-1:2011: E
worldwide for CEN national Members.
Contents Page
Foreword .3
Introduction .4
1 Scope .5
2 Normative references .5
3 Terms and definitions .5
4 Symbols and abbreviations .8
5 Basic principle of the method .8
6 Sampling .9
7 Culture-based analyses . 15
8 Performance characteristics and minimum requirements . 22
9 Quality assurance . 23
10 Trouble shooting during sampling and analyses . 23
Annex A (informative) Example for a validated sampling device . 26
Annex B (informative) Recovery of spores on gelatine filters in combination with polycarbonate
filters . 32
Annex C (informative) Examples for sampling and analyses reports . 35
Annex D (informative) Membrane filtration technique . 41
Annex E (informative) Calculation by weighted mean . 42
Bibliography . 44
Foreword
This document (CEN/TS 16115-1:2011) has been prepared by Technical Committee CEN/TC 264 “Air
quality”, the secretariat of which is held by DIN.
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 announce this Technical Specification: Austria, Belgium, Bulgaria, Croatia, Cyprus,
Czech Republic, Denmark, Estonia, Finland, France, Germany, Greece, Hungary, Iceland, Ireland, Italy,
Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia,
Spain, Sweden, Switzerland and the United Kingdom.
Introduction
Airborne particles of biological origin are called bioaerosols. Depending on the emission source bioaerosols
vary in composition; one component of ambient bioaerosols with possible ecological and health relevance can
be moulds. Natural and anthropogenic sources for mould spores are widely distributed in the environment.
Anthropogenic sources can for example be agriculture and construction activities or waste treatment.
Mould is a common name for filamentous fungi from different taxonomic groups (Zygomycetes, Ascomycetes,
Deuteromycetes). They form a mycelium (hyphae) and spores – namely conidiospores (conidia),
sporangiospores or ascospores – by which they become visible macroscopically. Most spores are in the size
range of 2 µm to 10 µm, some up to 30 µm and only few up to 100 µm. 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.
The procedure described in this document is based on VDI 4252 Part 2 [1], VDI 4253 Part 2 [2] and is related
to the ISO standards on indoor air ISO 16000-16 [3] and ISO 16000-17 [4].
1 Scope
This Technical Specification describes the measurement of moulds in ambient air in order to identify, quantify
and characterize bioaerosol pollution in ambient air resulting from emissions from different sources.
The method described specifies the sampling of moulds as part of the suspended particulate matter (SPM,
here particles with aerodynamic diameter up to ca. 30 µm) using a filter sampling system with gelatine/poly-
carbonate filter combination followed by the culture-based analyses on DG18 agar. The sampling duration can
be varied between 10 min to 24 h. The health effect of bioaerosols is not limited to any particle fraction,
therefore, this document describes the sampling of moulds as part of the suspended particulate matter as a
convention method.
NOTE The sampling method described in this document in principle is likely to be appropriate for the sampling of
actinomycetes and other spore-forming bacteria (resistant to desiccation). For these species a special analytical
procedure using different culture media should be applied, but this is not within the scope of this document.
The standard method set out in this Technical Specification is accepted by convention as reference method.
The measured quantity, here the number of colony forming units per cubic meter (CFU/m ), is determined by
the inlet design of the sampling head, the associated operational parameters and the analytical procedure.
Standardized methods for sampling, detection and enumeration of moulds including standards for sampling
strategies are important for comparative assessment of moulds in ambient air. Before doing any
measurements a plan for the measurement strategy is necessary (see CEN/TS 16115-2 [5]).
WARNING — The use of this Technical Specification may involve hazardous materials, operations and
equipment. This Technical Specification does not purport to address all the safety problems
associated with its use. It is the responsibility of the user of this standard to establish appropriate
safety and health practices and determine the applicability of regulatory limitations prior to use.
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.
EN ISO 8199:2007, Water quality ― General guidance on the enumeration of micro-organisms by culture
(ISO 8199:2005)
3 Terms and definitions
For the purposes of this document, the following terms and definitions apply.
3.1
aerodynamic diameter
diameter of a sphere of density 1 g/cm³ with the same terminal velocity due to gravitational force in calm air as
the particle, under the prevailing conditions of temperature, pressure and relative humidity
[ISO 7708:1995, 2.2 [6]]
3.2
ambient air
outdoor air in the lower troposphere excluding workplace air
[EN 14907:2005, 3.1.1 [7]]
3.3
analytical blank value
value determined by a blank sample covering the analytical procedure to ensure that no significant
contamination occurs during the complete analytical procedure including autoclaving, agar preparation,
suspension and extraction of the filters, dilution, incubation, counting, etc.
3.4
bioaerosol
airborne particles of biological origin
[EN 13098:2000, 3.3 [8]].
NOTE Bioaerosols in the sense of this document are all aggregations of particles in the atmosphere to which fungi
(spores, conidia, fragments of hyphae), bacteria, viruses and/or pollen as well as their cell membrane components and
metabolites (e.g. endotoxins, mycotoxins) are attached or that consist of the above mentioned components.
3.5
biological sampling efficiency
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
[EN 13098:2000, 3.4 [8]]
NOTE The biological sampling efficiency considers the sampling stress occurring during sampling and analysis in
addition to the physical sampling efficiency. It refers to the proportion (in percent) of collected organisms which have not
lost the ability to be cultured subsequently. It is strain- and species specific.
3.6
colony count
number of all visible colonies of microorganisms on a culture medium after incubation under the selected
conditions
3.7
Colony Forming Unit
CFU
unit by which the culturable number of microorganisms is expressed
[EN 13098:2000, 3.5 [8]].
NOTE 1 One Colony Forming Unit can originate from one single microorganism, an aggregate of many
microorganisms or from one or many microorganisms attached to one particle.
NOTE 2 The number of outgrowing colonies depends on cultivation conditions.
3.8
culture-based analyses
cultivation
growing of microorganisms on culture media
[ISO 16000-16:2008, 3.6 [3]]
NOTE The prerequisites for the detection are the abilities to grow and propagate.
3.9
face velocity
air flow rate divided by the face area
NOTE 1 The face velocity is expressed in metres per second.
[Adapted from EN 779:2002, 3.11 [9]]
NOTE 2 In this document, the face velocity is defined as the volume flow rate divided by the effective filter area.
3.10
field blank value
value determined by a blank sample covering the complete measurement procedure including preparation,
sampling, transport and analyses to ensure that no significant contamination has occurred during all steps of
measurement and to check that the operator can achieve a quantification level adapted to the task
NOTE A field blank sample is a sample taken in an identical manner as the real sample, but without sucking air
through the sampling device. The resulting blank represents the number of CFU entering the sample simply by handling
the filter during sampling. The results of the field blanks are not used for correction of measurement results but to detect
sampling errors.
3.11
filtration
sampling of particles suspended in gas or liquid by flow through a porous medium
[EN 13098:2000, 3.11 [8]]
NOTE In this document, filtration is understood as the separation of moulds from a defined volume of air by means of
filters.
3.12
indirect method
suspension of deposited microorganisms with subsequent plating of aliquots
...
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