EN 17359:2020

Stationary source emissions - Bioaerosols and biological agents - Sampling of bioaerosols and collection in liquids - Impingement method

This document contains specifications for active sampling of bioaerosols from exhaust air flowing through a defined cross-section of a stack. It defines general principles that have to be taken into account during an isokinetic sampling campaign for bioaerosols by bubbling the exhaust air through a specific impinger designed for emission measurements.
In this document the application with culturable organisms is specified but the same principle might be applicable for non-cultural based methods (e.g. molecular and/or enzyme-based methods).
The impinger is designed to allow a sample volume flow of 1 m3/h to 1,8 m3/h, or 16 l/min to 30 l/min, respectively, and has been tested with regard to various microorganisms within broad concentration ranges [1; 2; 3; 4]

Emissionen aus stationären Quellen - Bioaerosole und biologische Agenzien - Probenahme von Bioaerosolen und Abscheidung in Flüssigkeiten - Impinger-Methode

Dieses Dokument enthält Festlegungen zur aktiven Probenahme von Bioaerosolen aus der Abluft, die durch einen definierten Querschnitt eines Abluftkanals strömt. Sie legt allgemeine Grundlagen fest, die bei einer isokinetischen Probenahmekampagne für Bioaerosole berücksichtigt werden müssen, wobei die Abluft durch einen spezifischen Impinger, der auf Emissionsmessungen ausgelegt ist, geleitet wird.
In diesem Dokument wird die Anwendung mit anzüchtbaren Organismen festgelegt, jedoch kann dasselbe Prinzip auch auf Verfahren angewendet werden, die nicht auf Kulturen basieren (z. B. molekulare und/oder enzymbasierte Verfahren).
Der Impinger ist auf einen Teilvolumenstrom von 1 m3/h bis 1,8 m3/h bzw. 16 l/min bis 30 l/min ausgelegt, und wurde für verschiedene Mikroorganismen in breiten Konzentrationsbereichen geprüft [1; 2; 3; 4].

Émissions de sources fixes - Bioaérosols et agents biologiques - Prélèvement des bioaérosols et collecte dans les liquides - Méthode d’impaction par bullage

Le présent document contient des spécifications relatives au prélèvement actif des bioaérosols présents dans l’air rejeté à travers une section définie d’une cheminée. Il définit des principes généraux qui doivent être pris en compte au cours d’une campagne de prélèvement isocinétique de bioaérosols, en faisant passer des bulles d’air rejeté à travers un bulleur spécifiquement conçu pour les mesurages d’émissions.
Bien que le présent document spécifie l’utilisation d’organismes cultivables, le même principe peut s’appliquer aux méthodes hors cultures (méthodes moléculaires et/ou de type enzymatique, par exemple).
Le bulleur est conçu pour obtenir un débit volumique d’échantillon de 1 m3/h à 1,8 m3/h, soit de 16 l/min à 30 l/min, et a été soumis à essai avec divers micro-organismes sur des gammes de concentrations étendues [1 ; 2 ; 3 ; 4].

Emisije nepremičnih virov - Bioaerosoli in biološki agensi - Vzorčenje bioaerosolov in zajem v tekočini - Metoda z izpiranjem

General Information

• Status: Published
• Publication Date: 29-Nov-2020
• Withdrawal Date: 27-Feb-2021
• ICS: 13.040.20 - Ambient atmospheres
• Technical Committee: CEN/TC 264 - Air quality
• Drafting Committee: CEN/TC 264/WG 28 - Measurement of airborne microorganisms in ambient air
• Current Stage: 6060 - Definitive text made available (DAV) - Publishing
• Start Date: 12-Aug-2020
• Due Date: 22-Dec-2020
• Completion Date: 12-Aug-2020
• Ref Project: SIST EN 17359:2020 - Stationary source emissions - Bioaerosols and biological agents - Sampling of bioaerosols and collection in liquids - Impingement method

Standards Content (Sample)

SLOVENSKI STANDARD
SIST EN 17359:2020
01-oktober-2020
Emisije nepremičnih virov - Bioaerosoli in biološki agensi - Vzorčenje
bioaerosolov in zajem v tekočini - Metoda z izpiranjem
Stationary source emissions - Bioaerosols and biological agents - Sampling of
bioaerosols and collection in liquids - Impingement method
Emissionen aus stationären Quellen - Bioaerosole und biologische Agenzien -
Probenahme von Bioaerosolen und Abscheidung in Flüssigkeiten - Impinger-Methode
Émissions de sources fixes - Bioaérosols et agents biologiques - Prélèvement des
bioaérosols et collecte dans les liquides - Méthode d’impaction par bullage
Ta slovenski standard je istoveten z: EN 17359:2020
ICS:
13.040.40 Emisije nepremičnih virov Stationary source emissions
SIST EN 17359:2020 en,fr,de

2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

---------------------- Page: 1 ----------------------
SIST EN 17359:2020
---------------------- Page: 2 ----------------------
SIST EN 17359:2020
EN 17359
EUROPEAN STANDARD
NORME EUROPÉENNE
August 2020
EUROPÄISCHE NORM
ICS 13.040.20
English Version
Stationary source emissions - Bioaerosols and biological
agents - Sampling of bioaerosols and collection in liquids -
Impingement method

Émissions de sources fixes - Bioaérosols et agents Emissionen aus stationären Quellen - Bioaerosole und

biologiques - Prélèvement des bioaérosols et collecte biologische Agenzien - Probenahme von Bioaerosolen

dans les liquides - Méthode d'impaction par bullage und Abscheidung in Flüssigkeiten - Impinger-Methode

This European Standard was approved by CEN on 5 July 2020.

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, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway,

Poland, Portugal, Republic of North Macedonia, Romania, Serbia, 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: Rue de la Science 23, B-1040 Brussels

© 2020 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN 17359:2020 E

worldwide for CEN national Members.
---------------------- Page: 3 ----------------------
SIST EN 17359:2020
EN 17359:2020 (E)
Contents Page

European foreword ....................................................................................................................................................... 5

1 Scope .................................................................................................................................................................... 6

2 Normative references .................................................................................................................................... 6

3 Terms and definitions ................................................................................................................................... 7

4 Symbols and abbreviations ...................................................................................................................... 11

5 Principle of method ..................................................................................................................................... 13

6 Theoretical fundamentals ......................................................................................................................... 14

6.1 Isokinetic sampling ..................................................................................................................................... 14

6.2 Determination of concentration and load of the microorganisms ............................................. 14

7 Device and materials .................................................................................................................................. 16

7.1 General ............................................................................................................................................................. 16

7.2 Device and methods for measurement of the exhaust air parameters for the

calculation of the main volume flow ..................................................................................................... 16

7.2.1 General ............................................................................................................................................................. 16

7.2.2 Device for determination of the exhaust air velocity ...................................................................... 16

7.2.3 Device for determination of pressure, temperature and humidity ........................................... 17

7.3 Device for the sampling of bioaerosols ................................................................................................ 17

7.3.1 General ............................................................................................................................................................. 17

7.3.2 Material properties ..................................................................................................................................... 18

7.3.3 Entry nozzle, bend and sampling probe ............................................................................................... 18

7.3.4 Emission impinger ....................................................................................................................................... 18

7.3.5 Suction device and device for measurement of the gas volume or respectively the gas

volume flow .................................................................................................................................................... 20

8 Sampling .......................................................................................................................................................... 20

8.1 General ............................................................................................................................................................. 20

8.2 Preparation of the sampling equipment .............................................................................................. 20

8.2.1 General ............................................................................................................................................................. 20

8.2.2 Preparation of the emission impinger ................................................................................................. 20

8.2.3 Preparation of the entry nozzle and the sampling probe .............................................................. 21

8.2.4 Determination of appropriate sampling probe and sampling flow ........................................... 21

8.3 Performing bioaerosol sampling ............................................................................................................ 24

8.3.1 Leak test and sampling ............................................................................................................................... 24

8.3.2 Recovery of deposits upstream of the emission impinger ............................................................ 25

8.3.3 Determination of the mass of the sampling liquid ........................................................................... 26

8.3.4 Field blank value .......................................................................................................................................... 26

8.3.5 Analytical blank value ................................................................................................................................ 27

8.4 Transport and storage ................................................................................................................................ 27

9 Analysis ............................................................................................................................................................ 27

10 Evaluation ....................................................................................................................................................... 27

10.1 General ............................................................................................................................................................. 27

10.2 Transfer of the results by the analytical laboratory ....................................................................... 28

10.3 Sample gas volume during sampling..................................................................................................... 28

10.4 Microorganism number calculation ...................................................................................................... 30

10.5 Load calculation ............................................................................................................................................ 31

---------------------- Page: 4 ----------------------
SIST EN 17359:2020
EN 17359:2020 (E)

11 Performance characteristerics ................................................................................................................ 31

11.1 Measurement uncertainty ......................................................................................................................... 31

11.2 Parameters for the determination of measuring uncertainity in practice .............................. 32

12 Maintenance and quality assurance ...................................................................................................... 37

13 Sampling efficiency and limits of the method .................................................................................... 37

14 Interferences .................................................................................................................................................. 38

Annex A (informative) Practical example for moulds and bacteria ........................................................ 39

A.1 General ............................................................................................................................................................. 39

A.2 Determination of the measurement points ......................................................................................... 39

A.3 Devices and materials ................................................................................................................................. 39

A.3.1 General ............................................................................................................................................................. 39

A.3.2 Devices and methods for measurement of the exhaust air parameters for the

calculation of the main volume flow ...................................................................................................... 39

A.3.2.1 General ............................................................................................................................................................. 39

A.3.2.2 Devices for determination of the exhaust air velocity .................................................................... 39

A.3.2.3 Devices for determination of pressure, temperature and humidity.......................................... 40

A.3.3 Devices for sampling of bioaerosols ...................................................................................................... 40

A.3.3.1 General ............................................................................................................................................................. 40

A.3.3.2 Material properties ...................................................................................................................................... 40

A.3.3.3 Entry nozzle, bend and sampling probe ............................................................................................... 40

A.3.3.4 Emission impinger ....................................................................................................................................... 40

A.3.3.5 Measurement system for isokinetic sample volume flow abstraction ...................................... 40

A.4 Sampling process .......................................................................................................................................... 40

A.4.1 General ............................................................................................................................................................. 40

A.4.2 Preparation of the sampling equipment .............................................................................................. 40

A.4.2.1 General ............................................................................................................................................................. 40

A.4.2.2 Preparation of the emission impinger .................................................................................................. 41

A.4.2.3 Preparation of the entry nozzle, band and sampling probe .......................................................... 41

A.4.3 Measurement of the exhaust air parameters for isokinetic sampling ...................................... 41

A.4.4 Sampling .......................................................................................................................................................... 42

A.4.4.1 General ............................................................................................................................................................. 42

A.4.4.2 Recovery of deposits upstream of the emission impinger ............................................................ 42

A.4.4.3 Field blank value ........................................................................................................................................... 42

A.4.5 Transport and storage ................................................................................................................................ 43

Annex B (informative) Measurement uncertainty ......................................................................................... 49

B.1 General ............................................................................................................................................................. 49

B.2 Determination of measurement uncertainty ..................................................................................... 49

---------------------- Page: 5 ----------------------
SIST EN 17359:2020
EN 17359:2020 (E)

B.2.1 Moulds .............................................................................................................................................................. 49

B.2.2 Mesophilic bacteria ..................................................................................................................................... 50

B.2.3 Total cell count .............................................................................................................................................. 50

B.2.4 Measurements in the bioaerosol test channel ................................................................................... 50

B.3 Field blank value .......................................................................................................................................... 51

Annex C (normative) Summary of the requirements to the emission measurement ....................... 52

Annex D (informative) Sample protocol for sampling and analysis ....................................................... 54

D.1 Sampling .......................................................................................................................................................... 54

D.2 Analysis ............................................................................................................................................................ 55

Bibliography ................................................................................................................................................................. 56

---------------------- Page: 6 ----------------------
SIST EN 17359:2020
EN 17359:2020 (E)
European foreword

This document (EN 17359:2020) has been prepared 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 February 2021, and conflicting national standards shall

be withdrawn at the latest by February 2021.

Attention is drawn to the possibility that some of the elements of this document may be the subject of

patent rights. CEN shall not be held responsible for identifying any or all such patent rights.

According to the CEN-CENELEC Internal Regulations, the national standards organisations of the

following countries are bound to implement this European Standard: 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, Republic of North

Macedonia, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and the United

Kingdom.
---------------------- Page: 7 ----------------------
SIST EN 17359:2020
EN 17359:2020 (E)
1 Scope

This document contains specifications for active sampling of bioaerosols from exhaust air flowing

through a defined cross-section of a stack. It defines general principles that have to be taken into account

during an isokinetic sampling campaign for bioaerosols by bubbling the exhaust air through a specific

impinger designed for emission measurements.

In this document the application with culturable organisms is specified but the same principle might be

applicable for non-cultural based methods (e.g. molecular and/or enzyme-based methods).

3 3

The impinger is designed to allow a sample volume flow of 1 m /h to 1,8 m /h, or 16 l/min to 30 l/min,

respectively, and has been tested with regard to various microorganisms within broad concentration

ranges [1; 2; 3; 4].
2 Normative references

The following documents are referred to in the text in such a way that some or all of their content

constitutes requirements 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 1040:2005, Chemical disinfectants and antiseptics - Quantitative suspension test for the evaluation of

basic bactericidal activity of chemical disinfectants and antiseptics - Test method and requirements

(phase 1)

EN 13284-1:2017, Stationary source emissions - Determination of low range mass concentration of dust -

Part 1: Manual gravimetric method

EN 15259:2007, Air quality - Measurement of stationary source emissions - Requirements for measurement

sections and sites and for the measurement objective, plan and report

CEN/TS 16115-1, Ambient air quality - Measurement of bioaerosols - Part 1: Determination of moulds using

filter sampling systems and culture-based analyses

EN ISO 16911-1, Stationary source emissions - Manual and automatic determination of velocity and volume

flow rate in ducts - Part 1: Manual reference method (ISO 16911-1)

EN ISO 20988:2007, Air quality - Guidelines for estimating measurement uncertainty (ISO 20988:2007)

This method is accepted by convention as reference method for determination of total emissions under application

of an out stack configuration according to EN 13284-1.
---------------------- Page: 8 ----------------------
SIST EN 17359:2020
EN 17359:2020 (E)
3 Terms and definitions
For the purposes of this document, the following terms and definitions.

ISO and IEC maintain terminological databases for use in standardization at the following addresses:

— ISO Online browsing platform: available at https://www.iso.org/obp
— IEC Electropedia: available at http://www.electropedia.org/
3.1
waste gas
exhaust air
carrier gases with solid, liquid or gaseous constituents (emissions)

Note 1 to entry: The carrier gases can be natural air (e.g. from stable ventilation) or process gases.

3.2
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
Note 1 to entry: according to CEN/TS 16115-1
3.3
bacteria

large group of prokaryotic microorganisms with one chromosome in a nuclear region and which replicate

only asexually by cell division
[SOURCE: EN 13098:2019, 3.3]

Note 1 to entry: The current classification of bacteria, i.e. the grouping to genera and species, is done according

to molecular sequence-based, chemotaxonomic and physiological properties.
3.4
standard conditions

reference values for a dry gas at a pressure of 101,325 kPa rounded to 101,3 kPa and a temperature of

273,15 K rounded to 273 K

Note 1 to entry: Where a regulatory authority stipulates reference conditions, for example through a site Licence

or Permit, this shall override the conditions specified above.
3.5
bioaerosol-
airborne particles of biological origin
[SOURCE: EN 13098:2019, 3.1]

Note 1 to entry: Bioaerosols in the sense of this European Standard 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.
---------------------- Page: 9 ----------------------
SIST EN 17359:2020
EN 17359:2020 (E)
3.6
biological sampling efficiency

capacity of the sampler to maintain the viability of the airborne microorganisms during collection and

also to keep the microbial products intact
[SOURCE: EN 13098:2019, 3.4]

Note 1 to entry: The biological sampling efficiency considers the sampling stress occurring during sampling and

analysis in addition to the physical sampling efficiency. It is strain- and species specific.

3.7
endotoxin

constituent of the external membrane of Gram-negative bacteria (lipopolysaccharide), consisting of a

complex lipid, lipid A, which is covalently bound to a polysaccharide

Note 1 to entry: "Free endotoxin" is liberated after cell death and by budding from living cells. Lipid A is the active

(toxic) part and is a potent pro-inflammatory substance and may induce febrile, bronchial and other symptoms in

exposed workers. The composition and the toxicity of endotoxin differs between species.

[SOURCE: EN 13098:2019, 3.9]
3.8
load

product of measured concentration of bioaerosols and the volumetric flow rate of the stack of the plant

3.9
overall blank sample
field blank sample

sample taken at the plant site in an identical manner to the normal samples in the series, except that the

sampling system is not inserted into the duct and no gas is sampled during test duration

Note 1 to entry: In this standard the term field blank sample is used.
3.10
total cell count
total number of viable and dead cells in a given volume
Note 1 to entry: The total cell count can be determined e.g. by DAPI staining.

Note 2 to entry: When for example stained with DNA fluorescent dyes like DAPI (4,6-Diamidino-2-phenylindole-

dihydrochloride), living and dead cells are jointly counted and cannot be distinguished.

3.11
main volume flow
volumetric flow of the flue gas in the sampling plane
3.12
impingement

separation of airborne particles in liquids by different mechanisms, i.e. impaction, diffusion, interception

and sedimentation
---------------------- Page: 10 ----------------------
SIST EN 17359:2020
EN 17359:2020 (E)
3.13
Colony Forming Unit
CFU
unit by which the culturable number of microorganisms is expressed
[SOURCE: EN 13098:2019, 3.6 and 3.7]

Note 1 to entry: 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 to entry: The number of colonies which develop depends on cultivation conditions.

3.14
culture based analyses
cultivation
growing of microorganisms on culture media
[SOURCE: ISO 16000-16:2008, 3.6]
3.15
non-culture-based analyses

methods which do not rely on cultural methods including: microscopy, molecular methods, cytometry,

fluorescence etc.
3.16
indicator organisms

microorganisms which are characteristic for the emissions of a specific type of plant and can be detected

with currently available methods of sampling and analysis

Note 1 to entry: Indicator organisms which are characteristic of a specific source (process) can also occur in air

not influenced by the respective source, but mostly in minor concentrations. This is explained by the fact that many

microorganisms are found ubiquitously.
3.17
sampling line
measurement line

line in the sampling plane along which the sampling points are located, bounded by the inner duct wall

[SOURCE: EN 15259:2007, 3.15]
3.18
measurement site

place on the flue gas duct in the area of the sampling plane consisting of structures and technical

equipment, for example measurement platforms, measurement ports and energy supply

[SOURCE: EN 15259:2007, 3.11 – modified]
3.19
sampling point
measurement point

specific position on a sampling line at which a sample is extracted or the measurand directly determined

[SOURCE: EN 15259:2007, 3.16]
---------------------- Page: 11 ----------------------
SIST EN 17359:2020
EN 17359:2020 (E)
3.20
sampling plane
measurement plane
plane normal to the centreline of the duct at the sampling position
[SOURCE: EN 15259:2007, 3.13]

Note 1 to entry: In the case of emission measurements of area sources, the total surface corresponds to the

sampling plane.
3.21
microbial air pollution

concentrations of airborne microorganisms that exceed natural concentrations or compositions which

differ in type from those occurring naturally
3.22
microorganism

microbial entity, either cellular or non cellular, that is capable of multiplication or transfer of genetic

material, or entities that have lost these properties
[SOURCE: EN 13098:2019, 3.18]
3.23
physical sampling efficiency
capacity of the sampling device to collect particles suspended in air
[SOURCE: EN 13098:2019, 3.20]
3.24
fungi

unicellular yeasts and filamentous fungi from several taxonomic groups namely zygomycetes

(Mucormycotina and Entomophthoromycotina), ascomycetes (Ascomycota) and mitosporic fungi

(deuteromycetes, fungi imperfecti) [7]

Note 1 to entry: Moulds form different types of spores depending on the taxonomic group they belong to, namely

conidiospores (conidia), sporangiospores or ascospores.
3.25
sample volumetric flow

volumetric flow taken off from the main stream for determination of the measured component

[SOURCE: EN 15259:2007, 3.29]
3.26
sampling train
fully assembled sampling system as per Figure 1
---------------------- Page: 12 ----------------------
SIST EN 17359:2020
EN 17359:2020 (E)
4 Symbols and abbreviations
The symbols as outlined below are used throughout this standard:
Symbol Unit Meaning
A 2 sampling plane of the duct
A 2 area calculated on the basis of the diameter δ of the nozzle
S m S
c 3 3 number concentration of microorganisms in the sample flow
CFU/m ;TCC/m
C 3 3 number concentration of microorganisms in the exhaust air
CFU/m ;TCC/m
∑Cplate CFU sum of counted colonies on all evaluated plates
d dilution factor of the first evaluated dilution (for example d =
100 at a dilution degree 10 )
D additional dilution factor, which results from the fact that only
one aliquot of the dilutions has been outplated
f kg/m sample gas volume humidity at the volume meter (relative to
the standard conditions)
f kg/m humidity in the exhaust air
n number of plates at the lowest evaluable dilution (higher
concentration)
n number of plates at the nearest higher evaluable dilution (lower
concentration)
N number of duplicate analyses
N CFU number of bacteria in cfu
CFU
p pipetting factor
p Pa dynamic pressure
p kPa absolute exhaust air pressure
p kPa absolute pressure at the gas volume meter
p kPa standard pressure (101,3 kPa)
p kPa ambient air pressure
p kPa absolute pressure at the float-type flow meter inlet
p kPa static pressure in the duct
% volume fraction of water vapour in the humid exhaust air
H O, G
% volume fraction of water vapour in the humid exhaust air at the
H O, GM
gas volume meter
s 3 3 standard deviation under repeatability conditions
CFU/m ; TCC/m
...