1、BRITISH STANDARD Workplace atmosphere - Guidelines for measurement of airborne micro-organisms and endotoxin The European Standard EN 13098:2001 has the status of a British Standard ICs 13.040.30 BS EN 13098:2001 NO COPYING WITHOUT BSI PERMISSION EXCEPT AS PERMITTED BY COPYRIGHT LAW BS EN 13098:2001
2、 National foreword This British Standard is the official English language version of EN 13098:2001. The UK participation in its preparation was entrusted by Technical Committee EH/2, Air quality, to Subcommittee EH/2/2, Workplace atmospheres, which has the responsibility to: - - aid enquirers to und
3、erstand the text; present to the responsible European committee any enquiries on the interpretation, or proposals for change, and keep the UK interests informed; monitor related international and European developments and promulgate them in the UK. - A list of organizations represented on this subco
4、mmittee can be obtained on request to its secretary. Cross-references The British Standards which implement international or European publications referred to in this document may be found in the BSI Standards Catalogue under the section entitled “International Standards Correspondence Index”, or by
5、 using the “Find” facility of the BSI Standards Electronic Catalogue. A British Standard does not purport to include all the necessary provisions of a contract. Users of British Standards are responsible for their correct application. Compliance with a British Standard does not of itself confer immu
6、nity from legal obligations. This British Standard, having been prepared under the direction of the Environment Sector Committee, was published under the authority of the Standards Committee and comes into effect on 15 July 2001 Summary of pages This document comprises a front cover, an inside front
7、 cover, the EN title page, pages 2 to 24, an inside back cover and a back cover. The BSI copyright date displayed in this document indicates when the document was last issued. Amendments issued since publication Amd. No. /Date 1 Comments O BSI 07-2001 ISBN O 580 37873 X EUROPEAN STANDARD NORME EUROP
8、ENNE EUROPISCHE NORM O September 2000 ICs 07.100.99; 13.040.30 English version Workplace atmosphere - Guidelines for measurement of airborne micro-organisms and endotoxin Atmosphres des lieux de travail - Rgles pour le mesurage de micro-organismes et dendotoxine en suspension dans lair Arbeitsplatza
9、tmosphre - Leitlinien fr die Messung von Mikroorganismen und Endotoxin in der Luf This European Standard was approved by CEN on 17 August 2000. CEN members are bound to comply with the CENEENELEC Internal Regulations which stipulate the conditions for giving this European Standard the status of a na
10、tional standard without any alteration. Up-to-date lists and bibliographical references concerning such national standards may be obtained on application to the Central Secretariat or to any CEN member. This European Standard exists in three official versions (English, French, German). A version in
11、any other language made by translation under the responsibility of a CEN member into its own language and notified to the Central Secretariat has the same status as the official versions. CEN members are the national standards bodies of Austria, Belgium, Czech Republic, Denmark, Finland, France, Ger
12、many, Greece, Iceland, Ireland, Italy, Luxembourg, Netherlands, Norway, Portugal, Spain, Sweden, Switzerland and United Kingdom. EUROPEAN COMMEE FOR STANDARDIZATION COMITE EUROPEEN DE NORMALISATION EUROPISCHES KOMITEE FR NORMUNG Central secretariat: rue de Stassart, 36 B-i050 Brussels 0 0 2000 CEN A
13、ll rights of exploitation in any form and by any means reserved worldwide for CEN national Members. Ref. No. EN 13098:2000 E Page 2 EN 13098:2000 Contents Page Foreword 3 Introduction 4 1 Scope . 4 2 Normative references . 4 3 Terms and definitions 4 4 5 Measurement of micro-organisms and endotoxin
14、7 6 Sampling . 8 7 Analytical method . 10 8 Expression of results . 13 9 Report 13 Annex A (informative ) Recommendations for selection of measurement methods . 14 Annex B (informative) Example of sampling form . 20 Annex C (informative) List of general media . 21 Annex D (informative) Colony counti
15、ng - Formula and examples . 22 Bibliography . 24 Symbols and abbreviations . 7 . Page 3 EN 13098:2000 Foreword This European Standard has been prepared by Technical Committee CEN/TC 137 “Assessment of workplace exposure“, the secretariat of which is held by DIN. This European Standard shall be given
16、 the status of a national standard, either by publication of an identical text or by endorsement, at the latest by March 2001, and conflicting national standards shall be withdrawn at the latest by March 2001. According to the CENKENELEC Internal Regulations, the national standards organizations of
17、the following countries are bound to implement this European Standard: Austria, Belgium, Czech Republic, Denmark, Finland, France, Germany, Greece, Iceland, Ireland, Italy, Luxembourg, Netherlands, Norway, Portugal, Spain, Sweden, Switzerland and the United Kingdom. This European Standard has been p
18、repared by Technical Committee CEN/TC 137 “Assessment of workplace exposure“, the secretariat of which is held by DIN. Annexes A, B, C and D are informative. This standard includes a bibliography. Page 4 EN 13098:2000 Introduction Assessing occupational exposure to airborne microbial contaminants in
19、 a representative way is a challenging task. It is necessary however that information can be gathered to evaluate and to minimise exposure to biological agents. The sampling equipment used often introduces its own critical limitations, as in the assessment of aerosol fractions. Some sampling equipme
20、nt may be capable only of measuring culturable micro-organisms, while others allow the characterisation of both the total number of organisms and the culturable fraction. Analytical procedures may add further to the difficulties and the uncertainties, e.g. the method used may not allow the identific
21、ation of the biological agents present, or may cause unwanted interference between different biological agents. However, by adhering to the principles outlined in this standard for choice of sampling and analytical procedures, these uncertainties can be reduced and con- trolled, allowing comparable
22、and representative measurements to be made. i Scope including the determination of total number and culturable number of micro-organisms in the workplace atmosphere. T rn The European Standard provides guidelines for the assessment of workplace exposure to airborne micro-organis standard also provid
23、es methods for measurement of airborne endotoxin in the work environment. The European Standard does not apply to viruses, specific pathogenic micro-organisms and toxins other than endotoxin, although some of the measurement principles may be the same. 2 Normative references This European Standard i
24、ncorporates by dated or undated reference, provisions from other publications. These normative references are cited at the appropriate place in the text and the publications are listed hereafter. Dated references, subsequent amendments to or revisions of any of these publications apply to this Europ
25、ean Standard only when incorporated in it by amendment or revision. For undated references the latest edition of the publication referred to applies (including amendments). EN 481 , Workplace atmospheres - Size fraction definitions for measurement of airborne particles EN 482, Workplace atmospheres
26、- General requirements for the performance of procedures for the measurement of chemical agents EN 689, Workplace atmospheres - Guidance for the assessment of exposure by inhalation to chemical agents comparison with limit values and measurement strategy EN 1232, Workplace atmospheres - Pumps for pe
27、rsonal sampling of chemical agents - Requirements and test methods EN 1291 9, Workplace atmospheres - Pumps for the sampling of chemical agents with a volume flow rate of over 5 Vmin - Requirements and test methods IS0 7218, Microbiology of food and animal feeding stuffs - General rules for microbio
28、logical examinations 3 Terms and definitions For the purposes of this European Standard the following terms and definitions apply: 3.1 actinomycetes varied group of rod-shaped to filamentous Gram-positive bacteria. NOTE Filamentous actinomycetes form a branching network of thin filaments called a my
29、celium. Most actinomycetes replica. by conidia-like spores which can easily be made airborne. Page 5 EN 13098:2000 Leria large group of prokaryotic micro-organisms with one chromosome in a nuclear region and which replicate only asexu- ally by cell division. NOTE criteria divide into spheres (cocci)
30、 and rods. Some species produce endospores as survival units. Different cell-wall chemistry is used for the classification of Gram-positive and Gram-negative bacteria. Morphological 3.3 bioaerosol airborne particles with biological origin NOTE 1 In this standard, micro-organisms and endotoxin are co
31、nsidered. NOTE 2 The majority of bioaerosol particles are complex as far as size, shape and constituent elements are concerned 3.4 biological preservation efficiency the capacity of the sampler to maintain the viability of the airborne micro-organisms during collection and also to keep the microbial
32、 products intact. colony forming unit the unit by which the culturable number is expressed. 03.5 NOTE 1 One colony forming unit can originate from one single micro-organism, an aggregate of many micro-organisms or from one or many micro-organisms attached to one particle. NOTE 2 The number of outgro
33、wn colonies may depend on cultivation conditions. 3.6 culturable number the number of micro-organisms, single cells or aggregates able to form colonies on a solid nutrient medium. NOTE but are not necessarily culturable The viable number (see 3.21) includes all potentially metabolic active micro-org
34、anisms. Some micro-organisms are viable 3.7 elevated level a level above normal background level of micro-organisms in a specified environment. efdotoxin a constituent of the external membrane of Gram-negative bacteria (lipopolysaccharide), consisting of a complex lipid, lipid A, which is covalently
35、 bound to a polysaccharide. NOTE “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
36、 endotoxin differs between species. 3.9 endotoxin unit a unit standardized against the defined reference material (Reference standard endotoxin). 3.1 O exposure (by inhalation) a situation in which a chemical or biological agent is present in air which is inhaled by a person (see EN 482). 3.1 1 filt
37、ration collection of particles suspended in gas or liquid by flow through a porous medium. O Page 6 EN 13098:2000 3.1 2 fungi a diverse group of eukaryotic micro-organisms with membrane-bound nucleus comprising several chromosomes. NOTE Multiplication is mainly asexual but several groups replicate a
38、lso by sexual spores. Filamentous fungi (moulds) grow in lengthy hyphae and form compact tufts called mycelia. Asexual spores (conidia) are easily made airborne. Yeasts are usually unicellular, of spherical shape and their cells multiply sexually or asexually by budding. 3.13 impaction collection of
39、 airborne particles accelerated through a nozzle or orifice on a surface by inertia effect. 3.14 impingement combination of impaction onto a surface and subsequent dispersion into a liquid medium. 3.15 Limulus Amoebocyte Lysate enzymes extracted from the blood cells of the horse shoe crab (Limulus p
40、olyphemus) that are activated by endotoxin. 3.16 micro-organisms any microbiological entity, cellular or non cellular, capable of replication or of transferring genetic material, or entities that have lost these properties. NOTE The term micro-organism covers the term of biological agent, according
41、to the Directive 90/679/EEC: micro-organisms, including those which have been genetically modified, cell cultures and human endoparasites which may be able to provoke any infection, allergy or toxicity. 3.17 physical sampling efficiency the capacity of the sampler to collect particles with specific
42、sizes suspended in workplace air. 3.1 8 sieve sampler multi-orifice impactor. 3.19 total sampling efficiency the product of the physical sampling efficiency and the biological preservation efficiency. 3.20 total number of micro-organisms the number of micro-organisms determined as single organisms (
43、or a corresponding measure). NOTE Both the viable and the non-viable micro-organisms are included. 3.21 viable number of micro-organisms the number of micro-organisms having a potential for metabolic activity. NOTE underestimate the viable number. A viable micro-organism is not necessarily culturabl
44、e, which means that the number of culturable micro-organisms may 3.22 work pattern the definable series of activities from the periods under consideration (see EN 689). 3.23 workplace the workplace is the defined area or areas in which the work activities are carried out (see EN 689). O Page 7 EN 13
45、098:2000 4 Symbols and abbreviations O ATP Adenosine triphosphate CFU Colony forming unit CV Coefficient of variation DNA Deoxyribonucleic acid EU Endotoxin unit GSD Geometric standard deviation LAL Limulus Amoebocyte Lysate LPS Lipopolysaccharide (see 3.8) SEM Scanning electron microscope 05 Measur
46、ement of micro-organisms and endotoxin 5.1 Biological agents and biological properties Bioaerosols may contain different micro-organisms and/or different components originating from these. Micro-organisms may be classified in different taxonomic groups like Gram-positive and Gram-negative bacteria,
47、actinomycetes, fungi, protozoa, algae and viruses. These may be further classified to genus or species level. Immunologic reactions e.g. allergic and/or toxic reactions can result from exposure to micro-organisms irrespective of their viability. NOTE Species identification may be useful in studies o
48、f specific allergic and toxic reactions. 5.2 Aim of measurement The measurement of micro-organisms and endotoxin in the workplace air has two objectives: - to assess workers exposure; and/or - to assess the biological characteristics of air at different locations and/or at different times and over d
49、ifferent intervals of time. It is essential to state the purpose of the measurement and how the results will be interpreted. o NOTE Measurement tasks could be to locate the sources emitting micro-organisms, to measure a workers daily or work shift exposure, to identify peaks in exposure, to test the efficiency of control measures, or to control actions taken to diminish the exposure. It is essential to verify that the sampling and analysis corresponds to the assessment objectives and to the search for the targeted component of the bioaerosol (see 5.1). It can be appropriate to
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