1、July 2014Translation by DIN-Sprachendienst.English price group 17No part of this translation may be reproduced without prior permission ofDIN Deutsches Institut fr Normung e. V., Berlin. Beuth Verlag GmbH, 10772 Berlin, Germany,has the exclusive right of sale for German Standards (DIN-Normen).ICS 71
2、.100.35!%2V“2159751www.din.deDDIN EN 16438Chemical disinfectants and antiseptics Quantitative surface test for the evaluation of fungicidal or yeasticidalactivity of chemical disinfectants and antiseptics used in the veterinaryarea on non-porous surfaces without mechanical action Test method and req
3、uirements (phase 2, step 2);English version EN 16438:2014,English translation of DIN EN 16438:2014-07Chemische Desinfektionsmittel und Antiseptika Quantitativer Oberflchenversuch zur Bestimmung der fungiziden oder levurozidenWirkung chemischer Desinfektionsmittel und Antiseptika fr den Veterinrberei
4、ch aufnicht-porsen Oberflchen ohne mechanische Wirkung Prfverfahren und Anforderungen (Phase 2, Stufe 2);Englische Fassung EN 16438:2014,Englische bersetzung von DIN EN 16438:2014-07Antiseptiques et dsinfectants chimiques Essai quantitatif de surface pour lvaluation de lactivit fongicide ou levurici
5、de desantiseptiques et des dsinfectants chimiques utiliss dans le domaine vtrinaire sur dessurfaces non poreuses sans action mcanique Mthode dessai et prescriptions (phase 2, tape 2);Version anglaise EN 16438:2014,Traduction anglaise de DIN EN 16438:2014-07www.beuth.deIn case of doubt, the German-la
6、nguage original shall be considered authoritative.Document comprises 41 pages06.14 DIN EN 16438:2014-07 2 A comma is used as the decimal marker. National forewordThis document (EN 16438:2014) has been prepared by Technical Committee CEN/TC 216 “Chemical disinfectants and antiseptics” (Secretariat: A
7、FNOR, France), Working Group WG 2 “Veterinary use” (Secretariat: BSI, United Kingdom). The responsible German body involved in its preparation was the Normenausschuss Lebensmittel und landwirtschaftliche Produkte (Food and Agricultural Products Standards Committee), Working Committee NA 057-02-03 AA
8、 Desinfektionsmittel Tierhaltung/Lebensmittelbereich. ICS 71.100.35 English Version Chemical disinfectants and antiseptics - Quantitative surface test for the evaluation of fungicidal or yeasticidal activity of chemical disinfectants and antiseptics used in the veterinary area on non-porous surfaces
9、 without mechanical action - Test method and requirements (phase 2, step 2) Antiseptiques et dsinfectants chimiques - Essai quantitatif de surface pour lvaluation de lactivit fongicide ou levuricide des antiseptiques et des dsinfectants chimiques utiliss dans le domaine vtrinaire sur des surfaces no
10、n poreuses sans action mcanique - Mthode dessai et prescriptions (phase 2, tape 2) Chemische Desinfektionsmittel und Antiseptika - Quantitativer Oberflchenversuch zur Bestimmung der fungiziden oder levuroziden Wirkung chemischer Desinfektionsmittel und Antiseptika fr den Veterinrbereich auf nicht-po
11、rsen Oberflchen ohne mechanische Wirkung - Prfverfahren und Anforderungen (Phase 2, Stufe 2) This European Standard was approved by CEN on 30 November 2013. CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulate the conditions for giving this European Standard the
12、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
13、, 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, C
14、roatia, 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 an
15、d United Kingdom. CEN-CENELEC Management Centre: Avenue Marnix 17, B-1000 Brussels 2014 CEN All rights of exploitation in any form and by any means reserved worldwide for CEN national Members. Ref. No. EN 16438:2014 EEUROPEAN COMMITTEE FOR STANDARDIZATIONCOMIT EUROPEN DE NORMALISATIONEUROPISCHES KOM
16、ITEE FR NORMUNGEN 16438 February 2014 EUROPEAN STANDARDNORME EUROPENNEEUROPISCHE NORM2 Contents PageForeword 3 Introduction .4 1 Scope 5 2 Normative references 5 3 Terms and definitions .5 4 Requirements .5 5 Test method 6 5.1 Principle 6 5.2 Materials and reagents 7 5.2.1 Test organism 7 5.2.2 Cult
17、ure media and reagents .7 5.2.3 Test surface . 10 5.3 Apparatus and glassware 10 5.3.1 General . 10 5.3.2 Usual microbiological laboratory equipment . 10 5.4 Preparation of test organism suspensions and product test solutions . 11 5.4.1 Test organism suspension (test and validation suspension) 11 5.
18、4.2 Product test solutions 15 5.5 Procedure for assessing the fungicidal or yeasticidal activity of the product 16 5.5.1 General . 16 5.5.2 Test procedure (Dilution-neutralisation method) 17 5.5.3 Observation of the test surface agar 20 5.6 Experimental data and calculation 20 5.6.1 Explanation of t
19、erms and abbreviations 20 5.6.2 Calculation . 21 5.7 Verification of methodology 24 5.7.1 General . 24 5.7.2 Control of weighted mean counts . 24 5.7.3 Basic limits 24 5.7.4 Microscopic observation . 24 5.8 Expression of results and precision . 24 5.8.1 Reduction 24 5.8.2 Control of active and non-a
20、ctive product test solution (5.4.2) 25 5.8.3 Fungicidal or yeasticidal concentration . 25 5.8.4 Precision, repetitions . 25 5.9 Interpretation of results - conclusion . 25 5.9.1 General . 25 5.9.2 Fungicidal or yeasticidal activity for general purposes . 26 5.9.3 Qualification for certain fields of
21、application 26 5.10 Test report . 26 Annex A (informative) Referenced strains of national collections . 28 Annex B (informative) Examples of neutralisers of the residual antimicrobial activity of chemical disinfectants and antiseptics 29 Annex C (informative) Graphical representations of dilution-ne
22、utralisation method . 31 Annex D (informative) Example of a typical test report 35 Bibliography . 39 EN 16438:2014 (E) DIN EN 16438:2014-073 Foreword This document (EN 16438:2014) has been prepared by Technical Committee CEN/TC 216 “Chemical disinfectants and antiseptics”, the secretariat of which i
23、s held by AFNOR. 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 August 2014 and conflicting national standards shall be withdrawn at the latest by August 2014. Attention is drawn to the possibili
24、ty 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 organisations of the following countries are bo
25、und 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, Portug
26、al, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and the United Kingdom. EN 16438:2014 (E)DIN EN 16438:2014-074 Introduction This European Standard specifies a surface test for establishing whether a chemical disinfectant or antiseptic has or does not have fungicidal or yeasticida
27、l activity in the fields described in the scope. This laboratory test takes into account practical conditions of application of the product including contact time, temperature, test organisms and interfering substances, i.e. conditions which may influence its action in practical situations. The cond
28、itions are intended to cover general purposes and to allow reference between laboratories and product types. Each utilisation concentration of the chemical disinfectant or antiseptic, found by this test corresponds to the chosen experimental conditions. However, for some applications the instruction
29、s of use of a product may differ and therefore additional test conditions need to be used. EN 16438:2014 (E) DIN EN 16438:2014-075 1 Scope This European Standard specifies a test method and the minimum requirements for fungicidal or yeasticidal activity of chemical disinfectant and antiseptic produc
30、ts that form a homogeneous physically stable preparation in hard water or in the case of ready-to-use products with water. This European Standard applies to products for use in the veterinary area i.e. in the breeding, husbandry, production, transport and disposal of all animals except when in the f
31、ood chain following death and entry to the processing industry. EN 14885 specifies in detail the relationship of the various tests to one another and to “use recommendations”. NOTE 1 The method described is intended to determine the activity of commercial formulations or active substances under the
32、conditions in which they are used. NOTE 2 This method corresponds to a Phase 2 Step 2 test. 2 Normative references The following documents, in whole or in part, are normatively referenced in this document and are indispensable for its application. For dated references, only the edition cited applies
33、. For undated references, the latest edition of the referenced document (including any amendments) applies. EN 12353, Chemical disinfectants and antiseptics - Preservation of test organisms used for the determination of bactericidal (including Legionella), mycobactericidal, sporicidal, fungicidal an
34、d virucidal (including bacteriophages) activity EN 14885, Chemical disinfectants and antiseptics - Application of European Standards for chemical disinfectants and antiseptics ISO 4793, Laboratory sintered (fritted) filters Porosity grading, classification and designation 3 Terms and definitions For
35、 the purposes of this document, the terms and definitions given in EN 14885 apply. 4 Requirements The product shall demonstrate at least a 3 decimal log (lg) reduction from a water control, when tested in accordance with Table 1 and Clause 5 under simulated low level (3,0 g/l bovine albumin) or high
36、 level soiling (10 g/l yeast extract and 10 g/l bovine albumin) on a surface. EN 16438:2014 (E)DIN EN 16438:2014-076 Table 1 Obligatory and additional test conditions Test Conditions Fungicidal or yeasticidal activity on non-porous surfaces without mechanical action in the veterinary area Test organ
37、ism Fungicidal activity a) obligatory Aspergillus brasiliensis (conidiospores) and Candida albicans (vegetative cells) Test organism Yeasticidal activity a) obligatory Candida albicans (vegetative cells) b) additional any relevant test organism Test temperature a) obligatory 10 C 1 C b) additional 4
38、 C 1 C; 20 C 1 C; 40 C 1 C Contact time a) obligatory 60 min 10 s b) additional* 5 min 10 s; 30 min 10 s; 120 min 10 s Interfering substance a) obligatory low level soiling high level soiling 3,0 g/l bovine albumin 10 g/l yeast extract plus 10 g/l bovine albumin b) additional any relevant substance
39、The obligatory contact times for surface disinfectants stated in Table 1 were chosen to enable comparison of standard conditions. *The recommended contact time for the use of the product is within the responsibility of the manufacturer. NOTE For the additional conditions, the concentration defined a
40、s a result can be lower than the one obtained under the obligatory test conditions. Any additional specific fungicidal activity shall be determined in accordance with 5.2.1 and 5.5.1.1 in order to take into account intended specific use conditions. 5 Test method 5.1 Principle A test suspension of fu
41、ngal conidiospores or yeast and interfering substance is inoculated onto the test surface and dried. After a drying time, an aliquot of the product under test is transferred to the surface, in a manner which covers the dried film. The surface is maintained at a specified temperature for a defined pe
42、riod of time specified in Clause 4 and 5.5.1.1. At the end of that contact time the surface is transferred to a neutraliser so that the action of the disinfectant is immediately neutralised. The numbers of surviving organisms which can be recovered from the surface is determined quantitatively. The
43、number of fungi or yeast on a surface treated with water in place of the disinfectant is also determined and the reduction is calculated. The test is performed using the conidiospores of Aspergillus brasiliensis (formerly niger) and the vegetative cells of Candida albicans (fungicidal activity) or o
44、nly the vegetative cells of Candida albicans (yeasticidal EN 16438:2014 (E) DIN EN 16438:2014-077 activity) as test organisms (Clause 4, Table 1). Additional and optional contact times and temperatures are specified (Clause 4, Table 1). Additional interfering substances and test organisms may be use
45、d. 5.2 Materials and reagents 5.2.1 Test organism The fungicidal activity shall be evaluated using the following strains as the test organisms1): Aspergillus brasiliensis (formerly Aspergillus niger) ATCC 16404 Candida albicans ATCC 10231 The yeasticidal activity shall be evaluated using only Candid
46、a albicans. NOTE See Annex A for strain references in some other culture collections. The required incubation temperature for these organisms is 30 C 1 C (5.3.2.3). The same temperature shall be used for all incubations performed during a test and its control and validation. If additional test organ
47、isms are used, they shall be incubated under optimum growth conditions (temperature, time, atmosphere, media) noted in the test report. If the additional test organisms selected do not correspond to the specified strains, their suitability for supplying the required inocula shall be verified. If the
48、se additional test organisms are not classified at a reference centre, their identification characteristics shall be stated, in addition, they shall be held by the testing laboratory or national culture collection under a reference for five years. 5.2.2 Culture media and reagents 5.2.2.1 General All
49、 weights of chemical substances given in this European Standard refer to the anhydrous salts. Hydrated forms may be used as an alternative, but the weights required shall be adjusted to allow for consequent molecular weight differences. The reagents shall be of analytical grade and/or appropriate for microbiological purposes. They shall be free from substances that are toxic or inhibitory to the test organism. To improve rep
