1、BSI Standards PublicationBS EN 15457:2014Paints and varnishes Laboratory method fortesting the efficacy of filmpreservatives in a coatingagainst fungiBS EN 15457:2014 BRITISH STANDARDNational forewordThis British Standard is the UK implementation of EN 15457:2014. Itsupersedes BS EN 15457:2007 which
2、 is withdrawn.The UK participation in its preparation was entrusted to TechnicalCommittee STI/28, Paint systems for non-metallic substrates.A list of organizations represented on this committee can beobtained on request to its secretary.This publication does not purport to include all the necessaryp
3、rovisions of a contract. Users are responsible for its correctapplication. The British Standards Institution 2014. Published by BSI StandardsLimited 2014ISBN 978 0 580 84804 9ICS 87.040Compliance with a British Standard cannot confer immunity fromlegal obligations.This British Standard was published
4、 under the authority of theStandards Policy and Strategy Committee on 31 August 2014.Amendments issued since publicationDate Text affectedBS EN 15457:2014EUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORMEN 15457 August 2014 ICS 87.040 Supersedes EN 15457:2007English Version Paints and varnishes - La
5、boratory method for testing the efficacy of film preservatives in a coating against fungi Peintures et vernis - Mthode dessai en laboratoire permettant de dterminer lefficacit des prservateurs du feuil dun revtement contre les champignons Beschichtungsstoffe - Laborverfahren fr die Prfung der Wirksa
6、mkeit von Filmkonservierungsmitteln in einer Beschichtung gegen Pilze This European Standard was approved by CEN on 10 July 2014. CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulate the conditions for giving this EuropeanStandard the status of a national standar
7、d 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
8、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 Republ
9、ic, 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 CO
10、MMITTEE FOR STANDARDIZATION COMIT EUROPEN DE NORMALISATION EUROPISCHES KOMITEE FR 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 worldwide for CEN national Members. Ref. No. EN 15457:2014 EBS EN 15457
11、:2014EN 15457:2014 (E)2 Contents PageForeword3 Introduction .4 1 Scope 5 2 Normative references 5 3 Terms and definitions .5 4 Principle5 5 Apparatus and materials.6 6 Fungi .6 6.1 Fungi more likely to grow in an exterior environment.6 6.2 Fungi more likely to grow in an interior environment7 7 Samp
12、ling and preparation of test samples and of specimens .7 7.1 Sampling.7 7.2 Preparation of test samples (see Annex A) 7 7.3 Conditioning of the test samples.7 7.4 Preparation and number of specimens .7 8 Procedure .8 8.1 Preparation of the Petri dishes with the culture medium8 8.2 Preparation of sto
13、ck cultures and sub-cultures.8 8.3 Preparation of the spore suspension 8 8.4 Inoculation and incubation (see Annex A)8 8.5 Assessment8 9 Test report 9 Annex A (informative) Laboratory method for testing the efficacy of film preservatives in a coating against fungi 10 Annex B (informative) Designatio
14、n of the percentage area of disfigurements according to EN 16492:2014, Table A.3. 11 Bibliography . 12 BS EN 15457:2014EN 15457:2014 (E)3 ForewordThis document (EN 15457:2014) has been prepared by Technical Committee CEN/TC 139 “Paints and varnishes”, the secretariat of which is held by DIN.This Eur
15、opean 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 2015 and conflicting national standards shall be withdrawn at the latest by February 2015.Attention is drawn to the possibility that some of the el
16、ements 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.This document supersedes EN 15457:2007. According to the CEN-CENELEC Internal Regulations, the national standards organizations of the following
17、 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
18、, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and the United Kingdom.BS EN 15457:2014EN 15457:2014 (E)4 IntroductionThis document identifies criteria to assess efficacy of film preservatives in a coating against fungi. The results of the method allow evaluation
19、of an active substance with regard to its inclusion in Annex I of the Biocidal Products Directive 98/8/EC (Directive 98/8/EC of the European Parliament and of the Council of 16 February 1998 concerning the placing of biocidal products on the market BPD) or in the list of the Biocidal Product Regulat
20、ion (BPR, Regulation (EU) 528/2012).The characteristics of the biocide treated coating material should conform to national regulations with regard to health, safety and the environment.BS EN 15457:2014EN 15457:2014 (E)5 1 ScopeThis European Standard specifies a laboratory test method for determining
21、 the biocidal/biostatic efficacy of single active substances or combinations thereof used in film preservatives in a coating against fungal growth. This standard does not apply to coatings not susceptible to fungal growth. The test method comprises only active substances for film preservation, not t
22、he protection of the substrate itself, e.g. wood, which is dealt with in another standard. The test method is applicable for active substances used for wood and masonry coatings. It is not applicable to marine coatings.Safety, health and environmental aspects are not in the scope of this standard.De
23、termination of the performance of film preservatives in coatings by applying ageing procedures is not within the scope of this standard.2 Normative referencesThe following documents, in whole or in part, are normatively referenced in this document and are indispensable for its application. For dated
24、 references, only the edition cited applies. For undated references, the latest edition of the referenced document (including any amendments) applies.EN 12469, Biotechnology - Performance criteria for microbiological safety cabinetsEN 16492:2014, Paints and varnishes - Evaluation of the surface disf
25、igurement caused by fungi and algae on coatingsEN 23270, Paints and varnishes and their raw materials - Temperatures and humidities for conditioning and testing (ISO 3270)EN ISO 1513, Paints and varnishes - Examination and preparation of test samples (ISO 1513)3 Terms and definitionsFor the purposes
26、 of this document, the following term and definition applies.3.1active substancesubstance or micro-organism that has an action on or against harmful organismsSOURCE: Biocidal Product Regulation (BPR, Regulation (EU) 528/2012), Article 3.1 (c), modified the article “a“ between “or“ and “micro-organis
27、m“ was deleted 4 PrincipleTo determine the fungicidal efficacy of film preservatives in a coating, the coating material is applied to a substrate conditioned according to EN 23270, placed onto an agar surface, inoculated with a standard fungal spore suspension and incubated. Conclusions can be drawn
28、 to the fungicidal efficacy of the film preservatives in a coating from the intensity of the fungal growth on the surface of the specimen after incubation. The method described here is a semiquantitative, comparative method between coatings, with and without film preservatives.BS EN 15457:2014EN 154
29、57:2014 (E)6 5 Apparatus and materials5.1 Cutting device for preparing the specimens (coated filter paper with a diameter of 55 mm).5.2 Autoclave5.3 Incubator capable of maintaining (24 2) C.5.4 Pipette, in the range between 100 l to 1 000 l, with sterile tips or combi-tips of 12,5 ml.5.5 Filter pap
30、er without fungicidal effect (e.g. cellulose with a pore size of 0,45 m and a thickness of 650 m).5.6 Automatic welding apparatus to seal the bags.5.7 Sterilized glass bottles (100 ml).5.8 Laboratory balance, capable of weighing to an accuracy of 0,1 g.5.9 Microscope5.10 Device to determine cell cou
31、nt (commercially available counting chamber, e.g. Thoma chamber).5.11 Wetting agent (e.g. N-Methyltaurine).5.12 Device for applying the coating5.13 Sterilized test tubes or other sterilized glassware for preparing slant agar cultures.5.14 Sterile Drigalski spatula5.15 Sterile platinum loop5.16 Steri
32、le glass funnel with cotton wool5.17 Sterile Petri dishes (with a diameter of 94 mm, and a height of 16 mm).5.18 Sterile tweezers5.19 Sterile water5.20 Class 2 microbiological safety cabinet according to EN 12469. 6 Fungi6.1 Fungi more likely to grow in an exterior environmenta) Aureobasidium pullul
33、ans (DSM1)2404)b) Alternaria alternata (DSM 62010) 1)DSM = DSMZ = Deutsche Sammlung fr Mikroorganismen und Zellkulturen (German collection of micro organisms and cell cultures), Braunschweig, Germany.BS EN 15457:2014EN 15457:2014 (E)7 c) Cladosporium cladosporioides (DSM 62121) d) Phoma violaceae (I
34、MI2)49948ii)e) Ulocladium atrum (IMI 79906 or DSMZ 63068)6.2 Fungi more likely to grow in an interior environmenta) Aspergillus versicolor (DSM 1943) b) Aspergillus niger (DSM 12634) c) Stachybotrys chartarum (DSMZ 2144) d) Penicillium purpurogenum (DSM 62866) e) Rhodotorula mucilaginosa (DSM 70825)
35、 The spore suspension used for the test shall be a mixture containing two fungi selected from the first group (6.1) and two fungi selected from the second group (6.2).7 Sampling and preparation of test samples and of specimens7.1 SamplingTake a representative sample of the coating material or of the
36、 coating system for testing in accordance with EN ISO 1513. 7.2 Preparation of test samples (see Annex A)Coat a strip of filter paper without biocidal effect with the coating material to be tested. The application rate shall be that recommended by the coating manufacturer for normal use.7.3 Conditio
37、ning of the test samplesCondition the test sample in a horizontal position for at least 5 days at (23 2) C and (50 5) % relative humidity, in accordance with EN 23270. NOTE The conditioning time might vary according to the coating material and end use corresponding to information given by the manufa
38、cturer.7.4 Preparation and number of specimensAfter conditioning, three specimens each with a diameter of 55 mm shall be prepared from the test samples. The specimens shall be sealed in a plastics bag and sterilized using gamma radiation of 10 kGy. Other methods of sterilization may be agreed betwee
39、n the parties.For each test series three specimens coated with coating material containing the film preservative, three specimens coated with the same coating material without film preservative and three specimens of the uncoated substrate shall be tested.2)IMI = CABI = Bioscience Genetic Resource C
40、ollection, Egham, UK.BS EN 15457:2014EN 15457:2014 (E)8 8 Procedure8.1 Preparation of the Petri dishes with the culture mediumA malt (3 %)-agar(1,5 %)-medium shall be sterilized in the autoclave. After cooling the medium to 55 C to 60 C, 20 ml shall be poured into each sterile Petri dish under asept
41、ic conditions.8.2 Preparation of stock cultures and sub-culturesSub-cultures shall be obtained by inoculating spore material from a stock culture to freshly prepared agar slope culture media and shall be used for preparing the spore suspension. From these sub-cultures further sub-cultures can be der
42、ived in sufficient number. After the inoculation the sub-cultures shall be incubated at (24 2) C until good sporulation has been achieved. This might require 3 days to 7 days, depending on the fungal species used for testing. The sub-cultures can be stored satisfactorily at 3 C to 7 C for a period o
43、f 3 months.8.3 Preparation of the spore suspensionFor preparing the spore suspension a well-sporulating sub-culture is used. To this sub-culture 5 ml of sterile deionized water should be added (if required also add a surfactant e.g. 0,1 % N-Methyltaurine). The spores shall be carefully washed down f
44、rom the agar slope, using a platinum loop as an aid, filtered through a sterileglass funnel with cotton wool and collected in a sterile glass bottle.After counting the spore concentration by using a commercially available counting chamber and diluting each with sterile water to 106to 107spores/ml al
45、l spore suspensions intended for testing shall be mixed in equal parts.8.4 Inoculation and incubation (see Annex A)In addition to the coated and uncoated specimens (see 7.4) a further three Petri dishes containing the nutritive agar medium only shall be inoculated. The sterilized specimens shall be
46、placed centrally onto the surface of the culture media using sterile tweezers. The coated surface of the specimen shall be face up and there shall be full contact without air bubbles between the specimens and the surface of the culture medium.Should the coating lead to an undulation of the filter pa
47、per the paper should be kept even and in close contact with the agar by appropriate means. Otherwise a different substrate may be used. Check that the substrate does not inhibit growth of each selected test organism under the test conditions. If the substrate does inhibit growth it cannot be used.Un
48、der aseptic conditions in a safety cabinet the specimens shall be uniformly inoculated with 0,2 ml each of the mixed spore suspension using a suitable pipette. It is permissible to use up to an additional 0,8 ml of diluent to ensure an even distribution over the surface. Afterwards the suspension is
49、 spread out onto the agar plate with Drigalski spatula. The surface of the specimen shall remain free of agar medium during this operation.Following inoculation incubate the agar plates at (24 2) C.8.5 AssessmentAssess the fungal growth 7 days, 14 days and 21 days after the inoculation, using the assessment scale EN 16492:2014, Table A.3 (see also Annex B).The assessment is carried out visually macroscopically.NOTE 1 If required a microscope can be used, in order to exclude contamination by foreign materials.BS EN 15457:2014EN 15457:2014 (
