1、DEUTSCHE NORM November 1998 Feather and down English version of DIN EN 1884 Test methods - Determination of microbiological state DIN EN 1884 - ICs 59.040 Descriptors: Feather, down, microbiological state, testing. Federn und Daunen - Prfverfahren - Bestimmung des mikrobiologischen Zustandes Europea
2、n Standard EN 1884 : 1998 has the status of a DIN Standard. A comma is used as the decimal marker. National foreword This standard has been prepared by CEN/TC 222. The responsible German body involved in its preparation was the Normenausschu Textil und Textilma- schinen (Textiles and Textile Machine
3、ry Standards Committee), Technical Committee Federn und Daunen. EN comprises 12 pages. No pari of this standard may be reproduced without the prior permission of Ref. No. DIN EN 1884: 1998-1 . V Deutsches Institut fur Normung e VI Berlin Luth Verlag GmbH, D-10772 Berlin, has the exclusive right of s
4、ale for German Standards (DIN-Normen) English price group O7 Sales No 1107 04 99 EUROPEAN STANDARD NORME EUROPENNE EUROPISCHE NORM EN 1884 September 1998 ICs 59.040 Descriptors: Feather, down, microbiological state, testing. English version Feather and down Test methods - Determination of microbiolo
5、gical state Plumes et duvets- Mthodesdessais- Dtermination de ltat microbio- logique Zustandes Federn und Daunen - Prfverfahren - Bestimmung des mikrobiologischen This European Standard was approved by CEN on 1998-08-1 3. CEN members are bound to comply with the CENKENELEC Internal Regulations which
6、 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 stand- ards may be obtained on application to the Central Secretariat or to any CEN member. The European Stan
7、dards exist 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 Central Secretariat has the same status as the official versions. CEN members are the national stan
8、dards bodies of Austria, Belgium, the Czech Republic, Denmark, Finland, France, Germany, Greece, Iceland, Ireland, Italy, Luxembourg, the Netherlands, Norway, Portugal, Spain, Sweden, Switzerland, and the United Kingdom. CEN European Committee for Standardization Comit Europen de Normalisation Europ
9、isches Komitee fr Normung Central Secretariat: rue de Stassart 36, B-1050 Brussels O 1998. CEN - All rights of exploitation in any form and by any means reserved worldwide for CEN national members. Ref. No. EN 1884 : 1998 E Page 2 EN 1884 : 1998 Contents Page Foreword Introduction 1 Scope 2 Normativ
10、e references 3 Definitions 4 Principle 5 Dip slide method 6 Selective medium and count plate method 7 Repeatibility and reproducibility Annex A (informative) Bibliography 4 4 5 5 6 12 12 Foreword This European Standard has been prepared by Technical Committee CENITC 222 “Feather and down as filling
11、material for any article, as well as finished articles filled with feather and down”, the secretariat of which is held by UNI. 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 March 1999, and confl
12、icting national standards shall be withdrawn at the latest by March 1999. According to the CENKENELEC Intemal Regulations, the national standards organizations of the following countries are bound to implement this European Standard: Austria, Belgium, Czech Republic, Denmark, Finland, France, German
13、y, Greece, Iceland, Ireland, Italy, Luxembourg, Netherlands, Norway, Portugal, Spain, Sweden, Switzerland and the United Kingdom. Annex A is infomative. Page 3 EN 1884: 1998 Introduction Feather materials for filling which come from plucking of waterfowls and/or landfowls are, when raw, contaminated
14、 by pathogenic microorganisms of fecal and urinary origin (e.9. salmonellae, fecal streptococci, etc.). These are present in variable quantities, depending on the environment and to the hygienic conditions of breeding, plucking and storage packing. Fabrication processes always comprise the dusting,
15、washing and sanitization in order to eliminate the pathogenic micro-organisms and to ensure the protection of the consumer health. This European Standard specifies two methods to evaluate the microbiological state of feather materials for filling: the first one is used only as routine control, while
16、 the second one is used when it is necessary to have complete and specific information on the microbiological state. NOTE: Handling of microorganisms which are potentially hazardous requires a high degree of technical competence. Only personnel trained in microbiological techniques should carry out
17、the test. Code of practice for disinfection, sterilization and personal hygiene are strictly observed. It is recommended that workers should consult IS0 7218. 1 Scope 1.1 Dipslide method 1.1.1 This method describes the dip slide procedures, that uses two types of agar to test the presence of commens
18、al bacteria and coliforms (gram negative).This procedure is suitable when a manufacturer requires a simple test to screen finished filling material hygiene. 1.1.2 (gram positive) and salmonella (gram negative). This method cannot be used to test the presence of sulphito-reducing clostrides 1.2 This
19、method describes procedures that use different types of medium to verify the presence and quantity of: Selective medium and count plate method - mesophilic aerobic bacteria (see 6.4) - fecal streptococci (see 6.5) - sulphite-reducing clostridium (see 6.6) - salmonella (see 6.7). This method, used fo
20、r both raw and finished materials, gives more complete and specific information on the control of the microbiological state of the filling material than the dip slide method. Page 4 EN 1884 : 1998 2 Normative references This European Standard incorporates by dated or undated reference, provisions fr
21、om other publications. These normative references are cited at the appropriate places in the text and the publications are listed hereafter. For dated references, subsequent amendments or revisions of any of these publications apply to this European Standard only when incorporated in it by amendment
22、 or revision. For undated references the latest edition of the publication referred to applies. EN 1883 Feather and down - Sampling in view of tests EN 374-1 Protective gloves against chemicals and micro-organisms - Part 1 : Terminology and petformance requirements EN 374-2 Protective gloves against
23、 chemicals and micro-organisms - Part 2: Determination of resistance to penetration EN IS0 3696 Water for analytical laboratory use - Specification and test methods (IS0 3696: 1987) 3 Definitions For the purposes of this standard, the following definitions apply: 3.1 mesophilic aerobic bacteria cont
24、ent: Quantity of microorganisms, chiefly bacteria, which develop in the presence of oxygen at a temperature of (3012)“C. 3.2 The members of this kind are cocci (Gram positive). faecal streptococci: Kind of bacteria belonging to the family of Lactobacillaceae. 3.3 sulphite-reducing clostridium: Kind
25、of bacteria (clostridium) belonging to the family Bacillaceae. The members of this kind are rods (gram positive), anaerobic and sporigenic. 3.4 members of this kind are rods (Gram negative). salmonella: Kind of bacteria belonging to the family of Enterobacteriaceae. The 3.5 peptonic physiological so
26、lution in accordance with the conditions as prescribed. initial extract: Filtrate obtained from the treatment of the test specimen with the 3.6 decimal dilutions: Series of successive dilutions prepared from the initial extract (3.5) 3.7 colony forming units (CFU): Colony formed by millions of bacte
27、ria of the same species grown by multiplication of a single bacterial cell on a specific agar. This is visible to the naked eye and can have different shapes (lenticular, starry, etc.) of variable dimensions according to the species, the type of agar and the culture conditions. Page 5 EN 1884 : 1998
28、 4 Principle 4.1 Dip-slide method A dip slide (5.1.2) CLED (Cystine, Lactose, Electrolyte, Deficient) and MacConkey) is encased in a sterile cylindrical container, and submerged in an initial extract (3.5) and incubated at (3711)“C overnight. The count of microbial colonies grown on the two differen
29、t types of agar is indicative of the degree of microbiological state. NOTE: CLED medium is a selective medium for the cultivation of urinary and fecal pathogens. MacConkey agar preferentially supports the growth of coliforms. 4.2 4.2.1 is divided into two parts: 4.2.1.1 One part is diluted in a scal
30、ar manner using the procedure of decimal dilutions. The filtrate and the various dilutions are seeded, in groups of three, on each of three selective agars for the determination of the mesophilic aerobic bacterial charge, the faecal streptococci and the sulphite-reducing clostridium. 4.2.1.2 The sec
31、ond part is passed through a cellulose acetate membrane which prevents salmonellae from passing through. The membrane is then seeded on an enrichment broth. Finally a part of this broth is seeded on agar specific for salmonellae. 4.2.2 The count of the microbial colonies grown on each type of agar i
32、s indicative of the degree of contamination. Selective medium and count plate method The filtrate, obtained by mixing the initial extracts (3.5) of the two test specimens, 5 5.1 5.1.1 5.1.2 5.1.3 5.1.4 5.1.5 5.1.6 5.2 Dip slide method Apparatus Wide mouth sterile glass jars, of capacity 120 ml MacCo
33、nkey agar/CLED medium dip slides, 19 mm x 50 mm Incubator, capable of maintaining (37 f 1) “C Sterile and protective gloves (see EN 374-1 and EN 374-2) Sterile plastic bags Balance with a maximum permissible error of 0,Ol g Reagents Sterile saline solution, containing (8,5 f 0,2) g sodium chloride p
34、er litre Page 6 EN 1884 : 1998 5.3 Procedure 5.3.1 From a conditioned laboratory bulk sample (see EN 1883),using sterile techniques, take five test specimens of approximately 1,0 g, weighed to a maximum permissible error of 0,05 g 5.3.2 Place a test specimen in a sterile 120 ml wide mouth glass jar
35、(5.1.1) containing 100 ml of sterile saline solution (5.2). Close the jar and shake vigorously by hand for (6012) s 5.3.3 vertically in the shaken saline solution. Remove the dip slide from its sterile container and momentarily submerge it 5.3.4 container and incubate at (37 f 1) OC for (16 f 2) h R
36、emove the dip slide, allow to drain for a few seconds, replace it in the sterile 5.3.5 Repeat the procedure from 5.3.2 to 5.3.4 for the remaining four test specimens 5.3.6 Carry out a sterility control by submerging a dip slide in sterile saline solution only and incubating as for the test specimens
37、. The test is valid only if no bacterial growth is observed on this dip slide after incubation. 5.3.7 After incubation, remove the dip slide from the container and count, for each test specimen, the number of bacterial colony forming units (CFU) on both agar types. The result should be recorded as “
38、too many to count“ (TMTC) if the agar is completely covered by bacteria such that individual colonies cannot be distinguished or, if the colonies merge into one another making accurate counting difficult. Occasionally, bacteria growing at the edge of the dip slide will not form the normal disc shape
39、d colony seen in the centre of the slide, but will spread along the edge of the slide, sometimes up to a distance of a few centimetres. Such growth forms should be counted as single colonies. 5.4 Expression of results Express the results as the number of colony forming units (CFU) to one decimal pla
40、ce. 5.5 Test report Report the mean of the five results and the standard deviation. 6 Selective medium and count plate method 6.1 Apparatus 6.1.1 Autoclave for moist-heat sterilization at approximately (121 f 1)OC. 6.1.2 170C and 175C. Oven for dry sterilization of the glassware operating at a tempe
41、rature of between 6.1.3 Test-tube agitator for mixing the decimal solutions. 6.1.4 permissible error of f 1 “C. Thermostats adjustable to the required temperatures, with a maximum Page 7 EN 1884 : 1998 6.1.5 error of * 1C. Water baths adjustable to the required temperatures with a maximum permissibl
42、e 6.1.6 Colony count apparatus comprising an illuminated base with a dark backing suitable for a magnifying glass to be used with a magnification of 1,5 and a mechanical or numerical colony counter. 6.1.7 pH-meter 6.1.8 Balance with a maximum permissible error of 0,Ol g 6.1.9 6.1.1 O Glass test-tube
43、s with hermetic impermeable stoppers and a nominal capacity of 1 O ml and 1 ml. Wide-mouthed 2000 ml glass flasks with sealing stoppers 6.1.1 1 Glass pipettes for complete discharge, having a nominal capacity of 1 O ml and 1 ml. 6.1.1 2 Petri dishes of glass or plastic, 90 mm to 100 mm diameter. 6.1
44、.13 Sterile and protective gloves (see EN 374-1 and EN 374-2) 6.1.14 Sterile plastic bags 6.1.1 5 Sterile gauze 6.1.1 6 Cellulose acetate membrane with 0,45 pm pores. 6.1.17 Seitz filter 6.1.1 8 Mechanical vibratory agitator 6.1.19 Glass pellets 6.2 Reagents 6.2.1 Distilled water (grade 3 in accorda
45、nce with EN IS0 3696) or water of equivalent quality, .e. free from substances likely to inhibit or to influence the growth of microorganisms under the test conditions. If the distilled water is prepared from chlorinated water, neutralize the chlorine prior to the distillation. Page 8 EN 1884 : 1998
46、 6.2.2 0,l % sterile peptonic physiological solution prepared as follows: Composition: peptone lg sodium chloride 83 g distilled water 1000 ml Preparation: Dissolve this medium in 1 I of distilled water. Adjust the pH so that after sterilization it is 7,O at 25C. Sterilize in an autoclave at (12111)
47、“C for 15 min. 6.2.3 Standard agar medium for the plate count Composition: yeast extract 2,5 g Pancreatic digest of casein 5 g Glucose lg Agar 15 9 pH 7,O Preparation: Suspend this culture medium in 1 I of distilled water. Boil until completely dissolved. Divide among flasks and sterilize in an auto
48、clave at (12111)“C for 15 min. 6.2.4 Edwards culture medium Composition: Lab-Lemco (meat extract) 10 9 Peptone 10 9 Esculin lg Sodium chloride 5g Chrystal violet 0,001 3 g Thallium sulphate 03 9 Agar 15 9 PH 7,4 Preparation: Suspend this culture medium in 1 I of distilled water. Boil until completel
49、y dissolved. Cool to approximately 50C add 5% to 7% of sterile ox or sheep blood, mix well and pour onto plates. 6.2.5 Slanetz and Bartley agar medium for m-enterococci Composition: Tryptose 20 9 Yeast extract 59 Dextrose 29 Disodium phosphate 2H20 49 Sodium azide 0,4 Tetrazonium chloride 091 9 Agar 10 9 pH 72 Preparation: Suspend this medium in 1 I of distilled water. Boil until completely dissolved. Prolonged heating should be avoided. Divide among Petri dishes and leave to solidify. Page 9 EN 1884 : 1998 6.2.6 Tetrathionate broth according to Mller-Kauffmann Compo
