BS EN 1884-1999 Feather and down - Test methods - Determination of microbiological state《羽毛和羽绒 检验方法 微生物测定》.pdf

1、| | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | BRITISH STANDARD BS EN 1884:1999 The Europ

2、ean Standard EN 1884:1998 has the status of a British Standard ICS 59.040 NO COPYING WITHOUT BSI PERMISSION EXCEPT AS PERMITTED BY COPYRIGHT LAW Feather and down Test methods Determination of microbiological stateBS EN 1884:1999 This British Standard, having been prepared under the direction of the

3、Sector Committee for Materials and Chemicals, was published under the authority of the Standards Committee and comes into effect on 15 March 1999 BSI 03-1999 ISBN 0 580 30830 8 Amendments issued since publication Amd. No. Date Text affected National foreword This British Standard is the English lang

4、uage version of EN 1884:1998. The UK participation in its preparation was entrusted to Technical Committee TCI/73, Filling and filled articles, which has the responsibility to: aid enquirers to understand the text; present to the responsible European committee any enquiries on the interpretation, or

5、 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 committee can be obtained on request to its secretary. Cross-references The British Standards which implement

6、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 using the “Find” facility of the BSI Standards Electronic Catalogue. A British Standard does not purport to

7、 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 immunity from legal obligations. Summary of pages This document comprises a front cover, an inside front cover,

8、the EN title page, pages 2 to 7 and a back cover.CEN European Committee for Standardization Comite Europe en de Normalisation Europa isches Komitee fu r Normung Central Secretariat: rue de Stassart 36, B-1050 Brussels 1998 CEN All rights of exploitation in any form and by any means reserved worldwid

9、e for CEN national Members. Ref. No. EN 1884:1998 E EUROPEAN STANDARD EN 1884 NORME EUROPE ENNE EUROPA ISCHE NORM September 1998 ICS 59.040 Descriptors: stuffings, feathers, tests, microbiological analysis, bacteria, bacteria count methods, culture media English version Feather and down Test methods

10、 Determination of microbiological state Plumes et duvets Me thodes dessais De termination de le tat microbiologique Federn und Daunen Pru fverfahren Bestimmung des mikrobiologischen Zustandes This European Standard was approved by CEN on 13 August 1998. CEN members are bound to comply with the CEN/C

11、ENELEC 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 Central Secretariat or to an

12、y 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 Central Secretariat has the same status as the official versions. C

13、EN members are the national standards bodies of Austria, Belgium, Czech Republic, Denmark, Finland, France, Germany, Greece, Iceland, Ireland, Italy, Luxembourg, Netherlands, Norway, Portugal, Spain, Sweden, Switzerland and United Kingdom.Page 2 EN 1884:1998 BSI 03-1999 Foreword This European Standa

14、rd has been prepared by Technical Committee CEN/TC 222, Feather and down as filling 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 publica

15、tion of an identical text or by endorsement, at the latest by March 1999, and conflicting national standards shall be withdrawn at the latest by March 1999. According to the CEN/CENELEC Internal Regulations, the national standards organizations of the following countries are bound to implement this

16、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. Annex A is informative. Contents Page Foreword 2 Introduction 3 1 Scope 3 2 Normative

17、 references 3 3 Definitions 3 4 Principle 4 5 Dip slide method 4 6 Selective medium and count plate method 4 7 Repeatability and reproducibility 7 Annex A (informative) Bibliography 7Page 3 EN 1884:1998 BSI 03-1999 Introduction Feather materials for filling which come from plucking of waterfowls and

18、/or landfowls are, when raw, contaminated by pathogenic micro-organisms of faecal and urinary origin (e.g. salmonellae, faecal streptococci, etc.). These are present in variable quantities, depending on the environment and the hygienic conditions of breeding, plucking and storage packing. Fabricatio

19、n processes always comprise dusting, washing and sanitization in order to eliminate the pathogenic micro-organisms and to ensure the protection of consumer health. This European Standard specifies two methods to evaluate the microbiological state of feather materials for filling: the first one is us

20、ed only as routine control, while the second one is used when it is necessary to have complete and specific information on the microbiological state. NOTE The handling of micro-organisms which are potentially hazardous requires a high degree of technical competence. Only personnel trained in microbi

21、ological techniques should carry out the test. Codes of practice for disinfection, sterilization and personal hygiene are strictly observed. It is recommended that workers consult ISO 7218. 1 Scope 1.1 Dip-slide method 1.1.1 This method describes dip slide procedures that use two types of agar to te

22、st the presence of commensal 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 This method cannot be used to test the presence of sulfite-reducing clostrides (gram positive) and salmonella

23、(gram negative). 1.2 Selective medium and count plate method This method describes procedures that use different types of medium to verify the presence and quantity of: mesophilic aerobic bacteria (see 6.4); faecal streptococci (see 6.5); sulfite-reducing clostridium (see 6.6); salmonella (see 6.7).

24、 This method, used for 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. 2 Normative references This European Standard incorporates by dated or undated reference, provisions fro

25、m 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

26、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 performance requirements. EN 374-2, Protective gloves against

27、chemicals and micro-organisms Part 2: Determination of resistance to penetration. EN ISO 3696, Water for analytical laboratory use Specification and test methods. (ISO 3696:1987) 3 Definitions For the purposes of this standard, the following definitions apply: 3.1 mesophilic aerobic bacteria content

28、 quantity of micro-organisms, chiefly bacteria, which develop in the presence of oxygen at a temperature of (30 2)8C 3.2 faecal streptococci kind of bacteria belonging to the family of Lactobacillaceae: the members of this kind are cocci (gram positive) 3.3 sulfite-reducing clostridium kind of bacte

29、ria (clostridium) belonging to the family of Bacillaceae: the members of this kind are rods (gram positive), anaerobic and sporigenic 3.4 salmonella kind of bacteria belonging to the family of Enterobacteriaceae: the members of this kind are rods (gram negative) 3.5 initial extract filtrate obtained

30、 from treatment of the test specimen with peptonic physiological solution in accordance with the conditions as prescribed 3.6 decimal dilutions series of successive dilutions prepared from the initial extract (3.5)Page 4 EN 1884:1998 BSI 03-1999 3.7 colony forming units (CFU) colony formed by millio

31、ns of bacteria 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. 4 Principle

32、 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, submerged in an initial extract (3.5) and incubated at (37 1)8C overnight. The count of microbial colonies grown on the two different types of agar

33、is indicative of the degree of microbiological state. NOTE The CLED medium is a selective medium for the cultivation of urinary and faecal pathogens. The MacConkey agar preferentially supports the growth of coliforms. 4.2 Selective medium and count plate method 4.2.1 The filtrate, obtained by mixing

34、 the initial extracts (3.5) of the two test specimens, is divided into two parts. 4.2.1.1 One part is diluted in a scalar 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

35、the mesophilic aerobic bacterial charge, the faecal streptococci and the sulfite-reducing clostridium. 4.2.1.2 The second 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 b

36、roth is seeded on agar specific for salmonellae. 4.2.2 The count of the microbial colonies grown on each type of agar is indicative of the degree of contamination. 5 Dip slide method 5.1 Apparatus 5.1.1 Wide mouth sterile glass jars,o f capacity 120 ml. 5.1.2 MacConkey agar/CLED medium dip slides, 1

37、9 mm3 50 mm. 5.1.3 Incubator, capable of maintaining (37 1)8C 5.1.4 Sterile and protective gloves (see EN 374-1 and EN 374-2). 5.1.5 Sterile plastic bags. 5.1.6 Balance, with a maximum permissible error of 0,01 g. 5.2 Reagents Sterile saline solution, containing (8,5 0,2) g sodium chloride per litre

38、. 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 (5.1.1) containing 100 ml o

39、f sterile saline solution (5.2). Close the jar and shake vigorously by hand for (60 2) s. 5.3.3 Remove the dip slide from its sterile container and momentarily submerge it vertically in the shaken saline solution. 5.3.4 Remove the dip slide, allow to drain for a few seconds, replace it in the steril

40、e container and incubate at (37 1)8C for (16 2) h. 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. The test is valid only if n

41、o 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 “too many to count” (TMTC) if

42、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-shaped colony seen in the centre of

43、 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 place. 5.5 Test report Report the

44、 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 1)8C. 6.1.2 Oven, for dry sterilization of the glassware operating at a temperature of between 1708C and 1758C.Page 5 EN 1

45、884:1998 BSI 03-1999 6.1.3 Test-tube agitator, for mixing the decimal solutions. 6.1.4 Thermostats, adjustable to the required temperatures, with a maximum permissible error of 18C. 6.1.5 Water baths, adjustable to the required temperatures with a maximum permissible error of 18C. 6.1.6 Colony count

46、 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 0,01 g. 6.1.9 Wide-mouthed 2 000 ml glass flasks, wit

47、h sealing stoppers. 6.1.10 Glass test-tubes, with hermetic impermeable stoppers and a nominal capacity of 10 ml and 1 ml. 6.1.11 Glass pipettes, for complete discharge, having a nominal capacity of 10 ml and 1 ml. 6.1.12 Petri dishes, of glass or plastic, 90 mm to 100 mm diameter. 6.1.13 Sterile and

48、 protective gloves (see EN 374-1 and EN 374-2). 6.1.14 Sterile plastic bags. 6.1.15 Sterile gauze. 6.1.16 Cellulose acetate membrane, with 0,45mm pores. 6.1.17 Seitz filter. 6.1.18 Mechanical vibratory agitator. 6.1.19 Glass pellets. 6.2 Reagents 6.2.1 Distilled water, (grade 3 in accordance with EN

49、 ISO 3696) or water of equivalent quality, i.e. free from substances likely to inhibit or influence the growth of micro-organisms under the test conditions. If the distilled water is prepared from chlorinated water, neutralize the chlorine prior to the distillation. 6.2.2 0,1 % sterile peptonic physiological solution, prepared as follows. Composition: peptone 1 g sodium chloride 8,5 g distilled water 1 000 ml Preparation: Dissolve this medium in 1 l of distilled water. Adjust the pH so that after steril