1、BRITISH STANDARD BS ISO 17604:2003 Microbiology of food and animal feeding stuffs Carcass sampling for microbiological analysis ICS 07.100.30 ? +A1:2009 National foreword This British Standard is the UK implementation of ISO 17604:2003+A1:2009. It supersedes BS ISO 17604:2003 which is withdrawn. The

2、 UK participation in its preparation was entrusted to Technical Committee AW/9, Microbiology. A list of organizations represented on this committee can be obtained on request to its secretary. This publication does not purport to include all the necessary provisions of a contract. Users are responsi

3、ble for its correct application. Compliance with a British Standard cannot confer immunity from legal obligations. BS ISO 17604:2003+A1:2009 This British Standard was published under the authority of the Standards Policy and Strategy Committee on 16 September 2003 BSI 2009 Amendments/corrigenda issu

4、ed since publication Date Comments 30 September 2009 Implementation of ISO amendment 1:2009. Text amended in and added to clause 1 and Annex D added ISBN 978 0 580 60869 8 標(biāo)準(zhǔn)分享網(wǎng) w w w .b z f x w .c o m 免費下載 w w w . b z f x w . c o m Reference number ISO 17604:2003(E) INTERNATIONAL STANDARD ISO 17604

5、 First edition 2003-09-01 Microbiology of food and animal feeding stuffs Carcass sampling for microbiological analysis Microbiologie des aliments Prlvement dchantillons sur des carcasses en vue de leur analyse microbiologique BS ISO 17604:2003+A1:2009 w w w . b z f x w . c o m This page deliberately

6、 left blank 標(biāo)準(zhǔn)分享網(wǎng) w w w .b z f x w .c o m 免費下載 w w w . b z f x w . c o m iii Foreword ISO (the International Organization for Standardization) is a worldwide federation of national standards bodies (ISO member bodies). The work of preparing International Standards is normally carried out through ISO

7、 technical committees. Each member body interested in a subject for which a technical committee has been established has the right to be represented on that committee. International organizations, governmental and non-governmental, in liaison with ISO, also take part in the work. ISO collaborates cl

8、osely with the International Electrotechnical Commission (IEC) on all matters of electrotechnical standardization. International Standards are drafted in accordance with the rules given in the ISO/IEC Directives, Part 2. The main task of technical committees is to prepare International Standards. Dr

9、aft International Standards adopted by the technical committees are circulated to the member bodies for voting. Publication as an International Standard requires approval by at least 75 % of the member bodies casting a vote. Attention is drawn to the possibility that some of the elements of this doc

10、ument may be the subject of patent rights. ISO shall not be held responsible for identifying any or all such patent rights. ISO 17604 was prepared by Technical Committee ISO/TC 34, Food products, Subcommittee SC 9, Microbiology. This first edition of ISO 17604 cancels and replaces the second edition

11、 of ISO 3100-1:1991. ISO 3100-2:1988 is under revision as ISO 6887-2. BS ISO 17604:2003+A1:2009 w w w . b z f x w . c o m Introduction It is generally agreed that the determination of microbial counts and the prevalence of pathogenic microorganisms on carcasses is essential for monitoring and verifi

12、cation in risk-based slaughter hygiene assurance systems e.g. those employing the hazard analysis critical control points (HACCP) principles and quality assurance systems. Moreover, many institutes are involved in (international) surveillance programmes on the prevalence of pathogenic microorganisms

13、. The design of such monitoring and surveillance programmes will obviously benefit from the use of standardized and internationally accepted sampling procedures. iv BS ISO 17604:2003+A1:2009 標(biāo)準(zhǔn)分享網(wǎng) w w w .b z f x w .c o m 免費下載 w w w . b z f x w . c o m 1 Microbiology of food and animal feeding stuffs

14、 Carcass sampling for microbiological analysis 1 Scope This International Standard specifies sampling methods for the detection and enumeration of microorganisms on the carcass surface of freshly slaughtered meat animals. The microbiological sampling can be carried as part of the process control (an

15、d to verify process control) in slaughter establishments for cattle, horses, pigs, sheep, goats and game raised in captivity, risk-based assurance systems for product safety, and surveillance programmes for the prevalence of pathogenic microorganisms. This International Standard includes the use of

16、destructive and non-destructive techniques depending on the reason for the sample collection. It does not consider the use of sampling plans. When national legislation on the topic exists, this prevails over this International Standard. Annex A shows sampling sites on the carcass, and Annex B gives

17、requirements for microbiological examination. Annex C compares destructive and non-destructive methods. 2 Normative references The following referenced documents are indispensable for the application of this document. For dated references, only the edition cited applies. For undated references, the

18、latest edition of the referenced document (including any amendments) applies. ISO 4833, Microbiology of food and animal feeding stuffs Horizontal method for the enumeration of microorganisms Colony-count technique at 30 C ISO 5552, Meat and meat products Detection and enumeration of Enterobacteriace

19、ae without resuscitation MPN technique and colony-count technique ISO 6579, Microbiology of food and animal feeding stuffs Horizontal method for the detection of Salmonella spp. ISO 6887-1, Microbiology of food and animal feeding stuffs Preparation of test samples, initial suspension and decimal dil

20、utions for microbiological examination Part 1: General rules for the preparation of the initial suspension and decimal dilutions ISO 6887-2, Microbiology of food and animal feeding stuffs Preparation of test samples, initial suspension and decimal dilutions for microbiological examination Part 2: Sp

21、ecific rules for the preparation of meat and meat products out Annex D specifies methods for the sampling of poultry carcasses for microbiological analysis. BS ISO 17604:2003+A1:2009 w w w . b z f x w . c o m 2 ISO 7218, Microbiology of food and animal feeding stuffs General rules for microbiologica

22、l examinations. ISO 7251, Microbiology of food and animal feeding stuffs Horizontal method for the enumeration of presumptive Escherichia coli Most probable number technique ISO 10272, Microbiology of food and animal feeding stuffs Horizontal method for detection of thermotolerant Campylobacter ISO

23、10273, Microbiology of food and animal feedings stuffs Horizontal method for the detection of presumptive pathogenic Yersinia enterocolitica ISO 13720, Meat and meat products Enumeration of Pseudomonas spp. ISO 16654, Microbiology of food and animal feeding stuffs Horizontal method for the detection

24、 of Escherichia coli O157 3 Sampling procedure Both destructive and non-destructive methods may be used (see Annex C). Avoidance of adverse effects on the carcass value is the primary constraint on the use of destructive methods. Non-destructive techniques enable the examination of larger areas. Sma

25、ller areas targeted to proven areas of greatest contamination may be examined using either destructive or non-destructive methods (see C.2 and C.3). 4 Sampling frequency The time and frequency of sampling is governed by the slaughterhouse practices for each animal, the design of risk-based process c

26、ontrol assurance programmes, the production volume, and the epidemiological status of the region from where the animal originates. In the case of process control, the time and frequency of sampling shall relate to the level of slaughter hygiene. In the case of surveillance for pathogens, the samplin

27、g time, location on the carcass, and frequency should correspond to the greatest chance of isolating the pathogens sought. 5 Sampling points 5.1 Carcass selection Every carcass should have an equal chance of being selected for sampling. 5.2 Process control Sampling points in the slaughterhouse shoul

28、d relate to the slaughter practices used. They should be selected according to risk-based principles, and relate to identified problem areas in the process. Examples of control points are the following: after the carcass polishing machine (pig); BS ISO 17604:2003+A1:2009 標(biāo)準(zhǔn)分享網(wǎng) w w w .b z f x w .c o

29、m 免費下載 w w w . b z f x w . c o m 3 after the carcass washing machine (pig); after flaying (dehiding) (cattle, sheep, goat, game raised in captivity and others); after evisceration; in the chill room at least 12 h after slaughter (see C.4). 5.3 Detection of pathogenic microorganisms For the detection

30、 of pathogenic microorganisms, the following sampling points may be used for all species: immediately before chilling; in the chill room at least 12 h after slaughter (see C.4). 6 Sampling sites 6.1 Process control The sampling sites chosen depend on the slaughterhouse practices for different animal

31、s (see Figures A.1, A.2 and A.3). These sampling sites are not compulsory. Consistency in the choice of sampling sites over time is important. It is usually preferable to sample as many carcasses as possible at the expense of the number of sampling sites on the individual carcass. 6.2 Detection of p

32、athogenic microorganisms The sampling sites chosen depend on the slaughterhouse practices for different animals. The purpose is to examine the sites with the highest prevalence of contamination (see Table A.1). These sampling sites are not compulsory. Consistency in the choice of sampling sites over

33、 time is important. It is usually preferable to sample as many carcasses as possible at the expense of the number of sampling sites on the individual carcass. While prevalence determinations in surveillance programmes will generally benefit from larger sampling areas, sampling of smaller areas targe

34、ted to areas of greatest contamination may achieve the same result. 7 Sampling techniques 7.1 General For a given sampling situation, the same sampling technique should be used each time, to ensure that results are comparable. BS ISO 17604:2003+A1:2009 w w w . b z f x w . c o m 4 7.2 Destructive met

35、hods 7.2.1 Corkborer method 7.2.1.1 Reagents 7.2.1.1.1 Ethanol, 70 % and 90 % by volume. 7.2.1.2 Apparatus and materials 7.2.1.2.1 Sterile scalpels. 7.2.1.2.2 Sterile forceps. 7.2.1.2.3 Sterile corkborers, with a cutting area of 5 cm2. 7.2.1.2.4 Portable gas blow torch or portable Bunsen burner. 7.2

36、.1.2.5 Tissues or cotton wool. 7.2.1.2.6 Sterile plastic bags, for a peristaltic-type homogenizer of appropriate size for the area being sampled and the volume of diluent to be added. 7.2.1.3 Collection of samples At the relevant places on the carcass, holes are made in the surface with a sterile co

37、rkborer (7.2.1.2.3). The discs of skin or tissue (approximately 2 mm thick) are then cut loose with a sterile scalpel and forceps and put into a labelled sterile plastic bag (7.2.1.2.6). 7.2.1.4 Cleaning and sterilization of materials The corkborer (7.2.1.2.3), scalpel and forceps shall be cleaned a

38、nd sterilized after each sampling as follows. a) Clean with tissues or cotton wool dipped in 70 % ethanol (7.2.1.1.1). b) Dip in 70 % ethanol in a bottle. c) Burn the ethanol off; if the use of a naked flame is hazardous, then allow the ethanol to evaporate. d) Allow to cool. Due to the amount of ti

39、me needed to carry out the cleaning, it is best to use at least two sets of corkborer, scalpel and forceps. It is essential that these tools are not re-contaminated before use. As an alternative, the use of sterile disposable materials is allowed. 7.2.2 Template excision method 7.2.2.1 Apparatus and

40、 materials 7.2.2.1.1 Sterile scalpels. 7.2.2.1.2 Sterile forceps. 7.2.2.1.3 Sterile square templates, with hollow internal area of, for example, 10 cm2, 20 cm2 or 25 cm2. 7.2.2.1.4 Sterile plastic bags, for a peristaltic type homogenizer. BS ISO 17604:2003+A1:2009 標(biāo)準(zhǔn)分享網(wǎng) w w w .b z f x w .c o m 免費下載

41、w w w . b z f x w . c o m 5 7.2.2.2 Collection of samples At the relevant places of the carcasses, about 2 mm thick samples are cut delineated by sterile templates, using sterile scalpels and forceps. The instruments may be re-used as described under 7.2.1.4. 7.3 Non-destructive methods 7.3.1 Wet an

42、d dry swab method (see reference 1) 7.3.1.1 Reagents 7.3.1.1.1 Sterile peptone salt diluent, for general use (see ISO 6887-1), dispensed in 10,0 ml amounts in tubes or bottles. 7.3.1.2 Apparatus and materials 7.3.1.2.1 Sterile cotton wool swabs, large size with wooden shaft. 7.3.1.2.2 Sterile square

43、 templates, with hollow internal area of, for example, 50 cm2 or larger. 7.3.1.3 Collection of samples Moisten a swab in 10 ml peptone salt diluent (7.3.1.1.1). At each selected carcass test side, press the template (7.3.1.2.2) hard onto the surface. Rub the swab over the whole area using pressure,

44、moving first horizontally and turning the swab so that all sides are used. Place the swab into the diluent used to wet the swab, breaking off the wooden shaft against the inside of the bottle. Then, with a dry swab, sample the area again, as above and place this swab into the same container of dilue

45、nt. The instruments may be re-used as described under 7.2.1.4. 7.3.2 Sponge sampling method 7.3.2.1 Reagents 7.3.2.1.1 Sterile peptone salt diluent, for general use (see ISO 6887-1), dispensed in 25,0 ml amounts in bottles. 7.3.2.2 Apparatus and materials 7.3.2.2.1 Sterile specimen sponge (free of i

46、nhibitory substances), in a sterile plastic bag. 7.3.2.2.2 Sterile square template, with hollow internal area of 100 cm2 (10 cm 10 cm). 7.3.2.2.3 Sterile gloves. 7.3.2.3 Collection of samples Locate the sampling sites. Open the bag containing the sterile sponge (7.3.2.2.1) and add sufficient peptone

47、 salt diluent (7.3.2.1.1) to wet the sponge without excess fluid being visible. Massage the sponge from outside the bag to moisten it thoroughly. Put on a pair of sterile gloves and carefully remove the sponge from the bag. Place the template (7.3.2.2.2) over the location. Wipe the sponge over the e

48、nclosed sampling site (10 cm 10 cm) for a total of approximately 10 times in the vertical and 10 times in the horizontal direction. BS ISO 17604:2003+A1:2009 w w w . b z f x w . c o m 6 After swabbing, place the sponge back in the sponge sample bag. Add further diluent to the sample bag to make a to

49、tal of 25 ml. The instruments may be re-used as described under 7.2.1.4. 7.3.3 Gauze tampon method 7.3.3.1 Reagents 7.3.3.1.1 Sterile peptone salt diluent, for general use (see ISO 6887-1), dispensed in 25,0 ml amounts in bottles. 7.3.3.2 Apparatus and materials 7.3.3.2.1 Sterile gauze tampon. 7.3.3

50、.2.2 Sterile plastic bags, for a peristaltic-type homogenizer. 7.3.3.2.3 Sterile square template, with hollow internal area of 100 cm2 (10 cm 10 cm). 7.3.3.2.4 Sterile gloves. 7.3.3.3 Collection of samples At the sampling site, open the plastic bag containing the tampon (7.3.3.2.1) and add about 10

51、ml of peptone salt diluent (7.3.3.1.1). Squeeze and massage the tampon from outside the bag to thoroughly moisten it. Place the template (7.3.3.2.3) over the test area. Either hold the bag outside and turn inside out (use as a glove) or use a fresh pair of sterile gloves to wipe the tampon over the

52、test surface, 10 times in the horizontal direction then 10 times in the vertical direction. Place the tampon back in its plastic bag and add further diluent to make a total of 25 ml. The instruments can be re-used as described under 7.2.1.4. 8 Storage and transport of samples Transport the samples i

53、n an insulated cool box with frozen freezer blocks or a crushed melting ice cool box. Do not allow the samples to freeze or to come into contact with the frozen blocks of ice, if used. Either process the samples in the laboratory within 1 h of collection or store them at 2 C 2 C for a maximum of 24

54、h (see ISO 7218). BS ISO 17604:2003+A1:2009 標(biāo)準(zhǔn)分享網(wǎng) w w w .b z f x w .c o m 免費下載 w w w . b z f x w . c o m 7 Annex A (informative) Sampling sites The sampling sites to be chosen depend on the slaughterhouse practices for different animals. The purpose is to examine the sites with the highest prevalenc

55、e of contamination (see Table A.1). Figures A.1, A.2 and A.3 show examples of the sampling sites on the surface of the carcass of pig, beef and lamb respectively (see reference 2). Table A.1 Sites most consistently contaminated by high numbers of microorganisms Pig a Beef a Lamb a Distal hind limb (

56、trotter) (1) a Brisket (2) Abdomen (flank) (3) Hind limb, lateral (2) Forerib (3) Thorax, lateral (4) Abdomen, lateral (belly) (3) Flank (4) Crutch (6) Mid-dorsal region (mid-back) (4) Flank groin (6) Breast, lateral (7) Abdomen, medial (10) Round, lateral (8) a The numbers (x) indicate the sampling

57、 sites in Figures A.1 to A.3. a) Lateral b) Medial Figure A.1 Pig: Examples of sampling sites BS ISO 17604:2003+A1:2009 w w w . b z f x w . c o m 8 a) Lateral b) Medial Figure A.2 Beef: Examples of sampling sites BS ISO 17604:2003+A1:2009 標(biāo)準(zhǔn)分享網(wǎng) w w w .b z f x w .c o m 免費下載 w w w . b z f x w . c o m

58、9 a) Lateral b) Medial Figure A.3 Lamb: Examples of sampling sites BS ISO 17604:2003+A1:2009 w w w . b z f x w . c o m 10 Annex B (normative) Microbiological examination B.1 Preparation of test samples The preparation shall be performed in accordance with ISO 6887-2. For general rules for microbiological examinations, see ISO 7218. B.2 Process control Colony counts per square centimetre of carcass surface shall be performed in accordance with ISO 4833. Enumeration of Enterobacteriaceae shall be performed in accordance ISO 5552, presu