SIST EN ISO 13307:2013

SLOVENSKI STANDARD
SIST EN ISO 13307:2013
01-junij-2013
0LNURELRORJLMDåLYLOLQNUPH)D]DSULPDUQHSURL]YRGQMH7HKQLNHY]RUþHQMD,62

Microbiology of food and animal feed - Primary production stage - Sampling techniques
(ISO 13307:2013)
Mikrobiologie von Lebensmitteln und Futtermitteln - Primärproduktion -
Probenahmetechniken (ISO 13307:2013)
Microbiologie des aliments - Stade de production primaire - Techniques de prélèvement
(ISO 13307:2013)
Ta slovenski standard je istoveten z: EN ISO 13307:2013
ICS:
07.100.30 Mikrobiologija živil Food microbiology
SIST EN ISO 13307:2013 en
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

---------------------- Page: 1 ----------------------

SIST EN ISO 13307:2013

---------------------- Page: 2 ----------------------

SIST EN ISO 13307:2013


EUROPEAN STANDARD
EN ISO 13307

NORME EUROPÉENNE

EUROPÄISCHE NORM
March 2013
ICS 07.100.30
English Version
Microbiology of food and animal feed - Primary production stage
- Sampling techniques (ISO 13307:2013)
Microbiologie des aliments - Stade de production primaire - Mikrobiologie von Lebensmitteln und Futtermitteln -
Techniques de prélèvement (ISO 13307:2013) Primärproduktion - Probenahmetechniken (ISO
13307:2013)
This European Standard was approved by CEN on 1 March 2013.

CEN members are bound to comply with the CEN/CENELEC 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 CEN-CENELEC Management Centre or to any 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 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 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 and United
Kingdom.





EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION

EUROPÄISCHES KOMITEE FÜR NORMUNG

Management Centre: Avenue Marnix 17, B-1000 Brussels
© 2013 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN ISO 13307:2013: E
worldwide for CEN national Members.

---------------------- Page: 3 ----------------------

SIST EN ISO 13307:2013
EN ISO 13307:2013 (E)
Contents Page
Foreword . 3

2

---------------------- Page: 4 ----------------------

SIST EN ISO 13307:2013
EN ISO 13307:2013 (E)
Foreword
This document (EN ISO 13307:2013) has been prepared by Technical Committee CEN/TC 275 “Food
analysis - Horizontal methods”, the secretariat of which is held by DIN, in collaboration with Technical
Committee ISO/TC 34 "Food products".
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 September 2013, and conflicting national standards shall be
withdrawn at the latest by September 2013.
Attention is drawn to the possibility 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 organizations of the following
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, Poland, Portugal,
Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and the United Kingdom.
3

---------------------- Page: 5 ----------------------

SIST EN ISO 13307:2013

---------------------- Page: 6 ----------------------

SIST EN ISO 13307:2013
INTERNATIONAL ISO
STANDARD 13307
First edition
2013-03-01
Microbiology of food and animal
feed — Primary production stage —
Sampling techniques
Microbiologie des aliments — Stade de production primaire —
Techniques de prélèvement
Reference number
ISO 13307:2013(E)
©
ISO 2013

---------------------- Page: 7 ----------------------

SIST EN ISO 13307:2013
ISO 13307:2013(E)

COPYRIGHT PROTECTED DOCUMENT
© ISO 2013
All rights reserved. Unless otherwise specified, no part of this publication may be reproduced or utilized otherwise in any form
or by any means, electronic or mechanical, including photocopying, or posting on the internet or an intranet, without prior
written permission. Permission can be requested from either ISO at the address below or ISO’s member body in the country of
the requester.
ISO copyright office
Case postale 56 • CH-1211 Geneva 20
Tel. + 41 22 749 01 11
Fax + 41 22 749 09 47
E-mail copyright@iso.org
Web www.iso.org
Published in Switzerland
ii © ISO 2013 – All rights reserved

---------------------- Page: 8 ----------------------

SIST EN ISO 13307:2013
ISO 13307:2013(E)

Contents Page
Foreword .iv
1 Scope . 1
2 Normative references . 1
3 Terms and definitions . 1
4 General arrangement . 1
4.1 General . 1
4.2 Sampling personnel . 1
4.3 Packing and labelling of samples . 2
4.4 Preparation of a sampling form . 2
5 Diluents and disinfectants . 2
6 Apparatus and materials. 4
7 Sampling techniques — General recommendations . 5
8 Sampling techniques in the farm environment . 5
8.1 Samples taken after cleaning and disinfection . 5
8.2 Surface sampling . 6
8.3 Sampling floors . 6
8.4 Dust samples . 8
8.5 Water samples . 8
8.6 Moore’s drain swabs or tampon swab samples . 9
8.7 Rope swabs . 9
8.8 Disposable in-line milk filters . 9
9 Techniques for taking samples from animals . 9
9.1 Sampling from animals on the farm . 9
9.2 Sampling animals at slaughter .10
10 Sampling techniques in the hatchery .12
10.1 General .12
10.2 Hatcher basket liner samples .12
10.3 Broken eggshells .12
10.4 Hatcher fluff .12
10.5 Meconium .13
10.6 Swabs from hatcher baskets .13
10.7 Macerated hatchery waste .13
10.8 Dead-in-shell embryos .13
10.9 Cull chicks .14
10.10 Samples from chick deliveries on the farm .14
11 Techniques for sampling animal transport vehicles, modules, and crates .14
11.1 General .14
11.2 Vehicles for large animals (cattle, sheep, pigs, horses) .14
11.3 Transport crates and modules for poultry .14
11.4 Sampling transport vehicles after cleaning and disinfection .15
12 Storage and transport of samples .15
12.1 General recommendations.15
12.2 Recommendations for sensitive organisms .15
© ISO 2013 – All rights reserved iii

---------------------- Page: 9 ----------------------

SIST EN ISO 13307:2013
ISO 13307:2013(E)

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 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 closely 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. Draft 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 document may be the subject of
patent rights. ISO shall not be held responsible for identifying any or all such patent rights.
ISO 13307 was prepared by the European Committee for Standardization (CEN) in collaboration with
ISO Technical Committee TC 34, Food products, Subcommittee SC 9, Microbiology, in accordance with the
Agreement on technical cooperation between ISO and CEN (Vienna Agreement).
iv © ISO 2013 – All rights reserved

---------------------- Page: 10 ----------------------

SIST EN ISO 13307:2013
INTERNATIONAL STANDARD ISO 13307:2013(E)
Microbiology of food and animal feed — Primary
production stage — Sampling techniques
1 Scope
This International Standard specifies sampling techniques within the primary food-animal production stage,
for detection or enumeration of viable microorganisms with particular reference to food-borne pathogens.
This International Standard is not intended for use in diagnosis of animal disease.
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 latest edition of the referenced
document (including any amendments) applies.
ISO 6887-1, Microbiology of food and animal feeding stuffs — Preparation of test samples, initial suspension
and decimal dilutions for microbiological examination — Part 1: General rules for the preparation of the
initial suspension and decimal dilutions
ISO 7218, Microbiology of food and animal feeding stuffs — General requirements and guidance for
microbiological examinations
3 Terms and definitions
For the purposes of this document, the following terms and definitions apply.
3.1
primary production stage
includes all the stages of food production from farm until harvest or entry to the slaughterhouse
3.2
laboratory sample
sample prepared for sending to the laboratory and intended for inspection or testing
4 General arrangement
4.1 General
The parties concerned or their representatives may be given the opportunity to be present when
sampling is performed.
Whenever special, e.g statutory, requirements are given for the sampling and/or arise from a specific
analysis to be performed, these requirements shall be followed.
4.2 Sampling personnel
Sampling for microbiological examination shall always be undertaken by a person trained and
experienced in the technique of sampling for microbiological purposes.
© ISO 2013 – All rights reserved 1

---------------------- Page: 11 ----------------------

SIST EN ISO 13307:2013
ISO 13307:2013(E)

4.3 Packing and labelling of samples
Samples shall be packed in order to avoid cross-contamination and to prevent leakage or loss of moisture.
They shall be clearly identified.
The minimum details that shall accompany the samples are: the nature of the matrix, its identification,
the name or initials of the person responsible for taking the samples, as well as the date, time (if
appropriate), and place of sampling.
This information should be recorded on a form. One form can be used for several samples provided each
has unique identification and the samples are accompanied by the sampling form which lists the sample
details with their unique identifying codes.
4.4 Preparation of a sampling form
Samples shall be accompanied by a report, ideally completed on a standard form provided by the
laboratory, signed or initialled by the sampling personnel. The report shall give the following particulars:
— the place, date and time (if appropriate)of sampling;
— the names of the sampling personnel;
— the nature, number, and identity of samples constituting the consignment;
— the purpose of sampling and the microorganisms to be sought.
When appropriate, the report shall also include any relevant conditions or circumstances, and any special
information relating to the product being sampled, e.g. difficulty in achieving representative samples.
If any additives such as diluents, transport media or neutralizing agents are used, these shall be recorded.
5 Diluents and disinfectants
5.1 Diluents.
5.1.1 General. Diluent used for moistening all kind of swabs (bootswabs, stick swabs etc.):
— peptone salt solution prepared according to ISO 6887-1;
— buffered peptone water prepared according to ISO 6887-1;
— sterile water;
— potable water for samples where this would not interfere with the analysis, e.g. bootswabs.
5.1.2 Medium for transporting swabs for specific purposes. The general aim of these media is to
ensure survival of the target population, e.g. Campylobacter are particularly sensitive to drying.
Examples of transport media:
— buffered peptone water for Salmonella prepared according to ISO 6887-1;
— Cary–Blair transport medium or equivalent;
— Amies charcoal transport medium or equivalent.
In circumstances where the sample is acidic or alkaline, or may become so during transportation, it may
be useful to use a buffered diluent.
Consideration should be given, if enumeration is intended, to the possibility of multiplication of the
target or competing organisms before examination in the laboratory.
2 © ISO 2013 – All rights reserved

---------------------- Page: 12 ----------------------

SIST EN ISO 13307:2013
ISO 13307:2013(E)

5.2 Disinfectants for decontamination of packaging, instruments and surfaces of certain samples
5.2.1 Ethanol 70 % volume fraction.
5.2.2 Alcohol wipes.
5.3 Neutralizers for disinfectant residues
5.3.1 General. An appropriate neutralizer for all situations cannot be prescribed as each disinfectant is
optimally neutralized by a specific chemical substance (see Table 1). However, if the use of disinfectant is
suspected, but its composition is unknown, a neutralizer for general use (5.3.2) can be used.
Table 1 — Neutralizing agent components and neutralized constituents
Neutralizing agent components Neutralized constituents
Soya lecithin Quaternary ammonium
Sorbitan monooleate (polysorbate 80) Ethanol
L-Histidine Aldehydes
Sodium thiosulfate (Na S O•5H O) Halogen
2 2 3 2
Phenols
Disodium phosphate (Na HPO •12H O)
Acid or alkaline
2 4 2
5.3.2 Neutralizer for general use.
5.3.2.1 Composition.
Soya lecithin 3,0 g
Sorbitan monooleate (polysorbate 80) 30,0 g
l-Histidine 1,0 g
Sodium thiosulfate (Na S O ·5H O) 7,8 g
2 2 3 2
Disodium phosphate (Na HPO ·12H O) 100,8 g
2 4 2
Water 1 000 ml
5.3.2.2 Preparation. Dissolve the components in the water by heating. Sterilize for 15 min in the
autoclave maintained at 121 °C. The prepared medium can be stored at (5°± 3) °C for 3 months in a tightly
closed light-proof container.
This neutralizing liquid is normally used at 10 % volume fraction in diluents (5.1).
© ISO 2013 – All rights reserved 3

---------------------- Page: 13 ----------------------

SIST EN ISO 13307:2013
ISO 13307:2013(E)

6 Apparatus and materials
6.1 Sampling equipment and description.
6.1.1 General. Non-disposable sampling equipment should be sterilized before use, e.g. by moist heat
(autoclave) or dry heat (oven), according to ISO 7218. In certain situations, chemical decontamination
may be appropriate. After such treatment, the equipment should be clean, sterile and free of inhibitory
substances. If the equipment needs to be reused while taking samples, sterilization shall be done preferably
by flaming (see 6.1.10) or with 70 % volume fraction ethanol or with any other appropriate disinfectant
(refer to ISO 7218). Sealed packs containing multiple plastics disposable equipment (e.g. gloves, overboots,
plastics bags) are suitable for use during sampling of products from the primary production stage. On
each sampling occasion (e.g. each different farm), a fresh pack should be used. During sampling, every
precaution shall be taken to avoid contamination of the unused disposable materials/equipment.
6.1.2 Gloves, disposable, single use, impervious, used during sampling to protect the sampler and to
avoid cross-contamination. An alternative is to use plastics bags, inverted over the hand(s).
6.1.3 Overboots, strong clean plastics bags of appropriate size, or boot-shaped plastics covering made
specifically to put over boots or shoes, used for two purposes: as biosecurity while visiting a farm to avoid
introducing contamination; and to put over boots just before sampling with bootswabs (6.1.6).
6.1.4 Fabric swabs, normally large sterile portions of cloth, such as gauze, cellulose-based sponge,
woven or non-woven fabric which are used to swab large surface areas.
6.1.5 Stick swabs, cotton-bud swabs and all swabs comprising small pieces of cotton or synthetic
material fixed to the end of a wooden, metal or plastics stick. The swabs are often contained in sterile
tubes which may contain medium such as Amies charcoal transport medium. The material used should
be free of inhibitory substances unless these are specified for selecting the target agent.
6.1.6 Boot socks, also called bootswabs or sock swabs, adaptations of fabric swabs designed to be
worn over the feet so that samplers can take the swabs while walking around doing other things. Boot
socks used shall be sufficiently absorbent to soak up the moisture. They can be made from tubular elastic
bandage material, which is cut into suitable lengths and pulled over the shoes or boots. Alternatively,
commercial fabric overshoes (avoiding those with plastics soles) or other suitable and sterile material
which covers the sole of the foot can be used, such as sterile fabric mob-caps (hair covers). In order to
avoid possible contamination from the sampler’s footwear, the boot socks should be put on, over new
plastics overboots (6.1.3), after entering the area to be sampled.
6.1.7 Drag swabs, mostly used in the poultry industry, comprise a battery of four large moistened
cloths (e.g. absorbent pads without antimicrobial substances) attached to a bar which is dragged over
litter or slatted areas or pits containing faecal droppings. Small sponge drag swabs are also available
commercially, but have a limited surface area compared to the original design.
6.1.8 Moore’s drain swabs or tampon swabs, typically comprising large composite fabric swabs with
multiple layers of gauze or cotton wool encased in gauze. Sanitary towels or tampons (free of antimicrobial
substances), which are constructed in this way, are often used. Large cellulose sponges are also suitable.
6.1.9 Rope swab. A series of soft, sterile, manilla ropes of diameter 1 cm to 2 cm (e.g. seven ropes per
large feedlot-type pen) placed horizontally just above the feed and water troughs so that cattle in the pen
brush against them and are able to chew the ropes.
6.1.10 Portable gas burner or blow torch.
6.1.11 Sterile forceps, scalpels, scissors.
4 © ISO 2013 – All rights reserved

---------------------- Page: 14 ----------------------

SIST EN ISO 13307:2013
ISO 13307:2013(E)

6.1.12 Sterile spoons or spatulas.
6.1.13 Sterile stiff brushes.
6.1.14 Cool box, insulated, either with integral cooling system, or cold packs, capable of maintaining the
samples at low temperature (above 0 °C) during transportation to the laboratory.
6.2 Sample containers. Sample containers and closures shall be of materials and construction which
adequately protect the sample and which do not bring about a change in the sample which could affect
the results of subsequent analyses. These are usually plastics bags or rigid containers (plastics or glass
screw-capped bottles or jars). Containers and closures shall be dry, clean, leak-proof and sterile.
The shape and capacity of the containers shall be appropriate to the particular requirements of the
product to be sampled. Containers other than plastics bags shall be securely closed by means of suitable
stoppers or secure caps.
7 Sampling techniques — General recommendations
Samples may be taken from animals and their environment, including during transportation and in the
slaughterhouse, to monitor the carriage of zoonotic agents in the live animal.
Sampling shall be carried out in such a way as to obtain representative samples of the materials to be tested.
The samples shall be taken using aseptic techniques and equipment, materials, and containers specified
in Clause 6.
The precise method of sampling and the mass or volume of matrix to be taken varies with the nature
of the product and the purpose for which samples are required. For details of the requirements, see
Clauses 8 to 11. The sample container shall be closed immediately after sampling.
8 Sampling techniques in the farm environment
8.1 Samples taken after cleaning and disinfection
Taking samples from disinfected surfaces is problematic because residual disinfectant can be present,
and often the disinfectant used is not known. Specific or “universal” disinfectant neutralizers can be used,
but some of these have an unpredictable effect on the growth of stressed organisms and competitive
flora leading to false negative tests.
When sampling disinfected animal housing, it is best to sample after all surfaces have dried to minimize
the inhibitory effect of disinfectant gathered with samples. Examples of places to sample are wall and
floor surfaces, drinkers, feeders, nest boxes, partitions, movable equipment such as weighing machines,
ventilation ducting, beams and ledges, control panels, and floors of anterooms or service areas. Conveyor
systems passing through cage layer houses can also be sampled.
It is recommended, where practicable, to transfer the swab immediately after sampling into an excess
(at least 1 part by mass to 100 parts by volume) of specific pre-enrichment or enrichment broth (e.g. a
fabric swab into 225 ml BPW for Salmonella or other specific medium) which dilutes and/or inactivates
the disinfectant. In this case, laboratory samples shall be cultured on the day of collection.
If same-day examination is not possible, diluents with neutralizers shall be used for moistening the
swab before sampling.
If the disinfectant used is known, add the appropriate neutralizer (see Table 1) to the relevant diluent (5.1).
If the use of disinfectant is suspected but its identity is not known, add the “universal” neutralizer (5.3.2).
© ISO 2013 – All rights reserved 5

---------------------- Page: 15 ----------------------

SIST EN ISO 13307:2013
ISO 13307:2013(E)

8.2 Surface sampling
8.2.1 Sampling with fabric swabs
This type of swab (6.1.4) can be held with a new glove (6.1.2) for each sample or using an “inverted bag
technique”, in which a polyethylene bag (6.1.2) holding the fabric swab is inverted to expose the swab,
which is then used to sample the surface. Each swab is rubbed vigorously over selected surfaces so that
2
each surface is covered and the swab is visibly soiled, sampling a minimum area of 1 m . The bag is then
turned back the right way round
...