This part of ISO 23036 specifies a method that is applicable for the detection of Anisakidae L3 larvae commonly found in marine and anadromous fishes. The method can be applied to fresh fish and/or frozen fish, lightly processed fish products, such as marinated, salted or cold smoked.
This method allows quantifying parasitic infections by estimating the number of parasites in the fish musculature.
This method doesn’t allow determining species or genotype of detected parasites, which identification
is made by morphological and/or molecular methods

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This part of ISO 23036 specifies a method that is applicable for the detection of Anisakidae L3 larvae commonly found in marine and anadromous fishes. The method can be applied to fresh fish and/or frozen fish, lightly processed fish products, such as marinated, salted or smoked, and it’s also suitable for visceral organs as confirmatory method for visual inspection scheme.
The artificial digestion method allows quantifying parasitic infections by estimating the number of parasites in the fish musculature and, when applied to fresh fish or lightly processed fish products (never frozen before processing), determining the viability of Anisakidae L3, which may be present.
This method doesn’t allow determining species or genotype of detected parasites, which identification is made by morphological and/or molecular methods.

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This document gives guidelines for the establishment of a conversion relationship between the results
of an alternative method and an anchor method, and its verification for the quantitative determination
of the microbiological quality of milk.
NOTE The conversion relationship can be used a) to convert results from an alternative method to the anchor
basis or b) to convert results/limits, expressed on an anchor basis, to results in units of an alternative method.

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This document specifies a method for the detection of Anisakidae L3 larvae commonly found in marine and anadromous fishes. The method is applicable to fresh fish and/or frozen fish, as well as lightly processed fish products, such as marinated, salted or cold smoked. This method is applicable to quantifying parasitic infections by estimating the number of parasites in the fish musculature. This method does not apply to determining the species or genotype of detected parasites. Final identification is made by morphological and/or molecular methods.

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This document specifies a method for the detection of Anisakidae L3 larvae commonly found in marine and anadromous fishes. The method is applicable to fresh fish and/or frozen fish, as well as lightly processed fish products, such as marinated, salted or smoked. It is also suitable for visceral organs as a confirmatory method for a visual inspection scheme. The artificial digestion method[4][5][6] is applicable to quantifying parasitic infections by estimating the number of parasites in the fish musculature and, when applied to fresh fish or lightly processed fish products (never frozen before processing), determining the viability of Anisakidae L3, which can be present. This method does not apply to determining the species or genotype of detected parasites. Final identification is made by morphological and/or molecular methods.

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This document deals with verification of methods for the detection and/or the enumeration of microorganisms, with particular emphasis on the implementation of a reference/alternative method in the user laboratory and verification of a reference/alternative method using items included in the scope of the method and tested routinely but not tested in the original validation study

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This document gives guidelines for the establishment of a conversion relationship between the results of an alternative method and an anchor method, and its verification for the quantitative determination of the microbiological quality of milk. NOTE The conversion relationship can be used a) to convert results from an alternative method to the anchor basis or b) to convert results/limits, expressed on an anchor basis, to results in units of an alternative method.

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This document specifies the protocol for the verification of reference methods and validated alternative methods for implementation in the user laboratory. This document is applicable to the verification of methods used for the analysis (detection and/or quantification), confirmation and typing of microorganisms in: — products intended for human consumption; — products intended for animal feeding; — environmental samples in the area of food and feed production, handling; — samples from the primary production stage. This document is, in particular, applicable to bacteria and fungi. Some clauses can be applicable to other (micro)organisms or their metabolites, to be determined on a case-by-case basis. The technical protocols for the verification of validated qualitative methods and validated quantitative methods are described in Clauses 5 and 6. The technical protocol for the verification of validated alternative confirmation and typing methods is described in Clause 7. The protocols for the verification of non-validated reference methods are described in Annex F.

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This document specifies a method for the direct enumeration of potentially enteropathogenic V. parahaemolyticus (tdh and/or trh positive) and/or the enumeration of total V. parahaemolyticus in seafood.

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The proposed deliverable specifies an alternative technical protocol for the validation of mostly non-proprietary methods in the field of microbiological analysis of food, animal feed, and environmental and primary production stage samples.
It is closely related to ISO 16140-2. The latter specifies the technical protocol for the validation of proprietary methods, including a classical interlaboratory study and a method comparison study to be conducted in one laboratory. The realization of classical interlaboratory studies demands a sufficient number of participating laboratories (at least 8 laboratories are required). There are many occasions where a sufficient number of participating laboratories is not available (e.g. when a new method is required quickly after an outbreak of a new microorganism). In this case, the validation cannot be considered as reliable any longer.
The proposed deliverable uses a modified protocol based on orthogonal, factorial studies. By selection of suitable influencing factors (technician, nutrient media, sample preparation, temperature, duration) a high certainty of the determined method validation parameters is obtained, so that the number of required collaborating laboratories can be reduced up to a minimum of 4.
This validation protocol can be used in different ways. If the 4 collaborators can be considered a “random sample” of independent and competent laboratories and from different organizations, the test method can be considered as being validated in the sense that accurate and precise measurements are to be expected from any competent laboratory. If the 4 collaborators can be considered a “random sample” of independent and competent laboratories from one organization, the test method can be considered as being validated in the sense that accurate and precise measurements are to be expected from any competent laboratory in this organization.

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The proposed deliverable specifies the procedure for single-laboratory validation of mainly non-proprietary methods in the fields of microbiological analysis of food, feed, and environmental and primary production stage samples. Single-laboratory validation is required if an interlaboratory validation according to ISO 16140-2 is not appropriate, e.g. for in-house methods or when the required number of participating laboratories is not available. Single-laboratory validation is not part of the optimization of methods. It can be applied only for methods that are fully specified with regard to all relevant parameters (including tolerances on temperatures and specifications on nutrient media).
The proposed deliverable describes two protocols for single-laboratory validation, a conventional protocol, and a factorial protocol. The conventional protocol is a stepwise procedure; both the study design and the performance measures are derived from ISO 16140-2. The performance measures of the factorial protocol are also derived from ISO 16140-2; however, it is using an orthogonal, factorial study design. By selection of suitable influencing factors (technician, nutrient media, sample preparation, temperature, duration) a high certainty of the determined method validation parameters is obtained, so that the number of required individual tests can be reduced by more than 50 %.

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This document specifies the general principles and the technical protocols (based on orthogonal, factorial studies) for the validation of non-proprietary methods for microbiology of the food chain. This document is applicable to the validation of methods used for the analysis (detection or quantification) of microorganisms in: — products intended for human consumption; — products intended for animal feeding; — environmental samples in the area of food and feed production, handling; — samples from the primary production stage. This document is, in particular, applicable to bacteria and fungi. Some clauses can be applicable to other (micro)organisms or their metabolites, to be determined on a case-by-case basis. This document specifies protocols for the validation against a reference method for both quantitative and qualitative methods. This document also provides a protocol for the validation of quantitative methods without a reference method. Qualitative methods cannot be validated without a reference method in accordance with this document. NOTE ISO 16140-2 specifies the general principle and the technical protocol for the validation of alternative, mostly proprietary, methods against a reference method. This document is only applicable to the validation of methods that are fully specified with regard to all relevant parameters (including tolerances on temperatures and specifications on culture media) and that have already been optimized. Methods that have been validated in accordance with this document can be used by the laboratories of the specified population of laboratories.

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This document specifies the general principles and the technical protocols for single-laboratory validation of methods for microbiology in the food chain. The protocols in this document only validate the method for the laboratory conducting the study. This document is applicable to single-laboratory validation of: — methods used in the analysis (detection or quantification) of microorganisms in: — products intended for human consumption; — products intended for animal feeding; — environmental samples in the area of food and feed production, handling; — samples from the primary production stage; — methods for the confirmation or typing of microorganisms. This validation will replace only the confirmation or typing procedure of a specified method (see Annex G). This document is, in particular, applicable to bacteria and fungi. Some clauses can be applicable to other (micro)organisms or their metabolites, to be determined on a case-by-case basis. Single-laboratory validation is required if an interlaboratory validation in accordance with ISO 16140-2 is not appropriate. Possible applications are: — validation of an in-house method; — method evaluation study in the validation process of a reference method in accordance with ISO 17468; — extension of the scope of an ISO 16140-2 validated method, e.g. category extension or test portion size; — modifications of existing methods. Single-laboratory validation is the second step in the standardization of a reference method (see ISO 17468). It is only applicable to methods that are fully specified with regard to all relevant parameters (including tolerances on temperatures and specifications on culture media) and that have already been optimized.

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This document specifies rules for the preparation of samples of milk and milk products and their
suspensions for microbiological examination when the samples require a different preparation from
the general methods specified in ISO 6887-1.
This document excludes the preparation of samples for both enumeration and detection test methods
where preparation details are specified in the relevant International Standards.
This document is intended to be used in conjunction with ISO 6887-1.
This document is applicable to:
a) milk and liquid milk products;
b) dehydrated milk products;
c) cheese and cheese products;
d) casein and caseinates;
e) butter;
f) milk-based ice-cream;
g) milk-based custard, desserts and sweet cream;
h) fermented milks, yogurt, probiotics milk products and sour cream;
i) dehydrated milk-based infant foods, with or without probiotics.

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This document specifies rules for the preparation of samples of milk and milk products and their suspensions for microbiological examination when the samples require a different preparation from the general methods specified in ISO 6887-1. This document excludes the preparation of samples for both enumeration and detection test methods where preparation details are specified in the relevant International Standards. This document is intended to be used in conjunction with ISO 6887-1. This document is applicable to: a) milk and liquid milk products; b) dehydrated milk products; c) cheese and cheese products; d) casein and caseinates; e) butter; f) milk-based ice-cream; g) milk-based custard, desserts and sweet cream; h) fermented milks, yogurt, probiotics milk products and sour cream; i) dehydrated milk-based infant foods, with or without probiotics.

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This part of ISO 16140 specifies the general principle and the technical protocol for the validation of alternative, mostly proprietary, confirmation methods in the field of microbiological analysis of food, animal feed, and environmental and primary production stage samples. This procedure is limited to the validation of alternative (proprietary) confirmation methods that are intended to replace (partly or completely) the confirmatory procedure described in the standard method for the enumeration or detection of specific (group of) microorganisms. The "sample" to be used for confirmation shall be a suspected colony that has been obtained following the reference or alternative culture method procedure. It is however not intended for confirmation using a (pure) colony from an unknown origin. Validation studies according to this standard are intended to be performed by organizations involved in method validation.

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This International Standard gives requirements and guidance for the estimation and expression of measurement
uncertainty (MU) associated with quantitative results in microbiology of the food chain.
It is applicable to the quantitative analysis
 of products intended for human consumption or the feeding of animals, and
 of environmental samples in the area of food production and food handling,
 of samples at the stage of primary production.
The quantitative analysis is typically carried out by enumeration of microorganisms using a colony-count technique. It is
also generally applicable to other quantitative analyses, including Most Probable Number (MPN) techniques and
instrumental methods.
The uncertainty estimated by this International Standard does not include systematic effects (“trueness” or “bias”).

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This document specifies the general principle and the technical protocol for the validation of alternative confirmation methods for microbiology in the food chain. This document compares the result of the alternative confirmation method against the confirmation procedure of a reference method or, if needed, a reference confirmation method (e.g. whole genome sequencing).
This document is applicable to the validation of alternative confirmation methods used for the analysis (detection or quantification) of isolated microorganisms in:
— products intended for human consumption;
— products intended for animal feeding;
— environmental samples in the area of food and feed production, handling;
— samples from the primary production stage.
Validated alternative confirmation methods can be used to replace (partly or completely) the confirmation procedure described in:
— the reference method;
— an alternative method validated in accordance with ISO 16140-2 only if one of the isolation agars specified in the validation study of the alternative confirmation method is used.
This document is also applicable to the validation of alternative typing methods, where the reference method can be, for example, a serological method (e.g. serotyping of Salmonella) or a molecular method (e.g. typing of Shiga toxin-producing E. coli).
This document is, in particular, applicable to bacteria and fungi. Some clauses can be applicable to other (micro)organisms, to be determined on a case-by-case basis.
Validation studies in accordance with this document are primarily intended to be performed by organizations or expert laboratories involved in method validation, but can also be used by a single laboratory, especially when performing in-house validation under certain conditions (see ISO 16140-4).

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This document specifies requirements and gives guidance for the estimation and expression of measurement uncertainty (MU) associated with quantitative results in microbiology of the food chain.
It is applicable to the quantitative analysis of:
— products intended for human consumption or the feeding of animals;
— environmental samples in the area of food production and food handling;
— samples at the stage of primary production.
The quantitative analysis is typically carried out by enumeration of microorganisms using a colony-count technique. This document is also generally applicable to other quantitative analyses, including:
— most probable number (MPN) techniques;
— instrumental methods, such as impediometry, adenosine triphosphate (ATP) and flow cytometry;
— molecular methods, such as methods based on quantitative polymerase chain reaction (qPCR).
The uncertainty estimated by this document does not include systematic effects (bias).

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This document specifies the general principle and the technical protocol for the validation of alternative confirmation methods for microbiology in the food chain. This document compares the result of the alternative confirmation method against the confirmation procedure of a reference method or, if needed, a reference confirmation method (e.g. whole genome sequencing). This document is applicable to the validation of alternative confirmation methods used for the analysis (detection or quantification) of isolated microorganisms in: — products intended for human consumption; — products intended for animal feeding; — environmental samples in the area of food and feed production, handling; — samples from the primary production stage. Validated alternative confirmation methods can be used to replace (partly or completely) the confirmation procedure described in: — the reference method; — an alternative method validated in accordance with ISO 16140-2 only if one of the isolation agars specified in the validation study of the alternative confirmation method is used. This document is also applicable to the validation of alternative typing methods, where the reference method can be, for example, a serological method (e.g. serotyping of Salmonella) or a molecular method (e.g. typing of Shiga toxin-producing E. coli). This document is, in particular, applicable to bacteria and fungi. Some clauses can be applicable to other (micro)organisms, to be determined on a case-by-case basis. Validation studies in accordance with this document are primarily intended to be performed by organizations or expert laboratories involved in method validation, but can also be used by a single laboratory, especially when performing in-house validation under certain conditions (see ISO 16140-4).

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This document provides the overall framework for detection of sequences corresponding to species containing allergens using the polymerase chain reaction (PCR). It relates to the requirements for the specific amplification of target nucleic acid sequences (DNA) and for the confirmation of the identity of the amplified nucleic acid sequence.
Guidelines, minimum requirements and performance criteria laid down in this document are intended to ensure that comparable and reproducible results are obtained in different laboratories. This document has been established for food matrices.

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This document specifies a method for the detection of celery (Apium graveolens) in emulsion-type sausages (e.g. Frankfurter, Wiener).
Real-time PCR detection of celery is based on an 101 bp (base pair) sequence from the gene of the mannitol dehydrogenase (GenBank Acc. No. AF067082 ) of celery (Apium graveolens).
The method has been validated on emulsion-type sausages (Bavarian “Leberkäse”) spiked with celery. For this purpose meat batter containing mass fractions of 50 % pork meat, 25 % pork fat, 23 % crushed ice and 1,8 % of a mixture of sodium chloride, nitrite, nitrate, phosphates and ascorbates was prepared according to a standard procedure for emulsion-type sausage. The meat batter was spiked with either ground celery seeds or celery root powder to 1000 mg/kg. Lower spiking levels were obtained by diluting with celery-free meat batter. The batter was stuffed into casings and heated at 65 °C for 60 min [2].

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This document specifies requirements and gives guidance for the estimation and expression of measurement uncertainty (MU) associated with quantitative results in microbiology of the food chain. It is applicable to the quantitative analysis of: — products intended for human consumption or the feeding of animals; — environmental samples in the area of food production and food handling; — samples at the stage of primary production. The quantitative analysis is typically carried out by enumeration of microorganisms using a colony-count technique. This document is also generally applicable to other quantitative analyses, including: — most probable number (MPN) techniques; — instrumental methods, such as impediometry, adenosine triphosphate (ATP) and flow cytometry; — molecular methods, such as methods based on quantitative polymerase chain reaction (qPCR). The uncertainty estimated by this document does not include systematic effects (bias).

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EN-ISO 15216-2 specifies a method for detection of hepatitis A virus (HAV) and norovirus genogroups I (GI) and II (GII), from test samples of foodstuffs [(soft fruit, leaf, stem and bulb vegetables, bottled water, bivalve molluscan shellfish (BMS)] or surfaces using real-time RT-PCR.This method is not validated for detection of the target viruses in other foodstuffs (including multi-component foodstuffs), or any other matrices, nor for the detection of other viruses in foodstuffs, surfaces or other matrices.

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This document provides the overall framework for detection of sequences corresponding to species containing allergens using the polymerase chain reaction (PCR). It relates to the requirements for the specific amplification of target nucleic acid sequences (DNA) and for the confirmation of the identity of the amplified nucleic acid sequence.
Guidelines, minimum requirements and performance criteria laid down in European Standards are intended to ensure that comparable and reproducible results are obtained in different laboratories. This document has been established for food matrices.
This document is intended to be used in addition to EN 15842.

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This document specifies a method for the detection of celery (Apium graveolens) in emulsion-type sausages (e.g. Frankfurter, Wiener).
Real-time PCR (polymerase chain reaction) detection of celery is based on an 101 bp (base pair) sequence from the gene of the mannitol dehydrogenase (GenBank Acc. No. AF067082 ) of celery (Apium graveolens).
The method has been validated on emulsion-type sausages (Bavarian “Leberkäse”) spiked with celery. For this purpose meat batter containing mass fractions of 50 % pork meat, 25 % pork fat, 23 % crushed ice and 1,8 % of a mixture of sodium chloride, nitrite, nitrate, phosphates and ascorbates was prepared according to a standard procedure for emulsion-type sausage. The meat batter was spiked with either ground celery seeds or celery root powder to 1000 mg/kg. Lower spiking levels were obtained by diluting with celery-free meat batter. The batter was stuffed into casings and heated at 65 °C for 60 min [1].
This document is intended to be used in addition to EN 15842 and FprEN 15634 1.

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This document specifies a method for detection of hepatitis A virus (HAV) and norovirus genogroups I (GI) and II (GII), from test samples of foodstuffs [(soft fruit, leaf, stem and bulb vegetables, bottled water, bivalve molluscan shellfish (BMS)] or surfaces using real-time RT-PCR.
This method is not validated for detection of the target viruses in other foodstuffs (including multi-component foodstuffs), or any other matrices, nor for the detection of other viruses in foodstuffs, surfaces or other matrices.

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This document specifies a horizontal method for the enumeration of psychrotrophic microorganisms that are able to grow and form colonies on a solid agar culture medium after aerobic incubation at 6,5 °C. This document is applicable to — products intended for human consumption, — products intended for animal feeding, — environmental samples in the area of food and feed production, handling, and — samples from the primary production stage. NOTE Annex B specifies a rapid method for the estimated enumeration of psychrotrophic microorganisms in raw and pasteurized milk.

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This document specifies a method for detection of hepatitis A virus (HAV) and norovirus genogroups I (GI) and II (GII), from test samples of foodstuffs [(soft fruit, leaf, stem and bulb vegetables, bottled water, bivalve molluscan shellfish (BMS)] or surfaces using real-time RT-PCR. This method is not validated for detection of the target viruses in other foodstuffs (including multi-component foodstuffs), or any other matrices, nor for the detection of other viruses in foodstuffs, surfaces or other matrices.

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This document specifies protocols for conducting microbiological challenge tests for growth studies on
vegetative and spore-forming bacteria in raw materials and intermediate or end products.
The use of this document can be extended to yeasts that do not form mycelium.

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This document specifies protocols for conducting microbiological challenge tests for growth studies on vegetative and spore-forming bacteria in raw materials and intermediate or end products.
The use of this document can be extended to yeasts that do not form mycelium.

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This document specifies requirements and gives guidelines for the organization of proficiency testing
(PT) schemes for microbiological examinations of
a) foods and beverages,
b) feeding animals,
c) environmental samples from food and feed production and handling, and
d) primary production stages.
This document is also applicable to the microbiological examination of water where water is either used
in food production or is regarded as a food in national legislation.
This document relates to the technical organization and implementation of PT schemes, as well as the
statistical treatment of results of microbiological examinations.
This document is designed for use with ISO/IEC 17043 and ISO 13528, and deals only with areas where
specific or additional details are necessary for PT schemes dealing with microbiological examinations
for the areas specified in the first paragraph.

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This document specifies requirements and gives guidelines for the organization of proficiency testing (PT) schemes for microbiological examinations of
a) foods and beverages,
b) feeding animals,
c) environmental samples from food and feed production and handling, and
d) primary production stages.
This document is also applicable to the microbiological examination of water where water is either used in food production or is regarded as a food in national legislation.
This document relates to the technical organization and implementation of PT schemes, as well as the statistical treatment of results of microbiological examinations.
This document is designed for use with ISO/IEC 17043 and ISO 13528, and deals only with areas where specific or additional details are necessary for PT schemes dealing with microbiological examinations for the areas specified in the first paragraph.

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This document specifies protocols for conducting microbiological challenge tests for growth studies on vegetative and spore-forming bacteria in raw materials and intermediate or end products. The use of this document can be extended to yeasts that do not form mycelium.

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This document specifies requirements and gives guidelines for the organization of proficiency testing (PT) schemes for microbiological examinations of a) foods and beverages, b) feeding animals, c) environmental samples from food and feed production and handling, and d) primary production stages. This document is also applicable to the microbiological examination of water where water is either used in food production or is regarded as a food in national legislation. This document relates to the technical organization and implementation of PT schemes, as well as the statistical treatment of results of microbiological examinations. This document is designed for use with ISO/IEC 17043 and ISO 13528, and deals only with areas where specific or additional details are necessary for PT schemes dealing with microbiological examinations for the areas specified in the first paragraph.

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This standard specifies sampling techniques, using contact plates or swabs on surfaces, in the food industry environment (and food  processing plants), with a view of detecting or enumerating viable microorganisms.
NOTE The term “environment” means any item in contact with the food product or likely to represent a contamination or recontamination source, for example, material, premises, operators.

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