COMMISSION IMPLEMENTING REGULATION (EU) No 503/2013

of 3 April 2013

on applications for authorisation of genetically modified food and feed in accordance with Regulation (EC) No 1829/2003 of the European Parliament and of the Council and amending Commission Regulations (EC) No 641/2004 and (EC) No 1981/2006

(Text with EEA relevance)

THE EUROPEAN COMMISSION,

Having regard to the Treaty on the Functioning of the European Union,

Having regard to Regulation (EC) No 1829/2003 of the European Parliament and of the Council of 22 September 2003 on genetically modified food and feed (1), and in particular Articles 5(7), 11(5), 17(7) and 23(5) thereof,

After consulting the European Food Safety Authority,

Whereas:

Regulation (EC) No 1829/2003 lays down Union procedures for the authorisation and supervision of genetically modified food and feed, including rules for the labelling of such food and feed. That Regulation provides for a scientific evaluation to be carried out on the risks that the genetically modified food or feed may present for human and animal health and, as the case may be, for the environment. It also provides that a genetically modified food or feed must not mislead the consumer or the user and must not differ from the food or feed which it is intended to replace to such an extent that its normal consumption would be nutritionally disadvantageous for humans or animals.

Regulation (EC) No 1829/2003 provides, in particular, that applications for authorisation are to adequately and sufficiently demonstrate that the genetically modified food and feed satisfy the requirements laid down in that Regulation, in respect of their proposed uses.

In the interest of consistency of Union legislation, certain definitions laid down in Regulation (EC) No 178/2002 of the European Parliament and of the Council of 28 January 2002 laying down the general principles and requirements of food law, establishing the European Food Safety Authority and laying down procedures in matters of food safety (2) should also apply to this Regulation.

Commission Regulation (EC) No 641/2004 (3) on detailed rules for the implementation of Regulation (EC) No 1829/2003 provides for certain detailed rules concerning applications for authorisation submitted in accordance with Regulation (EC) No 1829/2003. To facilitate the preparation of applications and ensure that they contain all the information needed for their assessment, it is necessary to provide for more comprehensive and systematic rules concerning applications for authorisation, which should also be specific to each type of genetically modified organisms (GMO), namely plants, animals and micro-organisms.

The rules laid down in this Regulation should only cover applications concerning genetically modified plants for food or feed uses, food or feed containing or consisting of genetically modified plants and food or feed produced from such plants. Genetically modified plants, for which sufficient experience is available to date, constitute the vast majority of current applications.

The rules laid down in this Regulation should specify the general requirements for the presentation and preparation of applications, namely requirements to provide general and scientific information, including methods for detection, and identification, as well as reference material so as to ensure that applications comply with the conditions laid down in Articles 5, 17 and 30 of Regulation (EC) No 1829/2003.

The applicant should also take into consideration the scientific information to be provided in the application as regards the environmental risk assessment of GMOs or food and feed containing or consisting of GMOs, as set out in the principles for the environmental risk assessment in Annex II to Directive 2001/18/EC of the European Parliament and of the Council of 12 March 2001 on the deliberate release into the environment of genetically modified organisms and repealing Council Directive 90/220/EEC (4) , as well as the applicable guidance published by the European Food Safety Authority (EFSA) in this regard.

In addition to the general requirements for the presentation and preparation of applications, it is appropriate to provide for specific rules to ensure that the scientific information required in the application adequately and sufficiently demonstrates that the genetically modified food or feed satisfy the requirements laid down in Regulation (EC) No 1829/2003, in respect of their proposed uses.

These rules should therefore provide for a set of studies that should be included in all applications, as well as the test methods to be followed to perform such studies, whilst taking into account relevant international standards, such as the guideline of the Codex Alimentarius for the conduct of safety assessment of foods derived from the recombinant-DNA plant (5) .

In accordance with the applicable guidance of the EFSA (6) , the safety assessment of the genetically modified food or feed should include studies related to new components resulting from the genetic modification, the molecular characterisation of the genetically modified plant, the comparative analysis of the composition and the phenotype of the genetically modified plant compared to its conventional counterpart. Depending on the characteristics of the genetically modified plant and on the outcome of that first set of studies, the EFSA guidance indicates that it may be necessary to perform additional studies. In that respect, the EFSA considers that notwithstanding its limitations, a 90-day feeding study in rodents with whole food or feed is, when justified, the primary additional study to address uncertainties identified in the course of the safety assessment.

It has, however, not been proved possible to define with the necessary precision the level of uncertainties which would require the submission of 90-day feeding studies. In addition, some food and feed assessment bodies of Member States are of the opinion that such study should be performed in all applications for genetically modified plants containing single transformation events. Considering these diverging views, as well as to improve consumer confidence, such studies should be, for the time being, requested in all applications related to genetically modified plants with single transformation events and, where appropriate, on genetically modified plants containing stacked transformation events.

Studies to demonstrate that a genetically modified food or feed fulfils the requirements of Regulation (EC) No 1829/2003 involving the use of laboratory animals should be carried out in accordance with Directive 2010/63/EU of the European Parliament and of the Council of 22 September 2010 regarding the protection of the animals used for scientific purposes (7) , and should be kept to a minimum while ensuring an adequate demonstration of the safety of the genetically modified food or feed. The current uncertainties in relation to the need and design of 90-day feeding trials will be addressed by a large research project under the 2012 work programme of Theme 2 ‘Food, Agriculture and Fisheries, and Biotechnologies’ of the seventh Framework Programme for Research (FP7). The requirements regarding animal feeding trials in the context of GMO risk assessments should be reviewed in the light of the outcome of this project expected to be available by the end of 2015 at the latest. Other credible scientific knowledge which might be available at that time should also be taken into account.

While the rules laid down in this Regulation should be valid for all applications for genetically modified plants, the type and necessity of the studies to evaluate the characteristics and safety of genetically modified food or feed subject to an application may vary, depending on the nature of the genetic modification or of the product. For example, genetic modifications which have negligible impact on the composition of a genetically modified food or feed or highly refined products that may be proven to be identical to products produced from the conventional counterpart require different studies than a product resulting from complex genetic modification aiming to modify its nutritional characteristics.

The requirements laid down in this Regulation regarding the studies which have to be included in an application for authorisation under Regulation (EC) No 1829/2003 should not prevent the EFSA to request, where appropriate, the applicant to supplement the particulars accompanying the application in accordance with Articles 6(2) and 18(2) of Regulation (EC) No 1829/2003.

In order to ensure that studies are of high quality and documented in a transparent way, it is essential that they are performed under appropriate quality assurance systems and raw data should be provided in all cases and be in a suitable electronic format. Toxicological studies should be performed in accordance with the quality assurance principles laid down by Directive 2004/10/EC of the European Parliament and of the Council of 11 February 2004 on the harmonisation of laws, regulations and administrative provisions relating to the application of the principles of good laboratory practice and the verification of their applications for tests on chemical substances (8) . If such studies are carried out outside the Union, they should follow the state-of-the-art OECD Principles on Good Laboratory Practice (GLP). With regard to studies other than toxicological studies, they should be conducted under ISO or GLP standards.

It is also necessary to define the requirements regarding the submission of additional information related to the safety of the GMO and scientific peer-reviewed literature related to the potential effects on health and on the environment of the products covered by the application.

During the process of the genetic modification of plants and other organisms, marker genes are often used to facilitate the selection and identification of genetically modified cells, containing the gene of interest inserted into the genome of the host organism, among the vast majority of untransformed cells. Such marker genes should be carefully selected. In addition, it is now possible to develop GMOs without the use of antibiotic resistance marker genes. Against this background and in accordance with Article 4(2) of Directive 2001/18/EC, the applicant should therefore aim to develop GMOs without the use of antibiotic resistance marker genes.

The harvest of segregating genetically modified plants (segregating crops) containing stacked transformation events contains various subcombinations of transformation events. In addition, current control procedures do not allow identifying the origin of combinations of transformation events. Therefore, in order to ensure that authorisations are coherent with the products of which the placing on the market is unavoidable and for the feasibility of controls, the applications for genetically modified food and feed from segregating crops should include all subcombinations independently of their origin and not yet authorised.

Regulation (EC) No 1829/2003 provides that a proposal for post-market monitoring of the use of the genetically modified food or feed shall only be submitted by the applicant where it is appropriate. It is therefore necessary to set out the conditions under which such a proposal should, according to the outcome of the risk assessment, accompany the application. Post-market monitoring should only be considered in cases where, notwithstanding the fact that the safety of genetically modified food and feed has been demonstrated, it is appropriate to confirm the expected consumption, the application of conditions of uses or identified effects. This is for example the case when the genetically modified food or feed has altered nutritional composition or when that its nutritional value differs from the conventional food or feed that it would replace or when there is a likelihood of increased allergenicity due to the genetic modification.

This Regulation should take account of the international trade commitments of the Union and of the requirements of the Cartagena Protocol on Biosafety to the Convention on Biological Diversity (the Cartagena Protocol), approved by Council Decision 2002/628/EC of 25 June 2002 concerning the conclusion, on behalf of the European Community, of the Cartagena Protocol on Biosafety (9) as well as the provisions of Regulation (EC) No 1946/2003 of 15 July 2003 of the European Parliament and of the Council on transboundary movements of genetically modified organisms (10) .

In order to ensure that test methods included in the application are adequate to demonstrate that the food or feed complies with the requirements for authorisation set out in Regulation (EC) No 1829/2003, they should be carried out in accordance with the present Regulation, or internationally agreed guidelines such as those described by the OECD, when available. To ensure that applications for renewal meet the same standards as regards tests methods, it is appropriate that these requirements also apply to application for renewal of authorisation of GM food and feed.

In order to provide an accurate designation of the GM food or feed subject to an application under Regulation (EC) No 1829/2003, applications should include proposals for a unique identifier for each GMO concerned in accordance with Commission Regulation (EC) No 65/2004 of 14 January 2004 establishing a system for the development and assignment of unique identifiers for genetically modified organisms (11) .

This Regulation replaces certain provisions of Regulation (EC) No 641/2004 as regards genetically modified plants for food or feed uses, food or feed containing or consisting of genetically modified plants and food or feed produced from genetically modified plants. However, Regulation (EC) No 641/2004 should continue to apply as regards other types of genetically modified products, namely genetically modified animals and genetically modified micro-organisms. Moreover, certain provisions of that Regulation are obsolete. Regulation (EC) No 641/2004 should therefore be amended accordingly.

Commission Regulation (EC) No 1981/2006 of 22 December 2006 on detailed rules for the implementation of Article 32 of Regulation (EC) No 1829/2003 of the European Parliament and of the Council as regards the Community reference laboratory for genetically modified organisms (12) should be amended to include references to this Regulation.

Regulation (EC) No 1829/2003 provides that the Commission is to consult the EFSA before establishing implementing rules with regard to the applications for authorisation under that Regulation. The EFSA has been consulted on those rules accordingly.

This Regulation has been drawn up on the basis of current scientific and technical knowledge. Therefore, the Commission should monitor any developments in this field and the publication of new or additional guidance by the EFSA.

This Regulation applies to applications submitted after its entry into force. It is necessary to provide for transitional measures in order to enable the applicants to comply with those rules and for the current applications or the applications close to being submitted to proceed without unnecessary delays.

The measures provided for in this Regulation are in accordance with the opinion of the Standing Committee on the Food Chain and Animal Health,

HAS ADOPTED THIS REGULATION:

CHAPTER I

GENERAL PROVISIONS

Article 1

Scope

This Regulation shall apply to applications submitted under Articles 5, 11, 17 and 23 of Regulation (EC) No 1829/2003 for the authorisation of:

food produced from or containing ingredients produced from genetically modified plants or feed produced from such plants.

Article 2

Definitions

For the purposes of this Regulation, the definitions in Regulation (EC) No 1829/2003 apply.

The definitions of ‘risk’, ‘risk assessment’ and ‘hazard’ applicable for the purposes of this Regulation are those provided in Article 3 of Regulation (EC) No 178/2002.

CHAPTER II

GENERAL REQUIREMENTS

Article 3

Preparation and presentation of applications submitted under Articles 5(1) and 17(1)

1. The application submitted under Articles 5(1) and 17(1) of Regulation (EC) No 1829/2003 shall:

be submitted in accordance with the requirements for the preparation and presentation of applications set out in Annex I;

contain all the information required by Annex I, in accordance with the specific requirements of Articles 4, 5 and 6.

2. The application shall include, for each of the specific requirements laid down in Articles 4, 5 and 6:

the summaries and results of the studies referred to in the application;

3. The application shall contain a checklist demonstrating that the information required under Articles 4, 5 and 6 is complete.

4. Where an application is limited to either food or feed use, it shall contain a verifiable justification explaining why the authorisation shall not cover both uses in accordance with Article 27 of Regulation (EC) No 1829/2003.

5. The application shall, at the time of submission, clearly state which parts of the application are claimed to be confidential and provide verifiable justification in accordance with Article 30 of Regulation (EC) No 1829/2003.

Additional information submitted during the authorisation procedure shall, at the time of submission, clearly state which parts of this additional information is claimed to be confidential and provide verifiable justification in accordance with Article 30 of Regulation (EC) No 1829/2003.

6. When studies have been already submitted to the European Food Safety Authority (EFSA) for the purposes of an application and, where relevant, to the extent that they may be used by the applicant in accordance with Article 31 of Regulation (EC) No 1829/2003, a reference to such studies and the results of the EFSA’s assessment may, with the agreement of EFSA, be made in the framework of another application.

CHAPTER III

SPECIFIC REQUIREMENTS

Article 4

Requirements for the performance of studies for applications submitted under Articles 5(3) and 17(3)

1. Toxicological studies shall be conducted in facilities which comply with the:

The applicant shall provide evidence to demonstrate such compliance.

2. Studies, other than toxicological studies, shall:

3. Information on the study protocols and the results obtained from the studies referred to in paragraphs 1 and 2 shall be comprehensive and include the raw data in an electronic format, suitable for carrying out statistical or other analysis.

Article 5

Scientific requirements for the risk assessment of genetically modified food and feed for applications submitted under Articles 5(3) and 17(3)

1. Information, including studies, required to accompany the application as referred to in Article 5(3)(a) to (f) and (h) and in Article 17(3)(a) to (f) and (h) of Regulation (EC) No 1829/2003 shall be provided in accordance with the scientific requirements for the risk assessment of genetically modified food and feed set out in Annex II to this Regulation.

2. By way of derogation from paragraph 1, an application may be submitted that does not satisfy all the requirements of that paragraph provided that:

particular information is not necessary owing to the nature of the genetic modification or of the product; or

it is not scientifically necessary, or technically possible to supply such information.

The applicant shall submit reasoned justification for the derogation.

3. Paragraphs 1 and 2 shall not prevent the EFSA to request, where appropriate, the applicant to supplement the particulars accompanying the application as provided for in Articles 6(2) and 18(2) of Regulation (EC) No 1829/2003.

Article 6

Additional information related to the risk assessment of genetically modified food or feed for applications submitted under Articles 5(3) and 17(3)

1. In addition to the information required in accordance with Article 5 and Annex II, the application shall include a systematic review of studies published in the scientific literature and studies performed by the applicant within the period of 10 years prior to the date of submission of the dossier on the potential effects on human and animal health of the genetically modified food and feed covered by the application.

2. During the authorisation procedure, the applicant shall submit, without delay, to the EFSA additional information which might influence the risk assessment of the genetically modified food or feed generated following the submission of the application. In particular, the applicant shall submit to the EFSA information regarding any prohibition or restriction imposed by a competent authority of any third country on the basis of a risk assessment of the genetically modified food and feed.

Article 7

Requirements applicable for post-market monitoring of genetically modified food or feed for applications submitted under Articles 5(3) and 17(3)

1. The applicant shall submit a proposal for post-market monitoring regarding the use of the food and feed as referred to in Articles 5(3)(k) and 17(3)(k) of Regulation (EC) No 1829/2003 when the information provided in accordance with Articles 4, 5 and 6 demonstrates that the genetically modified food and feed comply with Articles 4(1) and 16(1) of Regulation (EC) No 1829/2003 and when, in accordance with the outcome of the risk assessment, it is appropriate to confirm:

that specific recommendations of uses are followed by the consumer/animal owner;

the relevance and intensity of effects and unintended effects detected during the pre-market risk assessment which can only be further characterised by post-market monitoring.

2. The applicant shall ensure that the post-market monitoring is:

developed to collect reliable information with respect to one or several of the aspects set out in paragraph 1. This information shall allow the detection of indications on whether any (adverse) effect on health may be related to genetically modified food or feed consumption;

based on strategies aiming at collecting relevant information from specific stakeholders including consumers and on a reliable and validated flow of information between the different stakeholders. More specific strategies shall be included when data on individual intakes of a specific food item or intakes of particular age groups have to be collected;

accompanied by adequate justification and a thorough description of the selected methodologies for the proposed post-market monitoring including aspects related to the analysis of the collected information.

Article 8

Requirements concerning the methods of detection, identification and quantification and for control samples and reference material of genetically modified food or feed for applications submitted under Articles 5(3), 11(2), 17(3) and 23(2)

1. Applications submitted under Articles 5(1) and 17(1) of Regulation (EC) No 1829/2003 shall comply with the following requirements, as referred to in Article 5(3)(i) and (j) and Article 17(3)(i) and (j) of that Regulation, and set out in Annex III to this Regulation, for:

samples of food or feed and their control samples, and information as to the place where the reference material can be accessed.

2. For applications submitted under Articles 11(1) and 23(1) of Regulation (EC) No 1829/2003, the requirements set out in Annex III to this Regulation for:

samples of food or feed and their control samples, and information as to the place where the reference material can be accessed,

shall apply only for the purposes of application of Articles 11(2)(d) and 23(2)(d).

CHAPTER IV

TRANSITIONAL AND FINAL PROVISIONS

Article 9

Transitional provisions

1. Until 8 December 2013, applicants may choose to submit applications falling under the scope of this Regulation under Regulation (EC) No 641/2004 in the version of that Regulation in force on 8 June 2013.

2. By way of derogation from Article 4(2), in the case of studies launched prior to the date of entry into force of this Regulation and conducted under quality assurance systems other than GLP and ISO, the applicant shall provide:

a thorough description of the quality assurance system under which such studies were performed; and

comprehensive information on the protocols and the results obtained from the studies including the raw data.

Article 10

Amendments to Regulation (EC) No 641/2004

Regulation (EC) No 641/2004 is amended as follows:

This chapter provides detailed rules concerning applications for authorisation submitted in accordance with Articles 5 and 17 of Regulation (EC) No 1829/2003 except for those applications covered by Commission Implementing Regulation (EU) No 503/2013 (13) .

Article 11

Amendments to Regulation (EC) No 1981/2006

Regulation (EC) No 1981/2006 is amended as follows:

the assessment of the precision and trueness of the method provided by the applicant.

in all other cases, in point 1(B) of Annex I to Regulation (EC) No 641/2004;

in the case of genetically modified plants for food or feed uses, food or feed containing or consisting of genetically modified plants and food produced from or containing ingredients produced from genetically modified plants or feed produced from genetically modified plants, in point 3.1.C.4 of Annex III to Commission Implementing Regulation (EU) No 503/2013 (14) ,

the assessment through a ring trial involving national reference laboratories of the method performance criteria set by the applicant as compliant with the document entitled “Definition of minimum performance requirements for analytical methods of GMO testing” referred to:

That amount shall be multiplied by the number of GMO events to be fully validated.’.

food produced from or containing ingredients produced from genetically modified plants or feed produced from such plants; or

‘2. The CRL shall request the applicant to pay an additional contribution of EUR 60 000 where a full validation procedure of a method of detection and identification for a single GMO event according to the requirements laid down in the following provisions is required:

in Article 3(2), the first and second subparagraphs are replaced by the following:

Article 12

Review

1. The Commission shall monitor the application of this Regulation, the developments in scientific knowledge on replacement, reduction and refinement of animal use in scientific procedures and the publication of new guidance from EFSA. The Commission shall in particular monitor the outcome of the research project called GRACE (GMO Risk Assessment and Communication of Evidence) under the 2012 work programme of the seventh Framework Programme for Research (FP7).

2. The Commission shall review the requirement to perform 90-day feeding studies in rodents with whole genetically modified food/feed (point 1.4.4.1 of Annex II) on the basis of new scientific information. The results of this review shall be published by 30 June 2016 at the latest.

Article 13

Entry into force

This Regulation shall enter into force on the twentieth day following that of its publication in the Official Journal of the European Union.

This Regulation shall be binding in its entirety and directly applicable in all Member States.

(1) OJ L 268, 18.10.2003, p. 1.

(2) OJ L 31, 1.2.2002, p. 1.

(3) OJ L 102, 7.4.2004, p. 14.

(4) OJ L 106, 17.4.2001, p. 1.

(5) Codex Alimentarius Commission, GL 45-2003.

(6) EFSA Journal 2011; 9(5):2150.

(7) OJ L 276, 20.10.2010, p. 33.

(8) OJ L 50, 20.2.2004, p. 44.

(9) OJ L 201, 31.7.2002, p. 48.

(10) OJ L 287, 5.11.2003, p. 1.

(11) OJ L 10, 14.1.2004, p. 5.

(12) OJ L 368, 23.12.2006, p. 99.

(13) OJ L 157, 8.6.2013, p. 1.’;

(14) OJ L 157, 8.6.2013, p. 1.’;

Results of the release with respect to any risk to human health and the environment

14.2. History of previous releases of the genetically modified plant carried out outside the Union by the same notifier

Results of the release with respect to any risk to human health and the environment, submitted to the competent authority in accordance with Article 10 of Directive 2001/18/EC

14.1. History of previous releases of the genetically modified plant notified under Part B of Directive 2001/18/EC or under Part B of Council Directive 90/220/EEC (3) by the same notifier

11.2. Potential changes in the interactions of the genetically modified plant with the biotic environment resulting from the genetic modification

Description of the experimental design (number of locations, growing seasons, geographical spread, replicates and number of commercial varieties in each location) and of the statistical analysis.

3.2.6. Any change to the ability of the genetically modified plant to transfer genetic material to other organisms (for environmental safety aspects)

3.2.5. Information (for environmental safety aspects) on how the genetically modified plant differs from the recipient plant in:

Information on developmental expression of the insert during the life cycle of the plant

In the case of modifications other than insertion or deletion, describe function of the modified genetic material before and after the modification, as well as direct changes in expression of genes as a result of the modification

3.2.1. Description of the trait(s) and characteristics which have been introduced or modified

Source of donor nucleic acid(s) used for transformation, size and intended function of each constituent fragment of the region intended for insertion

2.9. Other potential interactions, relevant to the genetically modified plant, of the plant with organisms in the ecosystem where it is usually grown, or used elsewhere, including information on toxic effects on humans, animals and other organisms (for environmental safety aspects)

2.8. In the case of plant species not normally grown in the Union description of the natural habitat of the plant, including information on natural predators, parasites, competitors and symbionts (for environmental safety aspects)

1.9. Measures suggested by the applicant to take in the case of unintended release or misuse of the product as well as measures for its disposal and treatment

In the case of products other than food and feed containing or consisting of genetically modified plants, a proposal for labelling which complies with the requirements of point A(8) of Annex IV to Directive 2001/18/EC must be included.

Any proposed labelling requirements in addition to those required by other applicable EU legislation than Regulation (EC) No 1829/2003 and when necessary a proposal for specific labelling in accordance with Article 13(2) and (3), Article 25(2)(c) and (d) and Article 25(3) of Regulation (EC) No 1829/2003

If applicable, geographical areas within the Union to which the product is intended to be confined under the terms of the authorisation applied for

Any specific instructions and recommendations for use, storage and handling, including mandatory restrictions proposed as a condition of the authorisation applied for

Types of products planned to be placed on the market according to the authorisation applied for and any specific form in which the product must not be placed on the market (such as seeds, cut-flowers, vegetative parts) as a proposed condition of the authorisation applied for

Name of the recipient or parental plant and the intended function of the genetic modification

In that case, specify the third country, the date of application and, where available, a copy of the risk assessment conclusions, the date of the authorisation and the scope of the application

1.7. Has the product been subject to an application and/or authorised in a third country either previously or simultaneously to this application?

1.6. Has the genetically modified plant or derived products been previously notified for marketing in the Union under Part C of Directive 2001/18/EC?

(in that case, provide risk analysis data on the basis of the elements of Part B of Directive 2001/18/EC)

1.4. Is the product or the uses of the associated plant protection product(s) already authorised or subject to another authorisation procedure within the Union?

Products other than food and feed containing or consisting of genetically modified plants with the exception of cultivation

Name and address of the representative of the applicant established in the Union (if the applicant is not established in the Union)

The summary shall not contain parts considered to be confidential in accordance with Article 30 of Regulation (EC) No 1829/2003.

Depending on the scope of the application, some of the requested information may not be applicable.

This Part specifies the standardised form, which the summary of the application dossier must follow.

The information required in the notification as set out in Annex III to Directive 2001/18/EC shall be provided where it is not covered by the requirements of other parts of the application.

ADDITIONAL INFORMATION TO BE PROVIDED FOR GENETICALLY MODIFIED PLANTS AND/OR FOOD OR FEED CONTAINING OR CONSISTING OF GENETICALLY MODIFIED PLANTS

The applicant shall follow the instructions for the preparation and the sending of the samples provided by the EU Reference laboratory (EURL) as referred to in Article 32 of Regulation (EC) No 1829/2003. These instructions are published on the following webpage: http://gmo-crl.jrc.ec.europa.eu/guidancedocs.htm

The application shall include information as to the place where the reference material can be accessed.

The application shall include a copy of the completed form for the submission of those samples to the EURL and proof of sending to the EURL.

The applicant shall provide methods for detection, sampling and identification, as well as samples of the food or feed and their controls samples to the European Union Reference Laboratory (EURL) as referred to in Article 32 of Regulation (EC) No 1829/2003.

when appropriate a proposal for labelling complying with the requirements of point A(8) of Annex IV to Directive 2001/18/EC.

either a reasoned statement that the food or feed does not give rise to ethical or religious concerns or a proposal for labelling in all official languages of the Union as required by Articles 5(3)(g) and 17(3)(g) of Regulation (EC) No 1829/2003;

a proposal for labelling in all official languages of the Union, where a proposal for specific labelling is required in accordance with Articles 5(3)(f) and 17(3)(f) of Regulation (EC) No 1829/2003;

Suggested methods for the safe handling, storage, transport and use, including packaging, labelling, documentation, disposal and contingency procedures, where appropriate.

Taxonomic status, common name, point of collection or acquisition, and characteristics of the donor organism or organisms related to biosafety;

Centres of origin and centres of genetic diversity, if known, of the recipient organism and/or the parental organisms and a description of the habitats where the organisms may persist or proliferate;

Taxonomic status, common name, point of collection or acquisition, and characteristics of recipient organism or parental organisms related to biosafety;

Description of the gene modification, the technique used, and the resulting characteristics of the GMO;

The name and contact details of the authority responsible for the decision;

The name and contact details of the applicant for a decision for domestic use;

The provided information shall contain as a minimum the information specified in Annex II to Regulation (EC) No 1946/2003 of the European Parliament and of the Council (2) :

The application shall provide the information required under Articles 5(3)(c) and 17(3)(c) of Regulation (EC) No 1829/2003 for the purpose of complying with Annex II to the Cartagena Protocol on Biosafety to the Convention on Biological Diversity.

Additional information which might influence the evaluation of the safety of the genetically modified food or feed generated following the submission of the application, as well as any information regarding any prohibition or restriction imposed by a competent authority of any third country on the basis of a safety assessment shall be provided by the applicant.

Where the information obtained from those studies is not coherent with the information obtained from the studies performed in accordance with the requirements set out in Annex II, the applicant shall provide a thorough analysis of the respective studies and provide plausible explanations for the observed discrepancies.

A systematic review of studies published in the scientific literature and studies performed by the applicant within the period of 10 years prior to the date of submission of the dossier on the potential effects on human and animal health of the genetically modified food and feed covered by the application shall be included in the application. This systematic review shall be carried out by taking into account the guidance of EFSA on application of systematic review methodology to food and feed safety assessments to support decision making (1) .

Other animal studies to examine the safety and the characteristics of genetically modified food and feed

Any change to the ability of the genetically modified plant to transfer genetic material to other organisms, namely:

Information on how the genetically modified plant differs from the recipient plant in reproduction, dissemination, survivability or other properties.

General description of the trait(s) and characteristics which have been introduced or modified

Source of donor nucleic acid(s) used for transformation, size and intended function of each constituent fragment of the region intended for insertion

other potential interactions of the genetically modified plant with organisms in the ecosystem where it is usually grown, or used elsewhere, including information on toxic effects on humans, animals and other organisms.

where a plant species is not grown in the Union, a description of the natural habitat of the plant, including information on natural predators, parasites, competitors and symbionts;

ways and extent of dissemination (to include, for example, an estimation of how viable pollen and/or seed declines with distance);

Data on the past and present use of the recipient plant, such as history of safe use for consumption as food or feed, including information on how the plant is typically cultivated, transported and stored, whether special processing is required to make the plant safe to eat, and the plant normal role in the diet (for example, which part of the plant is used as a food source, whether its consumption is important in particular subgroups of the population, what important macro- or micro-nutrients it contributes to the diet);

Information on the recipient or parental plants relevant to their safety, including any known toxicity or allergenicity;

All the requirements of Part II shall be provided in the application except where such requirements are not justified by the scope of the application (for example, where the application is limited to food or feed produced from GMOs).

Additional authorisation requirements provided for in Union law, related to the placing on the market of the food or feed or applicable ‘maximum residue level’ (MRL) where the food or feed is likely to contain residues of plant protection products.

Where applicable, the status of the food or feed or of related substances under other provisions of Union law.

Where appropriate, the conditions for the placing on the market of the genetically modified food(s) or feed(s), including specific conditions for use and handling.

This description would include, for example, a detailed description of specific methods of production of food or feed which would be due to the nature of the genetic modification or which would lead to food or feed with specific characteristics.

Products other than food and feed containing or consisting of genetically modified plants with the exception of cultivation

Name, qualification and experience of the responsible scientist(s) and contact details of the responsible person for all dealings with the European Food Safety Authority (EFSA).

ANNEX II

SCIENTIFIC REQUIREMENTS FOR THE RISK ASSESSMENT OF GENETICALLY MODIFIED FOOD AND FEED

I. INTRODUCTION

1. DEFINITIONS

For the purpose of this Annex, the following definitions shall apply:

1. ‘hazard identification’ means the identification of biological, chemical and physical agents capable of causing adverse health effects and which may be present in a particular food and feed or group of foods and feeds;

2. ‘hazard characterisation’ means the qualitative and/or quantitative evaluation of the nature of the adverse health effects associated with biological, chemical and physical agents which may be present in food and feed;

3. ‘risk characterisation’ means the qualitative and/or quantitative estimation, including attendant uncertainties, of the probability of occurrence and severity of known or potential adverse health effects in a given population based on hazard identification, hazard characterisation and exposure assessment.

2. SPECIFIC CONSIDERATIONS

2.1. Insertion of marker genes and other nucleic acid(s) sequences not essential to achieve the desired trait

In order to facilitate the risk assessment, the applicant shall endeavour to minimise the presence of inserted nucleic acid(s) sequences not essential to achieve the desired trait.

During the process of genetic modification of plants and other organisms, marker genes are often used to facilitate the selection and identification of genetically modified cells, containing the gene of interest inserted into the genome of the host organism, among the vast majority of untransformed cells. The applicant shall carefully select such marker genes and Article 4(2) of Directive 2001/18/EC shall be respected. Against that background, the applicant shall therefore aim to develop GMOs without the use of antibiotic resistance marker genes.

2.2. Risk assessment of genetically modified food and feed containing stacked transformation events

For the risk assessment of genetically modified food and feed containing stacked transformation events obtained by conventional crossing of genetically modified plants containing one or several transformation event(s), the applicant shall provide a risk assessment of each single transformation event or, in accordance with Article 3(6) of this Regulation, refer to already submitted application(s). The risk assessment of genetically modified food and feed containing stacked transformation events shall also include an assessment of the following aspects:

(a) stability of the transformation events;

(b) expression of the transformation events;

(c) potential synergistic or antagonistic effects resulting from the combination of the transformation events shall be subject to an assessment in accordance with Sections 1.4 (Toxicology), 1.5 (Allergenicity) and 1.6 (Nutritional assessment).

For genetically modified food and feed containing, consisting of or produced from genetically modified plants, whose cultivation is associated with the production of genetically modified material containing various subcombinations of transformation events (segregating crops), the application shall include all subcombinations independently of their origin which have not yet been authorised. In such a case, the applicant shall provide a scientific rationale justifying that there is no need to provide experimental data for the concerned subcombinations or, in the absence of such scientific rationale, provide the experimental data.

For genetically modified food and feed containing, consisting of or produced from genetically modified plants, the cultivation of which does not lead to the production of genetically modified material containing various combinations of transformation events (non-segregating crops), the application shall only cover the combination which is to be placed on the market.

The rules set out in this Section apply mutatis mutandis to transformation events which are combined by other means such as co- and retransformation.

II. SCIENTIFIC REQUIREMENTS

1. HAZARD IDENTIFICATION AND CHARACTERISATION

1.1. Information relating to the recipient or (where appropriate) parental plants

1.1.1. The applicant shall provide comprehensive information relating to the recipient or (where appropriate) the parental plants in order to: (a) evaluate all issues of potential concern, such as the presence of natural toxins or allergens; (b) identify the need for specific analyses.

1.1.2. For the purposes referred to in point 1.1.1, the applicant shall provide the following information: (a) Complete name: (i) family name; (ii) genus; (iii) species; (iv) subspecies; (v) cultivar/breeding line or strain; (vi) common name. (b) Geographical distribution and cultivation of the plant, including its distribution within the Union. (c) Information on the recipient or parental plants relevant to their safety, including any known toxicity or allergenicity. (d) Data on the past and present use of the recipient plant. This information shall include the history of safe use for consumption as food or feed, information on how the plant is typically cultivated, transported and stored, whether special processing is required to make the plant safe to eat, and describe the normal role of the plant in the diet (such as which part of the plant is used as a food or feed source, whether its consumption is important in particular subgroups of the population, what important macro- or micro-nutrients it contributes to the diet).

1.2. Molecular Characterisation

1.2.1. Information relating to the genetic modification

The applicant shall provide sufficient information on the genetic modification:

(a) to identify the nucleic acid(s) intended for transformation and related vector sequences potentially delivered to the recipient plant;

(b) to characterise the nucleic acid(s) actually inserted in the plant.

1.2.1.1. Description of the methods used for the genetic modification

The applicant shall provide information on the following elements:

(a) the method of genetic transformation including relevant references;

(b) the recipient plant material;

(c) the species and the strain of Agrobacterium and other microbes, if used during the genetic transformation process;

(d) the helper plasmids, if used during the genetic transformation process;

(e) the source of carrier nucleic acid(s), if used during the genetic transformation process.

1.2.1.2. Nature and source of vector used

The applicant shall provide the following information:

(a) a physical map of the functional elements and other plasmid/vector components together with the relevant information needed for the interpretation of the molecular analyses (such as restriction sites, the position of primers used in polymerase chain reaction (PCR), the location of probes used in Southern analysis). The region intended for insertion shall be clearly indicated;

(b) a table identifying each component of the plasmid/vector (including the region intended for insertion), its size, its origin and its intended function.

1.2.1.3. Source of nucleic acid(s) used for transformation, size and intended function of each constituent fragment of the region intended for insertion

The applicant shall provide information on the donor organism(s) and on the nucleic acid(s) sequence(s) intended to be inserted in order to determine whether the nature of the donor organism(s) or the nucleic acid sequence(s) may trigger any safety issue.

Information regarding the function of the nucleic acid region(s) intended for insertion shall comprise the following elements:

(a) the complete sequence of the nucleic acid(s) intended to be inserted, including information on any deliberate alteration(s) to the corresponding sequence(s) in the donor organism(s);

(b) the history of safe use of the gene product(s) arising from the regions intended for insertion;

(c) data on the possible relationship of the gene products with known toxins, anti-nutrients and allergens.

Information regarding each donor organism shall comprise:

— taxonomic classification;

— history of use regarding food and feed safety.

1.2.2. Information relating to the genetically modified plant

1.2.2.1. General description of the trait(s) and characteristics which have been introduced or modified

Information provided under this point may be limited to a general description of the introduced trait(s) and the resulting changes to the phenotype and metabolism of the plant.

For example, when the introduced trait is herbicide tolerance, the applicant shall provide information on the mode of action of the active substance and its metabolism in the plant.

1.2.2.2. Information on the sequences actually inserted/deleted

The applicant shall provide the following information:

(a) the size and copy number of all detectable inserts, both complete and partial; this is typically determined by Southern analysis. Probe/restriction enzyme combinations used for this purpose shall provide complete coverage of sequences that could be inserted into the genetically modified plant, such as any parts of the plasmid/vector or any carrier or foreign nucleic acid(s) remaining in the genetically modified plant. The Southern analysis shall span the entire transgenic locus(i) as well as flanking sequences and include all appropriate controls. For the determination of copy number of the insert, complementary methods may also be used (such as real-time PCR);

(b) the organisation and sequence of the inserted genetic material at each insertion site in a standardised electronic format, with the aim of identifying changes in the inserted sequences compared to the sequence intended for insertion;

(c) in the case of deletion(s), size and function of the deleted region(s), whenever possible;

(d) subcellular location(s) of insert(s) (nucleus, chloroplasts, mitochondria, or maintained in a non-integrated form) and methods for its/their determination;

(e) sequence information in a standardised electronic format for both 5’ and 3’ flanking regions at each insertion site, with the aim of identifying interruptions of known genes. Bioinformatic analyses shall be conducted using up-to-date databases with the aim of performing both intraspecies and interspecies similarity searches. In the case of genetically modified plants containing stacked transformation events, the safety of potential interactions between any unintended modifications at each insertion site shall be assessed;

(f) Open Reading Frames (hereafter referred to as ‘ORFs’ and defined as any nucleotide sequence that contains a string of codons that is uninterrupted by the presence of a stop codon in the same reading frame) created as a result of the genetic modification either at the junction sites with genomic DNA or due to internal rearrangements of the insert(s). The ORFs shall be analysed between stop codons, not limiting their lengths. Bioinformatic analyses shall be conducted to investigate possible similarities with known toxins or allergens using up-to-date databases. The characteristics and versions of the databases shall be provided. Depending on the information gathered, further analyses (such as transcription analysis) may be needed to complete the risk assessment.

1.2.2.3. Information on the expression of the insert(s)

The applicant shall provide information:

— to demonstrate whether the inserted/modified sequence results in intended changes at the protein, RNA and/or metabolite levels;

— to characterise the potential unintended expression of new ORFs identified under point 1.2.2.2(f) as raising a safety concern.

For those purposes, the applicant shall provide the following information:

(a) The method(s) used for expression analysis together with their performance characteristics;

(b) Information on developmental expression of the insert during the life cycle of the plant. The requirement for information on developmental expression shall be considered on a case-by-case basis taking into account the promoter used, the intended effect(s) of the modification and scope of the application;

(c) Parts of the plant where the insert/modified sequences are expressed;

(d) Potential unintended expression of new ORFs identified under point 1.2.2.2(f) which raise a safety concern;

(e) Protein expression data, including the raw data, obtained from field trials and related to the conditions in which the crop is grown. Data on expression levels from those parts of the plant used for food and feed purposes shall be provided in all cases. In addition, information shall also be provided on the expression of target genes in other parts of the plant when tissue-specific promoters have been used and when this is relevant for the safety assessment. The minimum requirement for protein expression shall be data provided from three growing sites or from one site over three seasons. Permutations of the sites and seasons shall be acceptable provided that the minimum requirement is met. When justified by the nature of the insert (such as silencing approaches or where biochemical pathways have been intentionally modified), specific RNA(s) or metabolite(s) shall be analysed. For silencing approaches by RNAi expression, potential ‘off target’ genes should be searched by in silico analysis to assess if the genetic modification could affect the expression of other genes which raise safety concerns;

(f) With regard to the stacking of transformation events by conventional crossing, expression data shall be provided to assess the potential interactions between the events which may raise any additional safety concerns over protein and trait expression compared with the single transformation events. The comparison shall be carried out with data obtained from plants grown in the same field trials. On a case-by-case basis, and where concerns arise, additional information may be necessary.

1.2.2.4. Genetic stability of the insert and phenotypic stability of the genetically modified plant

The applicant shall provide information:

(a) to demonstrate the genetic stability of the transgenic locus(i) and the phenotypic stability and inheritance pattern(s) of the introduced trait(s);

(b) in case of stacked transformation events, to establish that each of the transformation events stacked in the plant has the same molecular properties and characteristics as in the plants with the single transformation events.

For the purposes of that information, applicants shall provide data which demonstrate stability over multiple (normally five) generations or vegetative cycles for plant containing single transformation events. Data from the first and the last generations of vegetative cycles are sufficient. The source of the material used for the analysis shall be specified. Data shall be analysed using appropriate statistical methods.

For stacked transformation events comparisons between the original transformation events and the stacked transformation events shall be carried out using plant materials representative of those designed for commercial production. The applicant shall provide adequate justification for the plant materials used. Comparisons shall include comparisons of sequences of the inserts and the flanking regions obtained from GM plants containing single events and plants containing stacked transformation events.

To assess genetic stability of the transformation event(s), applicants shall use appropriate molecular approaches referred to in Section 1.2.2.2.

1.2.2.5. Potential risk associated with horizontal gene transfer

The applicant shall assess the probability of horizontal gene transfer from the product to humans, animals and micro-organisms and any potential associated risk when intact and functional nucleic acid(s) remains in the genetically modified food and feed.

1.2.3. Conclusions of the molecular characterisation

The molecular characterisation shall provide data on the structure and expression of the insert(s), and on the stability of the intended trait(s). This shall also apply to situations where transformation events have been stacked by conventional breeding.

It shall be specifically indicated whether the molecular characterisation of the genetic modification(s) raises safety concerns with regard to the interruption of endogenous genes or regulatory sequences.

The molecular characterisation shall also aim to identify whether the genetic modification(s) raise(s) any issues regarding the potential for producing proteins/substances other than those intended and in particular new toxins or allergens.

The potential unintended changes identified in this Section shall be addressed in the relevant complementary part(s) of the safety assessment.

1.3. Comparative analysis

The comparative analysis of composition and agronomic as well as phenotypic characteristics shall constitute, together with the molecular characterisation, the starting point to structure and conduct the risk assessment of a new genetically modified food and feed.

It shall aim at identifying similarities and differences:

(a) in composition, agronomic performance and phenotypic characteristics (intended and unintended alterations) between the genetically modified plant and its conventional counterpart;

(b) in composition between the genetically modified food and feed and its conventional counterpart.

Where no appropriate conventional counterpart can be identified, a comparative safety assessment cannot be made and consequently a safety and nutritional assessment of the genetically modified food or feed shall be carried out as for novel foods falling within the scope of Regulation (EC) No 258/97 of the European Parliament and of the Council (1) that do not have conventional counterparts (such as where the genetically modified food or feed is not closely related to a food or feed with a history of safe use or where a specific trait or specific traits are introduced with the intention of bringing complex changes in the composition of the genetically modified food or feed).

1.3.1. Choice of the conventional counterpart and additional comparators

In the case of vegetatively propagated crops, the conventional counterpart shall, in principle, be the near-isogenic variety used to generate the transgenic line.

In the case of crops that reproduce sexually, the conventional counterpart shall have a genetic background comparable to the genetically modified plant. When the GM plant has been developed using back-crossing, a conventional counterpart with a genetic background that is as close as possible to the genetically modified plant shall be selected.

In addition, the applicant may include a comparator having a closer genetic background to the genetically modified plant than the conventional counterpart (such as a negative segregant).

In the case of herbicide tolerant genetically modified plants and in order to assess whether the expected agricultural practices influence the expression of the studied endpoints, three test materials shall be compared: the genetically modified plant exposed to the intended herbicide; the conventional counterpart treated with conventional herbicide management regimes; and the genetically modified plant treated with the same conventional herbicide management regimes.

In the case of stacked transformation events, it is not always possible to use a conventional counterpart with a genetic background as close to the genetically modified plant as with conventional counterpart normally used for single transformation events. In such circumstances, the applicant shall provide reasoned justification on the choice of the conventional counterpart and assess its limitations for the risk assessment. In addition, single parental genetically modified lines or genetically modified lines containing a subcombination of the stacked transformation events for which an application has been submitted or negative segregants derived from these genetically modified lines may also be included as additional comparators. The applicant shall provide detailed information justifying the choice of additional comparators.

In all cases, the applicant shall provide information on the breeding scheme (pedigree) in relation to the genetically modified plant, the conventional counterpart and, where appropriate, additional comparators together with an adequate justification of their selection. The history of safe use of the conventional counterpart shall be adequately supported by both qualitative and quantitative data.

More detailed guidance for the application of the requirements of this Section is available in the EFSA scientific opinion ‘Guidance on selection of comparators for the risk assessment of genetically modified plants and derived food and feed’ (2).

1.3.2. Experimental design and statistical analysis of data from field trials for comparative analysis

1.3.2.1. Description of the protocols for the experimental design

(a) Principles of experimental design

Field trials used for the production of material for the comparative analysis shall be performed in order to determine whether the genetically modified plant and/or the genetically modified food and feed is different from its conventional counterpart and/or equivalent to non-genetically modified reference varieties with a history of safe use.

For each endpoint, the comparative analysis shall involve the following two approaches:

(i) a test of difference, to verify whether the genetically modified plant is different from its conventional counterpart and might therefore be considered a hazard depending on the type of the identified difference, and on the magnitude and type of exposure;

(ii) a test of equivalence to verify whether the genetically modified plant is equivalent or not to non-genetically modified reference varieties, apart from the introduced trait(s).

In testing for difference, the null hypothesis shall be that there is no difference between the GMO and its conventional counterpart against the alternative hypothesis that a difference exists.

Where additional comparator(s) are used for the risk assessment, a test of difference shall be carried out between the genetically modified plant and each of the additional comparator(s) following the requirements set out in Section 1.3.2.2 for the test of difference between the genetically modified plant and its conventional counterpart.

In testing for equivalence, the null hypothesis shall be that the difference between the GMO and the set of reference varieties is at least as great as a specified minimum size (see Section 1.3.2.2) against the alternative hypothesis that there is no difference or a smaller difference than the specified minimum between the GMO and the set of reference varieties.

Rejection of the null hypothesis shall be required in order to conclude that the GMO and the set of reference varieties are unambiguously equivalent for the endpoint considered. The equivalence limits used for the test of equivalence shall represent appropriately the range of natural variation expected for reference varieties with a history of safe use.

(b) Specific protocols for experimental design

Natural variation may have several sources: variation within a variety arises due to environmental factors and variation between varieties arises due to a combination of both genetic and environmental factors. In order to identify and estimate differences attributable only to genotypes, it is essential to control environmental variability. Therefore, non-genetically modified reference varieties shall be included in the experimental design of the field trials and in sufficient numbers to ensure an adequate estimate of the variability required to set the equivalence limits. All test materials consisting of genetically modified plants, conventional counterpart, reference varieties and, where appropriate, additional comparator(s) shall be randomised to plots within a single field at each site, usually in a completely randomised or randomised block experimental design. The different sites selected for the field trials shall reflect the different meteorological and agronomic conditions under which the crop is to be grown; the choice shall be explicitly justified. The choice of non-genetically modified reference varieties shall be appropriate for the chosen sites and shall be justified explicitly. In the case the sites cover a restricted range of growing conditions, the applicant shall replicate the field trials over more than one year.

Within each site the test materials consisting of genetically modified plants, conventional counterpart and, where appropriate, additional comparator(s) shall be identical for all replicates. In addition, unless there is explicit justification for not doing so, at each site there shall be at least three appropriate non-genetically modified reference varieties of the crop that have a known history of safe use, which shall also be identical between replicates. The replication at each site is the number of results obtained for each test material; the replication should never be less than four at any site. However, if only two appropriate reference varieties are available at a particular site, then the replication shall be six at that site; if only one is available then the replication shall be eight.

Each field trial shall be replicated at a minimum of eight sites, chosen to be representative of the range of likely receiving environments where the plant is to be grown. The field trials may be conducted in a single year, or spread over multiple years. The non-genetically modified reference varieties may vary between sites and at least six different reference varieties shall be used over the entire set of field trials.

When the genetically modified plant is tested together with other genetically modified plants of the same crop species (such as Zea mays) the production of material for the comparative assessment of these different genetically modified plant may be produced simultaneously at the same site and within the same field trial by the placing of the different genetically modified plants and their appropriate comparator(s) in the same randomised block. This shall be subject to the following two strict conditions:

(i) the conventional counterpart and, where appropriate, additional comparator(s) shall always occur together with the genetically modified plant in the same block;

(ii) all the different genetically modified plants and their comparator(s) and all the non-genetically modified reference varieties used to test equivalence with those genetically modified plants shall be fully randomised within each block.

If the number of plots per block required for such a field trial were to exceed 16, then a partially balanced incomplete block design may be used, to reduce the number of plots per block, by excluding some of the genetically modified plants and their appropriate comparator(s) from each block. This shall be subject to the following two strict conditions:

(i) the conventional counterpart shall always occur together with its particular genetically modified plant in the same block;

(ii) all of the non-genetically modified reference varieties shall appear in each of the incomplete blocks and be fully randomised with the plants and their comparator(s).

The field trials shall be adequately described, giving information on important parameters such as management of the field before sowing, date of sowing, soil type, herbicide use, climatic and other cultivation/environmental conditions during growth and time of harvest, as well as the conditions during storage of the harvested material.

More detailed guidance for the application of the requirements of this Section is available in the EFSA opinion on ‘Statistical considerations for the safety evaluation of GMOs’ (3).

1.3.2.2. Statistical analysis

Analysis of data shall be presented in a clear format, using standardised scientific units. The raw data and the programming code used for the statistical analysis shall be given in an editable form.

Data transformation may be necessary to ensure normality and to provide an appropriate scale on which statistical effects are additive. For many endpoint response variables, a logarithmic transformation is expected to be appropriate. In such cases, any difference between the genetically modified material and any other test material shall be interpreted as a ratio on the natural scale. However, when logarithmic transformation does not provide appropriate results, the natural scale or another scale shall be considered.

The total variability of each endpoint observed in the field trials shall be estimated and partitioned using appropriate statistical models in order to derive two sets of confidence limits and to set a lower and upper equivalence limit based on the variability observed among the reference varieties. One set of confidence limits shall be used in the test of difference; the other set and the equivalence limits shall be used in the test of equivalence.

A linear mixed statistical model shall be used for calculation of the confidence limits for both tests (that is to say, the difference and equivalence tests); a slightly different model shall be used to estimate the equivalence limits to be used in the equivalence test.

Denote by I an indicator variable (uncentered in the mixed model) such that I = 1 for a field plot having any of the non-GM reference varieties, and I = 0 otherwise. Then the random factors for model 1 should be, but not necessarily be restricted to, those representing the variation: (i) between the test materials (a set that includes the GM plant, its conventional counterpart, each of the non-GM reference varieties and any additional comparators); (ii) in the interaction between the test materials and I; (iii) between sites; and (iv) between blocks within sites. Model 2 should be identical to model 1 except that the random factor representing the interaction between the test materials and I is omitted.

The fixed factor for both models should have as many levels as there are test materials and represent the contrasts between the means of the test materials. The test materials are as defined above: the GM plant; its conventional counterpart; the set of non-GM reference varieties; and any additional test materials. The set of non-GM reference varieties is considered as a single level of the fixed factor. For the difference test, the component of the fixed factor of interest is the single degree-of-freedom contrast between the GM plant and its conventional counterpart. For the equivalence test, the component of the fixed factor of interest is the single degree-of-freedom contrast between the GM plant and the set of non-GM reference varieties.

Both the difference test and the equivalence test shall be implemented using the correspondence between hypothesis testing and the construction of confidence limits. In the case of equivalence testing, the approach used shall follow the two one-sided tests (TOST) methodology by rejecting the null hypothesis of non-equivalence when the both confidence limits fall between the equivalence limits. The choice of 90 % confidence limits corresponds to the customary 95 % level for statistical testing of equivalence.

The results of the difference and equivalence tests shall be represented visually for all the endpoints simultaneously, on a single graph or a few graphs.

The graph(s) shall show the line of zero difference between the genetically modified material and its conventional counterpart, and for each endpoint: the lower and upper adjusted equivalence limits; the mean difference between the genetically modified material and its conventional counterpart; and the confidence limits for this difference (see the set of possible outcomes for a single endpoint in the graph in Figure 1).

When, in addition to the conventional counterpart, another test material is used as comparator, the mean difference between the genetically modified material and that comparator, its confidence limits and its adjusted equivalence limits shall be displayed on the graph(s), for all such additional comparators, by referring this to the same zero baseline as defined by the conventional counterpart. The line of zero difference on the logarithmic scale corresponds to a multiplicative factor of unity on the natural scale. The horizontal axis shall be labelled with values that specify the change on the natural scale. In the case of logarithmic transformation, changes of 2x and ½x will appear equally spaced on either side of the line of zero difference.

Despite the expected proportion of spurious significant differences, the applicant shall report and discuss all significant differences observed between the genetically modified crop, its conventional counterpart and, where applicable, any other test material, focusing on their biological relevance (see Section 3 on Risk Characterisation).

For reporting, full details shall be given for each endpoint analysed, listing:

(a) the assumptions underlying the analysis;

(b) full specification of the mixed models chosen, including fixed and random effects;

(c) results of any test of interaction between the test materials and sites;

(d) fixed effects, together with the appropriate estimated residual variation with which they are compared, and variance components for the random factors;

(e) estimated degrees of freedom;

(f) any other relevant statistics.

A discussion on the likely impact of other growing conditions not tested in the field trial shall be provided.

Figure 1: simplified version of a graph for comparative assessment showing the seven outcome types possible for each single endpoint. After adjustment of the equivalence limits, a single confidence limit (for the difference) serves visually for assessing the outcome of both tests (difference and equivalence). Here, only the upper adjusted equivalence limit is considered. Shown are: the mean of the genetically modified crop on an appropriate scale (square); the confidence limits (whiskers) for the difference between the genetically modified crop and its conventional counterpart (bar shows confidence interval); a vertical line indicating no difference (for test of difference); and vertical lines indicating adjusted equivalence limits (for test of equivalence). For outcome types 1, 3 and 5, the null hypothesis of no difference cannot be rejected: for outcomes 2, 4, 6 and 7, the genetically modified crop is different from its conventional counterpart. Regarding interpretation of equivalence, four categories (i) to (iv) are identified: in category (i), the null hypothesis of non-equivalence is rejected in favour of equivalence; in categories (ii), (iii) and (iv), non-equivalence cannot be rejected.

A. Regarding test of difference, each outcome from the graph shall be categorised as follows and the respective appropriate conclusion shall be drawn. (i) Outcome types 1, 3 and 5: the confidence interval bar overlaps with the line of no-difference. The null hypothesis of no difference cannot be rejected and the appropriate conclusion is that there is insufficient evidence that the genetically modified crop and its conventional counterpart differ. (ii) Outcome types 2, 4, 6 and 7: the confidence interval bar does not overlap with the line of no-difference. The null hypothesis of no difference must be rejected and the appropriate conclusion is that the genetically modified crop is significantly different from its conventional counterpart.

B. Regarding test of equivalence, each outcome from the graph shall be categorised as follows, and the respective appropriate conclusion shall be drawn. (i) Outcome types 1 and 2 (category (i), Figure 1): both confidence limits lie between the adjusted equivalence limits and the null hypothesis of non-equivalence is rejected. The appropriate conclusion is that the genetically modified crop is equivalent to the set of non-genetically modified reference varieties. (ii) Outcome types 3 and 4 (category (ii), Figure 1): the mean of the genetically modified crop lies between the adjusted equivalence limits, but the confidence interval bar overlaps at least one of the adjusted equivalence limits on the graph. Non-equivalence cannot be rejected and the appropriate conclusion is that equivalence between the genetically modified crop and the set of non-genetically modified reference varieties is more likely to be the case than lack of equivalence. (iii) Outcome types 5 and 6 (category (iii), Figure 1): the mean of the genetically modified crop lies outside the adjusted equivalence limits, but the confidence interval bar overlaps with at least one of the adjusted equivalence limits. Non-equivalence cannot be rejected and the appropriate conclusion is that lack of equivalence between the genetically modified crop and the set of non-genetically modified reference varieties is more likely to be the case than equivalence. (iv) Outcome type 7 (category (iv), Figure 1): both confidence limits lie outside the adjusted equivalence limits. The appropriate conclusion is that there is lack of equivalence between the genetically modified crop and the set of non-genetically modified reference varieties. In the case of significant difference and/or lack of equivalence for any particular endpoint, further statistical analysis shall be carried out to assess whether there are interactions between any of the test materials and site, possibly using a simple standard ANOVA approach. Whatever approach is adopted, details shall be given, for each endpoint analysed, listing: (a) the assumptions underlying the analysis, and, when appropriate: (b) degrees of freedom; (c) the estimated residual variation for each source of variation, and variance components; (d) any other relevant statistics. These additional analyses are intended to aid the interpretation of any significant differences found and to study potential interactions between test materials and other factors. More detailed guidance for the application of the requirements of this Section is available in the EFSA opinion on ‘Statistical considerations for the safety evaluation of GMOs’ (4).

1.3.3. Selection of material and compounds for analysis

Analysis of the composition of plant material is crucial when comparing the genetically modified food and feed with its conventional counterpart. The material to be used for the comparative assessment shall be selected while taking into account the uses of the genetically modified plant and the nature of the genetic modification. In the case of herbicide tolerant genetically modified plants, three test materials shall be used: the genetically modified plant exposed to the intended herbicide; the conventional counterpart treated with conventional herbicide management regimes; and the genetically modified plant treated with the same conventional herbicide management regimes. Unless duly justified, analysis shall be carried out on the raw agricultural commodity, as this usually represents the main point of entry of the material into the food and feed production and processing chain. Additional analysis of processed products (such as food and feed, food ingredients, feed materials, food and feed additives or food flavourings) shall be conducted, where appropriate and on a case-by-case basis (see also Section 1.3.6). The sampling, analysis and preparation of the tested material shall be carried out according to appropriate quality standards.

1.3.4. Comparative analysis of composition

In addition to the analysis on the level of the newly expressed proteins (see Section 1.2.2.3), the compositional analysis shall be carried out on an appropriate range of compounds. In each case, the applicant shall provide at least analysis on proximates (including moisture and total ash), key macro- and micro-nutrients, anti-nutritional compounds, natural toxins, and already identified allergens, as well as other secondary plant metabolites characteristic for specific crop plant species, as referred to in the Organisation for Economic Cooperation and Development (OECD) consensus documents on compositional considerations for new plant varieties (OECD consensus documents) (5). The vitamins and minerals selected for analysis shall be those which are present at levels which are nutritionally significant and/or which make nutritionally significant contributions to the diet at the levels at which the plant is consumed. The specific analyses required shall depend on the plant species examined, but shall include a detailed assessment appropriate to the intended effect of the genetic modification, the considered nutritional value and use of the plant. The applicant shall pay particular attention to key nutrients such as proteins, carbohydrates, lipids/fats, fibre, vitamins and minerals. For example, a fatty acid profile shall be included for oil-rich plants (main individual saturated, mono-unsaturated and poly-unsaturated fatty acids) and an amino acid profile (individual protein amino acids and main non-protein amino acids) for plants used as an important protein source. Analysis of plant cell wall components are also required for the vegetative parts of plants used for feed purposes.

The applicant shall also provide an analysis on key toxins inherently present in the recipient plant which may adversely affect human/animal health depending on their toxic potency and levels. The concentrations of such compounds shall be assessed according to plant species and the proposed use of the food and feed product. Similarly, anti-nutritional compounds, such as digestive enzyme inhibitors, and already identified allergens shall be studied.

The characteristics of the introduced trait may trigger further analysis of specific compounds including metabolites of potentially modified metabolic pathways. The applicant shall consider when appropriate, the inclusion of compounds other than the key nutrients, key toxins, anti-nutrients and allergens identified by the OECD consensus documents and justify the selection of these compounds.

1.3.5. Comparative analysis of agronomic and phenotypic characteristics

The applicant shall provide a comparison between the genetically modified plant and its conventional counterpart. This comparison shall enable the applicant to identify unintended effects resulting from the genetic modification and shall address also plant biology and agronomic traits, including common breeding parameters (such as yield, plant morphology, flowering time, day degrees to maturity, duration of pollen viability, response to plant pathogens and insect pests, sensitivity to abiotic stress). The protocols of these field trials shall follow the specifications set out in Section 1.3.2.

Where transformation events are stacked by conventional crossing, there may also be changes to agronomic and phenotypic characteristics. Possible differences in phenotypic characteristics and agronomic properties of stacked transformation events shall be assessed in field trials. Where appropriate, the applicant shall provide additional information on agronomic traits of the stacked transformation events from additional field trials.

1.3.6. Effects of processing

The applicant shall assess whether or not the processing and/or preserving technologies applied are likely to modify the characteristics of genetically modified end products compared with their respective conventional counterpart. The applicant shall provide a description of the different processing technologies in sufficient detail, paying special attention to the steps which may lead to significant changes in the product content, quality or purity.

Genetic modification can target metabolic pathways resulting in changes in the concentration of non-protein substances or in new metabolites (such as in nutritionally enhanced foods). Processed products may be assessed together with the assessment of the genetically modified plant for the safety of the genetic modification, or a processed product may be assessed separately. The applicant shall provide the scientific rationale for the risk assessment of these products. On a case-by-case basis, the submission of additional experimental data shall be considered by the applicant.

When appropriate, depending on the product, information shall be necessary on the composition, level of undesirable substances, nutritional value and metabolism, as well as on the intended use.

When appropriate, depending on the nature of the newly expressed protein(s), it shall be necessary to assess the extent to which the processing steps lead to the concentration or to the elimination, denaturation and/or degradation of these protein(s) in the final product.

1.3.7. Conclusion

The conclusion of the comparative analysis shall clearly state:

(a) whether agronomic and phenotypic characteristics of the genetically modified plant are, except for the introduced trait(s), different to the characteristics of its conventional counterpart and/or equivalent to the reference varieties, taking into account natural variation;

(b) whether compositional characteristics of the genetically modified food and feed are, taking into account natural variation, different to the characteristics of its conventional counterpart and/or equivalent to the reference varieties, except for the introduced trait(s);

(c) characteristics for which the genetically modified plant or the genetically modified food and feed are different to the characteristics of its conventional counterpart and/or not equivalent to the reference varieties taking into account natural variation, which need further investigation;

(d) whether, in the case of transformation events stacked by conventional crossing, there are indications of interactions between the combined transformation events.

1.4. Toxicology

The toxicological impact of any changes on the whole genetically modified food/feed resulting from the genetic modification such as the introduction of new genes, gene silencing or over-expression of an endogenous gene shall be assessed.

Toxicological assessment shall be performed in order to:

(a) demonstrate that the intended effect(s) of the genetic modification has no adverse effects on human and animal health;

(b) demonstrate that unintended effect(s) of the genetic modification(s) identified or assumed to have occurred based on the preceding comparative molecular, compositional or phenotypic analyses, have no adverse effects on human and animal health;

(c) identify potential adverse effects of new constituents and determine their highest dose level that does not result in adverse effects. From data obtained from an appropriate animal study, an acceptable daily intake (ADI) of single compounds by humans may be derived by using uncertainty or safety factors that take into account differences between test animal species and humans, and inter-individual variations among humans;

(d) identify potential adverse effects on the whole genetically modified food/feed or address remaining uncertainties through the performance of 90-day feeding studies.

The applicant shall consider the nature of toxicological testing to be conducted on new constituents and whole genetically modified food/feed based on the outcome of the molecular and comparative analysis referred to in Sections 1.2 and 1.3, namely the differences identified between the genetically modified product and its conventional counterpart, including intended as well as unintended changes. The applicant shall also assess the results of the performed toxicological tests to consider the need to conduct additional testing on new constituents or on whole genetically modified food/feed as set out in Sections 1.4.4.2 and 1.4.4.3.

The applicant shall take into account the presence of: newly expressed proteins, the potential presence of other new constituents and/or possible changes in the level of natural constituents beyond normal variation. The specific information requirements and testing strategies are set out in Sections 1.4.1 to 1.4.4.

As regards applications of which the scope include or is restricted to genetically modified food and feed produced from genetically modified plants, toxicological studies with the processed products shall be provided except if the applicant provides a risk assessment of the genetically modified plant (or relevant parts of it) demonstrating its safety and there are no indications that the processed genetically modified food and feed would be any different from their respective conventional counterpart. The applicant shall provide adequate justification in this regard.

Toxicology studies designed to evaluate risks to human and/or animal health shall complement each other. Most studies required for the assessment of the safety of the genetically modified food are also valid for the assessment of genetically modified feed.

Besides the exposure of consumers and animals through intake of food and feed, any adverse effect(s) on individuals that could be due to their exposure to genetically modified food and feed material as part of their professional activities, for example, farming, seed processing, shall be reported by the applicant. Appropriate studies shall be performed to further characterise these indications of potential adverse effects.

The applicant shall use internationally agreed protocols and test methods for toxicity testing (see Tables 1 and 2 of Section 1.7). Adaptations of these protocols or use of any methods that differ from such protocols shall be justified in the application.

1.4.1. Testing of newly expressed proteins

The applicant shall provide an evaluation of all newly expressed proteins. The studies required to investigate the potential toxicity of a newly expressed protein shall be selected on a case-by-case basis, depending on the knowledge available with respect to the protein’s source, function or activity and history of human or animal consumption. As regards proteins expressed in the genetically modified plant, in the case where the history of safe use for consumption as food and/or feed of both the plant and the newly expressed proteins is duly documented, specific toxicity testing as provided for in this Section shall not be required. In such case, the applicant shall provide the necessary information regarding the history of safe use of the proteins.

Where specific testing is required, the tested protein shall be equivalent to the newly expressed protein as it is expressed in the genetically modified plant. If, due to the lack of sufficient amount of test materials from the plant, a protein produced by micro-organisms is used, the structural, biochemical and functional equivalence of this microbial substitute to the newly expressed plant protein shall be demonstrated. In particular, comparisons of the molecular weight, amino acid sequence, post-translational modification, immunological reactivity and, in the case of enzymes, the enzymatic activity are needed to provide evidence for the equivalence. In case of differences between the plant expressed protein and its microbial substitute, the significance of these differences for the safety studies shall be evaluated.

To demonstrate the safety of newly expressed proteins, the applicant shall provide:

(a) A molecular and biochemical characterisation of the newly expressed protein, including determination of the primary structure, molecular weight (for example using mass spectrometry), studies on post-translational modifications and a description of its function. In the case of newly expressed enzymes, information on the enzyme activities including the temperature and pH range for optimum activity, substrate specificity, and possible reaction products shall also be provided. The potential interaction with other plant constituents shall also be evaluated.

(b) An up-to-date search for homology to proteins known to cause adverse effects, such as toxic proteins. A search for homology to proteins exerting a normal metabolic or structural function may also contribute valuable information. The database(s) and the methodology used to carry out the search shall be specified.

(c) A description of the stability of the protein under relevant processing and storage conditions and the expected treatment of the food and feed. The influences of temperature and pH changes shall be examined and potential modification(s) of the proteins (such as denaturation) and/or production of stable protein fragments generated through such treatments shall be characterised.

(d) Data concerning the resistance of the newly expressed protein to proteolytic enzymes (such as pepsin), such as by in vitro investigations using appropriate and standardised tests. Stable breakdown products shall be characterised and evaluated with regard to the potential to cause adverse health effects linked to their biological activity.

(e) A repeated-dose 28-day oral toxicity study with the newly expressed protein in rodents. When appropriate depending on the outcome of the 28-day toxicity study, further targeted investigations shall be provided, including an analysis of immunotoxicity.

Acute toxicity testing of the newly expressed proteins of genetically modified plants is of little additional value for the risk assessment of the repeated human and animal consumption of genetically modified food and feed and shall not be provided as part of the studies performed under this point.

The applicant shall perform studies with combined administration of proteins when the genetic modification results in the expression of two or more proteins in the genetically modified plant and when, based on scientific knowledge, a possibility of synergistic or antagonistic interactions of safety concerns is identified.

1.4.2. Testing of new constituents other than proteins

The applicant shall provide a risk assessment of identified new constituents other than proteins. This shall include, on a case-by-case basis, an evaluation of their toxic potency and of the need of toxicological testing as well as a determination of their concentration in the genetically modified food and feed. To establish the safety of new constituents having no history of safe use for consumption in food and feed, the applicant shall provide information analogous to that described in the ‘Guidance for submissions for food additive evaluations by the EFSA Panel on Food Additives and Nutrient Sources added to Food’ of 16 August 2012 (6) and Commission Regulation (EC) No 429/2008 of 25 April 2008 on detailed rules for the implementation of Regulation (EC) No 1831/2003 as regards the preparation and the presentation of applications and the assessment and the authorisation of feed additives (7). This shall include the submission of information on a core set of studies such as on metabolism/toxicokinetics, subchronic toxicity, genotoxicity, chronic toxicity, carcinogenicity and reproduction and developmental toxicity, accompanied by any other appropriate type of study. For specific guidelines for animal tests, see Table 1 of Section 1.7 of this Annex. Genotoxicity test protocols are provided for in Table 2 of Section 1.7 of this Annex.

1.4.3. Information on altered levels of food and feed constituents

This Section shall apply only in the case where the intended or unintended effect of the genetic modification would result in an alteration of the levels of food and feed constituents beyond the natural variation.

To demonstrate the safety of the altered levels of food and feed constituents such as macro- and micro-nutrients, anti-nutrients, and natural toxins as well as other secondary plant metabolites, the applicant shall submit a detailed risk assessment based on the knowledge of the physiological function and/or toxic properties of these constituents.

The result of that risk assessment shall determine if, and to what extent, the applicant shall provide additional toxicological tests to the 90-day feeding study in rodents with whole genetically modified food/feed on selected food and feed constituents.

1.4.4. Testing of the whole genetically modified food and feed

The applicant shall primarily base its risk assessment of the genetically modified food and feed on molecular characterisation, comparative agronomic, phenotypic and comprehensive compositional analysis, and the toxicological evaluation of the identified intended and unintended effects, including a 90-day feeding study in rodents with whole genetically modified food/feed as specified in Section 1.4.4.1. Under the circumstances set out in points 1.4.4.2 and 1.4.4.3 of this Section, additional specific toxicological studies with the whole genetically modified food and feed shall be carried out.

1.4.4.1. 90-day feeding study in rodents with whole genetically modified food/feed

The applicant shall include a 90-day feeding study with whole food and feed in rodents for the assessment of food and feed containing, consisting of or produced from genetically modified plants with a single transformation event or with stacked transformation events which are not obtained by conventional crossing of genetically modified plants containing a single transformation event.

In the case of stacked transformation events obtained by conventional crossing of genetically modified plants containing one or several transformation event(s), a 90-day feeding study with whole food and feed in rodents shall be included for each of the genetically modified plant with a single transformation event which was used. An additional 90-day feeding study with whole food and feed in rodents with the genetically modified plant with the stacked transformation events shall be included where indications of potential adverse effects are identified during the assessment of: (i) the stability of the inserts; (ii) the expression of the inserts; and (iii) the potential synergistic or antagonistic effects resulting from the combination of the transformation events.

The design of the toxicity study with genetically modified food and feed should be performed according to the ‘subchronic oral toxicity test repeated-dose 90-day oral toxicity study in rodents’ (see Table 1) following an adapted protocol. In principle a minimum of two test doses and a negative control shall be used. The highest dose shall be the maximum achievable without causing nutritional imbalance; the lowest dose shall contain the tested food and/or feed in an amount always above the anticipated human/target animal intake level. The genetically modified food and feed analysed should be relevant to the product to be consumed. In the case of herbicide tolerant genetically modified plants, the tested material should come from the genetically modified plant exposed to the intended herbicide. Whenever possible, information on natural variation of test parameters shall be derived from historical background data rather than from the inclusion of reference varieties, consisting of commercially available food and feed derived from non-GM plants with a history of safe use, in the experiments. The statistical analysis shall focus on the detection of possible differences between the test material and its control. A power analysis to estimate a sample size capable of detecting a pre-specified biologically relevant effect size with a specified power and significance level should be used. More detailed guidance for performing this study is provided in the EFSA Guidance on conducting repeated-dose 90-day or