Application EFSA-GMO-NL-2005-13 (LLCotton25) · PDF fileis not completely integrated in LLCotton25 as 23 bp are ... the data performed with a 20% bioequivalence range was ... A randomised

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Application EFSA-GMO-NL-2005-13 (LLCotton25) ANNEX G Comments and opinions submitted by Member States during the three-month consultation period

Member State

Organisation Reference Comments EFSA GMO Panel Response

Comments from National Competent Authorities (NCA) under Directive 2001/18/EC

DK Danish Forest and Nature Agency

General comments The quantitative PCR-detection methods should be validated by the Joint Research Centre of the European Commission before placing on the market.

Outside the remit of the GMO Panel

FR Ministère de l'agriculture et de la pêche/DGAL

General comments L'évaluation des risques pour l'environement conduite par la Commission du génie biomoléculaire (avis du 16 novembre 2005) n'a pas mis en évidence de risques pour l'environnement liés à la mise sur le marché du coton LLCotton25, telle que décrite dans le dossier EFSA/GMO/NL/2005/13.

The GMO Panel agrees with the conclusions of the Commission du génie biomoléculaire.

FI Board for Gene Technology

General comments The comment of the Board for Gene Technology: Before marketing in certain areas the consent holder should ensure that a monitoring plan which takes into account the specific conditions of that area and the extent of the release is put into place.

See section 5.2.2 of the scientific opinion: (…). The scope of the monitoring plan provided by the applicant is in line with the intended uses for the GMO. Since the environmental risk assessment did not cover cultivation and identified no potential adverse environmental effects, no case-specific monitoring is necessary. The GMO Panel is aware that, due to the physical characteristics of cotton seeds and methods of transportation, accidental spillage is unavoidable. Therefore the GMO Panel recommends that specific measures are introduced to actively monitor the occurrence of feral cotton plants in areas where seed spillage is likely to occur as proposed in the EFSA Guidance document (EFSA, 2006a) and the opinion of the GMO Panel on post-market environmental monitoring (EFSA, 2006b). In other respects the GMO Panel is of the opinion that the general approaches and measures of the monitoring plan proposed by the applicant are in line with the EFSA opinion on post-market environmental monitoring (EFSA, 2006b) as well as with the intended uses of LLCotton25. Since the environmental risk assessment does not cover cultivation and identifies no potential adverse environmental effects, no case-specific monitoring is necessary.

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DE BVL A, 07 Where appropriate, the conditions for placing on the market the food(s) or…

The accompanying shipping documents should indicate that cotton seed “LLcotton” has not been approved for cultivation. Appropriate measures have to be taken during transport, storage and processing to avoid unintended release to the environment.

Outside the remit of the GMO Panel.

DE BVL D, 02 Information on the sequences actually inserted or deleted

Page 28, Statement about BglII-digestion: There is no >14kb fragment in lane 4 of the Southern blot experiment visible as stated by the authors (Fig.5, p.29). Instead, two (not three) bands are detected, one at the expected size at ~1.5 kb and one at ~12 kb. The latter might be a double band as the hybridization strength is in fact stronger than claimed in the description. The notifier should be asked to comment on this.

See section 2.2.2 of the scientific opinion: The nucleotide sequence of the insert introduced into LLCotton25 has been determined in its entirety. The DNA sequence of the LLCotton25 insert has been proven to be identical to the corresponding transforming plasmid pGSV71 sequences. PCR analysis of the terminal repeats of the vector plasmid confirmed that the right border (RB) terminal repeat is not completely integrated in LLCotton25 as 23 bp are missing. The left border (LB) terminal repeat sequence displays a deletion of 4 bp. The sequences of the plant genome adjacent to the 3’ and 5’ sequences of the insert were determined using TAIL-PCR. Comparison of the flanking sequences to the respective wild type target site revealed that upon integration of the T-DNA into the genomic DNA a 38 bp fragment of genomic DNA at the target site was deleted. There was no indication that the insert is integrated in a coding region or that the insert disrupts gene regulatory sequences. These data presented proof that the insert has been integrated in a single chromosome at a single locus as intended. The sequence information demonstrates the presence of two BgI II sites in the insert. Therefore it is possible that the >10 kb hybridisation signals is composed of two bands (Fig 5, page 29). This has been referenced in two different ways in the dossier either as >14 kb or >10 kb band.

DE BVL D, 04 Information on how the GM plant differs from the recipient plant in: …

Comments by the Federal Agency for Nature Conservation: With regard to a final assessment further information is required (see comments to D, 07.04).

See response to comments submitted under D, 07.04

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DE BVL D, 07.03 Selection of compounds for analysis

The notifier defines non-bioequivalency as follows: “In cases where the average value of the parameter in the GMO differs more than 20% of the average value of the parameter in the traditional comparator, further explanation and evaluation must be made” (Oberdörfer, 2003, p.16). In its opinion on the application C/BE/96/01 the GMO Panel stated, that this method is not internationally agreed on and, therefore, should not be used as a general principle: ”The statistical analysis of the data performed with a 20% bioequivalence range was provided by the applicant. In the absence of an international agreement on the use of this methodology, the GMO Panel does not agree with this approach to be applied as a general principle.” (EFSA Journal (2005) 281, p.12). Therefore the applicant is asked to justify the use of the 20% range for bioequivalence. Furthermore, the evaluation of the analysis results by the applicant appears not to be in agreement with the recommendation by the EFSA GMO panel as given in the EFSA guidance document. According to this document, the results should not be presented as pooled data only (as done by the applicant), but field trial data should be presented and be statistically analysed separately by location and growing season. (The EFSA Journal 2004-99, p. 24).

See section 7.2(b) statistical models for analysis, confidence intervals of the GMO Panel Guidance document (EFSA, 2006a): Experimental design should be rigorous and analysis of data should be presented in a clear format, using standardised scientific units. Field trial data should be presented separately, as well as pooled, and should be analysed statistically, using appropriate statistical tools. A randomised complete block design, for example, could indicate whether the experimental factors (location, year, climatic conditions, plant variety) interact with one another. The confidence intervals used for statistical analysis should be specified (normally 95%, with possible adjustment according to the hazard of the constituent to be compared). On 26 January 2006, the GMO Panel required from the applicant further clarifications with respect to the statistical analysis of the compositional data including statistical data analyses according to traditional methodology. The applicant responded on 18 May 2006 and the GMO Panel was satisfied with the response.

DE BVL D, 07.04 Agronomic traits

The studies of Chiavegato (2003) and Penna (2003) are in Portuguese. In order to assess these studies and their conclusions we request both to be fully translated into English. Additional comments by the Federal Agency for Nature Conservation: We do not share the applicant's opinion that the performed agronomic trials can prove the absence of pleiotropic effects. The studies by Freyssinet (2002d, 2002e), Kowite et al. (2001, 2002), Chiavegato (2003), Penna (2003) and Oberdörfer (2003) do not contain information about climate conditions during field trials or any statistical analysis for environmental interactions. Therefore the data give only insufficient evidence about gene-environment

The scope of the application is for import, processing as well as for food and feed uses and does not include cultivation. Therefore, there was no requirement for scientific information on possible environmental effects associated with the cultivation.

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interactions, unintended or pleiotropic effects. The detailed description of the test design and the climatic conditions of the Study "Comparison of the overwintering capacity of LLCotton25 and Cocker312 seed" are missing. Therefore, it is not possible to evaluate these results (Table 25, page 78).

DE BVL D, 07.08 Toxicology Comments by the Federal Agency for Nature Conservation: Testing of the whole GM food/feed is crucial to obtain the necessary information about any adverse unintended effects of LLcotton25 on human or animal health. One study with the GMO (Stafford 2004) has been carried out. The diets contained only 10% cotton meal. It must be stressed that chicken broiler studies are designed to test for effects on animal nutrition and are no model for toxicology. Therefore no valid toxicological study with the LLcotton25 was carried out. The applicant should be requested to test for unintended acute and chronic toxic effects of the LLcotton25 by carrying out a 90 day rat study. In addition we advise to carry out supplemental studies with especially ruminants which differ with respect to their digestive system and will be substantially exposed by feed derived from processing residues of LLcotton25.

See section 4.2.4 of the scientific opinion: The comparative compositional analysis and agronomic analyses showed that LLCotton25 is substantially equivalent to its non-transgenic counterpart Coker312 and other commercially grown cotton varieties except for the introduced trait. In addition, the analyses provided no indication for unintended effects of the genetic modification and therefore the GMO Panel concluded that no additional safety studies with laboratory animals are needed. The GMO Panel considers the feeding study on broilers as a study assessing the nutritional adequacy of the feed.

DE BVL D, 10 Potential changes in the interactions of the GM plant with the biotic…

Comments by the Federal Agency for Nature Conservation: Although the deliberate release into the environment of LLCotton25 is not within the scope of the application LLCotton25 might enter the environment at the processing facilities where sealed cargo containers are opened. Therefore, interactions of LLCotton25 with the biotic environment have to be considered.

The scope of the application is for import, processing as ell as for food and feed uses and does not include cultivation. Therefore, there was no requirement for scientific information on possible environmental effects associated with the cultivation. See section 5.2.2 of the scientific opinion: The GMO Panel is aware that, due to the physical characteristics of cotton seeds and methods of transportation, accidental spillage is unavoidable. Therefore the GMO Panel recommends that specific measures are introduced to actively monitor the occurrence of feral cotton plants in areas where seed spillage is likely to occur as proposed in the EFSA

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Guidance document (EFSA, 2006a) and the opinion of the GMO Panel on post-market environmental monitoring (EFSA, 2006b). In other respects the GMO Panel is of the opinion that the general approaches and measures of the monitoring plan proposed by the applicant are in line with the EFSA opinion on post-market environmental monitoring (EFSA, 2006b) as well as with the intended uses of LLCotton25. Since the environmental risk assessment does not cover cultivation and identifies no potential adverse environmental effects, no case-specific monitoring is necessary. See section 5.3 of the scientific opinion: LLCotton25 is being assessed for import, processing and food/feed uses and thus there is no requirement for scientific information on environmental effects associated with cultivation. The GMO Panel considered the environmental issues raised by Member States in the above sections of Chapter 5 and concludes as follows: Gossypium hirsutum L., which has no wild relatives in Europe, is a cultivated plant in Europe since the 19th century and occurs only occasionally as feral plants in Europe.

If accidental spillage and subsequent release into the environment of LLCotton25 seeds occurs, LLCotton25 plants will only be fitter in the presence of glufosinate-containing herbicides which are not currently used on cultivated cotton or in most areas where the GM cotton might be spilled. Therefore the GMO Panel is of the opinion that the likelihood of the establishment and spread of LLCotton25 is very low and that unintended environmental effects due to this GM cotton will be no different from that of conventional cotton varieties. Furthermore the scope of the monitoring plan provided by the applicant is in line with the intended uses of LLCotton25 since this does not include cultivation.

DE BVL D, 12 Environmental Comments by the Federal Agency for Nature

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Monitoring Plan Conservation: We do not share the applicant’s point of view towards the environmental monitoring plan. The following issues have to be considered: (1) Gossypium herbaceum is locally naturalized on disturbed ground (e.g. arable land and waste land) in various Southern European countries and G. hirsutum is perhaps locally naturalized in Southern Europe (Tutin et al. 1992, Polunin 1997). The term "naturalized" means that feral cotton does live outside cultivation and does not depend on repeated input of seeds from cultivated cotton plants. (2) It cannot be excluded that viable seed of the notified cotton is spilled at the processing facilities and might survive. (3) Gene flow through outcrossing from feral LLCotton25 into non-transgenic feral cotton or cultivated non-transgenic cotton may occur in cotton growing regions of Southern Europe. Since no precise information on the amount of spilled material and frequency of spillage events was given, spillage of LLCotton25 cannot be excluded. Thus, a case-specific monitoring plan is required to monitor the occurrence of spillage of LLCotton25 and of potential further effects on the environment at the places of spillage. Reference: Polunin, O. 1997: Flowers of Greece and the Balkans. Oxford University Press. Tutin, T. G., Heywood, V. H. et al. (eds.) (1992): Flora Europaea. Vol. 2., 5th reprint, Cambridge University Press, Cambridge: p. 469.

The GMO Panel comments on the scientific quality of the monitoring plan. EFSA has published guidance and opinion on PMEM (EFSA, 2006a,b) following a broad consultation with stakeholders, including national competent authorities. The information supplied by the applicant is in line with this guidance. See section 5.2.2 of the scientific opinion: (…). The scope of the monitoring plan provided by the applicant is in line with the intended uses for the GMO. Since the environmental risk assessment did not cover cultivation and identified no potential adverse environmental effects, no case-specific monitoring is necessary. The GMO Panel is aware that, due to the physical characteristics of cotton seeds and methods of transportation, accidental spillage is unavoidable. Therefore the GMO Panel recommends that specific measures are introduced to actively monitor the occurrence of feral cotton plants in areas where seed spillage is likely to occur as proposed in the EFSA Guidance document (EFSA, 2006a) and the opinion of the GMO Panel on post-market environmental monitoring (EFSA, 2006b). In other respects the GMO Panel is of the opinion that the general approaches and measures of the monitoring plan proposed by the applicant are in line with the EFSA opinion on post-market environmental monitoring (EFSA, 2006b) as well as with the intended uses of LLCotton25. Since the environmental risk assessment does not cover cultivation and identifies no potential adverse environmental effects, no case-specific monitoring is necessary. See section 5.2 of the PMEM opinion (EFSA, 2006b): Details of the specific plans and methods of monitoring in each country should not be included in the original application. The GMO Panel advises that the application should describe the general approaches and methods that the applicant would apply in different commercialisation sites,

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including the type of dialogue that would be established with risk managers in each Member State. (…) Thus detailed local arrangements will be developed by the applicant after the application has been accepted (…).

DE BVL D, 12.02 Case-specific GM plant monitoring

Comments by the Federal Agency for Nature Conservation: The case-specific monitoring plan has to be elaborated depending on the final conclusions drawn from an updated environmental risk assessment. The objective of the case-specific monitoring must be to monitor (1) any losses of LLCotton25, (2) the establishment of feral plants of the notified cotton on disturbed ground around the processing facilities, (3) gene flow through outcrossing from feral LLCotton25 into non-transgenic feral cotton or other cultivated non-transgenic cotton in the vicinity. The issues (2)–(3) of the case-specific monitoring plan need to be conducted only in case of substantial losses of LLCotton25. The case-specific monitoring plan has to meet the following requirements: – A fully specified list of monitoring parameters has to be defined. For each parameter a detailed statement of the parameter definition, of the observation methods (collection and analysis of samples with references), of the frequencies of observations (time and number of visits to collect data) and of the locations (habitat type, number, size and spatial arrangement of plots) have to be presented. – An operating schedule giving full details of points in time, places and monitoring parameters to be observed should be requested from the applicant. – Representativeness and local adaptation of the monitoring is required. The concept of sampling needs to be elaborated. Particularly, it must be explained how the necessary representativeness of the collected data in space and time is ascertained. – It must be stated, how the condition of the environment before the placing on the market of the notified cotton is described (determination of the baseline status of the receiving environment as a point of

The scope of the application is for import, processing as well as for food and feed uses and does not include cultivation. Therefore, there was no requirement for scientific information on possible environmental effects associated with the cultivation. See section 5.2.2 of the scientific opinion: (…). The scope of the monitoring plan provided by the applicant is in line with the intended uses for the GMO. Since the environmental risk assessment did not cover cultivation and identified no potential adverse environmental effects, no case-specific monitoring is necessary. The GMO Panel is aware that, due to the physical characteristics of cotton seeds and methods of transportation, accidental spillage is unavoidable. Therefore the GMO Panel recommends that specific measures are introduced to actively monitor the occurrence of feral cotton plants in areas where seed spillage is likely to occur as proposed in the EFSA Guidance document (EFSA, 2006a) and the opinion of the GMO Panel on post-market environmental monitoring (EFSA, 2006b). In other respects the GMO Panel is of the opinion that the general approaches and measures of the monitoring plan proposed by the applicant are in line with the EFSA opinion on post-market environmental monitoring (EFSA, 2006b) as well as with the intended uses of LLCotton25. Since the environmental risk assessment does not cover cultivation and identifies no potential adverse environmental effects, no case-specific monitoring is necessary. See section 5.2 of the PMEM opinion (EFSA, 2006b): Details of the specific plans and methods of monitoring in each country should not be included in the original

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reference). – The methods of data analysis including the statistical methods have to be elaborated in full detail. – Use of external people and existing networks: The monitoring expertise of external people involved in the monitoring activities and detailed information about participating networks (e.g. name, EU country, responsible authority, availability, scope and composition) have to be specified. In case that monitoring data are collected by external persons or institutions other than the applicant binding agreements/contracts with third parties are requested which clearly determine what data are provided and how these data are made available. – The submission of complete reports on the fully detailed monitoring results on an annual basis. All raw data have to be appended to the reports. – It has to be stated how the results of the monitoring will be published. – All monitoring activities need to focus primarily on the locations where sealed cargo containers are opened. In case of substantial losses and spread of LLCotton25, all receiving environments need to be monitored.

application. The GMO Panel advises that the application should describe the general approaches and methods that the applicant would apply in different commercialisation sites, including the type of dialogue that would be established with risk managers in each Member State. (…) Thus detailed local arrangements will be developed by the applicant after the application has been accepted (…).

DE BVL D, 12.03 General Surveillance of the impact of the GM plant

Comments by the Federal Agency for Nature Conservation: The general surveillance plan proposed by the applicant is not clearly defined and does not meet the requirements in Annex VII of Directive 2001/18/EC and in the supplementing guidance notes (Council Decision 2002/811/EC). For the general surveillance of unanticipated adverse effects of LLCotton25 it is an essential prerequisite to document the spread, persistence and accumulation of the inserted genetic sequences as well as of the corresponding proteins in organisms and environmental media (air, soil, water bodies). Therefore, an appropriate general surveillance plan is required (1) to observe the spread, persistence and accumulation of the inserted genetic sequences and the corresponding proteins in organisms and environmental media (air, soil, water bodies) and (2) to keep adverse long-term effects

The scope of the application is for import, processing as ell as for food and feed uses and does not include cultivation. Therefore, there was no requirement for scientific information on possible environmental effects associated with the cultivation. See section 5.2.2 of the scientific opinion: The GMO Panel is aware that, due to the physical characteristics of cotton seeds and methods of transportation, accidental spillage is unavoidable. Therefore the GMO Panel recommends that specific measures are introduced to actively monitor the occurrence of feral cotton plants in areas where seed spillage is likely to occur as proposed in the EFSA Guidance document (EFSA, 2006a) and the opinion of the GMO Panel on post-market environmental monitoring (EFSA, 2006b).

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on the diversity of organisms, food chains and habitats affected by the notified cotton under surveillance. The time-period of monitoring needs to be sufficient to detect delayed adverse effects of the notified cotton and its use. Therefore, it may be necessary to extend the monitoring of certain parameters beyond the period of the consent. The general surveillance plan has to meet the following requirements: – A fully specified list of monitoring parameters has to be defined. For each parameter a detailed statement of the parameter definition, of the observation methods (collection and analysis of samples with references), of the frequencies of observations (time and number of visits to collect data) and of the locations (habitat type, number, size and spatial arrangement of plots) have to be presented. – An operating schedule giving full details of points in time, places and monitoring parameters to be observed should be requested from the applicant. – Representativeness and local adaptation of the monitoring is required. The concept of sampling needs to be elaborated. Particularly, it must be explained how the necessary representativeness of the collected data in space and time is ascertained. – It must be stated, how the condition of the environment before the placing on the market of the notified cotton is described (determination of the baseline status of the receiving environment as a point of reference). – The methods of data analysis including the statistical methods have to be elaborated in full detail. – Use of external people and existing networks: The monitoring expertise of external people involved in the monitoring activities and detailed information about participating networks (e.g. name, EU country, responsible authority, availability, scope and composition) have to be specified. In case that monitoring data are collected by external persons or institutions other than the applicant binding agreements/contracts with third parties are requested

In other respects the GMO Panel is of the opinion that the general approaches and measures of the monitoring plan proposed by the applicant are in line with the EFSA opinion on post-market environmental monitoring (EFSA, 2006b) as well as with the intended uses of LLCotton25. Since the environmental risk assessment does not cover cultivation and identifies no potential adverse environmental effects, no case-specific monitoring is necessary. See section 5.2 of the PMEM opinion (EFSA, 2006b): Details of the specific plans and methods of monitoring in each country should not be included in the original application. The GMO Panel advises that the application should describe the general approaches and methods that the applicant would apply in different commercialisation sites, including the type of dialogue that would be established with risk managers in each Member State. (…) Thus detailed local arrangements will be developed by the applicant after the application has been accepted (…). See section 11.4.2 of the GMO Panel Guidance Document (EFSA, 2006a): Knowing the limitations of existing monitoring systems, it is important for the applicant to describe the processes and criteria that will be used for selecting and evaluating existing monitoring systems for supplying data related to the unanticipated adverse effects of GM plants in the general surveillance.

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which clearly determine what data are provided and how these data are made available. – The submission of complete reports on the fully detailed monitoring results (case-specific monitoring and general surveillance) on an annual basis. All raw data have to be appended to the reports. – It has to be stated how the results of the monitoring will be published. – All monitoring activities need to focus primarily on the locations where sealed cargo containers are opened. In case of substantial losses and spread of LLCotton25, all receiving environments need to be monitored.

DE BVL General comments The German CA is of the opinion that further information is required to conclude on the risk assessment of dossier EFSA/GMO/UK/2005/13. Diverging and additional comments from cooperating authorities (Federal Agency for Nature Conservation) are included for information and indicated.

See section ‘Conclusions and recommendations’ of the scientific opinion

ES Ministry of the Environment

D, 12 Environmental Monitoring Plan

Monitoring Plan The consent holder should provide further details of the arrangements of the monitoring plan, in particular for general surveillance, indicating which existing network programs could be used, the type of information that should be collected and a more detailed monitoring methodology in order to have a monitoring plan which could be implemented in a harmonised manner among the importer Member States.

The GMO Panel comments on the scientific quality of the monitoring plan. EFSA has published guidance and opinion on PMEM (EFSA, 2006a,b) following a broad consultation with stakeholders, including national competent authorities. The information supplied by the applicant is in line with this guidance. See section 5.2.2 of the scientific opinion: The GMO Panel is aware that, due to the physical characteristics of cotton seeds and methods of transportation, accidental spillage is unavoidable. Therefore the GMO Panel recommends that specific measures are introduced to actively monitor the occurrence of feral cotton plants in areas where seed spillage is likely to occur as proposed in the EFSA Guidance document (EFSA, 2006a) and the opinion of the GMO Panel on post-market environmental monitoring (EFSA, 2006b). In other respects the GMO Panel is of the opinion that the general approaches and measures of the monitoring plan

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proposed by the applicant are in line with the EFSA opinion on post-market environmental monitoring (EFSA, 2006b) as well as with the intended uses of LLCotton25. Since the environmental risk assessment does not cover cultivation and identifies no potential adverse environmental effects, no case-specific monitoring is necessary. See section 5.2 of the PMEM opinion (EFSA, 2006b): Details of the specific plans and methods of monitoring in each country should not be included in the original application. The GMO Panel advises that the application should describe the general approaches and methods that the applicant would apply in different commercialisation sites, including the type of dialogue that would be established with risk managers in each Member State. (…) Thus detailed local arrangements will be developed by the applicant after the application has been accepted (…).

MT Nature Protection Unit Environment Protection Directorate

General comments Since the classification and labelling of glufosinate ammonium is being reviewed under Directive 67/548/EEC, and there is the possibility that it is classified as a Toxic for Reproduction Cat 2 R61 and Cat 3 R 62, its use as a plant protection product is questioned. Consequently, its use in this GM product to confer resistance is also questioned. Even though the scope of this notification excludes cultivation of cotton in EU Member States, this cotton will still be cultivated in other countries around the world, and so any authorisations of products into the EU should take into consideration any global health and safety impacts. The analytical tests were all carried out on protein that was expressed in the bacterium Escherichia coli and hence also extracted from it. Analytical tests on protein expressed and extracted from the GM cotton in question should be have been also carried out. In the toxicity tests carried out on mice the applicant used only 5 mice in each group, whereas usually a group of 10 is used. In addition, the mice were only females and these are

Human and animal health issues related to plant-protection products are regulated by Directive 91/414/EEC and fall outside the remit of the GMO Panel.

See section 4.2.3.1 of the scientific opinion: Due to the low expression level of the PAT protein in LLCotton25 most of the safety studies were conducted with a PAT protein encoded in E. coli by the bar gene (PAT/bar protein). Examination of the structure and function of these plant and bacterial PAT proteins have shown a high degree of similarity, based on their size and sequence homology, enzymatic activity, immunoreactivity and absence of glycosylation. The PAT/pat and PAT/bar proteins have been shown to be structurally and functionally equivalent (Wehrmann et al., 1996; Herouet et al., 2005). Therefore the GMO Panel accepts the PAT/bar as well as the PAT/pat test material derived from E. coli for the safety assessment of PAT protein present in LLCotton25. With regard to the number and sex of animals used, the acute as well as the 14-day repeated dose toxicity studies were

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known to eat less than males. Thus, the females have ingested less of the GM product than would have their male counterparts. It is therefore debatable whether the results can be still considered as correct. As regards the Open Reading Frames, the notifier claims that none of them are located near promoters and therefore they are considered as biologically non-functional. However, the Northern Blots should be more accurate to determine if expression is occurring.

carried out in line with the respective OECD guidelines. See section D.3.(c) of the GMO Panel Guidance document (EFSA, 2006a): The potential creation of newly expressed fusion proteins should be investigated by bioinformatic analysis and the absence of any harmful fusion proteins demonstrated. An investigation of newly expressed transcripts is appropriate when a bioinformatic analysis identifies a putative fusion protein.

See section 2.2.2 of the scientific opinion: The nucleotide sequence of the insert introduced into LLCotton25 has been determined in its entirety. The DNA sequence of the LLCotton25 insert has been proven to be identical to the corresponding transforming plasmid pGSV71 sequences. PCR analysis of the terminal repeats of the vector plasmid confirmed that the right border (RB) terminal repeat is not completely integrated in LLCotton25 as 23 bp are missing. The left border (LB) terminal repeat sequence displays a deletion of 4 bp. The sequences of the plant genome adjacent to the 3’ and 5’ sequences of the insert were determined using TAIL-PCR. Comparison of the flanking sequences to the respective wild type target site revealed that upon integration of the T-DNA into the genomic DNA a 38 bp fragment of genomic DNA at the target site was deleted. There was no indication that the insert is integrated in a coding region or that the insert disrupts gene regulatory sequences. These data presented proof that the insert has been integrated in a single chromosome at a single locus as intended.

Molecular analysis of the insert and bioinformatic analysis of flanking regions do not suggest that transcriptional analysis is required.

NO Directorate for Nature

D, 12.03 General Surveillance of the

The Notifier states that “upon approval of the LLCotton25 in the EU, Bayer CropScience will ensure

See section 5.2 of the PMEM opinion (EFSA, 2006b): Details of the specific plans and methods of monitoring in

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Management impact of the GM plant that awareness of the GM crop is made widely available through stakeholders by providing key information, and will invite these stakeholders to participate in general surveillance”. There is, however, no guarantee that the invited stakeholders wish to participate and therefore more committed agreements between the Notifier and monitoring participants need to be in place (e.g. how and where the surveillance will be carried out and the responsibilities for carrying out the surveillance) before consent is given from the EU/EEC.

each country should not be included in the original application. The GMO Panel advises that the application should describe the general approaches and methods that the applicant would apply in different commercialisation sites, including the type of dialogue that would be established with risk managers in each Member State. (…) Thus detailed local arrangements will be developed by the applicant after the application has been accepted (…).

NO Directorate for Nature Management

D, 12.05 Implementing General Surveillance

The General Surveillance Plan proposed by the Notifier seems to be based on more or less voluntary surveillance and reporting carried out by a number of operators, networks etc. We are of the opinion that some representative processing plants should be chosen for a more active surveillance for unforeseen effects. This part of the general surveillance should be organised by the Notifier.

See section 5.2 of the PMEM opinion (EFSA, 2006b): Details of the specific plans and methods of monitoring in ach country should not be included in the original application. The GMO Panel advises that the application should describe the general approaches and methods that the applicant would apply in different commercialisation sites, including the type of dialogue that would be established with risk managers in each Member State. (…) Thus detailed local arrangements will be developed by the applicant after the application has been accepted (…).


D, 12.06 Reporting the results of monitoring

As stated in our comments submitted under D, 12.05-Implementing general surveillance, we are of the opinion that some representative processing plants should be chosen for a more active surveillance for unforeseen effects. This part of the general surveillance should be organised by the Notifier, and the results should be reported whether such effects were detected or not. In addition to the broad outline given by the Notifier we request more detailed information regarding the content of the form that will be used to facilitate post-market monitoring.

See section 5.2.2 of the scientific opinion: The GMO Panel is aware that, due to the physical characteristics of cotton seeds and methods of transportation, accidental spillage is unavoidable. Therefore the GMO Panel recommends that specific measures are introduced to actively monitor the occurrence of feral cotton plants in areas where seed spillage is likely to occur as proposed in the EFSA Guidance document (EFSA, 2006a) and the opinion of the GMO Panel on post-market environmental monitoring (EFSA, 2006b). In other respects the GMO Panel is of the opinion that the general approaches and measures of the monitoring plan proposed by the applicant are in line with the EFSA opinion on post-market environmental monitoring (EFSA, 2006b) as well as with the intended uses of LLCotton25. Since the

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environmental risk assessment does not cover cultivation and identifies no potential adverse environmental effects, no case-specific monitoring is necessary. See section 5.2 of the PMEM opinion (EFSA, 2006b): Details of the specific plans and methods of monitoring in each country should not be included in the original application. The GMO Panel advises that the application should describe the general approaches and methods that the applicant would apply in different commercialisation sites, including the type of dialogue that would be established with risk managers in each Member State. (…) Thus detailed local arrangements will be developed by the applicant after the application has been accepted (…). See section 11.4.2 of the GMO Panel Guidance Document (EFSA, 2006a): Knowing the limitations of existing monitoring systems, it is important for the applicant to describe the processes and criteria that will be used for selecting and evaluating existing monitoring systems for supplying data related to the unanticipated adverse effects of GM plants in the general surveillance.


D, 10.09 Impacts of the specific cultivation, management and harvesting…

The Notifier has provided an outline on the effects of the GM-cotton on agricultural practices in the US. We would, however, need more specified information on the application of herbicides throughout the growing season, from season to season and in all the intended LLCotton25 growing countries, in order to fully assess the impact of the GM-cotton on the environment.

The scope of the application is for import, processing as well as for food and feed uses and does not include cultivation. Therefore, there was no requirement for scientific information on possible environmental effects associated with the cultivation. Furthermore human and animal health issues related to plant-protection products are regulated by Directive 91/414/EEC and fall outside the remit of the GMO Panel.


A. General information The notification lacks information on possible contributions to a sustainable development, benefits to the society and other ethical considerations regarding the use of LLCotton25. These aspects will be addressed in

Costs-benefits related issues fall outside the remit of the GMO Panel. Furthermore human and animal health issues related to plant-protection products are regulated by Directive 91/414/EEC and fall outside the remit of the GMO Panel.

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the evaluation of the notification in Norway under the Norwegian Gene Technology Act. Amongst others, more information needs to be provided on how the GM-cotton affects agricultural practices, more specifically herbicide use, in the countries where it is grown. The Notifier has stated which countries that will grow the LLCotton25 (USA, Brazil, Mexico and Australia), but information on herbicide use has been submitted for the US only, and even here it would be of interest to have the changes in herbicide use quantified to a larger extent. The Notifiers’ claims that use of the cotton will lead to a reduced and more flexible use of herbicides for the grower, in all of the relevant climates and agricultural traditions, needs to be supported with more information from the countries Brazil, Mexico and Australia. An overall reduced use of herbicides would be regarded as positive both for human health and for the environment and therefore this type of information would be relevant.

Comments from other EFSAnet users

AT Ministry of Health

and Women D, 08 Post-market monitoring of GM food/feed

According to Art. 5 (3) k) of EU-Regulation 1829/2003 a post-market monitoring-plan should be added to the dossier.

See section 4.2.7 of the scientific opinion: The risk assessment concluded that no data have emerged to indicate that LLCotton25 is any less safe than its non-GM comparator. In addition, LLCotton25 is, from a nutritional point of view, substantially equivalent to conventional cotton. Therefore, and in line with the Guidance document (EFSA 2006a), the GMO Panel is of the opinion that post-market monitoring of the GM food/feed is not necessary. The GMO Panel is of the opinion that a Post-Market Monitoring (PMM) should be required only in specific cases where there is no traditional comparator. Those cases could include GM (functional) foods with altered nutritional composition and modified nutritional value and/or with specific health claims. See also section 7.11 of the GMO Panel Guidance document (2006a).

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AT Ministry of Health and Women

D, 07.08 Toxicology In one of its latest studies [Spök A., Hofer H., Lehner P., Valenta R., Stirn S. Gaugitsch H. (2005). Risk Assessment of GMO Products in the European Union. Umweltbundesamt Wien, Band 253 ,] the Austrian Federal Environment Agency developed recommendations for a standardized and harmonized approach to the generation, presentation and interpretation of data concerning allergenicity and toxicology of GM products. These recommendations are based on in depth scientific studies, performed by experienced scientists in the field. The proposed tests should be performed by the notifier and the resulting data provided in order to guarantee a high level of safety and public confidence in the approach taken. But in this notification neither allergenicity nor toxicology of this product has been assessed thoroughly in the risk assessment. For example: only a poultry feeding study and a nearly ten-year-old 14-day sub chronic oral toxicity study (Pfister et al., 1996) have been conducted. Therefore also an adequate risk assessment to evaluate possible long term effects is regarded as necessary.

See section 7.9 of the GMO Panel Guidance document (2006a): The specific allergy risk of GMOs is associated with i) exposure to newly expressed protein(s) that can be present in edible parts of the plants or in the pollen… and ii) with alterations to the allergenicity of the whole plant and derived products e.g. due to over-expression of natural endogenous allergens as an unintended effect of the genetic modification. i) Given this lack of complete predictability of allergenicity of a newly expressed protein, it is necessary to obtain, from several steps in the risk assessment process, a cumulative body of evidence which minimises any uncertainty with regard to the protein(s) in question. This approach is internationally accepted and has been conducted as described in section 4.2.5 of the scientific opinion. ii) The issue is to demonstrate that the GM crop will not be more allergenic than the non GM comparator because of an unintended effect of the insertion of the transgene. If the host of the inserted gene is known to be allergenic, any potential change in the allergenicity of the whole GM food should be tested by comparison of the allergen repertoire with that of the conventional non-GM variety. It should be pointed out that these approaches should be applied on a case-by-case basis depending on the available information on the allergenic potential of the host. See section 4.2.4 of the scientific opinion: The comparative compositional analysis and agronomic analyses showed that LLCotton25 is substantially equivalent to its non-transgenic counterpart Coker312 and other commercially grown cotton varieties except for the introduced trait. In addition, the analyses provided no indication for unintended effects of the genetic modification and therefore

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the GMO Panel concluded that no additional safety studies with laboratory animals are needed.


D, 07.09 Allergenicity In one of its latest studies [Spök A., Hofer H., Lehner P., Valenta R., Stirn S. Gaugitsch H. (2005). Risk Assessment of GMO Products in the European Union. Umweltbundesamt Wien, Band 253 , Spök et.al.: Suggestions for the Assessment of the Allergenic Potential of Genetically Modified Organisms; Int Arch Allergy Immunol 2005; 137: 167-180 ] the Austrian Federal Environment Agency developed recommendations for a standardized and harmonized approach to the generation, presentation and interpretation of data concerning allergenicity and toxicology of GM products. These recommendations are based on in depth scientific studies, performed by experienced scientists in the field. The proposed tests should be performed by the notifier and the resulting data provided in order to guarantee a high level of safety and public confidence in the approach taken. But in this notification neither allergenicity nor toxicology of this product has been assessed thoroughly in the risk assessment. The assessment of potential allergenic effects is mainly focusing on database-search. This can only be regarded as one component of a comprehensive risk assessment.

See section 7.9 of the GMO Panel Guidance document (2006a): The specific allergy risk of GMOs is associated with i) exposure to newly expressed protein(s) that can be present in edible parts of the plants or in the pollen… and ii) with alterations to the allergenicity of the whole plant and derived products e.g. due to over-expression of natural endogenous allergens as an unintended effect of the genetic modification. i) Given this lack of complete predictability of allergenicity of a newly expressed protein, it is necessary to obtain, from several steps in the risk assessment process, a cumulative body of evidence which minimises any uncertainty with regard to the protein(s) in question. This approach is internationally accepted and has been conducted as described in section 4.2.5 of the scientific opinion. ii) The issue is to demonstrate that the GM crop will not be more allergenic than the non GM comparator because of an unintended effect of the insertion of the transgene. If the host of the inserted gene is known to be allergenic, any potential change in the allergenicity of the whole GM food should be tested by comparison of the allergen repertoire with that of the conventional non-GM variety. It should be pointed out that these approaches should be applied on a case-by-case basis depending on the available information on the allergenic potential of the host.


D, 07.10 Nutritional assessment of GM food/feed

In chapter 7 of part I compositional analysis is described. Concerning the presented data it has to be stated that a for some parameters rather inconsistent data are presented (high zinc level, huge variations of Vitamin E

See section 7.2(b) statistical models for analysis, confidence intervals of the GMO Panel Guidance document : Experimental design should be rigorous and analysis of data should be presented in a clear format, using standardised

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content for e.g. delinted cotton seed or meal). The explanation is often “No values are exceeding the 20% bioequivalence range”. This can be regarded as appropriate to a certain extend but if the data are out of the natural range then the notifier writes e.g. “the measured anti-nutrient values (for crude cottonseed oil) are not falling into the literature range, but this is true for the results of all treatments, for the control samples and both transgenic samples.” This cannot be regarded as a state of the art risk assessment. Further studies are highly recommended

scientific units. Field trial data should be presented separately, as well as pooled, and should be analysed statistically, using appropriate statistical tools. A randomised complete block design, for example, could indicate whether the experimental factors (location, year, climatic conditions, plant variety) interact with one another. The confidence intervals used for statistical analysis should be specified (normally 95%, with possible adjustment according to the hazard of the constituent to be compared). On 26 January 2006, the GMO Panel required from the applicant further clarifications with respect to the statistical analysis of the compositional data including statistical data analyses according to traditional methodology. The applicant responded on 18 May 2006 and the GMO Panel was satisfied with the response.


D, 10 Potential changes in the interactions of the GM plant with the biotic…

The applicant writes in the dossier (p.19, part I) that “no wild relatives of cotton have been reported in Europe”. But it also has to be stated that even in Austria cotton-weeds have been found like Echinochloa crus-galli, Cynodon dactylon, Sorghum halepense or Abutilon theophrastii [Fischer M.: Exkursionsflora von Österreich, Stuttgart 1995] . Therefore this aspect should be taken into account for those member countries of the EC, where cotton is cultivated.

The scope of the application is for import, processing as well as for food and feed uses and does not include cultivation. Therefore, there was no requirement for scientific information on possible environmental effects associated with the cultivation.


D, 12 Environmental Monitoring Plan

Results of surveys and inspections in Austria [Heissenberger et al., 2003, Research Report 4/03, Federal Ministry of Social Security and Generations (now Federal Ministry of Health and Women), Vienna.] have shown that unintended or technically unavoidable contamination leads to the unintended release of seed or grains. This matter of fact is of great importance for those member countries of the EC, where cotton is cultivated. Therefore an adequate monitoring plan, focusing on spillage should be proposed by the applicant.

The scope of the application is for import, processing as well as for food and feed uses and does not include cultivation. Therefore, there was no requirement for scientific information on possible environmental effects associated with the cultivation. See section 5.2.2 of the scientific opinion: The GMO Panel is aware that, due to the physical characteristics of cotton seeds and methods of transportation, accidental spillage is unavoidable. Therefore the GMO Panel

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recommends that specific measures are introduced to actively monitor the occurrence of feral cotton plants in areas where seed spillage is likely to occur as proposed in the EFSA Guidance document (EFSA, 2006a) and the opinion of the GMO Panel on post-market environmental monitoring (EFSA, 2006b). In other respects the GMO Panel is of the opinion that the general approaches and measures of the monitoring plan proposed by the applicant are in line with the EFSA opinion on post-market environmental monitoring (EFSA, 2006b) as well as with the intended uses of LLCotton25. Since the environmental risk assessment does not cover cultivation and identifies no potential adverse environmental effects, no case-specific monitoring is necessary. See section 5.2 of the PMEM opinion (EFSA, 2006b): Details of the specific plans and methods of monitoring in each country should not be included in the original application. The GMO Panel advises that the application should describe the general approaches and methods that the applicant would apply in different commercialisation sites, including the type of dialogue that would be established with risk managers in each Member State. (…) Thus detailed local arrangements will be developed by the applicant after the application has been accepted (…).


D, 12.03 General Surveillance of the impact of the GM plant

The general surveillance should address the possibility of unexpected adverse effects. The proposed measures to facilitate this general surveillance have to take into account latest scientific research and results on potential ecological impacts and unintended effects/changes in plant metabolism, which occur frequently during development of conventional as well as genetically modified plants [“Expecting the unexpected”; oral presentation at “Biodiversity Implications of Genetically Modified Plants” (A.N.E. Birch et al., SCIR/Dundee, Scotland), international conference in Ascona, September

See the PMEM opinion (EFSA, 2006b) as well as section 11.4 of the GMO Panel Guidance document (EFSA, 2006a).

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2003.].

FR DGCCRF - Ministère de l'Economie, des Finances et de l'Industrie

D, 07.08 Toxicology (7.8) Toxicologie Les résultats d'une étude de toxicité aiguë, réalisée chez des souris à qui une dose unique de la protéine PAT (10 mg/kg p.c.) a été administrée par voie intraveineuse, n'ont pas mis en évidence d'effets néfastes ni de mortalité après 15 j d'observations ; il convient de noter que de nombreuses études de toxicité aiguë ont été réalisées par voie orale avec la protéine PAT et qu'il n'a jamais été mis en évidence d'effet néfaste (dose sans effet supérieure à 5000 mg/kg p.c.); La protéine PAT codée par le gène bar est dégradée en moins de 30 secondes dans un milieu gastrique simulé et en 5 minutes dans un milieu intestinal simulé. L'équivalence structurale et fonctionnelle de la protéine PAT codée par le gène bar extraite de la plante LLCotton25 et la protéine PAT extraite d'E.Coli a été vérifiée par analyse SDS-Page et Western blot, et qu'elle n'est pas glycosylée. La séquence du gène bar de LLCotton25 présente des homologies de séquences uniquement avec des séquences de gènes codant des acétyltransférases et que la séquence de la protéine PAT codée par le gène bar ne présente pas d'homologies de séquences avec des protéines répertoriées dans les bases de données connues pour avoir des propriétés toxiques; Cependant, il serait nécessaire de disposer d'une étude de toxicité subchronique chez le rat nourri avec des produits issus du cotonnier LLCotton25 pour exclure la survenue d'effets toxiques inattendus liés à la modification génétique. L'Agence française de sécurité sanitaire des aliments considère que pour s'assurer que les produits issus du cotonnier LLCotton25 présentent le même niveau de sécurité sanitaire que les produits issus d'un cotonnier conventionnel, il conviendrait de disposer une étude de toxicité subchronique de 90 jours chez le rat nourris avec de l'huile produite à partir de ce cotonnier génétiquement modifié.

See section 4.2.3.1 of the scientific opinion: Due to the low expression level of the PAT protein in LLCotton25 most of the safety studies were conducted with a PAT protein encoded in E. coli by the bar gene (PAT/bar protein). Examination of the structure and function of these plant and bacterial PAT proteins have shown a high degree of similarity, based on their size and sequence homology, enzymatic activity, immunoreactivity and absence of glycosylation. The PAT/pat and PAT/bar proteins have been shown to be structurally and functionally equivalent (Wehrmann et al., 1996; Herouet et al., 2005). Therefore the GMO Panel accepts the PAT/bar as well as the PAT/pat test material derived from E. coli for the safety assessment of PAT protein present in LLCotton25. Furthermore the GMO Panel did not consider the possible need for additional toxicity studies. See section 4.2.4 of the scientific opinion: The comparative compositional analysis and agronomic analyses showed that LLCotton25 is substantially equivalent to its non-transgenic counterpart Coker312 and other commercially grown cotton varieties except for the introduced trait. In addition, the analyses provided no indication for unintended effects of the genetic modification and therefore the GMO Panel concluded that no additional safety studies with laboratory animals are needed.

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GR Hellenic Food Authority

D, 02 Information on the sequences actually inserted or deleted

The data provided regarding the flanking 5’ region of the insert demonstrate a 38 bp deletion in the genome of the host plant (technical dossier, part 2.d, page 31). According to the dossier, bioinformatics analysis does not give evidence that a novel transcript might arise at either junction of the insert. Since this approach is theoretical, it is necessary to provide transcriptional analysis data such as RT-PCR and/ or northern blot analysis.

See section D.3.(c) of the GMO Panel Guidance document (EFSA, 2006a): The potential creation of newly expressed fusion proteins should be investigated by bioinformatic analysis and the absence of any harmful fusion proteins demonstrated. An investigation of newly expressed transcripts is appropriate when a bioinformatic analysis identifies a putative fusion protein.

See section 2.2.2 of the scientific opinion: The nucleotide sequence of the insert introduced into LLCotton25 has been determined in its entirety. The DNA sequence of the LLCotton25 insert has been proven to be identical to the corresponding transforming plasmid pGSV71 sequences. PCR analysis of the terminal repeats of the vector plasmid confirmed that the right border (RB) terminal repeat is not completely integrated in LLCotton25 as 23 bp are missing. The left border (LB) terminal repeat sequence displays a deletion of 4 bp. The sequences of the plant genome adjacent to the 3’ and 5’ sequences of the insert were determined using TAIL-PCR. Comparison of the flanking sequences to the respective wild type target site revealed that upon integration of the T-DNA into the genomic DNA a 38 bp fragment of genomic DNA at the target site was deleted. There was no indication that the insert is integrated in a coding region or that the insert disrupts gene regulatory sequences. These data presented proof that the insert has been integrated in a single chromosome at a single locus as intended.

Molecular analysis of the insert and bioinformatic analysis of flanking regions do not suggest that transcriptional analysis is required.


D, 07.08 Toxicology It is necessary to perform an additional subchronic feeding study in rats to further complete its safety assessment.

See section 4.2.4 of the scientific opinion: The comparative compositional analysis and agronomic analyses showed that LLCotton25 is substantially equivalent to its non-transgenic counterpart Coker312 and other commercially grown cotton varieties except for the introduced

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trait. In addition, the analyses provided no indication for unintended effects of the genetic modification and therefore the GMO Panel concluded that no additional safety studies with laboratory animals are needed.


General comments There is a discrepancy in part 2(b) of the application file which could be misleading. This discrepancy concerns the putative deletions occurred after the insertion of the target sequence. Although deletions are reported in the page 31 of the part 2(d) of the technical file, as mentioned below, these deletions are not described in the part 2(b) page 30. The deletions are: 1) one deletion of a 38 bp fragment from the wild type cotton DNA upon integration of the T-DNA 2)one deletion of a 23 bp fragment from the transforming plasmid DNA (right border terminal repeat sequence) 3)one deletion of a 4 bp fragment from the transforming plasmid DNA (left border terminal repeat sequence)

It is correct that the deletions are not mentioned on part 2(b) page 30. However, this section refers to intended deletions of DNA sequences, for example to deliberately knock out a function in the GM plant. The unintended deletions in the T-DNA borders in LLcotton25 and in the target DNA of the wild type cotton plant are therefore accurately described in the section 2(d) on page 31. See section 2.2.2 of the scientific opinion: The nucleotide sequence of the insert introduced into LLCotton25 has been determined in its entirety. The DNA sequence of the LLCotton25 insert has been proven to be identical to the corresponding transforming plasmid pGSV71 sequences. PCR analysis of the terminal repeats of the vector plasmid confirmed that the right border (RB) terminal repeat is not completely integrated in LLCotton25 as 23 bp are missing. The left border (LB) terminal repeat sequence displays a deletion of 4 bp. The sequences of the plant genome adjacent to the 3’ and 5’ sequences of the insert were determined using TAIL-PCR. Comparison of the flanking sequences to the respective wild type target site revealed that upon integration of the T-DNA into the genomic DNA a 38 bp fragment of genomic DNA at the target site was deleted. There was no indication that the insert is integrated in a coding region or that the insert disrupts gene regulatory sequences. These data presented proof that the insert has been integrated in a single chromosome at a single locus as intended.

NL Ministry of Agriculture

D, 07.03 Selection of compounds for analysis

According to the guidance of the EFSA GMO Panel, compositional analysis should include samples from

See section 7.2(b) statistical models for analysis, confidence intervals of the GMO Panel Guidance document (EFSA,

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multiple seasons and locations, taking these factors into account in the statistical analysis. Please note that the establishment of equivalence is in fact the starting point of the comparative safety assessment. Therefore the conclusions based upon the compositional data, including key nutrients and antinutrients, should be based on sufficient data. In the statistical analysis of compositional data, the results of the analysis of samples obtained from field trials carried out during two seasons have been combined. This does not allow for the analysis for consistency of differences between seasons, among others. In line with the guidance provided by EFSA, a statistical analysis of compositional data for each separate season should be provided. These data are essential in order to be able to identify issues that may need to be further addressed in the safety assessment of LLcotton25 for food and feed use.

2006a): Experimental design should be rigorous and analysis of data should be presented in a clear format, using standardised scientific units. Field trial data should be presented separately, as well as pooled, and should be analysed statistically, using appropriate statistical tools. A randomised complete block design, for example, could indicate whether the experimental factors (location, year, climatic conditions, plant variety) interact with one another. The confidence intervals used for statistical analysis should be specified (normally 95%, with possible adjustment according to the hazard of the constituent to be compared).

On 26 January 2006, the GMO Panel required from the applicant further clarifications with respect to the statistical analysis of the compositional data. The applicant responded on 18 May 2006 and the GMO Panel was satisfied with the response.

List of References

EFSA, 2006a. Guidance document of the Scientific Panel on Genetically Modified Organisms for the Risk Assessment of Genetically Modified Plants and Derived Food and Feed. The EFSA Journal 374, 1-115. http://www.efsa.europa.eu/etc/medialib/efsa/science/gmo/gmo_guidance/gmo_guidance_ej374.Par.0001.File.tmp/gmo_guidance_ej374_gmm.pdf

EFSA, 2006b. Opinion of the Scientific Panel on Genetically Modified Organisms on the Post Market Environmental Monitoring (PMEM) of genetically modified plants. The EFSA Journal 319, 1-27. http://www.efsa.europa.eu/science/gmo/gmo_opinions/1381/gmo_op_ej319_pmem_en1.pdf

http://www.efsa.europa.eu/etc/medialib/efsa/science/gmo/gmo_guidance/gmo_guidance_ej374.Par.0001.File.tmp/gmo_guidance_ej374_gmm.pdf

http://www.efsa.europa.eu/science/gmo/gmo_opinions/1381/gmo_op_ej319_pmem_en1.pdf

Documents

Application EFSA-GMO-NL-2005-13 (LLCotton25) · PDF fileis not completely integrated in LLCotton25 as 23 bp are ... the data performed with a 20% bioequivalence range was ... A randomised