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Maize, the second most important genetically modified (GM) crop, has the highest number of authorised GM events for food and feed in the EU. To provide consumer's information, labelling for food products containing more than 0.9% of GM material is demanded by the actual EU legislation. Analysis of foods is then essential to detect and quantify GM maize material and verify the compliance with labelling information. The aim of the present work was to assess the presence of GM maize in a range of processed foods commercialised in Portugal between 2007 and 2010. For this purpose, screening of GM material was carried out by qualitative PCR targeting the 35S promoter and the NOS terminator, followed by the specific detection of Bt11, MON810, Bt176, GA21, MON863, NK603, TC1507 (also known as DAS1507), DAS59122 and MIR604 events. The identified maize events were confirmed and quantified by real-time PCR with hydrolysis probes. The overall results of GMO screening were 30% for 35S promoter, 10% for NOS terminator and 25% for identified events. The most frequently detected events were MON810, TC1507 and NK603, with one sample containing GA21, while the other events were not detected in any of the analysed foods. The quantitative results suggest the need for a more severe control since 4% of the analysed foods contained more than the threshold for labelling and none of them declared the presence of GMO.

Monitoring of market products for detection of genetically modified organisms (GMO) is needed to comply with legislation in force in many regions of the world, to enforce traceability and to allow official control along the production and the distribution chains. This objective can be more easily achieved if reliable, time and cost-effective analytical methods are available. A GMO can be detected using either DNA-based or protein-based methods; both present advantages and disadvantages. The objective of this work was to assess the performance of a protein-based (lateral flow strips—LFT) and of a DNA-based (polymerase chain reaction—PCR) detection method for GMO analysis. One thousand five hundred samples of soybean, deriving from the sampling of 15 independent bulk lots in large shipments, were analysed to assess and compare the performance of the analytical methods and evaluate their suitability for GMO testing. Several indicators were used to compare the performance of the methods, including the percentage correlation between the PCR and LFT results. The GMO content of the samples ranged from 0 up to 100 %, allowing a full assessment of both analytical approaches with respect to all possible GMO content scenarios. The study revealed a very similar performance of the two methodologies, with low false-negative and false-positive results, and a very satisfactory capacity of both methods in detecting low amounts of target. While determining the fitness for purpose of both analytical approaches, this study also underlines the importance of alternative method characteristics, like costs and time.

free of charge pdf chapter available at: https://bdspublishing.com/_webedit/uploaded-files/All%20Files/Open%20Access/9781801462044.pdf ; International audience ; The growing knowledge in genetics, epigenomics, epitranscriptomics, and the 3D - or even 4D - genome structure provides an increasing number of detection targets that can be used to identify species or genetic lines, whether modified or not. Biotic and abiotic stresses also induce numerous unintentional genetic, epigenetic, and epitranscriptomic modifications. Those changes are transmissible and can be ordered in regions and classified. The detection target is characterised by the mutagenesis technique used. For instance, the detection of transgenic GMO or SDN3 modification of New Breeding Techniques (NBTs) will target their insertion's junction fragments into the genome. Each insertion induces epigenetic, and probably epitranscriptomic, changes which can also be targeted. In addition, one group of markers is linked to the trait(s) introduced or modified by the breeder whose sequence could be used in quantification and "screening". The other target will be a subset of the elements of a matrix approach (as described in the previous chapter). General selection markers, such as those used for plant breeding, together with mutagenesis techniques specific markers, could differentiate genetically modified organisms (GMOs) of any origin. They can be used to quantify and certify, through a global approach to the organism, that the trait modification is artefactual and not "natural." The growing mastery of single-cell sequencing techniques should soon make it possible to differentiate the modifications due, for example, to each step of a Crispr-Cas transformation of cells in culture. This chapter will focus mainly on detection targets based on nucleic acids, DNA, RNA, modified or unmodified, for their routine use in private and enforcement detection laboratories to comply with food labelling and European traceability rules. The chapter use the wording "hidden" ...

free of charge pdf chapter available at: https://bdspublishing.com/_webedit/uploaded-files/All%20Files/Open%20Access/9781801462044.pdf ; International audience ; The growing knowledge in genetics, epigenomics, epitranscriptomics, and the 3D - or even 4D - genome structure provides an increasing number of detection targets that can be used to identify species or genetic lines, whether modified or not. Biotic and abiotic stresses also induce numerous unintentional genetic, epigenetic, and epitranscriptomic modifications. Those changes are transmissible and can be ordered in regions and classified. The detection target is characterised by the mutagenesis technique used. For instance, the detection of transgenic GMO or SDN3 modification of New Breeding Techniques (NBTs) will target their insertion's junction fragments into the genome. Each insertion induces epigenetic, and probably epitranscriptomic, changes which can also be targeted. In addition, one group of markers is linked to the trait(s) introduced or modified by the breeder whose sequence could be used in quantification and "screening". The other target will be a subset of the elements of a matrix approach (as described in the previous chapter). General selection markers, such as those used for plant breeding, together with mutagenesis techniques specific markers, could differentiate genetically modified organisms (GMOs) of any origin. They can be used to quantify and certify, through a global approach to the organism, that the trait modification is artefactual and not "natural." The growing mastery of single-cell sequencing techniques should soon make it possible to differentiate the modifications due, for example, to each step of a Crispr-Cas transformation of cells in culture. This chapter will focus mainly on detection targets based on nucleic acids, DNA, RNA, modified or unmodified, for their routine use in private and enforcement detection laboratories to comply with food labelling and European traceability rules. The chapter use the wording "hidden" ...

free of charge pdf chapter available at: https://bdspublishing.com/_webedit/uploaded-files/All%20Files/Open%20Access/9781801462044.pdf ; International audience ; The growing knowledge in genetics, epigenomics, epitranscriptomics, and the 3D - or even 4D - genome structure provides an increasing number of detection targets that can be used to identify species or genetic lines, whether modified or not. Biotic and abiotic stresses also induce numerous unintentional genetic, epigenetic, and epitranscriptomic modifications. Those changes are transmissible and can be ordered in regions and classified. The detection target is characterised by the mutagenesis technique used. For instance, the detection of transgenic GMO or SDN3 modification of New Breeding Techniques (NBTs) will target their insertion's junction fragments into the genome. Each insertion induces epigenetic, and probably epitranscriptomic, changes which can also be targeted. In addition, one group of markers is linked to the trait(s) introduced or modified by the breeder whose sequence could be used in quantification and "screening". The other target will be a subset of the elements of a matrix approach (as described in the previous chapter). General selection markers, such as those used for plant breeding, together with mutagenesis techniques specific markers, could differentiate genetically modified organisms (GMOs) of any origin. They can be used to quantify and certify, through a global approach to the organism, that the trait modification is artefactual and not "natural." The growing mastery of single-cell sequencing techniques should soon make it possible to differentiate the modifications due, for example, to each step of a Crispr-Cas transformation of cells in culture. This chapter will focus mainly on detection targets based on nucleic acids, DNA, RNA, modified or unmodified, for their routine use in private and enforcement detection laboratories to comply with food labelling and European traceability rules. The chapter use the wording "hidden" ...

International audience ; The growing knowledge in genetics, epigenomics, epitranscriptomics, and the 3D - or even 4D - genome structure provides an increasing number of detection targets that can be used to identify species or genetic lines, whether modified or not. Biotic and abiotic stresses also induce numerous unintentional genetic, epigenetic, and epitranscriptomic modifications. Those changes are transmissible and can be ordered in regions and classified. The detection target is characterised by the mutagenesis technique used. For instance, the detection of transgenic GMO or SDN3 modification of New Breeding Techniques (NBTs) will target their insertion's junction fragments into the genome. Each insertion induces epigenetic, and probably epitranscriptomic, changes which can also be targeted. In addition, one group of markers is linked to the trait(s) introduced or modified by the breeder whose sequence could be used in quantification and "screening". The other target will be a subset of the elements of a matrix approach (as described in the previous chapter). General selection markers, such as those used for plant breeding, together with mutagenesis techniques specific markers, could differentiate genetically modified organisms (GMOs) of any origin. They can be used to quantify and certify, through a global approach to the organism, that the trait modification is artefactual and not "natural." The growing mastery of single-cell sequencing techniques should soon make it possible to differentiate the modifications due, for example, to each step of a Crispr-Cas transformation of cells in culture. This chapter will focus mainly on detection targets based on nucleic acids, DNA, RNA, modified or unmodified, for their routine use in private and enforcement detection laboratories to comply with food labelling and European traceability rules. The chapter use the wording "hidden" GMOs and "new" GMOs as defined by the French NGOs and farmers' union at the origin of the 2018 European Court of Justice ruling.

GMO testing of foodstuffs represents an important tool for assessing compliance with EU legislation on transgenic plants derived food and feed (Querci et al., 2005). This paper describes the main features of the analytical steps encompassed in the GMO testing procedures performed in our laboratory. Qualitative and quantitative determination of transformation event Bt176 in maize grains is used to exemplify how the methods are employed. Bt176 is no longer approved for commercialization within the EU but represents an adequate example for a case study in this field because of the availability of information and data on the subject. It is also well suited for tests using the instruments in our laboratory. The main analytical steps for GMO testing of raw and processed plant matrices are as follows: obtaining of a representative laboratory sample by employing special strategies; sample preparation (e.g. milling, separation of different fractions) in order to obtain the test sample; DNA extraction using the QIAamp DNA Stool Kit (Qiagen) was used; assessment of the extracted DNA by spectrophotometric readings and agarose gel electrophoresis; qualitative testing based on classical PCR aimed at detection taxon specific or transgenic specific DNA; quantitative testing based on real-time PCR using the RoundUp Ready Soya Qt KIT (Biotools). The analyses were conducted using experimental designs that satisfy all requirements and recommendations for this type of analyses (see ISO 21570:2006; Querci et al., 2005; Foti, 2004). The validation and interpretation of results is also presented.

The survey was conducted on a total of 100 non-labelled samples of feed and feed mixture containing maize, soybean and rapeseed, originating from countries with different legislation systems. Screening of all samples was performed using primers for Cauliflower Mosaic Virus 35S (CaMV35S) promoter, primers for the Agrobacterium tumefaciens nopaline synthase (NOS) terminator and event-specific primers for GT73 rapeseed. Roundup Ready soybean was found in 26 samples, with the amount of GM soybean above the limit of 0.9% in 9 of them. There was one maize seed sample positive for the presence of MON810 maize and no rapeseed meal samples contained GM rapeseed. The results found in this study clearly showed that imported maize and soybean and complete mixtures intended for animal feed on the Serbian market contain GMO. Monitoring plans are required to control the distribution of non-labelled feeds containing GMO in the Serbian market.

The European Union has adopted a very restrictive policy towards the dissemination and use of genetically modified organisms (GMOs), whose use in food is not well accepted by consumers. Although a maximum threshold exists for a food to be labelled "GM-free", only known GMOs are easily detectable. A GMO consists mainly of a host genome and a sequence inserted by a non-natural process that confers a particular property on the organism, such as resistance to certain diseases. In recent years, GMOs with an inserted sequence that is not known have been produced that are not detectable by approaches used until now (PCR-type). Hence the need to propose a tool for the detection of unknown GMOs, the subject of this thesis, based on recent advances in terms of high-throughput sequencing. Statistically, each organism has a specific frequency of nucleotide use in its genome. Any introduction of foreign genetic material will locally alter the nucleotide use frequencies in that region, resulting in different nucleotide use frequencies compared to those of the host organism. Based on this assertion, an unknown GMO detection tool has been developed from bacterial sequencing data when the GMO results from the insertion of a foreign gene, the truncation or fusion of a gene that may belong to the host genome. The tool has been tested on 4 GMO bacterial genomes, 7 wild bacterial genomes and 42 synthetic bacterial genomes. The results demonstrate the effectiveness of the method developed by presenting only one false positive gene and identifying more than 99% of the genes of GMO inserts. ; L'Union Européenne a adopté une politique très restrictive vis-à-vis de la diffusion et de l'utilisation des organismes génétiquement modifiés (OGM), dont l'utilisation dans l'alimentation est mal acceptée par les consommateurs. Bien qu'un seuil maximal existe pour qu'un aliment soit étiqueté « sans OGM », ne sont aisément détectables que les OGM connus. Un OGM est constitué principalement d'un génome hôte et d'une séquence insérée par un procédé ...

In the last years, the increase in the cultivated area of genetically modified (GM) maize has become a reality. GA21, MON810 and MON 863 maize crops are some of the authorized maize events for food and feed under the European Union (EU) regulations. These crops of transgenic maize bring profit towards the conventional ones, as they confer resistence to some plagues and/or herbices. Concerning the raise of production and consumption of foodstuffs derived from genetically modified organisms (GMO), the EU has established new demand levels, including the labeling requirements when the product has GMO in proportion higher than 0.9% (Regulation (EC) Nº. 1829/2003).

free of charge pdf chapter available at: https://bdspublishing.com/_webedit/uploaded-files/All%20Files/Open%20Access/9781801462037.pdf ; International audience ; The discovery in the 1980s of the pathogenesis' mechanisms of Agrobacterium tumefaciens led to transgenesis, a technique for increasing the diversity of traits that could be used in plant breeding. Various other means of plant transformation were then implemented. This new technique came when in vitro mutagenesis was stalled due to the lack of mutations' screening systems until the description in 2000 of the Tilling technique which was then followed by various developments.Consumers received the genetically modified organisms (GMO) products resulting from these artefactual transformations in different ways in different countries. In European countries with a long culinary tradition and numerous products under official quality labels, the precautionary principle, which had previously prevailed in third countries, was introduced in the face of these new techniques which at the time had lacked any history of safe use. From then on, these GMOs were only produced and marketed after a risk assessment. In addition, labelling and traceability, according to the farm-to-fork approach, are required with specific and general post-market environmental monitoring. This chapter describes the scientific, technical and regulatory framework of this European traceability system, which allows all European consumers to make informed choices about their food. Moreover, this traceability approach enables the coexistence of GM and non-GM supply chains and should thus make it possible to avoid mixing food products with those for pharmaceutical, functional food or industrial use.The framework we describe in this chapter must be used to deal with the traceability of "new" GMOs and "hidden" GMOs. GMOs resulting from in vitro mutagenesis of isolated cells and NBT techniques, so named by the non-governmental organisations (NGOs) and farmers' union that brought the dispute before the ...

free of charge pdf chapter available at: https://bdspublishing.com/_webedit/uploaded-files/All%20Files/Open%20Access/9781801462037.pdf ; International audience ; The discovery in the 1980s of the pathogenesis' mechanisms of Agrobacterium tumefaciens led to transgenesis, a technique for increasing the diversity of traits that could be used in plant breeding. Various other means of plant transformation were then implemented. This new technique came when in vitro mutagenesis was stalled due to the lack of mutations' screening systems until the description in 2000 of the Tilling technique which was then followed by various developments.Consumers received the genetically modified organisms (GMO) products resulting from these artefactual transformations in different ways in different countries. In European countries with a long culinary tradition and numerous products under official quality labels, the precautionary principle, which had previously prevailed in third countries, was introduced in the face of these new techniques which at the time had lacked any history of safe use. From then on, these GMOs were only produced and marketed after a risk assessment. In addition, labelling and traceability, according to the farm-to-fork approach, are required with specific and general post-market environmental monitoring. This chapter describes the scientific, technical and regulatory framework of this European traceability system, which allows all European consumers to make informed choices about their food. Moreover, this traceability approach enables the coexistence of GM and non-GM supply chains and should thus make it possible to avoid mixing food products with those for pharmaceutical, functional food or industrial use.The framework we describe in this chapter must be used to deal with the traceability of "new" GMOs and "hidden" GMOs. GMOs resulting from in vitro mutagenesis of isolated cells and NBT techniques, so named by the non-governmental organisations (NGOs) and farmers' union that brought the dispute before the ...