Food safety and quality
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OECD Unique Identifier details

BCS-GHØØ2-5
Commodity: Cotton
Traits: Glyphosate tolerance
Australia
Name of product applicant: Bayer CropScience Pty Ltd
Summary of application:
Glyphosate (N-phosphonomethylglycine) is a non-selective, broad spectrum herbicide. The mode of action of glyphosate is to specifically bind to, and block, the activity of a native plant enzyme, 5-enol-pyruvylshikimate-3-phosphate synthase (EPSPS). EPSPS is a key enzyme in the shikimate pathway in plants which links the metabolism of carbohydrates to the biosynthesis of ring-containing compounds including aromatic amino acids. Plant EPSPS enzymes are normally inactivated by glyphosate which leads to cellular deficiencies in certain amino acids resulting ultimately in the death of the plant.
In cotton line GHB614, tolerance to glyphosate is achieved through expression in the plant of a modified form of the EPSPS enzyme, 2mEPSPS, derived from corn. Two point (single nucleotide) mutations were introduced to the corn epsps gene to generate 2mepsps, using site directed mutagenesis. These changes significantly reduce the sensitivity of the 2mEPSPS enzyme to glyphosate, allowing it to continue to function in the presence of the herbicide.
Cotton line GHB614 has been developed for agriculture in major cotton producing countries worldwide, including Australia.
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Date of authorization: 17/09/2009
Scope of authorization: Food
Links to the information on the same product in other databases maintained by relevant international organizations, as appropriate. (We recommend providing links to only those databases to which your country has officially contributed.): OECD BioTrack Product Database
Summary of the safety assessment:
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Where detection method protocols and appropriate reference material (non-viable, or in certain circumstances, viable) suitable for low-level situation may be obtained:
Relevant links to documents and information prepared by the competent authority responsible for the safety assessment: Application A614 - Food Derived From Glyphosate-Tolerant Cotton Line GHB614
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Authorization expiration date:
E-mail:
janet.gorst@foodstandards.gov.au
Organization/agency name (Full name):
Food Standards Australia New Zealand
Contact person name:
Janet Gorst
Website:
Physical full address:
Boeing Building, 55 Blackall Street, Barton ACT 2600, Australia
Phone number:
+61 2 6271 2266
Fax number:
+61 2 6271 2278
Country introduction:
Food Standards Australia New Zealand (FSANZ) is the regulatory agency responsible for the development of food standards in Australia and New Zealand. The main office (approximately 120 staff) is located in Canberra (in the Australian Capital Territory) and the smaller New Zealand office (approximately 15 staff) is located in Wellington on the North Island. The Food Standards Australia New Zealand Act 1991 establishes the mechanisms for the development and variation of joint food regulatory measures and creates FSANZ as the agency responsible for the development and maintenance of a joint Australia New Zealand Food Standards Code (the Code). The Code is read in conjunction with corresponding NZ and State & Territory food legislation as well as other appropriate legislative requirements (e.g. Trade Practices; Fair Trading). Within the Code, Standard 1.5.2 deals with Foods produced using Gene Technology. Applicants seeking to have a GM food approved, request a variation to Std 1.5.2 to have the GM food (from a particular line) included in the Schedule to Std 1.5.2. Only those GM foods listed in the Schedule can legally enter the food supply. An Application Handbook provides information that is required to make an application to vary the Code. This Handbook is a legal document and therefore the specified mandatory information must be supplied. For GM foods, there is also a Guidance Document that, as the name suggests, provides applicants with further details and background information on the data needed for the safety assessment of GM foods. The assessment process must be completed within a statutory timeframe (9 - 12 months depending on the complexity of the application) and involves at least one public consultation period. All GM applications involve an Exclusive Capturable Commercial Benefit i.e. applicants are required to pay a fee (outlined in the Application Handbook). Following the last public consultation, an Approval Report is prepared and is considered by the FSANZ Board who make a decision about whether the requested variation to the Code should be approved or not. The Board's decision is then passed on to the Legislative and Governance Forum on Food Regulation (the Forum), a committee comprising senior goevernment Ministers from Australia and NZ. This Committee has approximately 2 months to review the Board's decision. If the Board's approval is accepted by the Forum, the approval is then gazetted and becomes law.
Useful links
Relevant documents
Stacked events:
FSANZ does not: Separately assess food from stacked event lines where food from the GM parents has already been approved; Mandate notification of stacked events by developers; Notify the public of stacked event ‘approvals’; List food derived from stacked event lines in the Code, unless the stacked event line has been separately assessed as a single line e.g. Application A518: MXB-13 cotton (DAS-21023-5 x DAS-24236-5)
Contact details of the competent authority(s) responsible for the safety assessment and the product applicant:
Food Standards Australia New Zealand (FSANZ) (http://www.foodstandards.gov.au)
Brazil
Name of product applicant: Bayer S.A
Summary of application:
Commercial release of of genetically modified herbicide tolerant cotton styled GHB614 (GlyTol® Cotton)
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Date of authorization: 09/12/2010
Scope of authorization: Food and feed
Links to the information on the same product in other databases maintained by relevant international organizations, as appropriate. (We recommend providing links to only those databases to which your country has officially contributed.): Center for Environmental Risk Assessment
Summary of the safety assessment:
Event GHB614 was obtained through transformation of a variety of Coker 312 cotton, the genetic modification of which took place through a system mediated by Agrobacterium tumefasciens for the insertion of gene 2mepsps, responsible for expression of enzyme 2mEPSPS (5-enolpyruvylshikimate-3-phosphate synthase) by changing just two amino acids in the original peptide sequence. The enzyme grants GlyTol® cotton selectivity to herbicides containing the active ingredient glyphosate, therefore enabling to control pest plants in the crop post-emergence without harming the culture of the genetically modified cotton. Safety assessments conducted with GHB614 cotton showed alimentary innocuity, similarly to the parent conventional plants (var. Coker312). Regarding safety studies, assessments were conducted to determine whether GlyTol® cotton is similar to other cotton varieties regarding its use in human/animal feeding. Several studies were carried out of molecular characterization, expression of 2mEPSPS protein indifferent tissues, nutritional/compositional analysis, animal nutrition, protein digestibility, protein acute toxicity, and protein homology with toxic/allergenic compounds. Results in each case indicated that GHB614 and its progeny are substantially equivalent to other cotton varieties. The inserted DNA, as well as the expressed 2mEPSPS protein, fails to offer significant risk to human/animal health comparatively to the use of conventional cotton and byproducts in food. Aspects of environmental safety were assessed in a number of studies conducted in the national territory on the fields where it remained proved that plants derived from event GHB614 display the same agronomic behavior and adaptability in comparison with conventional genotypes, with no change of characters regulating the species survival and reproduction. Comparative analyses of morphological, phenotypic, and agronomic characteristics showed that none of the variables was changed as a result of inserting 2mepsps gene. Besides, no character was recorded that could grant selective advantage to GlyTol® or atypical behavior to the species. Regarding possible risks to the environment, issues related to endemic distribution of the feral form of G. mustelinum were considered and assessed in the south of Rio Grande do Norte and northeast of Bahia, and sub-spontaneous forms of G. barbadense in the Amazon region, Pantanal, southeast of Piauí and west of Pernambuco, as well as in the Atlantic forest, corresponding to the states of RN, PB, AL, SE, BA, MG and ES. Specificity to tolerance to the glyphosate herbicide was tested in the fields when the plants were submitted to the action of ammonium gluphosinate, resulting in total destruction of the plants so treated. Analyses of morphological, phenotypic and agronomic characteristics failed to evidence any difference between the genetically modified and non-genetically modified cotton, and no parameters were recorded that could grant selective advantages to GHB614 cotton or atypical behavior to the species. Besides, CTNBio searched the independent scientific literature to assess occurrence of any unexpected effect from the crossing between the events. Under Article 14 of Law nº 11,105/05, CTNBio held that the request complies with the applicable rules and legislation aimed at securing safety of the environment, agriculture, and human and animal health, and concluded that GHB614 cotton (GlyTol® cotton) is substantially equivalent to conventional cotton, being its consumption safe for human and animal health. Regarding the environment, CTNBio concluded that farming of GHB614 cotton (GlyTol® cotton) is not a potential cause of significant degradation to the environment, keeping with the biota a relation identical to that of the conventional cotton. CTNBio TECHNICAL OPINION I. Identification of GMO Name of GMO: GlyTol® cotton, Event GHB614 Applicant: Bayer S.A. Species: Gossypium hirsutum L. Inserted Characteristics: Tolerance to glyphosate Method of insertion: Transformation mediated by Agrobacterium tumefaciens containing vector pTEM2 Proposed use: Production of fibers for the textile industry and grain for human and animal consumption from the GMO and its derivatives II. General Information Cotton belongs to genus Gossypium, Tribe Gossypiae, Family Malvaceae, order Malvales (FRYXELL, P.A. 1979; MUNRO, J.M. 1987). The genus is divided into four sub-genera (Gossypium, Sturtia, Houzingenia and Karpas) that, in turn, are subdivided into nine sections and several sub-sections (FRYXELL, P.A.; CRAVEN, L.A.; and STEWART, J.MCD. 1992). Genus Gossipyum currently encompasses 52 species distributed in Asia, Africa, Australia and America, of which only 4 are cultivated. Gossypium arboretum L., cultivated in India is commercially important, and Gossypium herbaceum L., which was more important in the past, is currently planted just in some dry regions of Africa and Asia. Finally, about 90% of the world population of cotton belongs to Gossypium hirsutum and 8% to Gossypium barbadense (LEE, 1984). Out of the over 50 species already listed, all have a basic chromosome number n= 13, in which one part is diploid (2n+ 26), such as the cotton plants of the Old World (Africa and Asia), endemic species of Australia (CRAVEN et al., 1994) and some species of America. Other cotton species are tetraploid (allotetraploid), with n= 2x13= 26 and 2n= 52, involving, according to FRYXELL (1984), six species: G. tomentosum, endemic in Hawaii, G. mustelinum, in the Brazilian northeast, G. darwinii, in the Galapagos archipelago, G. barbadense, which its origin centered in South America, G. laceolatum, in Mexico, and G. hirsutum, with origin centered in Mexico and south of the United States. Out of commercial fiber production, the species G. hirsutum and G. barbadense, tetraploid coming from the New World, G. herbaceum and G. arboretum are diploid of the Old World (GRIDI-PAPP, 1965). Brazil is the center of origin for G. mustelinum and an important center of diversity for G. barbadense and G. hirsutum r. maria galante. It has no diploid varieties. According to FREIRE et al. (1990), G. mustelinum, a genuinely Brazilian feral species, was never improved or commercially explored, despite an evident introgression of G. hirsutum alleles in its genome (WENDEL et al. 1994). Its center of origin is the Brazilian Northeast, where some populations may still be found in the municipalities of Caiacó, RN and Caraíba, BA (FREIRE, 2000). In Brazil, the species G. barbadense is widely distributed, in its feral and sub-spontaneous forms, from the Amazon basin to the tropical forest, from Rio Grande do Norte to Espírito Santo, at the Pantanal rim, in the Central West cerrados (MT, MS, GO) and the lowlands of the semiarid Maranhão and Piauí. Two species are found, G. barbadense var. barbadense and “Ox-kidney” G. barbadense var brasiliense, present in indigenous villages and backyards. The barbadense variety was probably introduced from the Caribbean islands, while the brasiliense came from the Amazon forest. The segregation of the two varieties in Brazil is the result of agricultural activities (BOULANGER and PINHEIRO, 1972), where the populations were maintained and cultivated as “local” varieties. Currently, the “modern” varieties of G. barbadense are seldom cultivated in Brazil. Gossypium hirsutum is represented by two biotypes in Brazil. The race latifolium, also known as “upland” cotton, is the main cotton farmed in Brazil. The maria galante race, known as “Mocó” or “arboreal” cotton, was very common in the Northeast during the seventies, and is currently restricted to very few areas. The chains of special, conventional and transgenic cottons have lived together in a satisfactory way, without records of coexistence problems. The cotton planted area in Brazil during the 2007/2008 crop was about a million and one hundred hectares, of which over 85% is concentrated in the Cerrado biome, especially in the states of Mato Grosso, Bahia, Goiás and Mato Grosso do Sul. Other tillage area is present in other Brazilian states, mainly in the Semi-Arid of the Northeast, Paraná, Minas Gerais and São Paulo (IBGE, 2008). Cotton plants are noticeable for their aspects of usefulness, including weaving fibers and oleaginous and proteinaceous seeds used in human and animal food. The species were improved by man since antiquity, both in Old and New Worlds. Cotton (Gossypium spp.) is a plant domesticated by man since the year 3000 B.C., and is cultivated in all continents. Its main use is the production of fibers and food, especially for animals. The cotton plant (Gossypium hirsutum L.) is one of four species cultivated in the world for the production of cotton fiber (PENNA, J.C.V., 2005) and is economically exploited in a wide tropical band and some subtropical regions. Cotton in Brazil is among the ten main agricultural cultures and occupies the sixth place in cultivated surface over the world. The cotton plant is held as a plant very sensitive to interferences caused by plant pests, due to internal aspects (photosynthesis type C3, high photorespiration rate, etc.) and external, such as plant architecture and for displaying very slow initial growth, endowing the plant with a low competitive capacity against invading rivals. With the employment of recombinant DNA technology, the development of a genetically modified cotton lineage, Event GHB614 has the purpose of improving control and management of pest plants in cotton culture through selective use of a glyphosate-based herbicide. The effect of the herbicide is given by inhibiting enzyme EPSPS (aromatic amino acids production pathway), being the active ingredient duly registered in Brazil, including for the cotton culture. III. Description of the GMO and Proteins Expressed GlyTol® - Event GHB614 cotton – was genetically modified by inserting gene 2mepsps, responsible for expressing enzyme 5-enolpyruvylshikimate-3-phosphate synthase (2mEPSPS) with mutation of 2 amino acids. This modified enzyme grants cotton GlyTol® selectiveness to the herbicide effect of the active component glyphosate, enabling therefore the control of pest plants in post-emergence in crops without harming the cotton culture. Event GHB614 was obtained by inserting the gene of interest and elements regulating the genome of the Coker 312 cotton variety. The genetic modification of the Event took place through a system mediated by Agrobacterium tumefasciens containing vector pTEM2. Gene 2mepsps originates from corn (Zea mays) epsps gene changed through a site-directed mutation in just 2 (two) amino acids in the original peptide sequence, resulting in a protein featuring better binding affinity to glyphosate, keeping its functionality (shikimate and synthesis of aromatic amino acids pathway) even under conditions of spraying with the herbicide. The elements present between the left and right borders in vector pTEM2 include, besides gene 2mepsps, those elements regulating the gene expression. Therefore, chimeric gene 2mepsps contains: Ph4a748At promoters and intron 1-h3At; optimized transit peptide – TPotp C – directing the protein to plastids and the signal sequence of polyadenylation 3’ histonAt. Glyphosate [N-(phosphonomethyl)glycine)] is a systemic, post-emergent and non-selective herbicide widely used in agriculture. Glyphosate way of action consists in changing different biochemical processes vital to plants, such as amino acids, proteins and nucleic acids biosynthesis (GLASS, 1984). The herbicide is absorbed by the living tissue and translocated, via phloem, through the plant by roots and rhizomes, and its action inhibits specific enzymes, such as enolpyruvylshikimate-3-phosphate synthase (EPSPS) suspending the synthesis of aromatic amino acids (AMRHEIN et al., 1980; COUTINHO and MAZO, 2005). However, gene 2mepsps, originally isolated from cells suspended in corn (Zea mays) (LEBRUN, et al., 1997), includes substitution of 2 (two) amino acids from its original sequence, resulting in the expression of a protein insensitive to the glyphosate activity. The gene expressing the attribute of selectivity to glyphosate in Event GHB614 is called 2mepsps, originated from gene epsps in corn (Zea mays) with a change through site-directed mutation of just 2 (two) amino acids in its peptide sequence in position 102 (replacing threonine by isoleucine) and position 106 (replacing proline by serine) (LEBRUN et al., 1997), resulting in a protein featuring less binding affinity to glyphosate, keeping its functionality even under conditions of spraying with the herbicide. Protein EPSPS (5-enolpyruvylshikimate-3-phosphate synthase, E.C. 2.5.1.19) is a key enzyme in the shikimate pathway. Though the pathway is present in plants and several microorganisms, it is absolutely absent in mammals, fish, birds, reptiles and insects. These forms of life do not depend on the shikimate pathway, since they rely on their diet to obtain the aromatic products they need. Plants, on the other hand, must produce these essential amino acids to survive and multiply (GRUYS & SIKORSKY, 1999). Glyphosate acts on EPSPS inhibiting the synthesis pathway of the aromatic amino acids such as phenilalanine, tryptophan and tyrosine, which are precursors of other products, such as lignin, alkaloids, flavonoids and benzoic acids (STEINORÜCKEN & AMRHEIN, 1980; BUSSE et al., 2001; ZABLOTOWICZ & REDDY, 2004). GlyTol® cotton expresses enzyme 2mEPSPS that, because of lack of sensitivity to the action of the herbicide, makes maintaining the shikimate pathway possible, therefore enabling full development of the plant, even when sprinkled with such herbicide. IV. Aspects Related to Human and Animal Health The expressed polypeptide chain has molecular mass of 47 kDa and 445 amino acids, highly similar to the corn protein sequence (> 99.5%). Expression of the nuclear gene results in mRNA translated in the cytoplasm, with the polypeptide imported to the cytoplasts interior, where the enzyme plays its role. Glycosilation sites indwell in asparagines 118 and 394. For acute toxicity studies, high quantities of protein are necessary and the identity was directly proven, in SDS-PAGE and by immunologic detection through Western blot of the plant material and the bacteria-expressed material. Chromatographic analysis, mass spectroscopy and N-terminal sequence of the polypeptide displayed identical results. Quantification of protein 2mEPSPS in different tissues of Event GHB614 were conducted through immunoassays (ELISA). Detailed data are shown in items 2.5.3. Below, there is a summary of the findings: a. Amount of 2mEPSPS in cored seeds of GlyTol® cotton: results show that the protein was detected in all fractions related to GM seeds. Over 95% of 2mEPSPS was found in the grain (smooth seed) an also in cored seed. The linter fraction contains less than 0.5% of 2mEPSPS. The protein level of expression varied among different locations and glyphosate treatments. Values ranged from 16.2 µg/g to 30.5 µg/g of fresh matter in GlyTol® samples submitted to application of glyphosate and from 15.8 µg/g to 25.5 µg/g in samples not sprayed with the herbicide representing, on average, 0.0093% and 0.01% of total protein, respectively. b. Tenor of protein 2mEPSPS in different tissues along the crop cycle. Tenor of 2mEPSPS in young leaves decreased along the cycle, coming from 11.16 µg/g + 3.73 to 0.45 + 0.22 µg/g of fresh matter when reaching stage 4. In the stalk, the tenor of protein kept relatively stable between stages 2 and 4, while in roots, 2mEPSPS increased. In general, the contents of protein 2mEPSPS in GlyTol® cotton were higher in leaves during stage 2 (7.94 + 2.87 MF) and lower in pollen (0.16 + 0.0 µg/g). There is a commercial device available for specific immune detection of the EPSPS protein in event GHB614. The detection technique of transgene by means of quantitative PCR is also validated and shall be at the disposal of those interested in case this request for commercial release is granted. Data on centesimal nutritional and compositional assessments of GlyTol® cotton and its equivalent to the non-modified parental, variety Coker312 and to other conventional varieties of cotton are summarized on Table 13 (OBERDORFER, 2010). The following analyses were conducted for this purpose: centesimal composition, fibers, micronutrients (minerals and vitamin E), total antinutrients, gossypol, cyclopropenoids, phytic acid, total amino acids and fatty acids. Cored seeds were used for compositional analyses of GlyTol® cotton (kernel + linter), coming from plants cultivated in 17 places of the United States during the 2005 and 2006 crops. In addition to this material, compositional analyses were carried out in linter, husk, delinted seeds, cotton meal (raw and roasted) and cotton oil (raw, refined and deodorized). Conventional cotton plants Coker312 and GlyTol® received typical cultural treatments with addition of glyphosate in parcels of GlyTol®. Additionally, one parcel of the same cotton was conducted with no use of the glyphosate herbicide. Cultures were conducted in typical conditions to cotton, except for the application of glyphosate, carried out in three parcels containing GlyTol® cotton. For the sake of comparison, 3 parcels of GlyTol® cotton were maintained in the same conditions, without application of glyphosate. Data showed that for the majority of the components examined there was no significant difference identified between GlyTol® cotton and its isoline. In cases where the change did occur, it failed to represent nutritional impact for the following reasons: - values found for GHB614 and coker312 samples were within the range reported for commercial products, especially for cyclopropenoids (malvalic, sterculic and dihydrosterculic fatty acids); - there was no trend in the change found. In the fiber (neutral) analysis, for instance, in some tissues higher values were found in GHB614 and lower in others, compared to Coker312. - differences in nutrient levels were found in just one product. Values measured for myristic fatty acid (C14:0) showed difference between genotypes only when analyzed in refined oil, what was not the case for cored seeds, delinted seeds and raw oil; - differences in nutrient levels were so small that fail to carry any nutritional relevance (variation of 0.1% for fatty acids measured in husk and fibers between genotypes). The mineral contents analyzed in GHB614 and Coker 312 seed was equivalent between genotypes (Table 20), except for Iron and Calcium that, for delinted seeds (Table 21) the tenors were higher in GHB614. Even so, the recorded values are within the average band described by other authors. Variations found for other chemical compounds (nutrients and anti-nutritional factors) were recorded on Tables 21 to 28 and are within the isoline equivalent values. Composition of amino acids analyzed in different parts of GHB614 and Coker 312 are shown in Tables 29 to 32. Samples of cotton meal tested displayed increased values of amino acids, despite the origin of the tested material in the preparation, as against the values found in the literature. The results may be a consequence of the preparation way or a specific characteristics of the parental variety used in the transformation. Differences between tested samples, GHB614 and Coker 312 were not statistically significant, which makes true a statement that they are equivalent regarding amino acid composition. Cotton is seldom consumed by mammals, birds and other species of sylvan animals present in the cultivation area. Non-target organisms, such as predators and preys of cotton pests present in cotton tillage will be exposed to lower levels of the 2mEPSPS protein through trophic transfer. However, there is no evidence of negative effects of the protein over such populations. Same as with GlyTol® cotton, the GHB614 cotton showed to be equivalent to the conventional cotton plant, except for its characteristic of tolerance to glyphosate, its basic interaction with microorganisms present on the environment failed to show differences of conventional cotton interactions. An experiment was conducted with poult during growth. One hundred and forty birds in two subgroups (males and females) with seven repetitions (10 birds each) were tested with commercial cotton, the isoline Coker 312 and event GHB614. The experiment lasted 40 days, when the following parameters were analyzed: animal performance, alimentary conversion and general health characteristics of the bird. There was no alteration that could be attributed to the presence of genetically modified cotton. There was no significant difference regarding mortality or pathological changes in different organs. Protein degradability was tested in a system that simulated human gastric fluid. One in vitro assay was conducted following the methodology described by THOMAS et al. (2004), describing the protocol using pH 1.2, addition of pepsin (Sigma), incubation at 37 ºC for 0.5 to 60 minutes and use of positive and negative controls with proteins peroxidase and ovalbumin. Solutions containing the peptides resulting from the essays were submitted to SDS-PAGE analysis (LAEMMLI, 1970) and stained with Comassie blue (NEUHOFF et al., 1988). Figures 19 and 20 illustrate the gels SDS-PAGE where protein 2mEPSPS was rapidly and completely degraded in such essay conditions. The peroxidase control protein was rapidly digested, while ovalbumin, slowly, as expected (HERUET-GUICHENEY et al., 2009). The simulation is probably a simplified imitation of the condition prevailing in the digestive tube while multiple enzymes act simultaneously and sequentially, leading to complete degradation. The result is an additional guarantee that the intact protein is not absorbed and absence of systemic interference, and already indicates low allergenic potential of such protein. In a acute toxicity assay, when high concentrations of proteins 2mEPSPS and BSA (bovine serum albumin) were administered per os in animals and after 15 days of essay, the individuals were sacrificed and submitted to necropsy for macroscopic analysis. Animals were anesthetized by isoflurane inhaling before the necropsy and later submitted to examination of thoracic and abdominal cavity and their main organs and tissues. The results showed no correlation between abnormalities found for both treatments. The changes are common due to the animal race, age and test conditions, therefore held as spontaneous variations, keeping no relation with effects caused by the 2mEPSPS protein. Besides, the assessment of clinical signs, data on corporeal weight and weight gain of animals coming from the same essay failed to evidence statistically significant differences, indicating that oral administration of highly concentrated 2mEPSPS protein does not result in any evidence of risk to human/animal health due to its use in food (ROUQUIE, 2006). Later, another acute toxicology study was conducted with rodents, injecting up to 10 mg of purified protein directly in the animals’ blood flow. The methodology applied followed the guidelines prescribed by US-EPA Health Effects Test Guidelines OPPTS 870.1100 and OECD Test Guideline 425, using components aprotinin and melitin as negative and positive controls, respectively. Animals treated with 1 mg and 10 mg of 2mEPSPS did not display mortality or symptoms of toxicity even after 15 days from the treatment. At the end of the experiment, when the rodents were submitted to necropsy and macroscopic examination of organs and tissues, no alteration or abnormality was recorded, even in individuals submitted to intravenous 10 mg of 2mEPSPS (ROUQUIE, 2008). In assessing the allergenicity potential, source of genes, historic of exposure, protein digestibility and a comparison with sequences of known allergenic proteins were taken into account. Gene 2mepsps was obtained from sources that are recognized as non causing allergic reactions and the proteins have a long history of safe use, including in other GMOs or derivates already approved for marketing in Brazil. As demonstrated in the scientific literature, a contributing factor towards allergenicity of proteins is its high concentration in food. The majority of allergenic proteins is present in high concentration in specific foods, generally representing from 2% to 3% or even up to 80% of total proteins (ASTWOOD & FUCHS, 1996). Conversely, protein 2mEPSPS is found in extremely low levels in the tissues analyzed, about 100 ng per mg of seed, with percentages ranging from 0.5% to 0.00016% of the plant protein content, depending on plant stadium and part analyzed. Besides, molecules produced by gene 2mepsps are readily degraded when ingested. As it is known, cotton products (kernel oil and short fibers) used for human and animal consumption are highly processed and therefore 2mEPSPS is not detected in such materials. Even considering immediate degradation of protein 2mEPSPS after ingestion and non-detection of such proteins in GlyTol® cotton derived products, existence of possible sequences of amino acids displaying similarities with already described allergens was investigated. Data presented in HEROUET et al. (2009) paper, it remains clear that protein 2mEPSPS displays similarity only with other EPSPS enzymes of different species. Besides, it fails to show any similarity with compounds recognizedly allergenic or toxic, and has no similarity with epitopes and glycolization sites. A result of sequence similarity is held positive when an identity higher than 35% among sequences of amino acids is recorded, or when there is identity in a sequence of 6 (six) contiguous amino acids. This number of amino acids has been a subject of discussion, and a number of 8 (eight) amino acids has been proposed to avoid false positive results (ILSI, 2001). In silico searches were conducted to identify all allergens known to feature an amino acid identity higher than 30% with protein 2mEPSPS in a window of 80 amino acids. A complete sequence of protein 2mEPSPS was compared with AllergenOnline (version 8.0, 1313 sequences). AllegenOnline (http://allergenonline.com) is a list of resources targeted to known allergens and putatives. This allergen databank was specifically developed to predict allergenicity of new proteins by the Food Allergy Research and Resource Program (FAR-RP. The databank is updated yearly through NCBI and International Union of Immunological Societies (IUIS) searches, as well as through assessment at the entry of candidates for evidence of allergenicity. Additions and deletions on the databank are conducted by a revision panel of international experts in allergy to assess whether the proteins are allergens or putative allergens based on predefined criteria. Criteria e references for evidence of allergenicity in groups are supplied on the website. In March, 2009, another in silico search was conducted in an attempt to determine whether protein 2mEPSPS shares the identities of an amino acid with known toxins. The full protein 2mEPSPS amino acid sequence was then contrasted with all sequences of the protein (including potential toxins) present in six wide databanks of public reference data: Unioprot-Swissprot (version 56.7, 2009; 408.099 sequences), Uniprot-trEMBL (version 39.7, 2009; 7,001,017 sequences), PIR (Protein Identification Resources, version 80, 2004; 283,416 sequences), Nri-3D (2007, 56,020 sequences), DAD (DEBJ Amino acid sequence Database, version 44.0, 2008; 2,561,319 sequences), and Genepept (version 169, 2009; 6,185.784 sequences). For all searches, the comparison algorithm used was BLASTP (basic local alignment search tool program for protein-basic protein), maintained by the National Center for Biotechnology Information (NCBI, ALTSCHUL et al., 1997). The BLOSUM62 punctuation matrix enabled comparison of sequences with no less than 62% divergence (HENIKOFF and HENIKOFF, 1998). The search for putative sites for N-glycosylation of protein 2mEPSPS was also examined based on its consensus known sequence. The sequences were Asparagine-Xaa-Serine/Treonin (where Xaa represents any amino acid, except Proline) and Asparagine-Xaa-Cysteine. The algorithm used to conduct analyses was the FindPatterns of the integrated software CGC. The final result of similarity BLAST, conducted in 2009, failed to show evidence of similarity between protein 2mEPSPS and any known allergen protein. The epitope similarity FindPatterns Analysis confirmed that there was no evidence of similarity with allergens, therefore featuring no identity with eight contiguous amino acids with known allergens, using the AllergenOnline databank. Results also showed lack of similarity withy known toxins, or any functional homology between the 2mEPSPS and known toxins. As expected, the BLAST results showed that protein 2mEPSPS displays high structural similarity only with other EPSPS proteins, including corn protein (>99%). The high similarity of 2mEPSPS proteins and the sylvan EPSPS indicates a safety profile. Finally, putative locals of N-glycosylation present in the sequence of 2mEPSPS, in positions 118 and 394, were identical to the locals of the EPSPS protein in corn, enabling a conclusion that the two putative locals fail do display safety problems (HEROUET et al., 2009). Summarizing, date submitted by Applicant support the conclusion that protein 2mEPSPS, under the conditions prevailing in derivatives of GlyTol® cotton fails to pose allergenicity risks to humans and animals higher than the threats posed by non-transgenic varieties. V. Environmental Aspects According to data surveyed by ABRAPA – Associação Brasileira de Produtores de Algodão (Brazilian Cotton Producers Association), Brazil cultivated 1.07 million hectares on the 2008/2009 crop, of which 300 thousand hectares were sowed with genetically modified cotton (SILVEIRA et al., 2009). The expectations of ABRAPA for the 2009/2010 crop is a reduction of about 23%, to 850 thousand hectares, with a production of 1.2 million of tons of cotton lint. Experimental results in Brazil generated by essays authorized by CTNBio and MAPA showed a significant reduction in the use of herbicides in tillage, which simplifies the system of pest plant control when compared with conventional cotton. An example of the high dissemination of the product are the adhesion rate to the technology attained in the United States and Australia: In 2006, nine years after the first commercial plantation of glyphosate tolerant cotton (Event MON1445), American farms showed that 65% of the planted area in the country was already using such Event. In Australia, the first commercial area was sowed in 2000 and, in just six years, the area so planted represented 75% of the cotton planted area in that country. These countries that feature, therefore, a long history of use in the environment, there is no record of adverse effects of environment degradation as an effect of the technology that enables the management of pest plants with glyphosate. In another study, CHIAVEGATO (2009b) assessed the Event response to different doses of the herbicide in different growth phases, making possible to evaluate whether the level of expression of gene 2mepsps was sufficient to protect the plant, as well as to assess the specificity of the gene to grant selectivity just to the glyphosate herbicide. In Annex II of the process, there is a complete description of the protocols and treatments used in the essays conducted in Brazil. The results obtained by analyzing phenologic data indicated that plants derived from Event GHB614 showed tolerant go glyphosate, including high doses of the herbicide, since the parameters assessed failed to statistically differ from data analyzed in plants managed without the herbicide. This indicates that protein 2mEPSPS was expressed in a level sufficient to grant due protection to the plant enabling maintenance of its growth and normal development. When the plants were submitted to the herbicide ammonium gluphosinate with the purpose of assessing specificity of the Event to selectivity of just the active ingredient glyphosate, both GM (GlyTol®) and nGM cotton varieties showed to be highly sensitive, leading to total injury of plants 14 days after application, showing that gene 2mepsps acts solely in glyphosate tolerance, with no additional survival or adaptive advantage attribute regarding GlyTol® cotton. The test demonstrated, besides that, should the need for eradication of Event GlyTol® derivative plants be the case, the use of other non-selective herbicides to the culture keeps being efficient for this purpose. According to FREIRE (2000), the likelihood of gene flow between GM and feral cottons is remote given the isolation of spatial distribution foreseen for commercial cultivars (distributed in high-technology tillage areas in the cerrado) in locations knowingly exempt from sylvan types. In case the transfer does take place, the adaptive advantage represented by a tolerance to a specific herbicide shall be null, since the cotton plants are cultivated in small areas and manually weeded. Besides, no biochemical/physiological change affecting the ability of natural survival of plants in the environment was detected. The new attribute that enables selective use of a wide ranged herbicide in post-emergence fails to insert any competitive or adaptive advantage to the individual, even in stress situations, such as pathogen infestations or competition with pest plants. The differentiated phenotype of plants expressing gene 2mepsps is only perceived under invading plants management conditions where the glyphosate herbicide has been used. Finally, under Brazilian conditions, the cotton cultivation regions fail to display any pest plat that may be sexually compatible with the cultivated species of Gossypium. Apparently, the possibilities of crossing between cotton and other cultivated Malvaceous species, (such as okra, Hibiscus sculentus, or Hibiscus cannabinus and the vinegar plant (Hibiscus sabdariffa) are quite remote. A significant change is not expected in organisms present in the farm ecosystem where cultivation of GlyTol® cotton is foreseen, since this GMO is only differentiated from the conventional parental for the presence of gene 2mepsps, which acts in the selectivity to glyphosate and fails to display any pleiotropic or epistatic effect. Protein 2mEPSPS expressed in Event GHB614 originates from the EPSPS of corn, containing two changes in its amino acid sequence: substitution of one threonine for a isoleucine in position 103, and proline for a serine in position 107. Even so, protein 2mEPSPS has 99.5% of homology with the corn EPSPS , 86% with rice, 79% with grape, 77% with rice and 75% with tomato (HEROUET-GUICHENEY et al., 2009). Up to now, no bibliographic reference suggests any adverse effects in simbionts, predators, pollinators, parasites or competitors caused by the EPSPS protein. Organisms interacting with cotton culture, such as pests, rodents and birds, may sustain some damage due to presence of the gossypol compound (BELL, 1986; ABOU-DONIA, 1989) that occurs in similar levels both in GlyTol® and its conventional parental, therefore keeping no relation with the genetic modification on screen. In Brazil, one of the treatments applied to GlyTol® cotton and its conventional isoline, cultivated in the environment, was biotic stress, where no pests and diseases were controlled along the cultivation cycle. Data collected and presented by CHIAVEGATO (2009a) show that pests, such as cotton worm, Spodoptera, cotton bollworm, aphis and whitefly, and phytopathogens ramularia, ramulose, and blue disease (virosis) infested both GlyTol® and the non modified lineage. This is an indication that no species of pest or phytopathogen displayed differential preference for any of the genotypes or had favored increase of its population. CURRIER (2006) made a study comparing morphology, viability and survival rate of cotton pollen of GlyTol® (genetically modified) and Coker 312 (conventional) genotypes. To this end, 14 plants derived from Event GHB614 and 10 coming from the conventional parental were cultivated in vases inside a vegetation house in the Bayer Cropscience premises (Research Triangle Park, Raleigh, NC, USA). Pollen samples were collected by hand and transferred to specific environments in order to conduct the study. Viability was measured by a method described by BARROW (1981), which analyzes the growth of initial structures of the pollinic tube in the microscope. This analysis was repeated for three days. Results showed that initial viability of GHB614 and Coker 312 cotton pollens exceeded 90%. Decrease of viability revealed a kinetics along of a similar term for the genotypes, without statistical differences. Based on the data and the bibliographic references, it is not likely that GlyTol® cotton has greater reproduction and survival ability in any environment that is in any way different from already marketed cultivars. Adaptive advantage of Event GHB614 is recorded only when the culture is sprayed with the glyphosate herbicide, where plants containing 2mepsps maintain their normal development, with no injuries or phytotoxicity. The use of GlyTol® cotton, event GHB614 has the purpose of improving the management of pest plants in the tillage of corn in post-emergence, using a herbicide that features wide spectrum of action. This characteristics of the Event does not result in tolerance to any environmental biotic element and therefore there is no organism that is a target of the technology. Invading plants normally occurring in cotton production areas that have caused significant damage and/or have reduced the intrinsic quality of the product harvested are: Bidens pilosa (Spanish needle), Cenchrus Echinatus (grass-bun), Ipomoea ssp. (morning glory) and perennial Malvaceae (Sida ssp.). Weed infestation in cotton culture results also in increased damages caused by pests and diseases that are hosts of such weeds; increase dispersion of invaders’ seeds; multiply the need for cultivation and chemical control practices and reduce soil structure and humidity due to the increased soil preparation (harrowing) in pre-sowing to reduce initial infestation. Use of minimum cultivation or direct sowing have been important measures to soil conservation in areas featuring greater risk of erosion. This way, control of pest plants through the use of glyphosate in post-emergence shall probably be an important component for systems aimed at better soil conservation, because of the reduced movement in pre-sowing and increased soil structure due to maintenance of roots and plant residues (FAWCETT & TOWERY, 2004; SANKULA, 2006). In field studies conducted abroad, Applicant’s technicians have frequently visited experimental areas to collect culture management data, and recorded infestation by pests, diseases and presence of natural enemies. In no case variations regarding increase or reduction of pests, diseases, pollinating insects, predators and any other beneficial organism were recorded in the experiments, when contrasted to the cultivation of conventional corn (Van DUYN, 2007). Results of an environmental study conducted by CHIAVEGATO (2009a) revealed that no pest species or phytopathogen had any differential preference for one of the genotypes (VGM or NM) nor has favored the population increase. One of the risk hypothesis attached to commercial release of a genetically modified culture, there is one according to which these VGM may turn into weed, with greater ability of surviving and invading natural habitats and, consequently, endanger their biodiversity. Therefore, it is important to assess whether the GM genotypes exhibit differentiated phenotypic characteristics that may make them more invasive (WILLIAMSON, 1993). Some important attributes of pest plants, such as dormancy, phenotypic plasticity, indeterminate growth, continuous flowering and ease seed dispersion (BAKER, ‘974) nave been eliminated from the majority of agriculture cultures along generations during genetic improvement. These characters are not pursued for gene transfer in cultivars, either by biotechnology (recombinant DNA) or by classical improvement, since such attributes drastically reduce the agronomic performance of modern varieties. Besides, these characteristics generally are commanded by different genes. CRAWLEY et al., (2001) discuss the results obtained in a long term study assessing the performance of GM cultures in natural habitats. Four cultures (canola, potato, corn and beet) showing the additional attributes of tolerance to herbicide or resistance to insects, were cultivated in 12 different habitats and monitored for 10 years. There was no record of in which the genetically modified plants were more invasive or more persistent than their conventional parental ones. One did not expect these additional characteristics of Events could increase the development of plants in such habitats. On the other hand, these results cannot discard the hypothesis that other genetic modifications could change the invading ability of a VGM, but also failed to indicated that such plants could survive for long periods of time outside culture cultivation conditions. LEMEAUX, (2009), in an extensive revision where he analyzed scientific publications related to risk analysis of genetically modified products, concluded that though no human activity could guaranty 100% safety, genetically modified cultivars and their products currently available in the market are as safe as those resulting from conventional methods. VI. Restrictions to the Use of the GMO and GMO Derivatives Technical Opinions related to agronomic performance concluded that there is equivalence between transgenic and conventional plants. Therefore, information indicates that transgenic plants are not fundamentally different from non-transformed cotton genotypes, except for the tolerance to the glyphosate herbicide. Besides, there is no evidence of adverse reactions to the use of GlyTol® cotton event GHB614. For these reasons, there are no restrictions to the use of such cotton or its derivatives in both human and animal food. According to Article 1 of Law no. 11,460, of March 21, 2007, “research and cultivation of genetically modified organisms are forbidden in indigenous and conservation unit areas”. VII. Considerations on Particulars from Different Regions of the Country (Subsidies to Monitoring Bodies) There are not Creole varieties of cotton and chains of special, conventional and transgenic cottons have lived side-by-side in a satisfactory way, with no record of coexistence problems. Zones where cultivating GM cotton, either of this event or other transformation, is restricted shall be strictly monitored by supervising bodies, both in the trade of seeds in such areas and in technical guidance and monitoring of the applying legal entity. Such areas are located in the south of Rio Grande do Norte and the Northeast of Bahia, all of the Amazon region, Pantanal, southeast of Piaui, west of Pernambuco and Atlantic Forest, encompassing the states of RN, PB, AL, SE, BA, MG and ES. VIII. Conclusion Whereas 1. Cotton event GHB614 is as safe for human and animal consumption as its conventional equivalent. 2. Protein 2mEPSPS has no homology with toxic or allergenic substance and has no glycosilation sites additional to those already existent in corn EPSPS. 3. Protein 2mEPSPS is rapidly degraded in the presence of digestive tract enzymes. 4. No adverse effect was recorded in acute toxicology studies and animal food containing protein 2mEPSPS. 5. Plants derived from Event GHB614 have the same survival and adaptation ability regarding the environment than conventional cotton. 6. Agronomic characteristics of GlyTol® cotton are only differentiated by selectivity to glyphosate, when compared to conventional cotton varieties. 7. The use of cotton and derivatives produced from GlyTol® cotton (Event GHB614) imply the same safety to human and animal health and to the environment as commercial conventional lineages, and even as genetically modified varieties, already approved and marketed, displaying selectivity to glyphosate. 8. The use of kernels or byproducts derived from Event GHB614, following the same recommendations and safety criteria already used for commercial cotton varieties, fail to represent greater exposure to any risk situation. CTNBio does not hold this activity a potential cause of significant degradation to the environment or hazardous to human and animal health. Restrictions to the use of the GMO in analysis and its derivatives are not conditioned to the provisions of the Ministry of Agriculture and Supply Directive 21/05. Under the provisions of Article 14 of Law nº 11,105/05, CTNBio holds that the request is in line with the legislation in force that guaranty environment, agriculture, human and animal health biosafety, and concluded that Cotton GHB614 (GlyTol®) is substantially equivalent to conventional cotton and that its consumption is safe for human and animal health. Regarding the environment, CTNBio concluded that Cotton GHB614 (GlyTol®) is not a potential cause of significant degradation to the environment, keeping with the biota a relation identical to that of the conventional cotton. Regarding the post-commercial release monitoring plan, CTNBio determines that the following instructions shall be attended and conducted the monitoring techniques mentioned below. (a) Monitoring must be conducted in commercial cultures and not experimental ones. Areas selected for monitoring shall not be separated from the others, be fenced or display any condition that may be a non-commercial standard. (b) Monitoring must be conducted in a model comparative between conventional cultivation and GMO cultivation systems, where data collection shall be made by sampling. (c) Monitoring must be conducted in biomes that are representative of the main GMO culture areas and, whenever possible, include different type of producers. (d) Monitoring must be conducted for a period of at least five years. (e) For all monitoring procedures, the applicant shall detail the data on all activities conducted in pre-sowing and sowing, on its performance, reporting all activities carried out in the monitoring area during the culture cycle, in harvesting activities and climatic conditions. (f) Any hazard to human and animal health shall be monitored through official adverse effects notification systems, such as SINEPS – Sistema de Notificação de Eventos Aversos Relacionados a Produtos de Saúde the Adverse Effects Related to Health Products Notification System as regulated by ANVISA. (g) Analytical methods, results attained and their interpretation must be developed in line with independence and transparence principles, except for commercial secrecy aspects previously justified and defined as such. (h) On technical and scientific grounds, CTNBio reserves the right to review this Opinion at any time. Technical Monitoring Actions to be Conducted (a) Nutritional state and sanity of GMO plants. (b) Chemical and physical soil attributes related to fertility and other basic pedologic characteristics. (c) Soil microbial diversity. (d) Soil dispersion bank. (e) Community of invading plants. (f) Development of resistance to herbicide in invading plants. (g) Residues of herbicide in the soil, kernels and aerial part. (h) Gene flow IX. Bibliographic References ABOU-DONIA, M. B. 1989. Gossypol. In: P. R. Cheeke (ed.) Toxicants of Plant Origin. vol. 4:Phenolics. pp 1–22. CRC Press, Boca Raton, FL. ALTSCHUL, S.F., MADDEN, T.L., SCHAFFER, A.A., ZHANG, J., ZHANG, Z., MILLER, W., LIPMAN, D.J., 1997. Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res. 25, 3389–3402. AMRHEIN, N.; DEUS, B.; GEHRKE, P.; STEINRUCKEN, H.C. The site of the inhibition of the shikimate pathway by glyphosate.II – Interference of glyphosate with chorismate formation in vivo and in vitro. Plant Physiol., v.66, p.830-834, 1980. ASTWOOD JD, FUCHS RL. Allergenicity of foods derived from transgenic plants. Monogr Allergy. 1996;32:105-20. BAKER, H.G. The evolution of weeds. Annu. Rev. Ecol. Syst. 5, 1–23. 1974. BATISTA R, SAIBO N, LOURENÇO T, OLIVEIRA MM (2008) Microarray analyses reveal that plant mutagenesis may induce more transcriptomic changes than transgene insertion. Proc Natl Acad Sci U S A. 105:3640-3645. BELL, A.A. 1986. Physiology of Secondary Products. Chapt. 38. Pages 597-621 in Cotton Physiology. J.R. Mauney and J. McD. Stewart, ed. The Cotton Foundation, Memphis, TN. BOULANGER J.; PINHEIRO D. 1972. Conseqüências genéticas da evolução da cultura algodoeira do Nordeste do Brasil. Pesquisas Agropecuárias no Nordeste, v.4, n.1, p.5-52. BUSSE, M. D. et al. Glyphosate toxicity and the effects of long-term vegetation control and soil on soil microbial communities. Soil Biology and Biochemistry, v. 33, p. 1777-1789, 2001. CANADIAN FOOD INSPECTION AGENCY, Animal Feed Division. Decision Document DD2008-72 Determination of the Safety of Bayer CropScience's GlyTol™ Cotton Event GHB614. CERA Center for Environmental Risk Assessement - http://cera-gmc.org/index.php?evidcode= GHB614&hstIDXCode=&gType=&AbbrCode=&atCode=&stCode=&coIDCode=&action=gm_crop_database&mode=Submit. Consulted on 11/12/2010. CERNY, R.E. et al. Development and characterization of a cotton (Gossypium hirsutum L.) event with enhanced reproductive resistance to glyphosate. Crop Sci. v.50, p. 1375–1384, 2010. CHIAVEGATO, E. J. Avaliação agronômica e de biossegurança do algodão Evento GHB614 – GlyTol. Relatório Técnico. Bayer Cropscience. 44p. 2009a. CHIAVEGATO, E. J. Avaliação da tecnologia e aspectos agronômicos do algodão Evento GHB614. Relatório Técnico. Bayer Cropscience. 39p. 2009b. COUTINHO, C.F.B.; MAZO, L.H. Complexos metálicos com herbicida glifosato: Revisão. Quim. Nova, Vol. 28, No. 6, 1038-1045, 2005. CRAVEN, L.A.; STEWART, J. MCD; BROWN, A.H.D.; GRACE, J.P. The Australian wild species of Gossypium. In: Proceedings of the world cotton research conference, 1. Brisbane, Australia. Challenging the future. P. 278-281. 1994. CRAWLEY, M.J.; BROWN, S.L.; HAILS, R.S.; KOHN, D.D.; REES, M. Transgenic crops in natural habitats. Nature, v.409, n.8, p.382-683, 2001. CURRIER, T. C. Pollen morphology, viability and survival rate of 2mEPSPS cotton event GHB614and its non-transgenic counterpart Coker 312. Bayer Cropscience. Internal Report. 27p. 2006. M-274210-01-1. EFSA European Food Safety Authority. EFSA Application: Part II Summary. EFSA European Food Safety Authority. Scientific Opinion: Application (Reference EFSA-GMO-NL-2008-51) for the placing on the market of glyphosate tolerant genetically modified cotton GHB614, for food and feed uses, import and processing under Regulation (EC) No 1829/2003 from Bayer CropScience. FAWCETT, R., TOWERY, D. 2004. Conservation Tillage and Plant Biotechnology: How New Technologies Can Improve the Environment By Reducing the Need to Plow. Conservation Technology Information Center. 1220 Potter Drive, Suite 170, West Lafayette, IN 47906. www.ctic.purdue.edu. 24 pages. #M-291021-01-1. FEDOROFF N E BROWN NM (2004) Mendel in the kitchen – a scientist’s view of genetically modified foods – Joseph Henry Press – Washington, DC. FOOD STANDARDS AUSTRALIA NEW ZEALAND. Final Assessment Report Application A614: Food Derived from Glyphosate-Tolerant Cotton Line GHB614. FREIRE, E.C.. Distribuição, coleta, uso e preservação das espécies silvestres de algodão no Brasil. Embrapa- CNPA. Documentos, 78. Campina Grande. 22p. 2000. FREIRE, E.C.; MOREIRA, J.A.N.; MIRANDA, A.R.; PERCIVAL, A.E. E STEWART, J.M. Identificação de novos sítios de ocorrência de Gossypium mustelinum no Brasil. Research in Progress, 10, 7p. 1990. FRYXELL, P.A., CRAVEN, L.A. E STEWART, J.MCD. 1992. A revision of Gossypium Sect. Grandicalyx (Malvaceae) including the description of six new species. Systematic Botany, v.17, n.1, p.91-114. FRYXELL, P.A. 1979. The natural history of the cotton Tribe Malvaceae (Tribe Gossypieae). Texas A&M University Press, College Station. FUNKE T, YANG Y, HAN H, HEALY-FRIED M, OLESEN S, BECKER A, SCHONBRUNN E (2009) Structural basis of glyphosate resistance resulting from the double mutation Thr97 Ile and Pro101 Ser in 5-Enolpyruvylshikimate -3-phosphate synthase from Escherichia coli. J. Biol. Chem. 284:9854-9860. GRIDI- PAPP, I.L. Botânica e genética. In: Instituto Brasileiro de Potassa. Cultura e adubação do algodoeiro. São Paulo. P.117-160. 1965. GRUYS, K. J.; SIKORSKI, J. A.; Inhibitors of Tryptophan, Phenylalanine and Tyrosine Biosynthesis as Herbicides, Dekker: New York, 1999. HAMMOND, B.G. et al. The feeding value of soybeans fed to rats, chickens, catfish and dairy cattle is not altered by genetic incorporation of glyphosate tolerance. J.Nutr., v.126, n.3, p.717-727, 1996. HARRIGAN, G.G. et al. Natural variation in crop composition and the impact of transgenesis. Nature Biotechnol., v. 28(5), 402-404, 2010. HARRISON, L.A. et al. The expressed protein in glyphosate synthase from Agrobacterium sp. Strain CP4, is rapidly digested in vitro and is not toxic to acutely gavaged mice. J.Nutr., v.126, n.3, p.728-740, 1996. HENIKOFF, S., HENIKOFF, J.G., 1992. Amino acid substitution matrices from protein blocks. Proc. Natl. Acad. Sci. USA 89, 10915–10919. HEROUET-GUICHENEY, C.; ROUQUIE, D.; FREYSSINET, M.; CURRIER, T.; MARTONE, A.; ZHOU, J.; BATES, E.E.M.; FERULLO, J.M.; HENDRICKX, K. ROUAN, D. Safety evaluation of the doublé mutant 5-enol pyruvylshikimate-3-phosphate synthase (2mEPSPS) from maize that confers tolerance to glyphosate herbicide in transgenic plants. Reg. Tox.Pharmacology, v.54, p.143-153, 2009. HEROUET-GUICHENEY, C.; ROUQUIE, D.; FREYSSINET, M.; CURRIER, T.; MARTONE, A.; ZHOU, J.; BATES, E.E.M.; FERULLO, J.M.; HENDRICKX, K. ROUAN, D. Safety evaluation of the doublé mutant 5-enol pyruvylshikimate-3-phosphate synthase (2mEPSPS) from maize that confers tolerance to glyphosate herbicide in transgenic plants. Reg. Tox.Pharmacology, v.54, p.143-153, 2009. HERRMANN, K.M. the shikimate pathway as an entry to aromatic secondary metabolism. Plant Physiol., v.107, p.7-12, 1995. HERRMANN, K.M.; WEAVER, L.M. The shikimate pathway. Ann. Rev. Plant Physiol Plant Mol. Bio. v.50, p.473-503, 1999. HORAK, M.J. et al. Characterization of Roundup Ready flex cotton, ‘MON 88913’, for use in ecological risk assessment: evaluation of seed germination, vegetative and reproductive growth, and ecological interactions. Crop Sci. v.47, p.268–277, 2007. ILSI. Allergenicity assessment for foods derive from genetically modified crops. Washington, DC, 2001, 17p. (HESI Codex comments, July 19, 2001). Instituto Brasileiro de Geografia e Estatística, IBGE, 2008. LAEMMLI, U.K., 1970. Cleavage of structural proteins during assembly of the head of the bacteriophage T4. Nature 227, 680–685. LEBRUN M., SAILLAND A., Freyssinet G. 1997. Mutant 5-enol pyruvylshikimate-3-phosphate synthase, gene encoding for said protein and transformed plants containing said gene. International patent publication W0 97/04103-A2. 06.02.97. 25 pages LEE, J.A Cotton as a world crop. In: RHOEL, R.J.; LEWIS, C.F. (eds). Cotton. Madison: American Society of Agronomy p.1-16. 1984. LEMAUX, P.G. Genetically Engineered plants and foods: a scientist´s analysis of the issues (part II). Annu. Rev. Plant Biol. V.60, p.511-559, 2009. LI, M. et al. Evaluation of cottonseed meal derived from genetically modified cotton as feed ingredients for channel catfish, Ictalurus punctatus. Aquacult. Nutr., v. 14, n. 6, 2008. MUNRO, J.M. 1987. Cotton. 2nd Ed. John Wiley & Sons, New York, NY. NEUHOFF, V., AROLD, N., TAUBE, D., EHRHARDT, W., 1988. Improved staining of proteins in polyacrylamide gels including isoelectric focusing gels with clear background at nanogram sensitivity using Coomassie Brilliant Blue G-250 and R-250. Electrophoresis 9, 255–262. OBERDORFER, R. Nutritional Impact Assessment reporto n glyphosate tolerant cotton transformation Event GHB614. Bayer Cropscience. Internal Report, 97p. 2010. M-289161-04-1. OECD. 2002. Task Force for the Safety of Novel Foods and Feeds. Draft Consideration for the Safety Assessment of Animal Feedstuffs Derived from Genetically Modified Plants. ENV/JM/FOOD(2001)8/REV1. France. PENNA, J.C.V. Melhoramento de algodão. Inc: Melhoramento de espécies cultivadas. Borém, A (Ed.) Viçosa; Ed. UFV.p. 15-53.2005.969p. ROUQIE, D. 2mEPSPS protein. Acute toxicity by intravenous injection in mice. Internal report, Bayer CropScience, 51p. 2008. ROUQUIE, D. 2mEPSPS protein. Acute toxicity by oral gavage in mice. Internal Report. Bayer CropScience, 64p. 2006. SANKULA S. 2006. Quantification of the impacts on US agriculture of biotechnology-derived crops planted in 2005. National Center for Food and Agricultural Policy. Washington, DC 20036. 110 pages. www.ncfap.org. # M-291016-01-1. SILVEIRA, D. et al. Fibras são saudáveis. In: Anuário brasileiro do algodão 2009. Ed. Gazeta Santa Cruz, 2009, 128p. STEINRÜCKEN H.C., AMRHEIN N. 1980. The herbicide glyphosate is a potent inhibitor of 5- enolpyruvylshikimic acid-3-phosphate synthase. Biochemical and Biophysical Research Communications. 94(4): 1207-1212. THOMAS, K., AALBERS, M., BANNON, G.A., BARTELS, M., DEARMAN, R.J., ESDAILE, D.J., FU, T.J., GLATT, C.M., HADFIELD, N., HATZOS, C., HEFLE, S.L., HEYLINGS, J.R., GOODMAN, R.E., HENRY, B., HEROUET, C., HOLSAPPLE, M., LADICS, G.S., LANDRY, T.D., MACINTOSH, S.C., RICE, E.A., PRIVALLE, L.S., STEINER, H.Y., TESHIMA, R., THOMAS, K., VAN REE, R., WOOLHISER, M., ZAWODNY, J., 2004. A multi-laboratory evaluation of a common in vitro pepsin digestion assay protocol used in assessing the safety of novel proteins. Regul. Toxicol. Pharmacol. 39, 87–98. USDA - U.S. Department of Agriculture, Animal and Plant Health Inspection Service. Petition for Determination of Non-regulated Status for Glyphosate-Tolerant cotton: GlyTol™ cotton Event GHB614. USDA - U.S. Department of Agriculture, Animal and Plant Health Inspection Service Federal Register Notification: Bayer CropScience; Determination of Non-regulated Status for Cotton Genetically Engineered for Glyphosate Herbicide Tolerance. U.S. E.P.A. (United States Environmental Protection Agency), 1998. Prevention, Pesticides and Toxic Substances (7101), Health Effects Test Guidelines OPPTS 870.1100, Acute Oral Toxicology, EPA 712-C-98-190, December 2002, 35 pages. Van DUYN, G. USDA Field termination reports for glyphosate tolerant cotton, 32p. 2007. WENDEL, J.F.; ROWLEY, R.; STEWART, J.M. Genetic diverstiy in and phylogenetic relationships of the Brazilian endemic cotton, Gossypium mustelinum (malvaceae). Plant Systematics and Evolution, v.192, p,49-59, 1994. WILLIAMSON, M. Invaders, weeds and the risks from GMOs. Experientia, 49, 219–224. 1993 ZABLOTOWICZ, R. M.; REDDY, K. N. Impact of glyphosate on the Bradyrhizobium japonicum symbiosis with glyphosate-resistant transgenic soybean: a minireview. J.Environ. Qual., v.33, n. 3, p.825-831, 2004. ZABLOTOWICZ, R. M.; REDDY, K. N. Impact of glyphosate on the Bradyrhizobium japonicum symbiosis with glyphosate-resistant transgenic soybean: a minireview. J.Environ. Qual., v.33, n.3, p.825-831, 2004. ZHANG, J. et al. Transgene integration and organization in Cotton (Gossypium hirsutum L.) genome. Transgenic Research, v. 17, n. 2, p. 293-306, 2008.
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The Brazilian National Biosafety Commission – CTNBio , is responsible to the technical decision on biological risk as a response to a request from the proponent. The technical decision is given on a definitive basis. Only the National Biosafety Council (CNBS) can revoke the decision (in case of commercial release), based on social-economical reasons and not on biosafety reasons. Once a decision is taken by CTNBio favorable to the commercial release of a new GMO (being it a plant or any other organism), CNBS has 30 days to issue a revoke. After these steps, the new product must be evaluated for conformity to the Brazilian standards by the registration and enforcement agencies (ANVISA – Ministry of Health, Ministry of Agriculture, Ministry of Environment and Ministry of Fisheries, according to the intended use of the product). If it conforms to the standards, it may be offered to the market. Every institution dealing with GMOs (including universities and public research institutes) has to have an Internal Biosafety Commission (CIBio), which is legally responsible of everything that may happen to be done or caused by the GMO
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At the discretion of, and upon consultation with, CTNBio, a new analysis and issuance of technical opinion may be released on GMOs containing more than one event, combined through classic genetic improvement and which have been previously approved for commercial release by CTNBio
Contact details of the competent authority(s) responsible for the safety assessment and the product applicant:
Dr. Edivaldo Domingues Velini (President of national Biosafety Commission)
United States of America
Name of product applicant: Bayer CropScience USA LP
Summary of application:
Cotton
Trait 1 Added Protein: Double-mutant 5-Enolpyruvylshikimate 3-phosphate synthase (2mEPSPS)
Source: Zea mays
Intended Effect: Tolerance to the herbicide glyphosate
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Date of authorization: 29/09/2008
Scope of authorization: Food and feed
Links to the information on the same product in other databases maintained by relevant international organizations, as appropriate. (We recommend providing links to only those databases to which your country has officially contributed.):
Summary of the safety assessment:
Please consult the FDA website links below.
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Where detection method protocols and appropriate reference material (non-viable, or in certain circumstances, viable) suitable for low-level situation may be obtained:
Relevant links to documents and information prepared by the competent authority responsible for the safety assessment: FDA's webpage regarding this variety
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Authorization expiration date:
E-mail:
jason.dietz@fda.hhs.gov
Organization/agency name (Full name):
Food and Drug Administration
Contact person name:
Jason Dietz
Website:
Physical full address:
5100 Paint Branch Parkway, College Park MD 20740
Phone number:
240-402-2282
Fax number:
Country introduction:
The United States is currently in the process of populating this database. The Food and Drug Administration regulates food and feed (food for humans and animals) from genetically engineered crops in conjunction with the Environmental Protection Agency (EPA). EPA regulates pesticides, including those that are plant incorporated protectants genetically engineered into food crops, to make sure that pesticide residues are safe for human and animal consumption and do not pose unreasonable risks of harm to human health or the environment. FDA In the Federal Register of May 29, 1992 (57 FR 22984), FDA published its "Statement of Policy: Foods Derived from New Plant Varieties" (the 1992 policy). The 1992 policy clarified the agency's interpretation of the application of the Federal Food, Drug, and Cosmetic Act with respect to human and animal foods derived from new plant varieties and provided guidance to industry on scientific and regulatory issues related to these foods. The 1992 policy applied to all foods derived from all new plant varieties, including varieties that are developed using genetic engineering (also known as recombinant deoxyribonucleic acid (rDNA) technology). In the 1992 policy, FDA recommended that developers consult with FDA about foods from genetically engineered plants under development and developers have routinely done so. In June 1996, FDA provided additional guidance to industry on procedures for these consultations (the consultation procedures). These procedures describe a process in which a developer who intends to commercialize food from a genetically engineered plant meets with the agency to identify and discuss relevant safety, nutritional, or other regulatory issues regarding the genetically engineered food and then submits to FDA a summary of its scientific and regulatory assessment of the food. FDA evaluates the submission and if FDA has questions about the summary provided, it requests clarification from the developer. At the conclusion of the consultation FDA responds to the developer by letter. The approach to the safety assessment of genetically engineered food recommended by FDA during consultations, including data and information evaluated, is consistent with that described in the Codex Alimentarius Guideline for the Conduct of Food Safety Assessment of Foods Derived from Recombinant-DNA Plants. EPA The safe use of pesticidal substances is regulated by EPA. Food from a genetically engineered plant that is the subject of a consultation with FDA may contain an introduced pesticidal substance, also known as a plant-incorporated protectant (PIP), that is subject to food (food for humans and animals) safety and environmental review by EPA. PIPs are pesticidal substances produced by plants and the genetic material necessary for the plant to produce the substance. Both the PIP protein and its genetic material are regulated by EPA. When assessing the potential risks of PIPs, EPA requires studies examining numerous factors, such as risks to human health, non-target organisms and the environment, potential for gene flow, and insect resistance management plans, if needed. In regulating PIPs, decisions are based on scientific standards and input from academia, industry, other Federal agencies, and the public. Before the first PIP product was registered in 1995, EPA required that PIP products be thoroughly tested against human safety standards before they were used on human food and livestock feed crops. EPA scientists assessed a wide variety of potential effects associated with the use of PIPs, including toxicity, and allergenicity. These potential effects were evaluated in light of the public's potential exposures to these pesticides, taking into account all potential combined sources of the exposure (food, drinking water, etc.) to determine the likelihood that a person exposed at these levels would be predisposed to a health risk. Based on its reviews of the scientific studies and often peer reviews by the Federal Insecticide, Fungicide and Rodenticide Scientific Advisory Panel, EPA determined that these genetically engineered PIP products, when used in accordance with approved label directions and use restrictions, would not pose unreasonable risk to human health and the environment during their time-limited registration.
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Relevant documents
Stacked events:
Contact details of the competent authority(s) responsible for the safety assessment and the product applicant:
Food and Drug Administration (premarkt@fda.hhs.gov); Environmental Protection Agency