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OECD Unique Identifier details

MON-89Ø34-3
Commodity: Corn / Maize
Traits: Lepidoptera resistance
Australia
Name of product applicant: Monsanto Australia Ltd
Summary of application:
MON 89034 corn is protected against a range of lepidopteran insect larvae including
European corn borer, Asian corn borer, southwestern corn borer, sugarcane borer, fall
armyworm and corn earworm.
Protection is achieved through expression in the plant of two insecticidal Cry proteins,
Cry1A.105 and Cry2Ab2, derived from Bacillus thuringiensis, a common soil bacterium.
Cry1A.105, encoded by the cry1A.105 gene, is a chimeric protein made up of different
functional domains derived from three wild-type Cry proteins from B. thuringiensis
subspecies kurstaki and aizawai. The Cry2Ab2 protein is encoded by the cry2Ab2 gene
derived from B. thuringiensis subspecies kurstaki.
The Cry proteins exert their effect on the host insect by causing lysis of midgut epithelial cells, which leads to gut paralysis, cessation of feeding and eventual death of the insect. The lysis of the midgut epithelial cells is mediated by the binding of the activated Cry protein to specialised receptors on these cells.
Hybrid corn lines containing the MON 89034 transformation event are intended for
cultivation in North America and are not intended to be grown in either Australia or New Zealand. Food from MON 89034 corn will therefore be entering the Australian and New Zealand food supply as imported, largely processed food products.
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Date of authorization: 04/12/2008
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.):
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 A595 - Food derived from insect-protected corn MON 89034
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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: Monsanto do Brasil Ltda.
Summary of application:
commercial release of genetically modified corn resistant to insects of the order Lepidoptera MON 89034
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Date of authorization: 15/10/2009
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:
The insect resistant MON 89034 corn was generated by transformation of hybrid LH172, mediated by Agrobacterium tumefaciens sp. with plasmid PV-ZMIR245 containing genes cr1A.105 and cry2Ab2. Proteins Cry1a.105 and Cry2Ab2 are codified by such genes, which are well known and characterized as insecticide proteins derived from soil bacterium Bacillus thuringiensis. Characterization of DNA inserted in MON 89034 corn was conducted through laboratory analyses showing that MON 89034 corn contains one single functional copy of genes cr1A.105 and cry2Ab2 inserted in its genome. Currently, the general mechanism of the Cry protein insecticide activity is well understood and Cry proteins comprehend different functional domains with highly conserved regions. Several field experiments for assaying agronomic and phenotypic characteristics of MON 89034 – contrasting with the control conventional corn of the same phenotype – were conducted during the 2007/2008 harvest in four representative locations of corn culture in Brazil: Cachoeira Dourada, State of Minas Gerais; Sorriso, State of Mato Grosso do Sul; Rolândia, State of Paraná; and Não-Me-Toques, State of Rio Grande do Sul. Phenotypic and agronomic characteristics of MON 89034 corn were assayed regarding its ecologic impact and potential as an invading plant. The studies covered several parameters, including: characteristics of plant growth and development, seed germination, pollen characteristics and observations for each plant-insect, plant-disease and plant-abiotic stress interaction. The results indicated that the MON 89034 corn has no characteristics that could pose risk as an invading plant or increase ecological risk when compared with conventional corn. Data on ecological interactions indicate that MON 89034 corn fails to grant greater susceptibility or tolerance to specific diseases, insects (except target-insects) and abiotic stress. The data ratify the idea that MON 89034 corn poses no risk of becoming an invading plant or of generating significant ecological impact when compared with conventional corn. Adverse effects in human and animal alimentary chain resulting from ingestion of MON 89034 corn and its derivatives are not expected based on an assay of alimentary safety of MON 89034 corn and its expressed proteins Cry1A.105 and Cry2Ab2. Allergic reactions to Cry proteins were not confirmed in application of microbial products derived from Bacillus thuringiensis for over four decades of use. Dietary safety of proteins Cry1A.105 and Cry2Ab2 present in food and rations derived from MON 89034 corn was assessed for risks posed to humans and animals and it was demonstrated that both proteins fail to show acute toxicity and do not cause adverse effect, even in high doses. Proteins Cry1A.105 and Cry2Ab2 are rapidly digested in simulated gastric juices, where 95% to 99% of proteins were digested in simulated gastric juices in a time not exceeding thirty seconds. Therefore the proteins, which are rapidly digested in the gastrointestinal tract of mammals, have negligible probability to cause allergies when consumed. Proteins Cry1A.105 and Cry2Ab2 do not share any similarity with amino acid sequences of known allergens or toxic proteins that may cause adverse effects in mammals. Dietary products and rations containing, or manufactured with, MON 89034 corn are as safe for human and animal consumption as the food manufactured with conventional corn. Potential adverse effects of MON 89034 corn to animal health were assayed in studies with mammals and birds, and the results corroborated the dietary safety of MON 89034 corn and proteins Cry1A.105 and Cry2Ab2 contained on this corn. Horizontal gene flow between MON 89034 corn and other species, even very closely related ones, have practically no probability of happening, since feral species related to corn do not occur in Brazil. Coexistence among corn cultivars (either improved or creole) and transgenic corn cultivars is possible from the agronomic viewpoint, and therefore the provisions of CTNBio Ruling Resolution nº4 shall be followed. Thus, analyses of the data supplied by applicant, as well as analysis of independent scientific literature, lead to a conclusion that proteins Cry1A.105 and Cry2Ab2 are innocuous and that MON 89034 corn is not potentially a cause of significant degradation of the environment; or of risks to human and animal health. These are the underlying reasons for the absence of restrictions to the use of MON 89034 corn and its derivatives. TECHNICAL OPINION I. GMO Identification GMO designation: MON 89034 corn. Applicant: Monsanto do Brasil Ltda. Species: Zea Mays L. Inserted characteristics: Resistance to insects. Method of insertion: MON 89034 corn, rated as Risk Class I, was produced though the methodology of transformation mediated by Agrobacterium tumefaciens sp. with plasmid PV-ZMIR245. Prospective use: Free registration, use, essays, tests, sowing, transport, storage, marketing, consumption, import, release into the environment and discarding. II. General Information Corn, (Zea mays L.) belongs to the Maydeae tribe, which belongs to subfamily Panicoideae of the Gramineae family. The Maydeae tribe includes genera Zea and Tripsacum in the Western Hemisphere and Coix, Polytoca, Chionachne, Schlerachne and Trilogachne in Asia. The Zea genus includes two sections: Luxiriantes and Zea. Corn (Zea Mays L.) is a separate species within subgenus Zea, together with three subspecies. Another genus included in the Maydeae tribe is the Tripsacum. Tripsacum includes sixteen species featuring a basic number of 18 chromosomes (n = 18), and the different Tripsacum species include multiples of 18 chromosomes within the interval of 2n = 36 to 2n = 108. Corn has a history of over eight thousand years in the Americas and is cultivated since the Pre-Columbian era. Among higher plants, corn is one of the best scientifically characterized and is currently the cultivated species that reached the highest degree of domestication and is unable to survive in nature but when cultivated by man(1). There are currently over 300 identified varieties of corn and, within each such variety there are thousands of cultivars. Corn is one of the most important food sources worldwide and used as input for a variety of food products, rations and industrial products. Brazil is the third largest world producer of corn, with a yield of about 35 million tons in 2005, following the United States of America (282 million tons) and China (139 million tons)(2). Corn is the second most planted culture in Brazil and is cultivated basically in two harvests practically all over the Brazilian territory. Regarding production, corn is second in a list of the largest Brazilian cultures, second only to soybeans(3). Brazil is a large world producer of corn, but it has also a significant consumption of this grain. National consumption is so high that an absence of the second crop would cause the need to import corn to meet national demand. Insect infestation in the tropics is higher than in temperate regions, where damages are higher. Among corn pests, importance of the fall armyworm (Spodoptera frugiperda) shall be underlined. Cruz et al.(4) estimated that the losses in Brazil caused by infestations of Spodoptera frugiperda reached 400 million Dollars each year. From 1999, occurrence of Spodoptera frugiperda increased and therefore, losses soared. Other species of the Lepidoptera order are important pests in the culture of corn, such as the corn earworm (Helicoverpa Zea) and stalk borer (Diatraea Saccharalis). Estimates are that the three species may damage up to 34% of corn production. The main controlling measure for insects in corn farming in Brazil has been the use of insecticides. In some of Brazilian Center-Western areas, for instance, tenths of insecticide spraying are needed for a single cycle of culture. Another measure to control pests would be the use of resistant cultivars. However, obtaining insect resistant corn cultivars by means of classic genetic improvement has not been as successful as desired. For Spodoptera frugiperda, several attempts have been made, with limited success(5). MON 89034 corn produces proteins Cry1A.105 and Cry2Ab2, derived from Bacillus thuringiensis, that are active against important pest lepidopterans in this culture. Compared with the corn containing the single event MON 810, MON 89034 corn controls a wider range of pests. MON 89034 corn provides efficient control against fall armyworm (Spodoptera frugiperda) over the whole harvest. Besides, MON 89034 corn gives significantly higher protection against damages caused by cotton bollworm (Helicoverpa Zea) when compared to genetically modified corn containing just one protein, and a high control of Ostrinia insects, such as European corn borer and Asian corn borer, and of Diatraea insects, such as stalk worm. The main corn pests in Brazil that are targets for corn containing event MON 89034 will be the fall armyworm, corn earworm and stalkworm. Brazil is held the world third larger consumer of agricultural pesticides, where currently there are 142 registered corn pesticides and, out of them, 107 have caterpillars as targets. There are several records of resistance for the constant and indiscriminate use of insecticides in corn culture in Brazil. Besides, one of the important factors affecting Brazilian farmers’ health is the use of agricultural pesticides, responsible for intoxicating a million individuals each year(6). III. Description of the GMO and Proteins Expressed MON 89034 corn was produced through the methodology of genetic transformation mediated by Agrobacterium tumefaciens, using the binary plasmid PV-ZMIR245. The plasmid consists of two separate T-DNA regions, each of them involved by the right and left border regions of the Ti plasmid(7). One T-DNA (T DNA I) contains genes cr1A.105 and cry2Ab2, while the other T-DNA (T DNA II) contains gene nptII, which grants resistance to the kanamycin antibiotic and was used in the initial process of selecting the transformed cells. Expression of genes cr1A.105 and cry2Ab2 is regulated by promoters e35S and FMV, respectively. The coding sequence of gene cr1A.105 produces protein Cry1A.105 that features insecticide action on corn farming pest lepidopterans. Protein Cry1A.105 is a modified Cry1A protein derived from Bacillus thuringiensis, that is 90.0%, 93.6% and 76.7% equivalent to proteins Cry1Ab, Cry1Ac and Cry1F, respectively. Gene cr2Ab2 coding sequence produces protein Cry2Ab2, a member of the Cry2Ab class of proteins with which has over 95% of the amino acid sequence in common(8,9). This is a variant of wild protein Cry2Ab2 isolated from Bacillus thuringiensis subspecies kurstaki. Bacillus thuringiensis, the donor organism of genes cr1A.105 and cry2Ab2, is a spore-forming gram-positive bacterium, naturally found in the soil. The bacterium produces proteins in the form of crystal or inclusion bodies that are selectively toxic for certain orders of pest insects. The technology has been used since 1958 to produce microbial formulations that are active as insecticides(10). This history or safe use shows that there is not harm associated to the use of Bacillus thuringiensis and to presence of Cry proteins in foodstuffs and rations, facts that have been corroborated by the lack of adverse effects for mammals in studies conducted with these products. The conclusion is supported by a revision on the history of safe use of Bacillus thuringiensis in agriculture(11). There are no known adverse effects to human beings that have been recorded over this long period of use of Bacillus thuringiensis microbial formulations(10). Betz et al(12) demonstrated the safety of several Cry proteins – expressed in genetically modified plants – for humans, animals and the environment. Gene nptII codifies enzyme neomycin phosphotransferase II (NPTII) that inactivates certain aminoglycosidic antibiotics such as kanamycin, neomycin and puromycin. The coding sequence of gene nptII derives from prokaryotic transposon Tn5(11). Enzime NPTII uses adenosine-triphosphate (ATP) to phosphorylate and inactivate aminoglycosidic antibiotics, which prevents the latter from injuring cells expressing NPTII when they are cultivated in a means that contain such selective agents. The only purpose of inserting gene nptII in MON 89034 corn was, therefore, selecting transformed cells containing genes cr1A.105 and cry2Ab2. Classic genetic improvement techniques were used to isolate plants that contained just the genes of interest cr1A.105 and cry2Ab2, and were devoid of gene nptII (T-DNA II), therefore producing plants free from the selection marker and containing just the characteristic of resistance to certain pest lepidopterans(14). Characterization of DNA inserted in MON 89034 corn took place through analyses of Southern blot, polymerase chain reaction (PCR) and DNA sequencing. Results show that MON 89034 corn contains a single functional copy of genes cr1A.105 and cry2Ab2 and that the complete insert contains: (1) The coding sequence of gene cr1A.105, whose transcription is controlled by the modified promoter e35S, the untranslated leading sequence Cab 5’ of the protein that bonds to wheat a/b chlorophyll, the introns of the rice actin gene (Ract1) and the termination transcription and polyadenylation sequence derived from the untranslated 3’ region of wheat (Hsp17) protein Hsp17.3 coding region; and (2) gene cry2Ab2 coding sequence whose transcription is controlled by figwort mosaic virus (FMV) promoter 35S, the first intron of protein Hsp70 gene (Hsp70), the DNA region containing the sequence of the transit peptide for the chloroplast of the smaller subunit of corn 1,5-biphosphate carboxylase ribulose and the first intron (SSU-CTP) and termination transcription and polyadenylation sequence derived from termination sequence 3’ of nopalyne synthase (nos 3’) of Agrobacterium tumefaciens. This T-DNA (T-DNA I) was inserted in the corn genome and resulted in the synthesis of Cry1A.105 and Cry2Ab2 proteins through expression of genes cr1A.105 and cry2Ab2. The transit peptide for chloroplast (CTP) is present do direct protein Cry2Ab2 to the corn plasmids. Analyses of PCR, sequencing of introduced and genomic DNA adjacent to the MON 89034 corn insert confirmed the organization of genetic elements within the genome. Results from sequencing amplified DNA fragments confirmed that the DNA sequence inserted in MON 89034 corn corresponds to the sequences that are present in plasmid PV-ZMIR245. However, promoter e35S that regulates gene cr1A.105 expression was modified by substitution of the right border present in plasmid PV-ZMIR245 by the left border sequence of MON 89034 corn. Protein Cry1A.105 consists of 1,177 amino acids with a molecular weight (MW) of 133 kDa, is a chimeric protein formed by domains I and II of proteins Cry1Ab or Cry1Ac1, a substantial portion of protein cry1F domain III and the C-terminal domain of protein Cry1aAc. The design strategy of protein Cry1A.105 used the change of domains to reach high levels of homology in several domains of protein Cry1A.105 with the respective domains in proteins Cry1Ab, Cry1Ac and Cry1F. Domains I and II of proteins Cry1A.105 are 100% identical to domains I and II of proteins Cry1Ab and Cry1Ac in amino acid sequences. Domain III of protein Cry1A.105 is 99% identical to domain III of protein Cry1F in amino acid sequence. The C-terminal portion is 100% homologous to the C-terminal portion of protein Cry1Ac. On average, sequence identity of protein Cry1A.105 with proteins Cry1Ac, Cry1Ab and CryF is 93.6%, 90.0% and 76.7%, respectively. According to the accepted phylogram for Cry proteins of Bacillus thuringiensis(8,9), protein Cry1A.105 may be grouped with proteins Cry1Ac and Cry1Ab due to their high degree of homology(15). Domain interchange is a well known mechanism that occurs in nature and increases the diversity of proteins Cry(16,17,18). The interchange, coupled with modern molecular biology tools, has been used to change functional domains of proteins Cry1 and develop commercial microbial biopesticides with greater specificity to pest lepidopterans. Microbial pesticides with the chimeric protein Cry1Ac/Cry1F have been used to control pest lepidopterans since 1997(19,20), and cotton expressing the chimeric protein containing domains of Cry1F, Cry1C and Cry1Ab also has been marketed(21). The general mechanism of Cry protein insecticide activity is well understood(22,23,24). The proteins encompass several functional domains possessing highly conserved regions among different classes. For instance, proteins Cry1A sequence of amino acids is highly conserved in domains I, II, and III. These functional domains determine the Cry proteins activity and specificity. Domain I is involved in inserting the membrane and formatting of pores. Domain II is involved in recognition and bonding to the specific receptor. Domain III keeps the structural integrity of the protein molecule(25) and contributes also to its specificity(16,26). C-terminal domain relates to crystal formation, which has no direct contribution to the protein activity(16). C-terminal domain is cut once within the insect guts or by certain in vitro proteases. Only insects possessing specific receptors are affected and there is no record of toxicity on other species that are devoid of such receptors(8,9). As in other proteins Cry1A, Cry1A.105 is active against important lepidopteran pests. The range of activities includes corn borers of genus Ostrinia (such as the European corn borer of 1 Cry1Ab and Cry1Ac share 100% of the amino acid sequence in domains I and II corn and the Asian corn borer) and Diatraea (corn stalk worm), fall armyworm (Spodoptera frugiperda sp.), cotton bollworm (Helicoverpa Zea) and black cutworm (Agrotis ipsilon). Protein cr1A.105 was purified from kernels of MON 89034 cotton and a recombinant lineage of Escherichia coli through fermentation. A panel of analytic tests was used to identify, characterize and compare protein cr1A.105 produced in the MON 89034 corn plant and the protein produced in Escherichia coli. Analysis of the purified and characterized cr1A.105 protein isolated in MON 89034 corn demonstrated its equivalence between the protein produced in the plant and the protein produced in Escherichia coli. Electrophoresis results in SDS-PAGE demonstrated that protein cr1A.105 produced in MON 89034 corn compared with protein produced in Escherichia coli indicate that both have equivalent molecular weight. The Western blot test showed that electrophoretic mobility and immunoreactivity of MON 89034 corn-produced Cry1A.105 protein are equivalent to that of Escherichia coli-produced protein. Mapping of the tryptic peptide by MALDI-TOF MS resulted in masses of peptides consistent with the expected tryptic peptides generated in silico based on the loci foreseen of trypsin breaking in cr1A.105 sequences. Besides, the two proteins were equivalent for functional activity and absence of glycosylation. Protein Cry2Ab2 produced in MON 89034 corn derives from Bacillus thuringiensis subspecies kurstaki and its amino acid sequence differs from that of the wild protein by a single amino acid. Protein Cry2Ab2 has 88% of identity in amino acid sequence with protein Cry2Aa that is present in commercial microbial products used to the control of pests. Proteins Cry2Ab2 produced in MON 89034 corn and in genetically modified cotton containing event MON 15985(27) share identical amino acid sequence. Protein Cry2Ab2 produced in MON 89034 corn is a variation of wild Cry2Ab2 of Bacillus thuringiensis. Its accumulation in MON 89034 corn is targeted to the chloroplast through the use of a transit peptide to the chloroplast (CTP) in the expression cassette. CTPs facilitate intracellular transport of proteins from the cytoplasm to plastids(28) and are typically eliminated from the mature protein once they are in the chloroplasts, where they are rapidly degraded. In order to enable accumulation of Cry2Ab2 in MON 89034 corn plastids, the DNA sequence that codes the CTP region of the smaller subunit of corn ribulose 1.5-biphosphate carboxylase was fused with the cry2Ab2 gene coding sequence. Protein Cry2Ab2 isolated from MON 89034 corn was purified, characterized, and the result was an equivalence between the plant produced protein and the protein produced in Escherichia coli. SDS-PAGE demonstrated that protein Cry2Ab2 produced in MON 89034 corn migrated in gel with protein produced in Escherichia coli, indicating that the proteins from the two sources have equivalent molecular weight. Western blot analysis showed that protein Cry2Ab2 electrophoretic mobility and immunoreactivity produced in MON 89034 corn are equivalent to that of the protein produced in Escherichia coli. The N-terminal of Cry2Ab2 protein in MON 89034 corn was blocked but mapping analysis by MALDI-TOF MS revealed masses of peptides consistent with tryptic peptides expected in loci foreseen for breaking by trypsin in Cry2Ab2 sequences, confirming the identity of the protein. Besides, the two proteins were equivalent in terms of functional activity and absence of glycosylation. The whole set of data supplied a detailed characterization of Cry2Ab2 protein isolated in MON 89034 corn and established its equivalence with the protein produced in Escherichia coli used in biosafety studies. IV. Aspects Related to Human and Animal Health A ninety day study with rats was conducted to assay potential adverse effects from MON 89034 corn to human and animal health(29). In the study, kernels of MON 89034 corn were fed to rats during ninety days. Results showed that there were no deaths or clinic observations related to the treatments, as well as changes in the parameters assayed: corporal weight; food consumption; hematology; serum chemistry; and urine tests. Besides, no changes in weight of organs were recorded that could have been attached to the administration of MON 89034 corn in the sample diet, nor macro or microscopic change associated to the treatment. Finally, the results enable the conclusion that the study with rats fed with MON 89034 corn kernels for ninety consecutive days failed to reveal adverse effects to growth and health of rats. This result corroborates the remaining conclusions on alimentary safety of MON 89034 corn and on proteins Cry1A.105 and Cry2Ab2 expressed in the plant. In addition, a forty-two day study was conducted to compare the nutritional value of MON 89034 corn kernels with the control conventional H1325023 and four commercial corn references when fed to fast-growing broiler chicken. Based on the study results, a conclusion was reached that the diet containing MON 89034 corn was as healthy as the control and commercial reference diet, considering the ability to promote fast growth in birds(30). A conclusion from the experiments was that weight measurements of the frozen carcass, fatty tissues, chest, thigh, drumsticks and wings were similar to the birds submitted to the different treatments. Similarly, there were no record of difference for humidity, proteins and fat rates in bird’s chest and drumsticks meat. Comparison were conducted between birds fed with the diet containing MON 89034 corn and birds fed on the control corn and the references failed to show differences regarding the parameters of performance, carcass and meat quality. Therefore, no differences were recorded in parameters measured in birds fed with MON 89034 when contrasted to birds fed with the control corn. One contributing factor to greater potential allergic oral sensitization to proteins is its stability for gastrointestinal digestion. Protein allergens tend to be stable to peptic and acid conditions of the digestive tract when they reach and pass through the intestinal mucosa to induce an allergic response(31,32,33,34). Rapidly digested proteins are highly correlated to significant decrease in the potential to cause allergic sensitization or reaction when consumed by humans. One aspect of this study includes analysis of protein in an essay with Simulated Gastric Fluid – SGF containing pepsin. The relation between digestibility in SGF and potential allergenicity was previously reported for a group of allergenic and non-allergenic proteins(31). The pepsin digestion essay was conducted to assess susceptibility of proteins Cry1A.105 and Cry2Ab2 to in vitro digestion in a fluid containing pepsin(35). Additionally to the SGF essay, an in vitro essay in Simulated Intestinal Fluid – SIF was also conducted to assay digestibility of alimentary components(36,37). In vitro susceptibility of Cry1A.105 and Cry2Ab2 proteins to pancreatin was assayed for SIF digestibility according to methods described in the literature(38). Protein Cry1A.105 digestibility was assayed by SDS-PAGE and Western blot. Digestion of protein Cry1A.105 was assayed by visual analysis in stained polycrylamide gel or by visual analysis in developed X-Ray films. Results of this study showed that protein Cry1A.105 was rapidly digested after incubation in SGF. Protein Cry1A.105 was digested below the 30 seconds LOD. Digestibility of Cry2Ab2 in SGF was assayed by SDS-PAGE and Western blot. Protein Cry2Ab2 digestion extension was assayed by visual analysis in stained polycrylamide gel or by visual analysis in developed X-Ray films. One gel or membrane was prepared in separate at the same time to determine the limit of detection (LOD) in each essay. Digestibility of Cry2Ab2 protein in SGF was assayed in SDS polycrylamide gel and the protein was rapidly digested. The results were consistent with reports from other Cry proteins that had its safety verified. The fact that Cry1A.105 protein was promptly digested in simulated gastric fluid makes its activity as an alimentary allergen unlikely. The same way, Cry2Ab2 is promptly digested in simulated gastric fluid and it also unlikely that it may act as an alimentary allergen. Adverse effects were no reported when mice received a total of 2,072 mg/kg of body weight of Cry1A.105 protein or 2,198 mg/kg of body weight of protein Cry2Ab2 in one day. Therefore, potential risks to health caused by acute ingestion in the diet of such proteins through consumption of MON 89034 corn was assayed by the exposure margin calculation based on no-observed effect level (NOEL) of the acute toxicity study with mice and estimates of the 95th percentile of acute exposure in their diet. Ingestion of Cry1A.105 and Cry2Ab2 in the diet of birds, piglings and fattening hogs was estimated using the daily consumption of cotton kernels and the highest levels of Cry1A.105 (7.0 μg/g of dry weight) Cry2Ab2 (2.1 μg/g of dry weight). A margin of exposure (MOE), defined as the ratio of NOEL with daily ingestion in the diet, was calculated for proteins Cry1A.105 and Cry2Ab2 for birds, piglings, young hogs, fattening hogs and dairy cattle. For birds and bovines, MOEs ranged from 1,930 to 13,500 and from 2,160 to 47,600 for proteins Cry1A.105 and Cry2Ab2, respectively. The high values of MOE indicate negligible risk to the health of birds and bovines exposed to rations containing MON 89034 corn in their diet. Ingestion of proteins Cry1A.105 and Cry2Ab2 in the diet of birds, piglings, and fattening hogs was estimated using a daily consumption of corn kernels and the highest levels of proteins Cry1A.105 (7.0 μg/g dry weight) and Cry2Ab2 (2.1 μg/g of dry weight). A margin of exposure (MOE), defined as the ratio of NOEL with daily ingestion in the diet, was calculated for proteins Cry1A.105 and Cry2Ab2 for birds, piglings, fattening hogs and daily cattle. For birds and bovines, MOEs were in the range from 1,930 to 13,500 and 2,160 to 47,600 for proteins Cry1A.105 and Cry2Ab2, respectively. The high values for MOE indicate negligible risk to the health of birds and bovines resulting from exposure to rations containing MON 89034 corn in their diet. Exogenous proteins produced in genetically modified cultures are not structural or functionally related to toxic or pharmacologically active proteins that may cause adverse effects on pregnant animals and their progeny. Besides, the likelihood that proteins expressed in genetically modified cultures resist to digestion in the gastrointestinal tract is minimal, as shown by in vitro studies performed as part of the alimentary safety assessment of such products (in this case, proteins Cry1A.105 and Cry2Ab2 of MON 89034 corn). Therefore, the likelihood that such exogenous proteins are absorbed intact, in amounts sufficient to gain access to the fetal circulation, is negligible. Normally, enzymes do not display evidence of producing toxic effects or being teratogenic to the reproductive system when supplied in food to rodents(39,40,41,42). Security studies conducted with a number of food enzymes produced by fermentation failed to record evidence that the proteins could trigger mutagenic or carcinogenic processes in laboratory animals(43). Some studies showed that food enzymes have no mutagenic nor teratogenic effects in animals and bacteria(44,45,46,47,48,49). Therefore, MON 89034 corn was assayed for its alimentary safety through available and internationally validated protocols, which, coupled with assaying guides for genetically modified cultures produced and/or updated by internationally renowned organizations will keep being used when necessary to secure that biotechnology derived food may be safely consumed(50). V. Environmental Aspects Corn is a monoic, allogamic and annual plant with anemochoric pollination, whereby distances that may be covered by pollen depend on wind patterns, humidity and temperature. Corn pollen disperses freely in areas located near the culture, and is able to reach stily-stigmas of the same or different genotypes and, under adequate conditions, starts germinating to originate the pollinic tube promoting ovule fecundation within an average period of 24 hours. Studies conducted on pollen dispersion demonstrated that pollen may travel long distances, though the major part of it is deposited close to the corn field, with a very low translocation rate, where over 95% of pollen may reach distances within sixty meters from its source. Luna et al.(51) examined the isolation and control distance for pollen and demonstrated that crossed pollination takes place within 200 meters, though no crossed pollination was recorded, under conditions of non-detasseling, for distances no lower than 300 meters from pollen sources. Results indicate that pollen viability is maintained for two hours and that crossed pollination was not recorded at a distance of 300 meters from the pollen source. By comparing concentrations at one meter from the source under low-to-moderate wind, one estimated that about 2% of the pollen are recorded at sixty meters, 1.1% at 200 meters, and 0.75-0.5% at 500 meters from the source. At a distance of ten meters from the source, on average, pollen grains by unit of soil is tenfold lower the pollen recorded at one meter from the border. Therefore, if separation distances established for corn seeds are observed, one expects that pollen transfer to adjacent varieties be minimal, being unlikely the presence of glyphosate-resistant genetic materials. Tesinte (Zea mays subspecies mexicana) and corn are species pollinated by wind, mutually compatible, highly variable and infertile(52,53). Both are genetically compatible and, in some areas of Mexico and Guatemala, cross freely when close to one another or under favorable conditions. Though corn can easily cross with teosinte, the species is not present in Brazil. Natural distribution of teosinte is limited to seasonally dry and sub-tropical zones with summer rains in Mexico and Guatemala and to the Mexican Central Plateau(53,54). With the introduction of genes cr1A.105 and cry2Ab2 one does not expect that the characteristics granted may give any competitive advantage, or greater aggressiveness to MON 89034 corn that would result in an invasive plant. The characteristics of resistance to some lepidopteran pest do not make MON 89034 corn a plant pest or a natural habitat invading plant, since the corn reproductive and development characteristics were not changed. Thus, the only advantage of MON 89034 corn is its resistance to damages caused by some insects of the Lepidoptera Order. The insect-resistant characteristics could even grant an adaptive advantage in the environment. However, this would only happen in a system where insects were agents limiting propagation of the plant, that is to say, if there was a strong pressure for selection by pests(55,56,57). This is not the case for MON 89034 corn, where genes cr1A.105 and cry2Ab2 grant protection only against some species of lepidopteran pests. A number of other insect species could cause impact in the survival of the plant in natural environments and Cry proteins produced in MON 89034 corn do not grant the plant any advantage regarding such non-target pests(56). Insecticide proteins Cry are extremely selective for insects of the Lepidoptera Order(58, 59, 60, 61, 62), with no poisonous effects to insects held as beneficial and non-target insects, among which predators, parasitoids, pollinating and other insects(63,64,65,66). An assay for both, target and non-target insects was conducted in the entomofauna during the 2007/2008 crop in four Monsanto of Brazil Experimental Stations (Cachoeira Dourada, State of Minas Gerais; Sorriso, State of Mato Grosso; Não-Me-Toques, State of Rio Grande do Sul; Rolândia, State of Paraná) located in areas of representative corn culture(67). The results showed that the insect species or families collected and identified in 235 comparisons failed to display a behavioral trend, with no records of increased or diminished insect visitation between plots with MON 89034 corn, control corn, or commercial references corn. Insects found represent species and/or families common to corn culture in different areas where the experiment was conducted. Similarity in visitation recorded in plots with MON 89034, control and commercial reference corn indicate that the introduction of the lepidopteran-resistant characteristics fails to interfere with insect visitation in the culture of corn in Brazil regarding the insects studied. The impact that may be caused, in general, by Cry proteins on non-target insects and soil organisms has been widely studied as part as an assessment of environmental safety of cultures with different events containing proteins Cry. Studies show that the tested terrestrial and soil non-target organisms were not affected by protein Cry1Ab, even though the level of the protein was above the maximum levels that may be verified in case of natural exposure(61,68,69). Additionally, a comparison of Cry proteins produced by Bacillus thuringiensis and the Cry1Ab protein produced by MON 89034 corn demonstrated that they may persist in tropical soils for a longer time due to its bonds to clay particles; however, effects on the soil microbiota were not recorded(70). Papers published in the scientific literature report that presence of Cry proteins do not affect in a significant way the microbiota and animals living in the soil(71,72). Additionally, Cry proteins of three subspecies of Bacillus thuringiensis failed to display microbiocide activity against a variety of bacteria, fungi and algae(73). Based on the experience with domesticated plant, the corn potential to be an invasive species to natural habitats, or to be persistent in a farm environment without human intervention is negligible. Corn plant is known to be a weak competitor, that when is outside cultivation areas has no significant impact in the environment. In assaying phenotypic, agronomic characteristics and ecologic interactions of MON 89034 corn, data were collected under five different categories: (1) germination, dormancy and emergence; (2) vegetative phase; (3) reproductive phase (including pollen characteristics); (4) retention of seeds in the plant; and (5) interactions of the plant with insects, diseases and abiotic stresses. Phenotypic, agronomic and ecologic interactions assays were based on combination of laboratory experiments and field studies conducted by experts in corn production and assay. Additionally, several commercial references were also used to provide a range3 of values common to corn commercial hybrids for each phenotypic, agronomic and ecologic interaction characteristic assayed. Completed the experiments, a conclusion was reached that MON 89034 corn is equivalent to conventional corn regarding its propagation and reproduction structures dispersion ability beyond cultivation areas and mechanisms for dispersion in the air, including that pollen viability of MON 89034 corn is equivalent to that of conventional corn. VI. Restrictions to the Use of the GMO and its Derivatives Studies submitted by applicant demonstrated that there was no significant difference between corn hybrids derived from non-modified lineages and MON 89034 corn regarding agronomic characteristics, reproduction, dissemination and survival ability. All evidence submitted in the proceedings and bibliographic references ratify that the risk level of transgenic variety is equivalent to non-transgenic varieties before soil microbiota, as well as that of other plants and to human and animal health. Thus, farming and consumption of MON 89034 corn are not potentially causes of significant environment degradation and fail to pose any risk to human and animal health. For these reasons, there are no restrictions to the use of MON 89034 corn and its derivatives, except in places mentioned by Law nº 11460, of March 21, 2007. Gene flow to local varieties (so-called creole corns) of open pollination is possible and poses the same risk caused by commercial genotypes available in the market (80% of conventional corn planted in Brazil comes from commercial seeds that underwent genetic improvement). Coexistence between conventional corn (either improved or creole) and transgenic corn cultivars is possible from the agronomic viewpoint(41,42) and shall comply with the provisions of CTNBio Ruling Resolution nº 4. After the years of use in different countries, there is no record of problems for human and animal health that could be attributed to transgenic corns. It shall be stressed that the lack of negative effects from cultivation of transgenic corn plants do not entail that such effects are free from occurring. Zero risk and absolute safety do not exist in a biological world, although there is a mass of reliable scientific information and a safe history of ten years of use that enable us to assert that MON 89034 corn is as safe as conventional corn versions. Therefore, applicant shall conduct post commercial-release monitoring under CTNBio Ruling Resolution nº 3. VII. Consideration on the Particulars of Different Regions of the Country (Information to supervisory agencies) As established by Article 1 of Law nº 11,460, of March 21, 2007 “research and cultivation of genetically modified organisms may not be conducted in indigenous lands and areas of conservation units.” VIII. Conclusion Considering that MON 89034 corn (Zea mays) variety belongs to a well characterized species with a solid safety record for human consumption and that cr1A.105 and cry2Ab2 genes introduced in this variety codify proteins that are ubiquitous in nature and are present in plants, fungi and microorganisms that are part of the alimentary diet of humans and animals; Considering that the insertion of the genotype took place though classical genetic improvement, resulting in insertion of one stable and functional copy of genes cr1A.105 and cry2Ab2 that granted resistance to insects; Considering that data on centesimal composition fail to show significant differences between genetically modified varieties and conventional varieties, suggesting nutritional equivalence between them; Considering that CTNBio granted approval do different corns containing event with protein Cry and that even Whereas: 1. studies showed that, as well as other Cry proteins, Cry1A.105 and Cry2Ab2 proteins are not potential causes of adverse effects at the exposure levels in fields on representative terrestrial beneficial invertebrate species, among them bees (larvae and adults); 2. expression of Cry1A.105 and Cry2Ab2 proteins fails to change the morphology and viability of MON 89034 corn pollen when compared to the pollen of conventional corn; 3. Cry1A.105 and Cry2Ab2 proteins do not share similarities with known allergen amino acid sequences, gliadins, gluteins and toxic proteins that have adverse effects on mammals; 4. laboratory studies with indicator species showed that proteins Cry1A.105 and Cry2Ab2 fail to cause adverse effects on tested non-target organisms; 5. molecular comparative analysis of MON 89034 corn ratified insertion of a single functional copy of genes cr1A.105 and cr1A.105 expression cassettes in a single locus of the corn genome; 6. comparative biochemical studies indicate that Cry1A.105 and Cry2Ab2 proteins have an important difference in their mode of action, namely in the form in which they bond to receptors in lepidopteran insects midgut and therefore the likelihood of crossed resistance between the two proteins is low; 7. the two proteins belong to the Cry family of proteins, derived from Bacillus thuringiensis, an organism that has been commercially used for over forty years in producing microbial insect controlling formulations; 8. the history of safe use and data from a number of studies support the conclusions on safety of MON 89034 corn and proteins Cry1A.105 and Cry2Ab2; 9. agronomic and efficacy assays of MON 89034 corn indicate that the event failed to express any other characteristics besides the expected ones; 10. phenotypic, agronomic and ecologic interaction assays indicate that MON 89034 corn is comparable to conventional corn and carries no higher risk to change into an invading plant. Therefore, considering internationally accepted criteria in the process of analyzing risks in genetically modified raw-material it is possible to conclude that MON 89034 corn is as safe as its conventional equivalent. CTNBio considers the activity free from being a potential cause of significant degradation to the environment, or harm to human and animal health. Restrictions to the use of this GMO and its derivatives are conditioned to the provisions of Law nº 11460, of March 21, 2007, CTNBio Ruling Resolution nº 03, and CTNBio Ruling Resolution nº 04. CTNBio analysis took into consideration opinions of the Commission members; ad hoc consultants; documents delivered to CTNBio Executive Secretary; results of planned releases into the environment; and lectures, texts and debates of the public hearing held on 03.20.2007. Also considered and consulted were applicant’s independent studies and scientific literature conducted by third parties. According to Annex I to Ruling Resolution nº 5, of March 12, 2009, the applicant shall have a term of thirty (30) days from publication of this Technical Opinion to adjust its proposal to the post-commercial release monitoring plan. VIII. Bibliography 1. BAHIA FILHO, A. F. C.; GARCIA, J. C. 2000. Análise de avaliação do mercado brasileiro de sementes de milho In: UDRY C. V.; DUARTE, W. F. (org) Uma história brasileira do milho: o valor de recursos genéticos. Brasilia: Paralelo 15,167-172. 2. FOOD AND AGRICULTURE ORGANIZATION OF THE UNITED NATIONS - FAO. 2007. FAOSTAT. Disponível em http://faostat.fao.org/site/340/default.aspx. 3. Companhia Nacional de Abastecimento – CONAB. 2007 Milho total (1a e 2a safra) Brasil – Série histórica de área plantada: safra 1976-77 a 2006-07. http://www.conab.gov.br/conabweb/download/safra/milhototalseriehist.xls. 4. CRUZ, I.; FIGUEIREDO, M. L. C.; OLIVEIRA, A. C.; VASCONCELOS, C. A. 1999. Damage of Spodoptera frugiperda (Smith) in different maize genotypes cultivated in soil under three levels of aluminium saturation. International Journal of Pest Management 45:293-296. 5. WAQUIL, J. M.; VILLELA, F. M. F.; FOSTER, J. E. 2002. Resistência do milho (Zea mays L.) transgênico (Bt) à lagarta-do-cartucho, Spodoptera frugiperda (Smith) (lepidoptera: Noctuidae). Revista Brasileira de Milho e Sorgo 1 (3): 1-11. 6. ALVES FILHO, J. P. 2001. Agrotóxicos e Agenda 21: sinais e desafios da transição para uma agricultura sustentável. In: II SINTAG Anais. II Simpósio Internacional de Tecnologia de Aplicação de Agrotóxicos: Eficiência, Economia e Preservação da Saúde Humana e do Ambiente. Jundiaí, SP, 17/07/2001 a 20/07/2001. 7. Komari, T. Y.; Hiei Y.; Saito, N.; Murai, and T. Kumashiro. 1996. Vectors carrying two separate T-DNAs for cotrasformation of higher plants mediated by Agrobacterium tumefaciens and segregation of transformants free from selection markers. The Plant J. 10:165-174. 8. Crickmore, N.; D. R. Zeigler; J. Feitelson; E. Schnepf; J. van Rie; D. Lereclus, J. Baum; and D. H. Dean. 1998a. Revision of the nomenclature for the Bacillus thuringiensis pesticidal crystal proteins. Microbiol Mol Biol Rev 62:807-13. 9. Crickmore, N.; D. R. Zeigler; J. Feitelson; E. Schnepf; J. van Rie; D. Lereclus; J. Baum, and D. H. Daen. 1998b. Revision of the nomenclature for the bacillus thuringiensis pesticidal crystal proteins. Microbiology and molecular biology reviews 62:807-813. 10. U.S. EPA. 1998. Bacillus thuringiensis (B.t.) plant-pesticides and resistance management EPA 738-F-98-001. United States Environmental Protection Agency. 11. McClintock, J. T.; C.R. Schaffer; and R.D. Sjobald. 1995. A comparative review of the mammalian toxicity of Bacillus thuringiensis-based pesticides. Pest. Sci. 45:95-105. 12. Betz, F. S., B. G. Hammond, and R. L. Fuchs. 2000. Safety and advantages of Bacillus thuringiensis-protected plants to control insect pests. Reg. Toxicol. and Pharmacol. 32:156-173. 13. Beck, E.; G. Ludwig; E. A. Auerswald; B. Reiss, and H. Schaller. 1982. Nucleotide sequence and exact localization of the neomycin phosphotrasferase gene from trasposon tn5. Gene 19:327-36. 14. Monsanto do Brasil. 2008. Relatório Técnico Liberação Comercial milho MON 89034. 15. Crickmore, N., D. R. Zeigler; E. Schnepf; J. van Rie; D. Lereclus; J. Baum, A. Bravo; and D. H. Dean. 2004. Bacillus thuringiensis toxin nomenclature. http//www.biols.susx.ac.uk/home/neilcrickmore/bt/index.html. 16. De Maagd, R. A.; A. Bravo, and N. Crickmore. 2001. How Bacillus thuringiensis has evolved specific toxins to colonize the insect world. Trends Genet. 17:193-9. 17. De Maagd, R. A.; A. Bravo; C. Berry; N. Crickmore; and H. E. Schnepf. 2003b. Structure, diversity, and evolution of protein toxins from spore-forming entomopathogenic bacteria. Annu. Rev. Genet. 37:409-433. 18. Masson, L.; A. Mazza; S. Sangadala; M. J. Adang; and R. Brusseau. 2002. Polydispersity of Bacillus thuringiensis Cry1 toxins in solution and its effect on receptor binding kinetics. Bichim biophys Acta 1594:266-75. 19. Baum, J.A 1998. Transgenic Bacillus thuringiensis. Phytopotection 79:127-130. 20. Baum, J. A., T. B. Johnson, and B.C. Carlton. 1999. Bacillus thuringiensis. Natural and recombinant bioinsecticide products, p. Pp 189-209, In F. R. Hall and J. J. Menn, eds. Methods in Biotechnology. Pesticides: Use and Delivery, vol. 5. Humana Press, Inc., Totowa, New Jersey. 21. Gama, M. I. C. S. 1998. Identificação de plantas transgênicas por PCR. Manual de transformação genética de plantas. Ed. Brasileiro, A.C.M. and Carneiro, V.T.C. Brasília - EMBRAPA-SPI / EMBRAPA CENARGEN.pp.309. 22. Gill, S.S.; E. A. Cowles, and P. V. Pietrantonio. 1992. The mode of action of Bacillus thuringiensis endotoxins. Ann. Rev. entomol. 37:615-636. 23. Schnepfe, E.; N. Crickmore; J. van Rie; D. Lereclus; J. Baum; J. Feitelson; D. R. Zeigler; and D. H. Dean. 1998. Bacillus thuringiensis and its pesticidal crystal proteins. Microbial. Mol. Biol. Rev. 62:775-806. 24. Zuang, M.; and S.S. Gill. 2003. Mode of action of Bacillus thuringiensis toxins., p. Pp 213-236, In G. Voss and G. Ramos, eds. Chemistry of crop protection, progress and prospects in Science and Regulation. Wiley-VCH, Weinheim, Germany. 25. Li, J.D.; J. Carroll; and D. J. Ellar. 1991. Crystal structure of insecticidal delta-endotoxin from Bacillus thuringiensis at 2.5 a resolution. Nature 353:815-21. 26. De Maagd, R. A. ; M. Weemen-Hendriks; W. Stiekema, and D. Bosch. 2000. Bacillus thuringiensis deltaendotoxin Cry1c domain III can function as a specificity determinant for Spodoptera exigua in different, but not all, Cry1-cry1c hybrids. Appl Environ Microbiol 66:1559-63. 27. Comissão Técnica Nacional de Biossegurança. 2009. Parecer Técnico 1.832/2009. D.O.U 22/05/2009, Seção 01 pág.10. 28. Bruce, B. D. 2000. Chloroplast transit peptide: structure, function and evolution. Trends cell biol. 10:440-447. 29. Kirkpatrick, J. B. 2007. A 90-day feeding study in rats with MON 89034. Monsanto Technical Report MSL0020649. 30. Taylor, M.; G. Hartnell; M. Nemeth; D. Lucas, and S. Davis. 2007. Comparison of broiler performance when fed diets containing grain from second-generation insect-protected and glyphosate-tolerant, conventional control or commercial reference corn. Poultry Science 86:1972-1979. 31. Astwood, J.; J. J. Leach; and R. L. Fuchs. 1996. Stability of food allergens to digestion in vitro. Nature Biotechnology 14:1269-1273. 32. Astwood, J.D.; and R. L. Fuchs. 1996a. Allergenicity of foods derived from transgenic plants. Monogr Allergy 32:105-20. 33. Fuchs, R. L.; and J. D. Astwood. 1996. Allergenicity assessment of foods derived from genetically modified plants. Food technology 50:83-88. 34. Metcalfe, D. J.; Astwood T. R.; S. H.; T.M. L.; and F. R. 1996. Assessment of the allergenic potential of foods derived from genetically engineered crop plants. Critical Reviews in Food Science and Nutrition 36:165-186. 35. Thomas, K.; M. Aalbers; G. A. Bannon; M. Bartels; R.J. Dearman; D. J. Esdaile; T.J. Fu; C. M. Glatt; N. Hadfield; C. Hatzos; S. L. Hefle; J.R. Heylings; R. E. Goodman; B. Hhenry; c. Herouet; M. Holsapple; G. S. Ladics; T. D. Landry; S. C. Macintosh; E. A. Rice; L. S. Privalle; H. Y. Steiner; R. Teshima; R. van Ree; M. Woolhiser; and J. Zawodny. 2004. A multi-laboratory evaluation of common in vitro pepsin digestion assay protocol used in assessing the safety of novel proteins. Regul.t Toxicol. Pharmacol.39:87-98. 36. Okunuki, H.; Techima, R.; Shigeta, T.; Sakushima; J.; Akiyama, H.; Yukihiro, G.; Toyoda, M.; and Sawada, J. J. 2002. Increased digestibility of two products in genetically modified food (CP4-EPSPS and Cry1ab) after preheating. Food Hyg. Soc. Japan 43:68-73. 37. Yagami, T.; Hashima Y.; Nakamura, A.; Hiroyuki, O.; and Ikesawa, Z. 2000. Digestibility of allergens extracted from natural rubber latex and vegetable foods. J. Allergy Clin. Immunol.106:52-762. 38. Pharmacopeia, U.S. 1995. The National Formulary Mark Printing Co., Easton, PA. (ed) 2000. Proceedings of the 6th International feed production conference, Piacenza, Italy. 39. Ashby, R.; R. K. Hjortkjaer; M. Stavnsbjerg; H .Gurtler; P. B. Pedersen; J. Bootman; G. Hodson-Walker; J. M. Tesh; C.R. Willoughby; H. West; and J.P. Finn. 1987. Safety evaluation of Streptomyces murinus glucose isomerase. Toxicol. Lett. 36:23-35. 40. Flood, M.T.; and M. Kondo. 2004. Toxicity evaluation of a ƒÒ-galactosidase preparation produced by Penicillium multicolor. Regul. Toxicol. Pharmacol. 40:281. 41. Greenough, R. J.; and D. J. Everett. 1991. Safety evaluation of alkaline cellulose. Food Chem. Toxicol. 29:781. 42. Hjortkjaer, R.K. 1993. Safety evaluation of esperase. Food Chem. Toxicol. 31:999. 43. Pariza, M. W.; and E. M. Foster. 1983. Determining the safety of enzymes used in food processing. J. Food Protection 46:453-468. 44. Bergmam, A.; and A. Broadmeadow. 1997. An overview of the safety evaluation of the Thermomyces lanuginosus xylanase enzyme (SP 628) and the Aspergillus aculeatus xylanase enzyme (SP 578). Food additives and contaminants 14:389-398. 45. Dean, S. 1997. Industrial genotoxicology group (IGG): The use of historical data in data interpretation and genotoxicity testing of biotechnology products, Royal Society of Medicine, London, UK, December 1995. Mutagenesis 12:49-50. 46. Kondo, M.; T. Ogawa; Y. Matsubara; A. Mizutani; S. Murata and M. Kitagawa. 1994. Safety evaluation of lipase G from Penicillium camembertii. Fd. Chem. Toxic. 32:685-696. 47. Mackenzie, K. M.; S. R. W. Petsel; R. H. Weltman; and N. M. Zeman. 1989a. Subchronic toxicity studies in dogs and in utero rats fed diets containing Bacillus stearothermophilus alpha-amylase from a natural or recombinat DNA host. Fd. Chem. Toxic. 27:599-606. 48. Mackenzie, K. M.; S. R. W. Petsel; R. H. Weltman; and N.W. Zeman. 1989b. Subchronic toxicity studies in dogs and in utero – exposed rats fed diets containing Bacillus megaterium amylase derived from a recombinant DNA organism. Fd. Chem. Toxic. 27:301-305. 49. Stavnsbjeg, M.; R. K. Hjorkjaer; V. Bille-Hansen; B. F. Jensen; R.J. Greenough; M. McConville; M. Holmstroem, and K. P. Hazelden. 1986. Toxicologic safety evaluation of a Bacillus acidopullulyticus pullulanase. J. Food Protect. 49. 50. Hammond, B. and A. Cockburn. 2008. The safety assessment of proteins introduced into crops developed through agricultural biotechnology: a consolidated approach to meet current and future needs. In: Hammond, B. (2008) Food Safety of Proteins in Agricultural Biotechnology, CRC Press, Taylor and Francis Group, LLC.:259-288. 51. LUNA, S. V.; FIGUEROA, J. M.; BALTAZAR, M. B.; GOMEZ, L. R.; TOWNSEND, R.; and SCHOPER, J. B. 2001. Maize pollen longevity and distance isolation requirements for effective pollen control. Crop Sci. 41:1551-1557. 52. Wilkes, G. 1989. Maize: domestication, racial evolution, and spread. Foraging and farming - the evolution of plant exploration: 440-455. 53. Wilkes, H.G. 1972. Maize and its wild relatives. Science 177:1071-1077. 54. Gonzalez, J.; and J. Corral. 1997. Teosinte distribution in Mexico. Proceedings of a forum: gene flow among maize landraces, improved maize, varieties and teosinte: implications for transgenic maize:18-39. 55. Borém, A . 2001. Escape gênico & transgênicos. Viçosa, UFV:206 p. 56. Glover, J. 2002. Gene flow study: implications for GM crop release in Australia. Bureau of rural Sciences, Canberra, Australia http://www.affa.gov.au/brs : 71p. 57. Nilsen, K. M.; J. D. van Elsas; and K. Smalla. 2000b. Safety issues in antibiotic resistance marker genes in transgenic crops. Proc. of the 6th International Feed production Conference:146-162. 58. MENDELSOHN, M.; KOUGH, J.; VAITUZIS, Z.; MATTEWS, K. Are Bt crops safe? Nature Biotechnology, v.21, n. 9, p. 1003-1009, 2003. 59. DULMAGE, H. T. Microbial control of pests and plant diseases 1970 – 1980. In: BURGES, H. D. (ed). London: Academic Press, 1981. p. 1993-222. 60. KLAUSNER, A. Microbial insect control. Bio/technology, v.2, p. 408-419, 1984. 61. ARONSON, A. I. BACKMAN, W.; DUNN, P. Bacillus thuringiensis and related insect pathogens. Microbiol. rev., v. 50, p. 1-24,1986. 62. MACINTOSH, S. C.; STONE, T. B.; SIMS, S. R.; HUNST, P.; GREENPLATE, J. T.; MARRONE, P. G.; PERLAK, F. J.; FISCHHOFF, D.A.; FUCHS, R. L. Specificity and efficacy of purified Bacillus thuringiensis proteins against agronomically important insects. J.Insect Path., v. 56, p. 258-266, 1990. 63. WHITELEY, H. R.; SCHNEPF, H. E. The molecular biology of parasporal crystal body formation in Bacillus thuringiensis. Ann. Rev. Microbiol., v.40, p. 549-576, 1986. 64. CANTWELL, G. E.; LEHNERT, T.; FOWLER, J. Are biological insecticides harmful to the honey bee. Am. Bee J., v. 112, p. 294-296, 1972. 65. KRING, A.; LANGENBRUCH, G. A. Susceptibility of arthropod species to Bacillus thuringiensis. In: Microbial Control of Pests and plant Diseases. BURGES, H. D. (Ed). London: Academic Press, 1981. p. 837-896. 66. FLEXNER, J. L.; LIGHTHART, B.; CROFT, B. A. The effects of microbial pesticides on non-target beneficial arthropods. Agric. Ecosys. Environ., v. 16, p. 203-254,1986. 67. Oliveira, W. 2008a. Agronomic and phenotypic evaluation of corn MON 89034 in Brazil field trials during 2007/2008 season. Monsanto Technical Report MSL-013. 68. FERNANDES, O. D. Efeito do milho geneticamente modificado (MON 810) em Spodoptera frugiperda (J. E. Smith,1797) e no parasitóide de ovos Trichogramma spp. 164 f. Tese (Doutorado em Entomologia), Departamento Entomologia, ESALQ, Universidade de São Paulo, Piracicaba, 2003. 69. SIMS, S. R. Bacillus thuringiensis var. kurstaki (Cry1Ac) protein expressed in transgenic cotton: effects on beneficial and other non-target insects. Southwestern Entomol., v. 20, p. 493-500, 1995. 70. SANDERS, P. R.; LEE, T. C.; GROTH, M. E.; ASTWOOD, J. D.; FUCHS, R. L. Safety assessment of insect-protected corn. In: THOMAS, J. A. Biotechnology and Safety Assessment. 2 ed. Taylor and Francis, 1998. p. 241-256. 71. MUCHAONYERWA, P.; WALADDE, S.; NYAMUGAFATAR, P.; MPEPEREKI, S.; and RISTORI, G. G. Persistence and impact on microorganisms of Bacillus thuringiensis proteins in some Zimbabwean soils. Plant and soil, v. 266, p. 41-46, 2004. 72. STOTZKY,G. Clays and humic acids affect the persistence and biological activity of insecticidal proteins from Bacillus thuringiensis in soil. In: Developments in Soil Science 28B (Soil Mineral - Organic Matter Microorganism Interactions and Ecosystem Health), p: 1-16, 2002. 73. STOTZKY, G. Persistence and biological activity in soil of the insecticidal proteins from Bacillus thuringiensis, especially from transgenic plants. Plant and Soil, v. 266, p. 77-89, 2004.
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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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Stacked events:
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)
Canada
Name of product applicant: Monsanto Canada Inc.
Summary of application:
Genetically modified MON 89034 corn was developed using recombinant DNA techniques to introduce two Bacillus thuringiensis (Bt) derived novel genes: the cry1A.105 sequences encoding Cry1A.105, a synthetic chimeric protein; and the cry2Ab2 gene encoding the Cry2Ab2 protein. The Cry1A.105 protein is a chimeric protein comprised of various domains from Cry1Ac, Cry1Ab, and Cry1F. The Cry2Ab2 protein as expressed in MON 89034 corn differs by a single amino acid from the wild type protein equivalent. All cry sequences are originally derived from Bt subsp. kurstaki with the exception of domain III sequences for cry1F, which are originally derived from Bt subsp. aizawai

The safety assessment performed by Food Directorate evaluators was conducted according to Health Canada's Guidelines for the Safety Assessment of Novel Foods. The assessment considered: how corn event MON 89034 was developed; how the composition and nutritional quality of corn grain derived from plants containing this event compare to non-modified corn; and what the potential is for food products derived from plants containing this event to be toxic or cause allergic reactions.

The Food Directorate has a legislated responsibility for pre-market assessment of novel foods and novel food ingredients as detailed in Division 28 of Part B of the Food and Drug Regulations (Novel Foods). Foods derived from corn lines containing event MON 89034 are considered novel foods under the following part of the definition of novel foods: "c) a food that is derived from a plant, animal or microorganism that has been genetically modified such that

i.the plant, animal or microorganism exhibits characteristics that were not previously observed in that plant, animal or microorganism".
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Date of authorization: 18/05/2008
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.): BioTrack Product Database
Summary of the safety assessment:
Please see decision document weblinks
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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: Novel Foods Decision
Novel Feeds Decision
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Authorization expiration date:
E-mail:
luc.bourbonniere@hc-sc.gc.ca
Organization/agency name (Full name):
Health Canada
Contact person name:
Luc Bourbonniere
Website:
Physical full address:
251 Sir Frederick Banting Driveway, Tunney's Pasture, PL 2204A1
Phone number:
613-957-1405
Fax number:
613-952-6400
Country introduction:
Federal responsibility for the regulations dealing with foods sold in Canada, including novel foods, is shared by Health Canada and the Canadian Food Inspection Agency (CFIA). Health Canada is responsible for establishing standards and policies governing the safety and nutritional quality of foods and developing labelling policies related to health and nutrition. The CFIA develops standards related to the packaging, labelling and advertising of foods, and handles all inspection and enforcement duties. The CFIA also has responsibility for the regulation of seeds, veterinary biologics, fertilizers and livestock feeds. More specifically, CFIA is responsible for the regulations and guidelines dealing with cultivating plants with novel traits and dealing with livestock feeds and for conducting the respective safety assessments, whereas Health Canada is responsible for the regulations and guidelines pertaining to novel foods and for conducting safety assessments of novel foods. The mechanism by which Health Canada controls the sale of novel foods in Canada is the mandatory pre-market notification requirement as set out in Division 28 of Part B of the Food and Drug Regulations (see Figure 1). Manufacturers or importers are required under these regulations to submit information to Health Canada regarding the product in question so that a determination can be made with respect to the product's safety prior to sale. The safety criteria for the assessment of novel foods outlined in the current document were derived from internationally established scientific principles and guidelines developed through the work of the Organization for Economic Cooperation and Development (OECD), Food and Agriculture Organisation (FAO), World Health Organisation (WHO) and the Codex Alimentarius Commission. These guidelines provide for both the rigour and the flexibility required to determine the need for notification and to conduct the safety assessment of the broad range of food products being developed. This flexibility is needed to allow novel foods and food products to be assessed on a case-by-case basis and to take into consideration future scientific advances.
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Stacked events:
Food: Consistent with the definition of "novel food" in Division 28 of the Food and Drug Regulations, the progeny derived from the conventional breeding of approved genetically modified plants (one or both parents are genetically modified) would not be classified as a novel food unless some form of novelty was introduced into such progeny as a result of the cross, hence triggering the requirement for pre-market notification under Division 28. For example, notification may be required for modifications observed in the progeny that result in a change of existing characteristics of the plant that places those characteristics outside of the accepted range, or, that introduce new characteristics not previously observed in that plant (e.g. a major change has occurred in the expression levels of traits when stacked). In addition, the use of a wild species (interspecific cross) not having a history of safe use in the food supply in the development of a new plant line may also require notification to Health Canada. However, molecular stacks are considered new events and are considered to be notifiable as per Division 28.

Feed:
Contact details of the competent authority(s) responsible for the safety assessment and the product applicant:
Luc Bourbonniere, Section Head Novel Foods
Philippines
Name of product applicant: Monsanto Philippines
Summary of application:
Monsanto has developed a biotechnology derived product corn MON 89034 through Agrobacterium mediated transformation to express the Bacillus thuringiensis insecticidal proteins Cry1A.105 and Cry2Ab2 using the plasmid vector PV-ZMIR245, which is a binary vector containing two separate transfer DNA’s (2T-DNA). The first T-DNA, designated as T-DNA I, contains the cry1A.105 and the cry2Ab2 expression cassettes. The second T-DNA designated as T-DNA II contains the nptII (neomycin phosphotransferase II) expression cassette, as a selectable marker. Traditional breeding was used to isolate those plants only contained the cry1A.105 and cry2Ab2 expression cassettes (T-DNA I) and did not contain the nptII expression cassette (T DNA II), thereby, producing marker-free corn MON 89034. The introduction of the second generation product MON 89034 is expected to provide enhanced benefits for the control of lepidopteran insects pests such as Ostrinia furnacalis (ACB) and Spodoptera frugiperda (FAW) and Helicoverpa zea (CEW) compared to existing products.
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Date of authorization: 29/04/2009
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:
Monsanto Philippines, has filed an application with attached technical dossiers to the Bureau of Plant Industry (BPI) for a biosafety permit for direct use as food, feed and for processing under Administrative Order (AO) No. 8 Part 5 for Corn MON 89034, a second generation product, which has been genetically modified for insect resistance. Corn MON 89034 has been evaluated according to BPI’s safety assessment by concerned agencies of the Department of Agriculture, such as [Bureau of Animal Industry (BAI), Bureau of Agriculture, Fisheries and Product Standards (BAFPS), and a Scientific and Technical Review Panel (STRP) members]. The process involves an intensive analysis of the nature of the genetic modification together with the consideration of safety assessment paradigm which includes molecular characterization, protein characterization, and food/feed composition. The petitioner/applicant published the said application in two widely circulated newspapers for public comment/review. During the 30-day comment period, BPI had not received comments on the said application. The STRP and agencies’ assessment and review of results of evaluation by the BPI Biotech Core Team completed the approval process.
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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:
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Authorization expiration date:
E-mail:
bpibiotechsecretariat@yahoo.com
Organization/agency name (Full name):
Bureau of Plant Industry
Contact person name:
Thelma L. Soriano
Website:
Physical full address:
San Andres St., Malate, Manila
Phone number:
632 521 1080
Fax number:
632 521 1080
Country introduction:
The Philippines is the first ASEAN country to establish a modern regulatory system for modern biotechnology. The country's biosafety regulatory system follows strict scientific standards and has become a model for member-countries of the ASEAN seeking to become producers of agricultural biotechnology crops. Concerns on biosafety in the Philippines started as early as 1987 when scientists from the University of the Philippines Los Banos (UPLB) and International Rice Research Institute (IRRI), the Quarantine Officer of the Bureau of Plant Industry (BPI) and the Director for Crops of the Philippine Council for Agriculture, Forestry and Natural Resources Research and Development (PCARRD) recognized the potential for harm of the introduction of exotic species and genetic engineering. The joint committee formed the biosafety protocols and guidelines for genetic engineering and related research activities for UPLB and IRRI researchers. This proposal was eventually adapted into a Philippine Biosafety policy by virtue of Executive Order No 430, Series of 1990, issued by then President Corazon C. Aquino on October 15, 1990, which created the National Committee on Biosafety of the Philippines (NCBP). The NCBP formulates, reviews and amends national policy on biosafety and formulates guidelines on the conduct of activities on genetic engineering. The NCBP comprised of representative from the Department of Agriculture (DA); Department of Environment and Natural Resources (DENR); Health (DOH); and Department of Science and Technology (DOST), 4 scientists in biology, environmental science, social science and physical science and 2 respected members of the community. The Philippines’ Law, Executive Order No.514 (EO514), Series of 2006 entitled “Establishing the National Biosafety Framework (NBF), Prescribing Guidelines for its Implementation, Strengthening the National Committee on Biosafety of the Philippines, and for Other Purposes was also issued. This order sets the establishment of the departmental biosafety committees in the DA, DENR, DOH and DOST. The mandates jurisdiction and other powers of all departments and agencies in relation to biosafety and biotechnology is guided by the NBF in coordination with the NCBP and each other in exercising its power. The Department of Agriculture (DA) issued Administrative Order No 8, Series of 2002, (DA AO8, 2002), which is part of EO 514, for the implementation of guidelines for the importation and release into the environment of plants and plant products derived from the use of modern biotechnology. The DA authorizes the Bureau of Plant Industry (BPI) as the lead agency responsible for the regulation of agricultural crops developed through modern biotechnology. The BPI has adopted a protocol for risk assessment of GM crops for food and feed or for processing based on the Codex Alimentarius Commission’s Guideline for the Conduct of Food Safety assessment of Foods Derived from Recombinant-DNA plants and a protocol for environmental risk assessment in accordance with the Cartagena Protocol on Biosafety and with the recommendation of the Panel of Experts of the Organization for Economic Cooperation and Development (OECD). DA AO8, 2002 ensures that only genetically food crops that have been well studied and found safe by parallel independent assessments by a team of Filipino scientists and technical personnel from the concerned regulatory agencies of the Department are allowed into our food supply and into our environment. The DA AO 8, 2002 has a step by step introduction of GM plant into the environment. The research and development phase would require testing the genetically modified (GM) crop under controlled conditions subject to regulation by the government agencies. The first stage of evaluation for GM crops is testing under contained facilities such as laboratories, greenhouses and screenhouses. After satisfactory completion of testing under contained facilities, confined environmental release or field trial is done. Confined field trial (CFT) is the first controlled introduction of the GM crop into the environment. The approval for field trial shall be based on the satisfactory completion of safety testing under contained conditions. Unconfined environmental release or commercialization of the product would follow after the safe conduct of the CFT. Approval for propagation shall only be allowed after field trials and risk assessment show no significant risk to human and animal health and the environment.
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Stacked events:
Gene stacking in plants can be conferred either through genetic engineering or conventional breeding A full risk assessment as to food and feed or for processing shall be conducted to plant products carrying stacked genes conferred through genetic engineering or conventional breeding, where the individual traits have no prior approval for direct use as food and feed or processing from the Bureau of Plant Industry (BPI) A desktop or documentary risk assessment on the possible or expected interactions between the genes shall be conducted for stacked gene products with multiple traits conferred through conventional breeding and individual events granted prior approval by the Bureau of Plant Industry.
Contact details of the competent authority(s) responsible for the safety assessment and the product applicant:
Bureau of Plant Industry 692 San Andres St, Malate, Manila 1004