Decision Document DD2013-99
Determination of the Safety of Monsanto Canada Inc.'s Soybean (Glycine max (L.) Merr.) Event MON 87712

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This Decision Document has been prepared to explain the regulatory decisions reached under Directive 94-08 (Dir94-08) - Assessment Criteria for Determining Environmental Safety of Plants with Novel Traits, its companion biology document BIO1996-10 - The Biology of Glycine max (L.) Merr. (Soybean), and Section 2.6 - Guidelines for the Assessment of Novel Feeds: Plant Sources, of Chapter 2 of the RG-1 Regulatory Guidance: Feed Registration Procedures and Labelling Standards.

The Canadian Food Inspection Agency (CFIA), specifically the Plant Biosafety Office of the Plant Health and Biosecurity Directorate, the Plant and Biotechnology Risk Assessment Unit of the Plant Health Science Directorate, and the Animal Feed Division of the Animal Health Directorate, has evaluated information submitted by Monsanto Canada Inc. This information concerns soybean event MON 87712, a soybean with increased yield potential. The CFIA has determined that this plant with a novel trait (PNT) does not present altered environmental risk nor, as a novel feed, does it present livestock feed safety concerns when compared to soybean varieties currently grown and permitted to be used as livestock feed in Canada.

Taking into account these evaluations, unconfined release into the environment and use as livestock feed of soybean event MON 87712 is therefore authorized by the Plant Biosafety Office of the Plant Health and Biosecurity Directorate and the Animal Feed Division of the Animal Health Directorate, respectively, as of September 11, 2013. Any soybean lines derived from soybean event MON 87712 may also be released into the environment and used as livestock feed, provided that

  1. no inter-specific crosses are performed,
  2. the intended uses are similar,
  3. it is known based on characterization that these plants do not display any additional novel traits and are substantially equivalent to soybean varieties that are currently grown and permitted to be used as livestock feed in Canada, in terms of their potential environmental impact and livestock feed safety, and
  4. the novel gene is expressed at a level similar to that of the authorized line.

Soybean event MON 87712 is subject to the same phytosanitary import requirements as unmodified soybean varieties. Soybean event MON 87712 is required to meet the requirements of other jurisdictions, including but not limited to, the Food and Drugs Act.

Please note that the livestock feed and environmental safety assessments of novel feeds and PNTs are critical steps in the potential commercialization of these plant types. Other requirements, such as the evaluation of food safety by Health Canada, have been addressed separately from this review.

September 11, 2013

Table of Contents

  1. Brief Identification of the Modified Plant
  2. Background Information
  3. Description of the Novel Traits
    1. Development Method
    2. Increased Yield Potential
    3. Stable Integration into the Plant Genome
  4. Criteria for the Environmental Assessment
    1. Potential of Soybean Event MON 87712 to Become a Weed of Agriculture or Be Invasive of Natural Habitats
    2. Potential for Gene Flow from Soybean Event MON 87712 to Sexually Compatible Plants Whose Hybrid Offspring May Become More Weedy or More Invasive
    3. Altered Plant Pest Potential of Soybean Event MON 87712
    4. Potential Impact of Soybean Event MON 87712 on Non-Target Organisms
    5. Potential Impact of Soybean Event MON 87712 on Biodiversity
  5. Criteria for the Livestock Feed Assessment
    1. Potential Impact of Soybean Event MON 87712 on Livestock Nutrition
    2. Potential Impact of Soybean Event MON 87712 on Animal Health and Human Safety as it Relates to the Potential Transfer of Residues into Foods of Animal Origin, and Worker/Bystander Exposure to the Feed
  6. New Information Requirements
  7. Regulatory Decision

I. Brief Identification of the Modified Plant

Designation of the Modified Plant: Soybean event MON 87712, OECD Unique Identifier MON-87712-4
Applicant: Monsanto Canada Inc.
Plant Species: Soybean (Glycine max L.)
Novel Trait: Increased yield potential
Trait Introduction Method: Agrobacterium-mediated transformation
Intended Use of the Modified Plant: Soybean event MON 87712 is intended to be grown for traditional soybean human food and livestock feed uses. Soybean event MON 87712 is not intended to be grown outside the normal production area for soybean in Canada.

II. Background Information

Monsanto Canada Inc. has developed soybean event MON 87712, which has increased yield potential. Soybean event MON 87712 was developed using recombinant deoxyribonucleic acid (DNA) technology, resulting in the introduction of the BBX32 gene. The BBX32 gene is derived from the plant Arabidopsis thaliana (A. thaliana) and encodes a B-Box type zinc finger protein that interacts with endogenous transcription factors to control the plant's response to dark to light transition.

Monsanto Canada Inc. has provided information on the identity of soybean event MON 87712, a detailed description of the transformation method, information on the gene insertion site, gene copy number, and levels of gene expression in the plant, and the role of the inserted genes and regulatory sequences. The novel protein was identified and characterized. Information was provided for the evaluation of the potential toxicity of the novel protein to livestock and non-target organisms and potential allergenicity of the novel protein to humans and to livestock.

Soybean event MON 87712 was field tested at 19 sites in the United States (US) in 2009. The locations of these trials share similar environmental and agronomic conditions to soybean production areas in Canada and were considered representative of major Canadian soybean growing regions.

Agronomic characteristics of soybean event MON 87712, such as seed germination and seed dormancy, early stand count, seedling vigour, plant height, days to 50% flowering, days to 50% senescence, lodging, susceptibility to various soybean pests and pathogens, and yield, were compared to those of an unmodified control soybean variety, which shares the same genetic background as soybean event MON 87712 but has not been modified. Several commercial control soybean varieties were also included in these trials to establish a reference range for typical soybean behaviour.

Nutritional components of soybean event MON 87712 grain and forage, such as protein, crude fat, ash, moisture, amino acids, fatty acids, vitamins, minerals, and anti-nutrients were compared with those of the unmodified control soybean variety and conventional control soybean varieties.

The Plant and Biotechnology Risk Assessment (PBRA) Unit of the Plant Health Science Directorate, CFIA, has reviewed the above information, in light of the assessment criteria for determining environmental safety of PNTs, as described Directive 94-08 (Dir94-08) - Assessment Criteria for Determining Environmental Safety of Plants with Novel Traits. The PBRA Unit has considered:

  • the potential of soybean event MON 87712 to become a weed of agriculture or to be invasive of natural habitats;
  • the potential for gene flow from soybean event MON 87712 to sexually compatible plants whose hybrid offspring may become more weedy or more invasive;
  • the potential for soybean event MON 87712 to become a plant pest;
  • the potential impact of soybean event MON 87712 and its gene products on non-target organisms, including humans; and
  • the potential impact of soybean event MON 87712 on biodiversity.

The Animal Feed Division (AFD) of the CFIA has also reviewed the above information with respect to the assessment criteria for determining the safety and efficacy of livestock feed, as described in Section 2.6 - Guidelines for the Assessment of Novel Feeds: Plant Sources, of Chapter 2 of the RG-1 Regulatory Guidance: Feed Registration Procedures and Labelling Standards.

The AFD has considered both intended and unintended effects and similarities and differences between soybean event MON 87712 and unmodified soybean varieties relative to the safety and efficacy of feed ingredients derived from soybean event MON 87712 for their intended purpose; including:

  • the potential impact of soybean event MON 87712 on livestock nutrition; and
  • the potential impact of soybean event MON 87712 on animal health and human safety, as it relates to the potential transfer of residues into foods of animal origin and worker/bystander exposure to the feed.

The AFD has also considered whether feeds derived from soybean event MON 87712 meet the definitions and requirements of feeds as listed in Schedule IV of the Feeds Regulations.

Monsanto Canada Inc. has provided the CFIA with a method for the detection and identification of soybean event MON 87712.

III. Description of the Novel Trait

1. Development Method

Soybean event MON 87712 was developed through Agrobacterium-mediated transformation of conventional soybean meristems using two transfer DNAs (T-DNA I and T-DNA II). T-DNA I contains the BBX32 expression cassette. T-DNA II contains the cp4 epsps expression cassette, encoding the 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) protein from Agrobacterium spp. strain CP4, known as CP4 EPSPS. The CP4 EPSPS protein confers tolerance to the herbicide glyphosate and was only used as a selectable marker. During plant transformation, T-DNA I and T-DNA II were inserted into two unlinked loci within the soybean genome. Transformants that contained both T-DNAs were selected based on their tolerance to glyphosate then selfed to segregate the unlinked insertions of T-DNA I and T-DNA II. The progeny that contained T-DNA II were eliminated from subsequent breeding, while the progeny that only contained T-DNA I were maintained. Soybean event MON 87712 was identified as a successful transformant based on molecular analyses and agronomic evaluations and was thus chosen for further development.

2. Increased Yield Potential

Soybean event MON 87712 contains the BBX32 gene from A. thaliana which encodes BBX32, a zinc finger protein of the B-Box zinc finger family. In A. thaliana, the BBX32 forms a protein complex with endogenous transcription factors to modulate light signal transduction. In soybean event MON 87712, the BBX32 protein functions within a similar signalling pathway to control the plant's response to dark to light transition. This modulation of diurnal metabolism in the soybean plant translates into increased yield potential of soybean event MON 87712 compared to the unmodified control soybean.

BBX32 protein expression in soybean event MON 87712 is driven by a constitutive promoter. Samples of soybean tissues were collected from plants from eight field trials in the US. The BBX32 protein expression levels in nanograms of protein per gram of dry weight tissue (ng/g dwt) were determined using a highly sensitive Western blot method. The average BBX32 protein expression levels ranged from below the detection limit to 35 ng/g dwt in leaves. The average BBX32 protein expression level was 3.9 ng/g dwt in roots of plants that had reached the physiological maturity (R6) stage. In some tissues, including seed and forage, the BBX32 protein expression levels were below the detection limit.

The potential allergenicity and toxicity of the BBX32 protein to livestock and non-target organisms were evaluated. The weight of evidence indicates that the BBX32 protein is unlikely to be allergenic. The source of the BBX32 gene protein, A. thaliana, is not known to produce allergens, the BBX32 protein amino acid sequence lacks relevant similarities to known allergens, and the microbial BBX32 protein was shown experimentally to be rapidly degraded in simulated gastric fluid. It was also concluded that the BBX32 protein is unlikely to be toxic to livestock or non-target organisms because it lacks a mode of action to suggest that it is intrinsically toxic to livestock or non-target organisms, the BBX32 protein amino acid sequence lacks relevant similarities to known toxins, and no observed adverse effects were observed when microbial BBX32 protein was ingested by mice at doses of 29 milligrams protein per kilogram body weight (mg/kg bwt). For a more detailed discussion of the potential allergenicity and toxicity of the BBX32 protein, see Section V, part 2: Potential Impact of Soybean Event MON 87712 on Animal Health and Human Safety as it Relates to the Potential Transfer of Residues into Foods of Animal Origin, and Worker/Bystander Exposure to the Feed.

3. Stable Integration into the Plant Genome

Molecular characterization by Southern blot analysis demonstrated that soybean event MON 87712 contains one intact copy of the gene cassette containing the BBX32 gene and its regulatory elements inserted at a single site in the soybean genome. No additional elements, including intact or partial DNA fragments of the gene cassette, T-DNA II sequences, or backbone sequences from the plasmid vector, linked or unlinked to the intact insert, were detected in soybean event MON 87712.

The stability of the insert within soybean event MON 87712 was verified by Southern blot analysis over five generations. The inheritance pattern of the insert across three segregating generations of soybean event MON 87712 showed that the insert segregates according to Mendelian rules of inheritance for a single genetic locus.

IV. Criteria for the Environmental Assessment

1. Potential of Soybean Event MON 87712 to Become a Weed of Agriculture or be Invasive of Natural Habitats

The biology of soybean, as described in the CFIA biology document BIO1996-10 - The Biology of Glycine max (L.) Merr. (Soybean), is such that unmodified plants of this species are not invasive of unmanaged habitats in Canada. Soybean does not possess an intrinsic potential to become weedy in Canada due to traits such as the lack of seed dormancy and the poor competitive ability of seedlings. According to the information provided by Monsanto Canada Inc., soybean event MON 87712 was determined not to be significantly different from unmodified soybean varieties in this respect.

The CFIA evaluated data submitted by Monsanto Canada Inc. on the reproductive biology and life history traits of soybean event MON 87712. As previously mentioned, soybean event 87712 was field tested in the US at 19 locations in the 2009 growing season. It was determined that the US locations share similar environmental and agronomic conditions to southwestern Ontario and were considered to be representative of major Canadian soybean growing regions. During the field trials, soybean event MON 87712 was compared to the unmodified control soybean variety. Eighteen commercial control soybean varieties were also included in these trials to establish ranges of comparative values that are representative of currently grown soybean varieties. Phenotypic and agronomic traits were evaluated, covering a broad range of characteristics that encompass the entire life cycle of the soybean plant. The traits included early stand count, seedling vigour, days to 50% flowering, flower color, days to 50% end of flowering, days to 50% senescence, days to physiological maturity, plant height, lodging, pod shattering, final stand count, grain moisture, 100 seed weight, and yield. Combined site comparison of soybean event MON 87712 to the unmodified control soybean variety showed a trend occurring in which soybean event MON 87712 had a higher early stand count, increased days to 50% senescence, increased days to physiological maturity, higher final stand count, and higher yield. Increased days to 50% senescence and increased days to physiological maturity are indicative of an extended period of photosynthetic activity, which is consistent with the putative mode of action of the BBX32 protein. Increased yield is consistent with the expected novel trait. Although statistically significant differences between soybean event MON 87712 and the unmodified control soybean variety were observed for the five above characteristics, the values for soybean event MON 87712 were within the reference range established from the commercial soybean varieties grown in the same field trials. Therefore, the statistical analysis of these observations supports a conclusion of phenotypic and agronomic equivalence of soybean event MON 87712 to currently grown soybean varieties.

Monsanto Canada Inc. evaluated the germination of soybean event MON 87712 seed under six different temperature regimes. Seed germination characteristics were evaluated including percent germinated seed (normal and/or abnormal), percent viable hard seed (an indicator of seed dormancy), percent dead seed, and percent viable firm swollen seed. Soybean event MON 87712 was compared to the unmodified control soybean variety. Ten commercial control soybean varieties were also included to establish ranges of comparative values that are representative of currently grown soybean varieties. Seed lots were produced at three sites in the US. Although instances of statistically significant differences were observed between soybean event MON 87712 and the unmodified control soybean variety for percent germinated seed and percent dead seed in the individual-site analyses, these differences were small in magnitude or within the reference range established from the commercial control soybean varieties and therefore not considered to be biologically meaningful. Therefore the introduction of the novel trait did not impact the germination of the soybean seed and did not confer dormancy to the soybean seed.

The volunteer potential of soybean event MON 87712 was assessed at four sites in the US Soybean event MON 87712 seeds were seeded in the fall of 2009. A small number of volunteer plants were detected at two sites in the following spring; however there was no significant difference between soybean event MON 87712 and the unmodified control soybean variety.

Soybean event MON 87712 was exposed to cold, soil compaction, drought, flood, frost, hail, mineral toxicity, nutrient deficiency, and wind during the agronomic characteristic studies. No trend in increased or decreased susceptibility to these abiotic stressors was observed in soybean event MON 87712 compared to the unmodified control soybean variety.

The susceptibility of soybean event MON 87712 to soybean pests and pathogens was evaluated in the field at the same locations as the agronomic characteristic studies (further detail provided below in Section IV, part 3: Altered Plant Pest Potential of Soybean Event MON 87712). No trend in increased or decreased susceptibility to pests and pathogens was observed in soybean event MON 87712 compared to the unmodified control soybean variety.

The novel trait has no intended or observed effects on weediness or invasiveness. No competitive advantage was conferred to plants of soybean event MON 87712, as the reproductive and growth characteristics, and tolerance to abiotic and biotic stresses of soybean event MON 87712 were comparable to those of the unmodified control soybean variety or commercial control soybean varieties.

The CFIA has therefore concluded that soybean event MON 87712 has no altered weediness or invasiveness potential in Canada compared to currently grown soybean varieties.

2. Potential for Gene Flow from Soybean Event MON 87712 to Sexually Compatible Plants Whose Hybrid Offspring May Become More Weedy or More Invasive

Natural hybridization between cultivated soybean and the wild annual species Glycine soja (G. soja) can occur. However, G. soja is not naturalized in North America, and although this species is occasionally grown in research plots, there are no reports of its escape to unmanaged habitats nor of it becoming a weed in Canadian agro-ecosystems. The biology of soybean, as described in the CFIA biology document BIO1996-10 - The Biology of Glycine max (L.) Merr. (Soybean), shows that soybeans exhibit a high degree of self-fertilization. Cross pollination is usually less than one percent, suggesting that any pollen flow from cultivated soybeans to related species is minimal.

Monsanto Canada Inc. provided information on the viability, diameter, and morphology of pollen of soybean event MON 87712. No statistically significant differences were detected between soybean event MON 87712 and the unmodified control soybean variety for percent viable pollen or pollen diameter. Furthermore, no visual differences in general morphology were observed between soybean event MON 87712 and the unmodified control soybean variety. Therefore the introduction of the novel trait did not impact the viability and morphology of soybean pollen.

This information, together with the fact that the novel trait has no intended effects on soybean reproductive biology, led the CFIA to conclude that there is minimal potential for gene flow from soybean event MON 87712 to related species in Canada.

3. Altered Plant Pest Potential of Soybean Event MON 87712

Soybean is not considered a plant pest in Canada and the novel trait introduced into soybean event MON 87712 (increased yield potential) is unrelated to plant pest potential (e.g. the potential for the plant to harbour new or increased populations of pathogens or pests).

The susceptibility of soybean event MON 87712 to various soybean pests and pathogens was evaluated in the field at the same locations as the agronomic characteristic studies. The stressors observed included aphids, armyworms, bean leaf beetles, cabbage loopers, grasshoppers, green cloverworms, japanese beetles, leafhoppers, lygus bug, silver spotted skippers, soybean loopers, soybean stem borer, stink bugs, three-cornered alfalfa hoppers, thrips, velvetbean caterpillars, white fly, wireworm, Anthracnose, aster yellow phytoplasma, bacterial blight, bacterial leaf spot, brown stem rot, Cercospora, charcoal rot, downy mildew, frogeye leaf spot, Fusarium, leaf spots (Septoria and Alternaria), Phytophtora, powdery mildew, Pythium, Rhizoctonia, seedling blight, Septoria brown spot, southern blight, soybean mosaic virus, soybean rust, stem canker, sudden death, and white mold. Evaluations of soybean event MON 87712 did not show any increase or decrease in susceptibility to pathogens compared to the unmodified control soybean variety for any of the 198 observations. A total of eight statistically significant differences involving bean leaf beetle, green cloverworms, Japanese beetles, leafhoppers, soybean loopers, and velvet caterpillars were observed between soybean event MON 87712 and the unmodified control soybean variety out of 137 observations relating to susceptibility of pest insects. The mean damage ratings for soybean event MON 87712 were outside the reference ranges established from the commercial control soybean varieties grown in the same trials. However the differences between soybean event MON 87712 and the unmodified control soybean variety were small in magnitude and were not consistently detected across observation times or sites. These results support that the detected differences in susceptibility to pest insects are not associated with the novel trait and are not biologically meaningful.

The CFIA has therefore concluded that soybean event MON 87712 does not display any altered plant pest potential compared to currently grown soybean varieties.

4. Potential Impact of Soybean Event MON 87712 on Non-Target Organisms

The novel trait introduced into soybean event MON 87712 (increased yield potential) is unrelated to a potential impact on non-target organisms.

Detailed characterization of the BBX32 protein expressed in soybean event MON 87712 led to the conclusion that this protein does not display any characteristics of a potential toxin or allergen (see Section III: Description of the Novel Trait). Therefore, no negative impacts resulting from exposure of organisms to the BBX32 protein expressed in soybean event MON 87712 are expected.

Composition analyses showed that the levels of key nutrients and anti-nutrients in grain and forage from soybean event MON 87712 are comparable to those in conventional soybean varieties (see Section V, part 1: Potential Impact of Soybean Event MON 87712 on Livestock Nutrition). Therefore, it is very unlikely that the introduction of the novel trait may have caused unintended changes to the composition of soybean event MON 87712 tissues that would negatively impact organisms interacting with soybean event MON 87712.

Soybean event MON 87712 and the unmodified control variety were compared for key characteristics of soybean's symbiotic association with the nitrogen-fixing soil bacteria Bradyrhizobium japonicum. No statistically significant differences were observed for any characteristics, including nodule number, shoot percent total nitrogen, shoot total nitrogen, and dry weight of nodules, shoots, and roots. Therefore, the symbiotic relationship between B. japonicum and soybean has not been altered by the introduction of the novel trait.

The abundance of pest and beneficial arthropods was compared in plots of soybean event MON 87712 and plots of the unmodified control soybean variety grown at the same locations. The pest and beneficial arthropods assessed included aphids, bean leaf beetles, corn earworms, green cloverworms, garden flea hoppers, leafhoppers, soybean loopers, stink bugs, three-cornered alfalfa hoppers, tarnished plant bugs, yellow-stripped armyworms, yellow woollybear caterpillars, spiders, big-eyed bugs, carabid beetles, lacewings, ladybird beetles, parasitic wasps, damsel bugs (Nabis spp.), and Orius spp. A total of 123 observations of arthropod abundance were made between soybean event MON 87712 and the unmodified control soybean variety, including 59 observations of pest arthropods and 56 observations of beneficial arthropods. A statistical analysis was not generated for eight out of the 123 observations because of lack of variability in the data. Although five statistically significant differences were detected out of 59 observations for pest arthropod abundance involving aphids, bean leaf beetles, green cloverworms, and stink bugs, these differences were small in magnitude and were not consistently detected across collection times or sites. Furthermore, a total of four statistically significant differences were detected out of 56 observations for beneficial arthropod abundance involving big-eyed bugs, ladybird beetles, micro-parasitic hymenoptera, and Orius spp. These differences were small in magnitude and were not consistently detected across collection times or sites. The results support that the detected differences in beneficial and pest arthropod abundance between soybean event MON 87712 and the unmodified control soybean variety are not associated with the novel trait and are not biologically meaningful.

Furthermore, field evaluations of soybean event MON 87712 did not show any increased resistance to pest insects or pathogens compared to the unmodified control soybean variety (see Section IV, part 3: Altered Plant Pest Potential of Soybean Event MON 87712).

Collectively, these information elements indicate that the interactions between soybean event MON 87712 and the populations of animals and microorganisms interacting with soybean will be similar compared to currently grown soybean varieties.

The CFIA has therefore determined that the unconfined release of soybean event MON 87712 in Canada will not result in altered impacts on non-target organisms, including humans, compared to currently grown soybean varieties.

5. Potential Impact of Soybean Event MON 87712 on Biodiversity

Soybean event MON 87712 expresses no novel phenotypic characteristics that would extend its range beyond the current geographic range of soybean production in Canada. Soybean's only sexually compatible wild relative in Canada (G. soja) does not occur in unmanaged habitats, and the possibility of soybean outcrossing to G. soja is very low. Soybean event MON 87712 is unlikely to cause adverse effects on non-target organisms and does not display increased weediness, invasiveness, or plant pest potential. The introduction of soybean event MON 87712 is not likely to affect current soybean cultivation practices or crop rotation practices. It is therefore unlikely that soybean event MON 87712 will have any direct or indirect effects on biodiversity, in comparison to the effects that would be expected from the cultivation of the soybean varieties that are currently grown in Canada.

The CFIA has concluded that the introduced gene and its corresponding novel trait do not confer to soybean event MON 87712 any characteristic that would result in unintended environmental effects following unconfined environmental release. The CFIA has therefore concluded that the potential impact on biodiversity of soybean event MON 87712 is unlikely to be different from that of the soybean varieties that are currently grown in Canada.

V. Criteria for the Livestock Feed Assessment

The AFD considered nutrient and anti-nutrient profiles, the safety of feed ingredients derived from soybean event MON 87712, including the presence of gene products, residues, and metabolites in terms of animal health and human safety as it relates to the potential transfer of residues into foods of animal origin, and worker/bystander exposure to the feed, and whether feeds derived from soybean event MON 87712 meet the definitions and requirements of feeds as listed in Schedule IV of the Feeds Regulations.

1. Potential Impact of Soybean Event MON 87712 on Livestock Nutrition

Nutrient and Anti-nutrient Composition:

The nutritional equivalence of soybean event MON 87712 to its non-transgenic unmodified control soybean (A3525) variety and 16 conventional soybean varieties was determined from eight replicated field sites in the US during the 2009 growing season. At each field site, soybean event MON 87712, the unmodified control, and the conventional soybean varieties were planted in a randomized complete block design with four plots of each soybean type. Forage and harvested soybeans were analysed for protein, crude fat, ash, moisture, acid detergent fibre (ADF), and neutral detergent fibre (NDF). Harvested soybeans were further analysed for amino acids, fatty acids, vitamin E, isoflavones (diadzein, genistein, and glycitein), and anti-nutrients (lectin, phytic acid, trypsin inhibitor, raffinose, and stachyose), as recommended by the OECD consensus document for new varieties of soybean (OECD, 2001). Composition data was analysed statistically using a mixed model of analysis of variance, and statistical differences among soybean treatments were identified (P<0.05). The biological relevance of any statistical differences between soybean event MON 87712 and the unmodified control was assessed within the 99% tolerance interval calculated from the 16 conventional soybean varieties. Statistical significant differences between soybean event MON 87712 and the unmodified control were also assessed within the context of the natural variability of conventional soybean composition data published in scientific literature; OECD (2001) and in the ILSI Crop Composition Database (ILSI 2010).

The crude fat content of soybean event MON 87712 forage was statistically significantly lower than that of the unmodified control, but the mean values were within the natural variation observed for the conventional soybean varieties and published literature. No statistical significant differences were observed between soybean event MON 87712 and the unmodified control forage for protein, ash, moisture, ADF and NDF. All means were within the range for the conventional varieties and published literature. The crude protein content in soybean event MON 87712 was statistically significantly higher than the unmodified control, however the means were within the range of the conventional soybean varieties and the differences was therefore not considered biologically relevant. No statistically significant differences were observed between soybean event MON 87712 and unmodified control for crude fat, ash, moisture, ADF and NDF. Statistically significant differences were observed between soybean event MON 87712 and the unmodified control for amino acids: alanine, arginine, aspartic acid, cystine, glutamic acid, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, and valine. However, all means were within the range for the conventional soybean varieties and published literature values and therefore the differences were not considered biologically relevant. Palmitic and stearic acids in soybean event MON 87712 were statistically significantly lower than the levels in the unmodified control, but the means were within the range of the conventional soybean varieties, and therefore the differences were not considered biologically relevant. No statistically significant differences were observed between soybean event MON 87712 and the unmodified control for oleic, linoleic, linolenic, arachidic, eicosenoic and behenic acids. No statistically significant differences were observed between soybean event MON 87712 and the unmodified control of the isoflavones (diadzein, genistein, and glycitein), and vitamin E. For anti-nutrients (phytic acid, trypsin inhibitor, lectin, raffinose, and stachyose), no statistically significant differences were observed between soybean event MON 87712 and the unmodified control. All means were within the range of the conventional soybean varieties.

Conclusion:

It was concluded based on the evidence provided by Monsanto Canada Inc. that the nutritional composition of soybean event MON 87712 is similar to that of the conventional control soybean varieties grown in the trials and to that reported for other soybeans in the published scientific literature.

2. Potential Impact of Soybean Event MON 87712 on Animal Health and Human Safety as it Relates to the Potential Transfer of Residues into Foods of Animal Origin, and Worker/Bystander Exposure to the Feed

Soybean event MON 87712 expresses the BBX32 gene derived from A. thaliana, which affects DNA transcription and light signalling pathways that result in increased yield potential for the soybean plant. The assessment of soybean event MON 87712 evaluated the impact of the following potential hazards relative to the safety of feed ingredients derived from these events:

  • the presence of the novel BBX32 protein.

Novel BBX32 Protein:

To obtain sufficient quantities of BBX32 protein for evaluation of environmental and feed safety studies, it was necessary to express the BBX32 gene in a microbial production system.

Equivalency was demonstrated between soybean event MON 87712-produced BBX32 protein and the microbial-produced BBX32 protein that was used in the safety studies based on similar molecular weights and immunoreactivities.

The potential allergenicity and toxicity of the BBX32 protein to livestock was evaluated. With respect to its potential allergenicity, no single experimental method yields decisive evidence, thus a weight-of-evidence approach was taken, taking into account information obtained with various test methods. The source of the BBX32 gene, A. thaliana, is not known to produce allergens, and a bioinformatics evaluation of the BBX32 protein amino acid sequence confirmed the lack of relevant similarities between the BBX32 protein and known allergens. Studies with the microbial BBX32 protein indicated that it is rapidly degraded in simulated gastric fluid and is not heat stable. The weight of evidence thus indicates that the BBX32 protein is unlikely to be allergenic.

In terms of its potential toxicity to livestock, the BBX32 protein lacks a mode of action to suggest that it is intrinsically toxic to livestock and a bioinformatics evaluation of the BBX32 protein amino acid sequence confirmed the lack of relevant similarities between the BBX32 protein and known toxins. In addition, microbial BBX32 protein safety studies indicated that no adverse effects were observed when the BBX32 protein was ingested by CD-1 mice at doses of 29 milligrams protein per kilograms body weight (mg/kg bwt). This information indicates that the BBX32 protein is unlikely to be toxic to livestock.

The livestock exposure to the BBX32 protein is expected to be negligible as the protein is expressed at very low levels in soybean event MON 87712, is rapidly degraded under conditions which simulate the mammalian digestive tract, and is unstable under heating conditions expected to be encountered during processing of some soybean products.

The effects in soybean event MON 87712 resulting from the insertion of the BBX32 gene were considered in detail. In characterizing the mode of action, it was determined that novel BBX32 protein functions analogously to native BBX proteins, within the existing framework of soybean light signalling processes. The expression of BBX32 influences the transcription of DNA, and other targets which control the soybean responses to dark to light transition, to produce an outcome in those light responses that is different to the native BBX protein functions. Evidence was supplied from which it was determined that the novel BBX32 makes use of existing signalling pathways; and this leads to the conclusion that there is limited scope for unintended effects within the event MON 87712.

The lack of performance effects or significant health observations in the broiler study using soybean meal derived from soybean event MON 87712 supports the safety of the BBX32 protein.

Conclusion:

Feed ingredients derived from soybean event MON 87712 are considered to meet present ingredient definitions for soybean and as such are approved for use as livestock feed in Canada.

VI. New Information Requirements

If at any time Monsanto Canada Inc. becomes aware of any new information regarding risk to the environment, livestock, or human health, which could result from release or livestock feed use of soybean event MON 87712 or lines derived from it, Monsanto Canada Inc. is required to immediately provide such information to the CFIA. On the basis of such new information, the CFIA will re-evaluate the potential impact of soybean event MON 87712 on the environment, livestock, and human health, and may re-evaluate its decision with respect to the livestock feed use and unconfined environmental release authorizations of soybean event MON 87712.

VII. Regulatory Decision

Based on the review of the data and information submitted by Monsanto Canada Inc., and input from other relevant scientific sources, the Plant and Biotechnology Risk Assessment Unit of the Plant Health Science Directorate, CFIA, has concluded that the unconfined environmental release of soybean event MON 87712 does not present altered environmental risk when compared to soybean varieties that are currently grown in Canada.

Based on the review of the data and information submitted by Monsanto Canada Inc., and input from other relevant scientific sources, the Animal Feed Division of the Animal Health Directorate, CFIA, has concluded that the novel BBX32 protein-based increased yield trait will not confer to soybean event MON 87712 any characteristic that would raise any concerns regarding the safety or nutritional composition of soybean event MON 87712. Grain soybean, its by-products, and soybean oil are currently listed in IV of the Feeds Regulations and are therefore approved for use in livestock feeds in Canada. Soybean event MON 87712 has been found to be as safe as and as nutritious as currently and historically grown soybean varieties. Soybean event MON 87712 and its products are considered to meet present ingredient definitions and are approved for use as livestock feed ingredients in Canada.

Unconfined release into the environment and use as livestock feed of soybean event MON 87712 is therefore authorized by the Plant Biosafety Office of the Plant Health and Biosecurity Directorate and the Animal Feed Division of the Animal Health Directorate, respectively, as of September 11, 2013. Any soybean lines derived from soybean event MON 87712 may also be released into the environment and used as livestock feed, provided that

  1. no inter-specific crosses are performed,
  2. the intended uses are similar,
  3. it is known based on characterization that these plants do not display any additional novel traits and are substantially equivalent to soybean varieties that are currently grown and permitted to be used as livestock feed in Canada, in terms of their potential environmental impact and livestock feed safety, and
  4. the novel gene is expressed at a level similar to that of the authorized line.

Soybean event MON 87712 is subject to the same phytosanitary import requirements as unmodified soybean varieties. Soybean event MON 87712 is required to meet the requirements of other jurisdictions; including but not limited to, the Food and Drugs Act.

Please refer to Health Canada's Decisions on Novel Foods for a description of the food safety assessment of soybean event MON 87712.

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