Decision Document DD2011-85 Determination of the Safety of Monsanto Canada Inc.'s Soybean (Glycine max (L.) Merr.) Event MON 87769

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October 13, 2011

This Decision Document has been prepared to explain the regulatory decision reached under Directive 94-08 (Dir94-08), entitled "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 Chapter 2.6 of the Regulatory Guidance: Feed Registration Procedures and Labelling Standards, entitled "Guidelines for the Assessment of Novel Feeds: Plant Sources".

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 is in regard to the improved fatty acid profile in soybean oil event MON 87769. 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 currently commercialized soybean varieties in Canada.

Taking into account these evaluations, unconfined release into the environment and use as livestock feed of soybean event MON 87769 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 as of October 13, 2011. Any soybean lines derived from soybean event MON 87769 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. based on characterization that these plants do not display any additional novel traits and are substantially equivalent to currently grown soybean in Canada, in terms of their potential environmental impact and livestock feed safety; and
  4. the novel genes are expressed at a level similar to that of the authorized line.

Soybean event MON 87769 is subject to the same phytosanitary import requirements as its unmodified counterparts.

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.

This bulletin is published by the Canadian Food Inspection Agency. For further information, please contact the Plant Biosafety Office or the Animal Feed Division at:

Plant Biosafety Office
Plant Health and Biosecurity Directorate
59 Camelot Drive
Ottawa ON  K1A 0Y9
Telephone: 613-225-2342
Animal Feed Division
Animal Health Directorate
59 Camelot Drive
Ottawa ON  K1A 0Y9
Telephone: 613-225-2342

Table of Contents

  1. Brief Identification of the Modified Plant
  2. Background Information
  3. Description of the Novel Trait
    1. Modified fatty acid profile
    2. Development Method
    3. Stable Integration into the Plant Genome
  4. Criteria for the Environmental Assessment
    1. Potential of Soybean Event 87769 to become a Weed of Agriculture or be Invasive of Natural Habitats
    2. Potential for Gene Flow from Soybean Event 87769 to Wild Relatives whose Hybrid Offspring May Become More Weedy of More Invasive
    3. Altered Plant Pest Potential of Soybean Event 87769
    4. Potential Impact of Soybean Event MON 87769 on Non-Target Organisms
    5. Potential Impact of Soybean Event MON 87769 on Biodiversity
  5. Criteria for the Livestock Feed Assessment
    1. Potential Impact of Soybean Event MON 87769 on Livestock Nutrition
    2. Potential Impact of Soybean Event MON 87769 on Livestock and Workers/Bystanders
  6. New Information Requirements
  7. Regulatory Decision

I. Brief Identification of the Modified Plant

Designation of the Modified Plant: Soybean event MON 87769, OECD Unique Identifier MON-87769-7
Applicant: Monsanto Canada Inc.
Plant Species: Soybean (Glycine max (L.) Merr.)
Novel Traits: Modified fatty acid profile in soybean oil (Increased levels of stearidonic acid (SDA) (20-30% by weight of total fatty acids) and gamma linolenic acid (GLA) (~7% by weight of total fatty acids)
Trait Introduction Method: Agrobacterium-mediated transformation
Intended Use of the Modified Plant: Human consumption of the oil and livestock feed use of the meal. Soybean event MON 87769 is not intended to be grown outside the normal production area for soybean in Canada.

II. Background Information

Monsanto Canada Inc. has developed, through the use of recombinant DNA techniques, a soybean with a modified fatty acid profile. The soybean event, designated as MON 87769, was developed to provide an alternate source of an omega-3 fatty acid to help meet the need for increased dietary intake of long chain omega-3 fatty acids. Soybean event 87769 is expected to provide oil that is more stable to oxidation than other oils such as fish oils.

Event MON 87769 was produced by Agrobacterium-mediated transformation of soybean with plasmid vector PV-GMPQ1972 containing two T-DNAs (T-DNA I and T-DNA II). The T DNA I region contains two expression cassettes: the Pj.D6D expression cassette coding for Primula juliae Δ6 desaturase (PjΔ6D) and the Nc.Fad3 gene expression cassette coding for Neurospora crassa Δ15 desaturase (NcΔ15D). The introduction of these two genes results in the seed-specific production of the PjΔ6D and NcΔ15D desaturase proteins. These proteins desaturate certain endogenous fatty acids resulting in the production of Stearidonic acid at approximately 20-30% of total fatty acids and gamma linolenic acid (GLA) at approximately 7%. The T DNA II region contains the cp4 epsps gene expression cassette, encoding the 5 enolpyruvylshikimate-3-phosphate synthase (EPSPS) protein from Agrobacterium sp. strain CP4, known as CP4 EPSPS, which confers tolerance to glyphosate. The cp4 epsps gene expression cassette was used for early event selection, and was segregated away from T-DNA I by conventional breeding.

Monsanto Canada Inc. has provided a wide variety of studies to determine the safety of soybean event MON 87769, including detailed description of the transformation method and the role of the inserted genes, gene segments and regulatory sequences, as well as data and information on the insertion site and gene copy number. Data was also provided for the evaluation of the potential toxicity and allergenicity of the novel proteins to humans and to livestock.

Soybean event MON 87769 was field tested in the United States at a total of 21 locations during 2006 and 2007. Some of the locations (Illinois, Iowa, Michigan, and Ohio) of these trials share similar environmental and agronomic conditions to soybean production areas in Southern Ontario and were considered representative of major Canadian soybean growing regions.

Agronomic characteristics of soybean event MON 87769 such as early stand count, seedling vigour, plant growth stage, days to 50% flowering, flower colour, plant pubescence, plant height, lodging, pod shattering, final stand count, seed moisture, 100 seed weight, test weight, and yield were compared to those of the unmodified control.

Nutritional components of soybean event MON 87769, such as protein, fat, carbohydrates, fiber, ash, moisture, amino acids, fatty acids, vitamins, and anti-nutrients were compared with those of the unmodified control.

The Plant and Biotechnology Risk Assessment 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 in the Directive 94-08 (Dir94-08), entitled "Assessment Criteria for Determining Environmental Safety of Plants With Novel Traits". The PBRA Unit has considered the:

  • potential of soybean event MON 87769 to become a weed of agriculture or be invasive of natural habitats;
  • potential for gene flow from soybean event MON 87769 to wild relatives whose hybrid offspring may become more weedy or more invasive;
  • potential for soybean event MON 87769 to become a plant pest;
  • potential impact of soybean event MON 87769 or its gene products on non-target species, including humans; and
  • potential impact of soybean event MON 87769 on biodiversity.

The Animal Feed Division of the Animal Health Directorate, 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 Chapter 2.6 of the Regulatory Guidance: Feed Registration Procedures and Labelling Standards, entitled "Guidelines for the Assessment of Novel Feeds: Plant Sources" . The Animal Feed Division has considered the:

  • impact of soybean event MON 87769 on livestock nutrition; and
  • potential impact of soybean event MON 87769 on livestock and workers/bystanders.

Monsanto Canada Inc. has provided the CFIA with a method for the detection and identification of soybean containing the soybean event MON 87769.

III. Description of the Novel Trait

1. Modified fatty acid profile

Stearidonic acid is a metabolic precursor to the long chain omega-3 fatty acids, eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) from alpha linolenic acid, a common dietary constituent. Stearidonic acid is found in products such as fish and fish/algal oils. An alternative approach to increase omega-3 fatty acid intake is to provide a wider range of foods that are enriched in omega-3 fatty acids so that people can choose foods that suit their usual dietary habits. Soybean event MON 87769 contains approximately 20 to 30% stearidonic acid. Stearidonic acid soybean oil is expected to be more stable to oxidation thereby expanding the potential food formulation options for food companies and consumers and serving as an alternative to fish oils where they are currently used in food production.

Soybean event MON 87769 was created by introducing a Δ6 desaturase gene from Primula juliae and a Δ15 desaturase gene from Neurospora crassa, into conventional soybean, A3525. The introduction of these two genes results in the seed-specific production of the PjΔ6D and NcΔ15D proteins. These proteins desaturate certain endogenous fatty acids resulting in the production of stearidonic acid at approximately 20-30% of total fatty acids. Since soybean lacks a Δ6 desaturase, the minimal requirement for production of stearidonic acid in soybean would be the introduction of a gene encoding Δ6 desaturase. However, Δ6 desaturase may also convert linolenic acid to alpha linolenic acid. Addition of a Δ15 desaturase with temporal expression similar to the Δ6 desaturase increases alpha linolenic acid levels, allowing greater flux to stearidonic acid accumulation in soybean event MON 87769. The Δ15 desaturase also lowers linolenic acid levels, and hence lowers the substrate pool for GLA production.

Protein characterization in soybean event MON 87769 confirms that both PjΔ6D and NcΔ15D proteins are safe for human and animal consumption. The proteins share sequence similarity to proteins that are ubiquitous in the human diet and are directly consumed in many common foods. The proteins are expressed at low levels in the seed. Both proteins lack biologically-relevant amino acid sequence similarities to known allergens, toxins and anti-nutritional proteins known to have adverse effects on mammals. Additionally, both proteins are rapidly digested in simulated gastric and intestinal fluids and do not exhibit any signs of toxicity when administered to mice via oral gavage.

The 2 T-DNAs system was used in the creation of soybean event MON 87769 to generate marker-free soybean plant. The 2 T-DNAs system allowed for insertion T-DNA I (encoding the desaturase proteins) and T-DNA II (encoding the CP4 EPSPS selectable marker) into two independent loci within the genome of the soybean plant. Following selection of the transformants that contain both T-DNAs, T-DNA II was segregated in progeny through subsequent selfing of plants and genetic selection, while progeny with the T-DNA I were maintained.

2. Development Method

Soybean event MON 87769 was developed using 2T-DNA vector transformation and selection techniques. Vectors and methods were selected for transformation to achieve high probability of obtaining the trait of interest and integration of the introduced DNA into a single locus in the plant genome. This helped ensure that only the intended DNA encoding the desired trait(s) was integrated into the plant genome and facilitates the molecular characterization of the product. Soybean event MON 87769 was developed through Agrobacterium-mediated transformation of soybean meristem tissue using plasmid vector PV-GMPQ1972. The vector contains two separate T-DNAs. The first T-DNA (T-DNA I) contains two expression cassettes: the Pj.D6D gene expression cassette and the Nc.Fad3 gene expression cassette. The second T-DNA region (T-DNA II) contains the cp4 epsps gene expression cassette that encodes the CP4 EPSPS protein (5-enolpyruvyl shikimate-3-phosphate synthase protein from Agrobacterium sp. strain CP4) that provides tolerance to the action of glyphosate, which is the active ingredient in Roundup® agricultural herbicide. The use of the 2 T-DNAs system was the basis for an effective approach to generate marker-free plants.

3. Stable Integration into the Plant Genome

Molecular characterization of soybean event MON 87769 confirmed the presence of a single insertion site for each genetic element present in T-DNA I, the lack of plasmid backbone elements and absence of T-DNA II, and the insert stability across generations. In addition, DNA sequencing analyses confirmed the nucleotide sequence and the expected organization of the insert in soybean event MON 87769. Insert segregation analysis showed that the insert segregated as expected according to Mendel's laws of genetics. These results indicated a single detectable chromosomal insertion of T-DNA I in soybean event MON 87769.

Generational stability analysis demonstrated that the expected Southern blot fingerprint of soybean event MON 87769 has been maintained across four generations of breeding, thereby confirming the stability of the DNA insert over multiple generations. These generations were also shown not to contain any detectable T-DNA II or backbone sequence from plasmid PV-GMPQ1972. These results also confirmed the organization of the genetic elements within the Pj.D6D and Nc.Fad3 expression cassettes of soybean event MON 87769, which was identical to those cassettes in plasmid PV-GMPQ1972. Finally, heritability, stability, and protein expression of the stearidonic acid production phenotype were as expected across multiple generations, which corroborates the molecular insert stability analysis and establishes the genetic behaviour of the DNA insert at a single chromosomal locus.

IV. Criteria for the Environmental Assessment

1. Potential of Soybean Event 87769 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 87769 was determined not to be significantly different from their conventional counterparts in this respect.

The Plant and Biotechnology Risk Assessment Unit of the Plant Health Science Directorate evaluated data submitted by Monsanto Canada Inc. on the reproductive biology and life history traits of soybean event MON 87769. This event was field tested in the US in 17 locations in the 2006 growing season and in 4 locations in the 2007 growing season. The US locations share similar environmental and agronomic conditions to south western Ontario and were considered to be representative of major Canadian soybean regions. During the field trials, several phenotypic characteristics were evaluated: early stand count, seedling vigour, plant growth stages, days to 50% flowering, plant height, lodging, pod shattering, final stand count, seed moisture, 100 seed weight, test weight and yield. For the majority of phenotypic characteristics observed, no statistical difference was detected between soybean event MON 87769 and the unmodified parental control. The few statistical differences that were detected between soybean event MON 87769 and the unmodified parental control, showed no consistent trends in the data across locations or years. In addition, in the combined site analysis, no statistical difference was detected between soybean event MON 87769 and the unmodified parental control, for any of the phenotypic characteristics evaluated. The statistical analysis of these observations showed no biologically meaningful differences between soybean event MON 87769 and the unmodified parental control, and supports a conclusion of phenotypic equivalence to currently commercialized soybean varieties.

The seed dormancy and germination of soybean event MON 87769 were compared with the unmodified parental control. No significant differences were detected in percent germinated seed, percent dead seed and percent viable firm, swollen seed and viable hard seed.

The susceptibility of soybean event MON 87769 to various soybean pests and pathogens and various abiotic stresses was evaluated in the field at the same locations as the phenotypic characteristic studies. The stressors observed included: cold stress, soil compaction, drought, frost, hail, heat, mineral toxicity, nutrient deficiency, wind, alternaria leaf spot, anthracnose, Asian rust, bacterial blight, bacterial pustule, brown spot, brown stem rot, Cercospora leaf blight, charcoal rot, downy mildew, Fusarium, Phytophthora, powdery mildew, Pythium, Rhizoctonia, Sclerotinia, Septoria, soybean cyst nematode, soybean mosaic virus, soybean rust, stem canker, sudden death, white mold, aphids, bean leaf beetles, blister beetles, corn rootworm, flea beetles, grasshoppers, green cloverworm, Japanese beetles, leafhoppers, leafrollers, Mexican bean beetles, seed corn maggots, soybean stem borers, spider mites, stink bugs, thistle caterpillars, thrips, velvetbean caterpillar, white flies, wireworms and woolybear caterpillar. No qualitative differences were observed for 701 of 703 observations. For two of the observations where qualitative differences were observed, excess moisture stress and frog eye leaf spot, there was no trend observed across locations and both were within the range of reference cultivars and not biologically meaningful.

The introduction of the Pj.D6D and Nc.Fad3 gene cassettes did not make soybean event MON 87769 weedy or invasive of natural habitats since none of the soybean's reproductive or growth characteristics were modified, and soybean event MON 87769's tolerance to abiotic and biotic stresses was unchanged as well.

This information, together with the fact that the novel traits have no intended effects on soybean weediness or invasiveness, led the Plant and Biotechnology Risk Assessment Unit of the Plant Health Science Directorate to conclude that soybean event MON 87769 has no altered weed or invasiveness potential compared to conventional soybean varieties.

2. Potential for Gene Flow from Soybean Event 87769 to Wild Relatives whose Hybrid Offspring May Become More Weedy of More Invasive

Natural hybridization between cultivated soybean and the wild annual species Glycine 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 agroecosystems. The biology of soybean, as described in the CFIA Biology Document (BIO1996-10), 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 regarding floral development in MON 87769 soybean, including days to 50% flowering and pollen characteristics. No significant differences in time to 50% flowering, pollen viability or pollen diameter were detected between MON 87769 soybean and the unmodified control variety, nor was any difference in pollen morphology observed. This indicates that the reproductive characteristics of soybean event MON 87769 are unaltered with respect to those of unmodified soybean.

This information, together with the fact that the novel traits have no intended effects on soybean reproductive biology, led the Plant and Biotechnology Risk Assessment Unit of the Plant Health Science Directorate to conclude that there is minmal potential for gene flow from soybean event MON 87769 to related species in Canada.

3. Altered Plant Pest Potential of Soybean Event 87769

Soybean is not considered a plant pest in Canada. The novel traits are unrelated to plant pest potential. Ecological evaluations of soybean event MON 87769 did not show any increase or decrease in susceptibility to any insect or disease stressor compared to lines with similar genetic background and commercial soybean varieties grown at the same locations. These stressors included 22 diseases and 17 pest insects.

The Plant and Biotechnology Risk Assessment Unit of the Plant Health Science Directorate therefore concludes that soybean event MON 87769 does not display any altered pest potential compared to conventional soybean varieties.

4. Potential Impact of Soybean Event MON 87769 on Non-Target Organisms

The detailed characterizations of the novel proteins PjΔ6D and NcΔ15D (as summarized in Part III: Description of the Novel Trait) has led to the conclusion that the these proteins are not potential toxins or allergens.

Compositional analyses showed that as a result of the expression of the PjΔ6D and NcΔ15D desturases, MON 87769 soybean seeds contain four fatty acids which are not present in conventional soybean grain, namely stearidonic acid, GLA and two minor fatty acids, trans-stearidonic acid and trans-alpha linolenic acid. Soybean event MON 87769 seeds contain lower levels of linolenic acid than conventional soybean seeds. This reduction was expected as linolenic acid is a precursor of both stearidonic acid and GLA. The levels of palmitic, stearic, oleic, linolenic, arachidic and behenic acids in MON 87769 seeds were within the range of levels found in conventional soybean seeds.

Stearidonic acid and GLA fatty acids are already present in the environment. stearidonic acid and GLA are produced in seeds of many plant species at levels greater than 1% of their total fatty acid composition. Stearidonic acid and GLA are intermediate in mamalian fatty acid metabolism. Many marine fish species contain stearidonic acid. The safety of stearidonic acid soybean oil is supported by the results of a published 28-day gavage study and 90-day/one generation reproductive toxicity study in rats. GLA is present in meats, fish and human breast milk. Trans-stearidonic acid and trans-alpha linolenic acid represent only 0.18% and 0.44% of total fatty acids in MON 87769 seeds, respectively. These two fatty acids in MON 87769 are unlikely to pose any risk to wildlife or bystanders.

As natural components of the plant and animal kingdoms, GLA, stearidonic acid and other polyunstaturated fatty acids are not expected to be detrimental to the environment. Therefore, the modified fatty acid profile of MON 87769 soybean seeds is not expected to negatively impact organisms interacting with soybean.

There were no biologically meaningful differences noted for levels of proximates (fat, carbohydrates, protein, moisture, ash), fiber, amino acids, vitamin E, isoflavones and antinutrients in seeds when MON 87769 soybean was compared to conventional soybean varieties, confirming that the compositional change in MON 87769 soybean seeds is limited to certain fatty acids. Compositional analyses showed that the levels of proximates and fiber in MON 87769 forage are comparable to those in conventional soybean varieties, confirming that the gross composition of the plant green matter is unchanged.

Collectively, this information indicates that exposure to MON 87769 soybean is unlikely to cause negative impacts on organisms interacting with soybean.

Ecological evaluations confirmed that the abundance of beneficial arthropods in soybean event MON 87769 soybean plots was equivalent to that in conventional soybean grown at the same locations. The beneficial organisms assessed included the big-eyed bug, ladybird beetles, micro parasitic hymenoptera, Nabis sp., Orius sp., and spiders. In addition, ecological evaluations of soybean event MON 87769 did not show any increase in resistance to pest insects or pathogens compared to commercial soybean varieties.

Soybean interacts with a plant-associated nitrogen-fixing bacterium (Bradyrhizobium japonicum), which results in the fixation of atmospheric nitrogen to a plant-available form inside root nodules. Soybean event MON 87769 was compared to an unmodified parental control and a negative isoline with respect to nitrogen fixation and nodulation. In addition, reference ranges were established for 6 commercially available soybean varieties. Nodule number and dry weight, shoot dry weight, root dry weight, shoot total nitrogen in percentage and shoot total nitrogen in grams were observed in a greenhouse study. No biologically meaningful differences between soybean event MON 87769 and the controls were observed for any of these characteristics, indicating that the interaction with B. japonicum and nitrogen fixation are not affected by the novel trait.

The Plant and Biotechnology Risk Assessment Unit of the Plant Health Science Directorate has therefore determined that the unconfined release of soybean event MON 87769 will not result in altered impacts on non-target organisms, including humans, compared to conventional soybean varieties.

5. Potential Impact of Soybean Event MON 87769 on Biodiversity

Soybean event MON 87769 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. The novel traits are unlikely to increase the plant pest potential of soybean or to cause adverse effects on non-target organisms. It is therefore unlikely that soybean event MON 87769 will have any direct effects on biodiversity, in comparison to the effects that would be expected from the cultivation of conventional varieties.

The CFIA has therefore concluded that the potential impact on biodiversity of soybean event MON 87769 is unlikely to be different from that of conventional soybean varieties.

V. Criteria for the Livestock Assessment

1. Potential Impact of Soybean Event MON 87769 on Livestock Nutrition

Nutritional Composition of soybean event MON 87769

The compositional equivalence of soybean event MON 87769 was determined from 5 replicated field sites across the US during the 2006 growing season. At each field site, soybean event MON 87769 (test), A3525 (control) and three commercial soybean varieties were planted. Forage samples were collected and analysed for proximate and fibre. Sharvested seed samples were analysed for proximate, fibre, amino acids, fatty acids, vitamins, isoflavones (diadzein, genistein and glycitein) and anti-nutrients (lectin, phytic acid, raffinose, stachyose and trypsin inhibitor). In another field trial conducted with soybean event MON 87769, control (A3525) and eight conventional soybean varieties at two US sites, seed samples were processed (defatted toasted soybean meal, refined soybean oil, protein isolates and crude lecithin) and analysed for amino acids, fatty acids and phophatides. No statistically significant differences were observed between soybean event MON 87769 and A3525 soybean forage for moisture, ash, total fat, protein, acid detergent fibre (ADF) and neutral detergent fibre (NDF). Protein was significantly higher in soybean event MON 87769 soybean seed than the control, however all means were within the commercial tolerance interval and literature values.

Soybean event MON 87769 soybean was developed to contain stearidonic acid and to a lesser extent GLA. Stearidonic acid in soybean event MON 87769 soy seed ranged from 16.8% to 33.9% (average 26.1%), while GLA ranged from 6.07-8.03% (avg. 7.1%) of the total fatty acids. Low levels of trans-stearidonic acid (0.2%) and trans-alpha linolenic acid (trans-alpha linolenic acid; 0.4%) were also observed in soybean event MON 87769 soybean seed. Fatty acids, stearidonic acid, GLA, trans-alpha linolenic acid and trans-stearidonic acid were not detected in the conventional soybean control seed. Trans-alpha linolenic acid was present in refined, bleached and deodorized oil (RBD) at levels ranging from 0.1 to 0.16 weight percentage of total fatty acids. The formation of trans-alpha linolenic acid and trans-stearidonic acid is due to the known spontaneous trans-isomerization of unsaturated fatty acids at rates that increase with increased degree of unsaturation. As stearidonic acid and alpha linolenic acid represent a significant portion (35-40%) of total fatty acids in MON 87769 , trans alpha linolenic acid and trans-stearidonic acid are expected to be present. Fatty acids, stearidonic acid, GLA and trans-stearidonic acid were absent in the control or commercial soybean varieties. The presence and levels of stearidonic acid, and GLA which are in vivo intermediates in the metabolism of long-chain omega-3 and 6 fatty acids in mammals, were shown to pose no detrimental risk to humans and livestock consuming soybean event MON 87769. Given the intended shift in fatty acid metabolism to produce stearidonic acid, the fatty acid profile in soybean event MON 87769 was expected to differ from the conventional soybean. Oleic, linolenic, arachidic, palmitic and behenic acids were significantly different statistically in soybean event MON 87769 compared to control, but the means were within commercial tolerance interval and literature values. Linoleic acid, the starting fatty acid from which stearidonic acid and GLA are produced was significantly lowered statistically in soybean event MON 87769 compared to the control soybean seed. The mean linolenic acid values in the modified crop were outside the tolerance interval and literature values. This difference in linolenic acid levels observed between the test and control crop was however shown not to have any negative nutritional impact on humans and livestock requirements of linolenic acid or usage in ration formulation.

Statistically significant differences were observed between soybean event MON 87769 and A3525 soybean seed for all amino acids analyzed, but the means and range values of test crop were within the commercial tolerance interval and literature values. No statistically significant differences between test and control seed were shown for Vitamin E. Glycitein, diadzein and genistein were significantly lower in soybean event MON 87769 than A3525 soybean, however all means were within the commercial tolerance interval and literature values. There were no significant differences in the levels of lectin, phytic acid, raffinose, stachyose and trypsin inhibitor, between soybean event MON 87769 and the control.

With regards to co-products, statistically significant differences were observed between the test and control soybean meal for, ADF, aspartic acid, glutamic acid, histidine, and tryptophan, but the means were within the tolerance interval of the commercial varieties and/or literature values. In soybean event MON 87769 oil, the levels of stearidonic acid, GLA, trans-stearidonic acid and trans-alpha linolenic acid were comparable to the levels found in the seed. Fatty acid, trans-alpha linolenic acid was present in commercial soybean oil (0.1 to 0.16% of total fatty acids) as a result of the oil refining process. Palmitic, stearic, and trans-alpha linolenic acid fatty acids were higher in soybean event MON 87769 than control, while lignoceric acid was lower in test soybean than control soybean oil. All means except for trans-alpha linolenic acid were within outside the commercial tolerance interval., as found in the seed. Vitamin E was significantly higher statistically in soybean event MON 87769 oil than the control, but the level was within the commercial interval and literature values. Except for leucine, all amino acids in the protein isolates were not significantly different between the test and control soybean varieties. All means were within commercial ranges. All phosphatides in crude lecithin were similar for soybean event MON 87769 and A3525 control soybeans.

Broiler Performance

800 Ross birds were distributed randomly to 80 pens to examine the health and performance effects of feeding soybean event MON 87769 soybean meal, a control A3525 diet and six commercial varieties for 42 days. Bird mortality ranged from 2-8% across all treatment groups from day 7-42, mortality of birds on MON87769 soybean was 2%. Performance of birds on soybean event MON 87769 diets were not statistically different than those of control and conventional soybean varieties over the entire test period. No statistically significant differences were observed between soybean event MON 87769 and the control and commercial diets for average daily gain, feed intake and feed:gain ratio. Carcass yield measurements were not different for broilers fed diets containing soybean event MON 87769 compared to those fed control and conventional control soybean meal. No unexpected effects on broiler performance were observed when broilers were fed diets from soybean event MON 87769 soybean meal, control and reference conventional varieties.

Conclusion

The evidence provided by Monsanto supports the conclusion that except for the elevated levels of stearidonic acid and GLA and lowered levels of linoleic in soybean event MON 87769 soybean, the nutritional composition of soybean event MON 87769 is within the range of conventional soybean varieties. No detrimental effects were shown on the performance and health of broilers on diets containing soybean event MON 87769 meal.

2. Potential Impact of Soybean Event MON 87769 on Livestock and Workers/Bystanders

Soybean event MON 87769 has two novel proteins and a novel nutritional profile. The proteins PjΔ6D and NcΔ15D produce a changed nutritional fatty acid profile, resulting in the production of stearidonic acid, and GLA that are not normally found in soybean. Because linolenic acid is the starting material from which stearidonic acid and GLA are produced, the linolenic acid levels were reduced in MON 87769 compared to conventional soybean. Due to spontaneous trans-isomerizaton of unsaturated fatty acids, small levels of trans-stearidonic acid is also present in MON 87769 and oil compared to control soybean and oil.

The expressed proteins PjΔ6D and NcΔ15D are neither known toxins nor allergens and they do not have homology with any known allergens or toxins. The proteins are also heat labile and rapidly degraded under conditions similar to those encountered in the gastrointestinal tract. This information suggests that PjΔ6D and NcΔ15D proteins are unlikely to be novel toxins or allergens, and that intact desaturase enzymes will not be absorbed by livestock.

No adverse effects from the PjΔ6D and NcΔ15D proteins were observed in acute oral toxicity studies in male and female mice at levels expected to be several order of magnitude higher than the highest predicted livestock dose level per kg body weight. No adverse effects on nutrition or health were observed in a broiler feeding trial comparing soybean event MON 87769 soybean grain to several conventional varieties of soybean grain in the diet.

We note at this time that the soybean event MON 87769 is not permitted to be used for the purpose of supplementing the livestock diet with SDA, or increasing SDA/GLA/DHA/EPA deposition in foods of animal origin, either through oil alone or a meal-oil combination in the livestock diet. Such purposes would require a supplementary approval from the CFIA.

The evidence provided by Monsanto Inc. indicates there is no potential impact of soybean event MON 87769 on livestock and workers/bystanders when compared to commercialized soybean lines.

VI. New Information Requirements

If at any time, Monsanto Canada becomes aware of any information regarding risk to the environment, including risk to human or animal health, which could result from release of soybean event MON 87769 in Canada or elsewhere, Monsanto Canada will 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 87769 on the environment, livestock and human health, and may re-evaluate its decision with respect to the livestock feed use and environmental release authorizations of soybean event MON 87769.

VII. Regulatory Decision

Based on the review of the data and information submitted by Monsanto Canada, and through comparisons of soybean event MON 87769 with unmodified soybean counterparts, the Plant and Biotechnology Risk Assessment Unit of the Plant Health Science Directorate, CFIA, has concluded that the trait does not confer to soybean event MON 87769 any characteristic that would result in unintended environmental effects following unconfined release.

Based on the review of submitted data and information by Monsanto Canada, including comparisons of soybean event MON 87769 with its unmodified soybean counterparts, the Animal Feed Division of the Animal Health Directorate, CFIA has concluded that the trait will not confer to soybean event MON 87769 any characteristic that would raise any concerns regarding the safety or nutritional composition of soybean event MON 87769. Grain soybean, its byproducts and soybean oil are currently listed in Schedule IV of the Feeds Regulations and are, therefore approved for use in livestock feeds in Canada. Soybean event MON 87769 has been found to be as safe as and as nutritious as traditional soybean varieties. Soybean event MON 87769 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 87769 is therefore authorized by Plant Biosafety Office of the Plant Health and Biosecurity Directorate and the Animal Feed Division of the Animal Health Directorate as of October 13, 2011. Any soybean lines derived from soybean event MON 87769 may also be released into the environment and used as livestock feed, provided that no inter-specific crosses are performed, the intended uses are similar, and it is known based on characterization, that these plants do not display any additional novel traits and are substantially equivalent to currently grown soybean varieties in Canada, in terms of their potential environmental impact and livestock feed safety.

Soybean event MON 87769 is subject to the same phytosanitary import requirements as its unmodified counterpart.

Please refer to Health Canada's Decisions on Novel Foods for a description of the safety assessment for soybean MON 87769, as it pertains to foods for human consumption.

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