Determination of the Safety of Monsanto Canada Inc.'s Corn (Zea mays ...

Tag : Aquatic Product
II . Background Information
Monsanto Canada Inc. developed, through the use of recombinant DNA techniques, a cornevent resistant to lepidopteran insect pests of corn in Canada. Thecorn event, designated as MON 89034, was developed to provide amethod to control yield losses from insect feeding damage caused bythe European Corn Borer ( Ostrinia nubilalis ) and other lepidopteran insect pests.
Corn event MON 89034 was developed using recombinant DNA technology, resulting in the introduction of the synthetic cry1A.105 gene and the cry2Ab2 gene. The cry1A.105 gene is derived from the cry1Ab , cry1Ac and cry1F genes from Bacillus thuringiensis and encodes a synthetic δ-endotoxin Cry1A.105 protein. Thecry2Ab2 gene is also derived from Bacillus thuringiensis and encodes a variant of the wild-type δ-endotoxin Cry2Ab2protein. The combination of the Cry1A.105 and Cry2Ab2 insecticidalproteins in a single plant provides insect control against a rangeof lepidopteran insect pests. In addition, a selectable marker gene nptII was used in the initial transformation and early event selectionprocess but was removed from the final event selected. The nptII gene from Escherichia coli encodes the neomycin phosphotransferase II protein conferring resistance to the antibiotic kanamycin. Thistrait is of no agronomical interest but was used to selecttransformed from non-transformed plants during the earlydevelopment phase of corn event MON 89084. Once the transgenicplants were identified, the selectable marker gene nptII was no longer needed. Traditional breeding was used to isolateplants that contain the cry1A.105 and cry2Ab2 genes but do not contain the nptII gene, therefore producing marker-free corn event MON 89034.
Monsanto Canada Inc. has provided data on the identity of corn event MON 89034, adetailed description of the transformation method, data andinformation on the insertion site, gene copy number and levels ofgene expression in the plant and the role of the inserted genes andregulatory sequences. The novel proteins were identified andcharacterized. Data was provided for the evaluation of thepotential toxicity of the novel proteins to livestock andnon-target organisms and potential allergenicity of the novelproteins to humans and to livestock.
Corn event MON 89034 has been field tested in the United States andCanada and data for trial years 2004 and 2005 were submitted.
Agronomic characteristics of corn event MON 89034 such as seedlingemergence, plant height, time to reproduction, lodging,susceptibilities to various corn pests, pathogens and abioticstressors and yield parameters were compared to those of unmodifiedcorn counterparts.
Nutritional components of corn event MON 89034, such as proximates,amino acids and fatty acids were compared with those of unmodifiedcorn counterparts.
The Biotechnology Environmental Release Assessment (BERA) Unit ofthe Science Strategies Directorate, CFIA , has reviewed the above information, in light of the assessmentcriteria for determining environmental safety of PNTs , as described in the Directive 94-08 (Dir94-08), entitled " Assessment Criteria for Determining Environmental Safety of PlantsWith Novel Traits ". The BERA Unit has considered: potential of corn event MON 89034 to become a weed of agricultureor be invasive of natural habitats; potential for gene flow from corn event MON 89034 to wild relativeswhose hybrid offspring may become more weedy or more invasive; potential for corn event MON 89034 to become a plant pest; potential impact of corn event MON 89034 or its gene products onnon-target species, including humans; and potential impact of corn event MON 89034 on biodiversity.
The Animal Feed Division of the Animal Health Directorate, CFIA , has also reviewed the above information with respect to theassessment criteria for determining the safety and efficacy oflivestock feed, as described in Directive 95-03 (Dir95-03),entitled " Guidelines for the Assessment of Novel Feeds: Plant Sources ". The Animal Feed Division has considered: potential impact of corn event MON 89034 on livestock nutrition;and potential impact of corn event MON 89034 on livestock andworkers/bystanders
Monsanto Canada Inc. has provided the CFIA with a method for the detection and identification of cornproducts containing the corn event MON 89034 III . Description of the Novel Traits 1. Resistance to Lepidopteran Pests of Corn
Bacillus thuringiensis is a common gram-positive soil-borne bacterium. In the sporeforming stage, it produces several insecticidal protein crystals,including the δ-endotoxin Cry1Ab, Cry1Ac, Cry2Ab and Cry1Fproteins which are active against certain lepidopteran insects,such as the European Corn Borer. These proteins are insecticidal tosusceptible lepidopteran insects after cleavage by proteases in theinsect's gut, forming a protease-resistant active fragment (coretoxin) that is the bio-active form of the protein. Insecticidalactivity is believed to depend on the binding of the activefragment to specific receptors present in susceptible insects onmidgut epithelial cells, forming pores which disrupt osmoticbalance and eventually result in cell lysis and insect death. Cornevent MON 89034 produces two Bt proteins, namely Cry1A.105 and Cry2Ab2. The Cry1A.105 protein is achimeric protein that consists of different domains from Cry1Ab,Cry1Ac and Cry1F. It was designed by Monsanto using domain exchangestrategy to achieve a high level of activity against targetlepidopteran pests. The overall amino acid sequence identity ofCry1A.105 to Cry1Ac, Cry1Ab and Cry1F is 93.6%, 90.0% and 76.7%,respectively. The other Bt protein produced in MON 89304, Cry2Ab2, is identical to Cry2Ab2protein expressed in Lepidopteran-resistant cotton Bollgard II which has been authorized by the CFIA for livestock feed use. This protein has been shown to benon-toxic to humans and other vertebrates. Except for oneamino-acid, the amino acid sequence of the Cry2Ab2 protein producedin Bollgard II cotton and MON 89034 corn is identical to that of the wild-type Bt protein. The cry1A.105 and cry2Ab2 coding sequences were optimized to accommodate the preferred codonusage for maize.
The cry1A.105 and cry2Ab2 genes are expressed in event MON 89034 using promoters whichconfer constitutive expression of proteins. Tissue samples werecollected at various growth stages from a MON 89034 hybrid grown atfive representative US field trial sites. Levels of Cry1A.105 and Cry2Ab2 proteins wereevaluated by enzyme-linked immunosorbent assays. Average Cry1A.105levels across all stages and all sites, expressed in micro-gramsCry1A.105 protein per gram dry weight tissue (µg/g dwt),ranged from ca . 75-520 in leaf, 11-79 in root and 42-380 in whole plant. Thelevels of Cry1A.105 protein tended to decline over the growingseason. The mean Cry1A.105 protein levels across all sites were 5.9 µg/g dwt in grain and 12 µg/g dwt in pollen.
Average Cry2Ab2 levels across all stages and all sites, expressedin micro-grams Cry2Ab2 protein per gram dry weight tissue(µg/g dwt), ranged from ca . 130-180 in leaf, 21-58 in root and 38-130 in whole plant. Thelevels of Cry2Ab2 protein tended to decline over the growingseason. The mean Cry2Ab2 protein levels across all sites were 1.3 µg/g dwt in grain and 0.64 µg / g dwt in pollen.
The Cry1A.105 and Cry2Ab2 proteins were shown to degrade readily inthe soil. Soil degradation experiments used three different soiltypes representative of corn growing regions. Depending on the soiltype, the time to degrade 50% and 90% of the initial spikedconcentration of Cry1A.105 protein ranged from 2 to 7 days, and 7to 19 days, respectively, and the time to degrade 50% and 90% ofthe initial spiked concentration of Cry2Ab2 protein ranged from 0.5to 3 days, and 3 to 13 days, respectively.
The Cry1A.105 and Cry2Ab2 proteins were purified from the grain ofcorn event MON 89034 and characterized. The identity of thepurified proteins was confirmed by Western immunoblot analysis,tryptic peptide mass mapping, N-terminal characterization andinsecticidal activity.
The levels of Cry1A.105 and Cry2Ab2 proteins in corn MON 89034tissues were too low to extract sufficient amounts for evaluationof environmental and feed safety. To obtain sufficient quantitiesof the proteins for safety studies, it was necessary to express the cry1A.105 and cry2Ab2 genes in an E. coli production system. The equivalency of the plant-produced proteinsto the E. coli - produced proteins was evaluated by comparing their molecularweight, immunological reactivity, tryptic peptide mass map,insecticidal activity and glycosylation status. Based on theresults, Cry1A.105 and Cry2Ab2 proteins produced in corn MON 89034were found to be equivalent to their respective E. coli -produced counterparts.
The potential mammalian toxicity and allergenicity of the Cry1A.105and Cry2Ab2 proteins were evaluated. Both proteins lack sequencesimilarity to known allergens and protein toxins which have adverseeffects to mammals. No adverse effects were observed when theCry1A.105 protein was ingested by mice at a dose of 2,072 mg / kg body weight, or the Cry2Ab2 protein at a dose of 2,198 mg / kg body weight. In vitro digestive fate studies have shown that both Cry1A.105 and Cry2Ab2proteins are rapidly degraded in simulated gastric fluid, unlikeprotein allergens which are normally resistant to digestion. TheCry1A.105 and Cry2Ab2 proteins expressed in event MON 89034 are notglycosylated, unlike many known allergens, providing additionalevidence that Cry1A.105 and Cry2Ab2 proteins do not have theproperties of known allergens. 2. Development Method
Corn event MON 89034 was developed through Agrobacterium -mediated transformation of immature embryos derived from corn lineLH172 using a transformation plasmid vector that includes twoseparate T-DNAs . 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 expression cassette. Following cultivation with Agrobacterium , the embryos were transferred to a selection medium containingparomomycin to eliminate cells that were not transformed and selectcells containing T-DNA - II or T-DNA I + T-DNA II . During subsequent breeding the unlinked insertions of T-DNA I and T-DNA II were segregated. The plants containing only the cry1A.105 and the cry2Ab2 expression cassettes were selected using molecular analysis whilethe plants containing the nptII expression cassette were eliminated from subsequent breeding.Event MON 89034 was identified as a successful transformant and waschosen for further development. 3. Stable Integration into the Plant Genome
Molecular characterization by Southern blot analysis demonstratedthat corn event MON 89034 contains one copy of the cry1A.105 and cry2Ab2 gene cassettes inserted at a single site in the corn genome. Noadditional elements, including intact or partial DNA fragments of the cry1A.105 or cry2Ab2 cassette, T-DNA II sequences or backbone sequences from the plasmid vector, linked orunlinked to the intact insert, were detected in corn event MON89034. Sequencing of the introduced DNA confirmed the organization and sequence of the genetic elementswith two exceptions. Some elements of the promoter that regulatesthe expression of the cry1A.105 gene have been deleted and the right border sequence present inthe transformation vector has been replaced by a left border in MON89034 corn. These modifications have no effect on the functionalityof the DNA insert.
The stability of the inserted DNA was demonstrated by Southern blot analysis across sevengenerations in the breeding history of MON 89034 corn. Analysis ofthe inheritance pattern of the cry1A.105 and cry2Ab2 genes and the Cry1A.105 and Cry2Ab2 proteins across fivegenerations of event MON 89034 confirmed the stability of theinserted DNA and the stability of Cry1A.105 and Cry2Ab2 protein expression. Theresults of the analysis are consistent with the finding of a singlesite of insertion that segregates according to the Mendelian law ofgenetics. IV . Criteria for the Environmental Assessment 1. Potential of Corn event MON 89034 to Become a Weed ofAgriculture or be Invasive of Natural Habitats
The biology of corn, described in the CFIA Biology Document BIO1994-11, states that unmodified plants of thisspecies are not invasive of unmanaged habitats in Canada. Corn doesnot possess the potential to become weedy due to the lack of seeddormancy, the non-shattering nature of corn cobs and the poorcompetitive ability of seedlings. According to the informationprovided by Monsanto Canada Inc. , corn event MON 89034 was determined to be similar to unmodifiedcorn in this respect. The introduction of novel traits did notalter the dormancy, germination and volunteer potential of the seedof MON 89034 compared to conventional corn seed.
MON 89034 corn hybrids were tested in the United States corn beltat 9 locations in 2004 and 9 locations in 2005, and in Canada at 5locations in 2005. A total of 14 phenotypic and agronomic traitswere evaluated. These agronomic traits covered a broad range ofcharacteristics that encompass the entire life cycle of the maizeplant and included data assessing seedling emergence, vegetativevigour, time to reproduction and yield characteristics. For themajority of agronomic traits, no statistically significantdifferences between MON 89034 corn hybrids and theirnon-transformed counterpart were observed. Although instances ofstatistically significant differences between MON 89034 and controlhybrids were observed for some traits, there were no consistenttrends in the data across locations or years that would indicatethat any of these differences were due to the genetic modification.
In addition to the phenotypic and agronomic characteristics,susceptibility of MON 89034 corn to abiotic stressors and variouscorn pests and pathogens was evaluated. Differences insusceptibility to wind damage or excess moisture were observedbetween MON 89034 and control corn at a few trial sites in 2004;however, they were not considered biologically meaningful since noconsistent trends were observed across sites and years. Theseobservations indicated that MON 89034 corn had no alteredsusceptibility or tolerance to the biotic and abiotic stressorscompared to control hybrids, except for the introducedlepidopteran-resistance trait.
The results showed that no competitive advantage was conferred tocorn event MON 89034 by the expression of the Cry1A.105 and Cry2Ab2proteins, other than that conferred by resistance to lepidopteraninsect pests. None of the corn's reproductive or growthcharacteristics were modified, and MON 89034's tolerance to abioticand biotic stresses was unchanged except for the intended trait ofresistance to lepidopteran insect pests. As feeding damage bylepidopteran larvae is not known to be a major factor restrictingthe establishment or distribution of corn in Canada, theintroduction of this novel trait does not make MON 89034 corn weedyor invasive of natural habitats.
The above considerations led the CFIA to conclude that corn event MON 89034 has no increased weedinessor invasiveness potential compared to currently commercialized cornvarieties. 2. Potential for Gene Flow from Corn event MON 89034 to WildRelatives Whose Hybrid Offspring May Become More Weedy or MoreInvasive
The biology of corn, as described in CFIA Biology Document BIO1994-11, indicates that there are no wildrelatives in Canada that can hybridize with corn. None of the datasubmitted by Monsanto Canada Inc. on corn MON 89034 indicated any change in sexual compatibility asa result of the genetic modification.
The CFIA has therefore concluded that gene flow from corn event MON 89034to wild relatives is not possible in Canada. 3. Altered Plant Pest Potential of Corn event MON 89034
The expression of the Cry1A.105 and Cry2Ab2 proteins is unrelatedto plant pest potential, and corn is not considered a plant pest inCanada. Field evaluation of MON 89034 corn did not show anyincrease or decrease in susceptibility to disease or insectstressors, other than to some lepidopteran insects, compared tounmodified corn counterparts. Feeding damage by lepidopteran larvaeis not known to be a major factor restricting the establishment ordistribution of corn in Canada.
The CFIA has, therefore, determined that the corn event MON 89034 does notdisplay any altered pest potential compared to currentlycommercialized corn varieties. 4. Potential Impact of Corn event MON 89034 on Non-Target Organisms
The history of use and available literature indicate that Bacillus thuringiensis δ-endotoxins are active only against specific insect groups,and are not toxic to other organisms including humans and othervertebrates. The Cry1A.105 expressed in MON 89034 corn is achimeric protein derived from the well characterized Cry1Ab, Cry1Acand Cry1F proteins. The spectra of activity of these three proteinsare restricted to some lepidopteran insects. The Cry2Ab2 proteinexpressed in MON 89034 corn is identical to the Cry2Ab2 proteinexpressed in Bollgard II cotton and has been shown to be specific to certain lepidopteranspecies (see DD2003-45).
Monsanto Canada Inc. provided studies on the insecticidal activities of the Cry1A.105and Cry2Ab2 proteins on a range of insects from three taxa. Insectspecies tested included four lepidopterans (European corn borer,corn earworm, fall armyworm and black cutworm), two coleopterans(boll weevil, southern corn rootworm) and two hemipterans (westerntarnished plant bug and green peach aphid). The results confirmedthat Cry1A.105 and Cry2Ab2 proteins have activity against all fourlepidopteran species but no activity against the coleopteran orhemipteran species.
Consequently, the endangered species assessment focused on larvaeof the order Lepidoptera. The habitat of the Karner blue butterfly( Lycaeides melissa samuelis ) has the potential to occur in proximity of corn fields; however,the Karner blue butterfly is listed by the Committee on the Status of Endangered Wildlife as extirpated in Canada (Please refer tohttp://www.cosewic.gc.ca/index.htm for more information). Evenshould Karner blue butterfly populations be recovered in Canada,Karner blue larvae would have low likelihood of exposure to cornpollen because corn pollen shed typically occurs after Karner bluelarvae have finished feeding. The lepidopteran species currentlylisted by the Committee on the Status of Endangered Wildlife inCanada as being a threatened or endangered species are usually notfound in proximity of corn fields. Therefore the potential exposureof threatened or endangered lepidopteran species in Canada toCry1A.105 or Cry2Ab2 protein expressed in MON 89034 corn will beminimal.
Among the butterfly species of special concern in Canada, themonarch butterfly ( Danaus plexippus ) is the only species that may be exposed to significant amounts ofthe Cry1A.105 and Cry2Ab2 proteins expressed in MON 89034. Asmonarch butterfly larvae feeding on milkweed plants may be exposedto corn pollen drifting from adjacent MON 89034 corn plants, a riskassessment was conducted for quantification of the potential impactof corn pollen from MON 89034 corn on populations of monarchbutterflies in the North American corn belt. Data from an extendeddietary assay conducted on newly-hatched monarch larvae indicatethat pollen from MON 89034 corn has no adverse effect on survivaland development of monarch larvae to adult at pollen densitieslower or equal to 147 grains/ cm 2 on milkweed leaves. An estimation of monarch exposure to MON 89034corn pollen was conducted using published procedures and parametersdescribed in Sears et al. (2001), Pleasants et al. (2001) and Dively et al. (2004). These assessment results indicate that MON 89034 cornwould affect approximately 0.6% of the monarch population acrossthe North American corn belt, which includes Ontario. Similarlevels are anticipated for the other Canadian corn growing areas,including Quebec. Such small population effects are unlikely tohave any significant impact on the sustainability of monarchbutterfly populations in Canada.
An assessment of the potential for interaction between Cry1A.105and Cry2Ab2 proteins using two sensitive lepidopteran speciesdemonstrated that the proteins only showed additive insecticidalactivity and no synergistic or antagonistic activities. The lack ofinteraction between Cry1A.105 and Cry2Ab2 proteins allows for eachprotein to be tested independently.
Monsanto Canada Inc. has submitted data from dietary toxicity studies on the effect ofCry1A.105 or Cry2Ab2 protein on non-target invertebrates, includingthe honeybee larvae and adult ( Apis mellifera ), minute pirate bug ( Orius insidiosus ), ladybird beetle ( Coleomegilla maculata ), a parasitic wasp ( Ichneumon promissorius ), and earthworm ( Eisenia foetida ). Collembola ( Folsomia candida ) were fed an artificial diet containing 50% of MON 89034 leaftissue. In all cases, the Cry1A.105 and Cry2Ab2 proteins weredemonstrated to be safe to these indicator species at doses equalto or exceeding 14 times the estimated environmental concentrationof Cry1A.105 and Cry2Ab2 proteins in the diet of non-targetinvertebrates feeding on MON 89034 tissues or exposed to MON 89034corn via their preys. In addition, no adverse effects were observedwhen the aquatic invertebrate Daphnia magna was exposed to MON 89034 corn pollen at a concentration of 100 mg / L , which indicates that no hazard is anticipated to aquaticinvertebrates from exposure to MON 89034 corn pollen.
Data was also submitted on non-target vertebrates including themouse, the bobwhite quail and broiler chicken. No adverse effectswere detected when mice were exposed to a single oral dose of 2,072 mg Cry1A.105 protein/ kg body weight or 2,198 mg Cry2Ab2 protein / kg body weight. These doses represent several thousand times theworst-case daily dose of Cry1A.105 and Cry2Ab2 proteins to humansor livestock feeding on MON 89034 grain. No adverse effects weredetected when bobwhite quail or broiler chicken were fed a dietcontaining 50% MON 89034 corn grain for 8 days and 42 days,respectively.
Composition analyses showed that the levels of key nutrients andanti-nutrients in corn MON 89034 grain and forage are comparable tothose in commercial corn varieties.
Based on the above, the CFIA has determined that, compared to current commercial cornvarieties, the unconfined release of corn event MON 89034 will notresult in altered impacts on non-target organisms, includinghumans. 5. Potential Impact of Corn event MON 89034 on Biodiversity
Corn event MON 89034 has no novel phenotypic characteristics thatwould extend its range beyond the current geographic range of cornproduction in Canada. Since corn has no wild relatives with whichit can outcross in Canada, there will be no transfer of the noveltraits to other species in unmanaged environments. In addition thenovel traits were determined to pose minimal risks to non-targetorganisms.
At present, the use of chemical insecticides and Bt corn hybrids to control corn pests is common practice in Canada.Therefore, the reduction in local pest species as a result of therelease of MON 89034 corn does not present a significant changefrom existing agricultural practices.
The CFIA has therefore concluded that the potential impact on biodiversityof corn event MON 89034 is equivalent to that of currentlycommercialized corn varieties. 6. Potential for Development of European Corn Borer Resistance toCorn event MON 89034
In order to significantly minimize the likelihood of thedevelopment of insect pest resistance to modified plants expressingnovel insect resistance, the CFIA requires that an insect resistance management (IRM) plan beimplemented for these products. Lepidopteran insects have asignificant ability to develop resistance to conventional chemicalinsecticides. Therefore, it is reasonable to expect that resistanceto the insecticidal properties of corn event MON 89034 may develop.The following IRM design is intended to reduce or delay European corn borer (ECB)resistance to the Cry1A.105 and Cry2Ab2 proteins. A component ofthe IRM strategy that will be used with MON 89034 corn is theestablishment of a refuge of ECB -susceptible corn within or near the MON 89034 corn field. Shouldresistant insects occur, they would then be able to mate withsusceptible insects from the refuge to keep the frequency ofresistance genes diluted in the insect population.
As Cry1A.105 and Cry2Ab2 proteins are highly active against ECB larvae, each protein individually achieves 95% or greater controlof ECB at the concentrations produced in MON 89034 corn leaves. Inaddition, as Cry1A.105 and Cry2Ab2 proteins have different primarystructures, share only 14% of amino acid sequence identity and binddifferently to distinct proteins in the midgut of ECB , the likelihood of cross-resistance between the two proteins isvery low. In view of the dual effective dose delivered by MON 89034corn and the lack of cross-resistance between the two proteins, thelikelihood of ECB resistance development to MON 89034 corn is significantly reducedcompared to corn events expressing a single Bt protein. Therefore, MON 89034 corn cultivation is expected to besustainable using a reduced refuge. The current refuge sizerequirement for single Bt protein corn products is 20% of total corn acres. Based onconservative mathematical models, a 5% refuge will preserve thedurability of MON 89034 corn.
CFIA believes that sound management practices and IRM strategies can significantly reduce and delay the development of ECB populations resistant to Cry1A.105 protein and/or Cry2Ab2 protein.However, the ECB populations must be monitored for the development of resistance ina regular and consistent manner.
CFIA understands that Monsanto Canada Inc. has developed, and will implement, an IRM plan that includes the following key components: The use of structured refugia to provide a population of Europeancorn borers that have not been exposed to the Cry1A.105 and Cry2Ab2proteins and are available to reproduce with potentially resistant ECB individuals which may emerge from the MON 89034 crop. The early detection of ECB populations resistant to MON 89034 corn-expressed insecticidalproteins is extremely important. Close monitoring for the presenceof such populations, in ECB -resistant corn fields and surrounding areas, is thereforewarranted. Monitoring includes the development of appropriatedetection tools such as visual field observations and laboratorybioassays, education of growers, reporting schedules, andmitigation procedures in case of resistance development. Education tools will be developed and provided to all growers,district managers and field managers. These will includeinformation on product performance, resistance management,monitoring procedures and timetables, detection protocols forresistant ECB individuals, instructions to contact Monsanto Canada Inc. and strategies to be followed if unexpected levels of ECB damage occur. Monsanto Canada Inc. will have documented procedures in place for responding to thesereported instances of unexpected ECB damage. These procedures will include, where warranted, thecollection of plant tissue and ECB individuals and use of appropriate bioassays to evaluate suspectedresistant individuals, and a protocol for immediate action tocontrol resistant individuals. Detection of confirmed resistant ECB populations and mitigation measures will be immediately reportedto CFIA . Integrated Pest Management practices will be promoted, such asprediction of infestation problems from field histories.
Note: The Plant Biosafety Office periodically audits compliancewith the IRM requirements. V . Criteria for the Livestock Feed Assessment 1. Potential Impact of Corn event MON89034 on Livestock Nutrition
Nutritional Composition:
The compositional equivalence of MON 89034 to its isogenicnon-transgenic control (LH198 x LH172) was assessed from five sitesin the US during the 2004 growing season. Forage and grain samples werecollected from replicated plots and analyzed for proximate, aciddetergent fibre (ADF), neutral detergent fibre (NDF), totaldetergent fibre (TDF), minerals, amino acids, fatty acids, vitaminsand secondary metabolites. There were no statistically significantdifferences between MON 89034 and control across locations forcrude protein, total fat, ash, moisture, carbohydrates, ADF and NDF , TDF in forage and grain. Crude protein and NDF were higher in test than control at one site, but means werewithin the normal variation of commercial corn. Phosphorus inforages was significantly higher in MON 89034 than control;however, the means were within the tolerance interval forcommercial reference corn and literature values. No statisticallysignificant differences were observed between MON 89034 and controlgrain for all minerals across sites. Significant differencesbetween test and control were observed for Cu and Fe at two sites and Ca and Mn at one site, but all means were within literature ranges. Exceptfor stearic and arachidic acids, all other fatty acids in grainwere similar between MON 89034 and control across locations.Stearic and arachidic acid levels were within the commercialtolerance interval and literature values. Linolenic, palmitoleic,oleic, linoleic and eicosenoic acids were significantly differentbetween test and control at one site, but all means were within thetolerance interval for commercial corn. No statisticallysignificant differences were observed between test and control foramino acids across locations, while significant differences wereobserved at one site; however, no consistent trend was observed.All vitamin levels were statistically similar between test andcontrol. Ferulic and p-coumaric acid in MON 89034 were notstatistically significantly different from the control in thecombined analyses. All means were within the commercial toleranceinterval and literature values.
Anti-nutritional factors:
Phytic acid was analyzed in MON 89034 grain and compared to its nontransgenic control (LH198 x LH172). No statistically significantdifferences were observed between MON 89034 and control for phyticacid. All means were within the conventional tolerance interval andliterature values.
Nutrient Bioavailability:
A 42-d broiler study was performed to evaluate the effects oftransgenic MON 89034 on broiler performance of 600 birds on sixdiets (including diets of MON 89034; near isogenic control and fourreference commercial corn hybrids). Broiler mortality averaged 4%and was not related to any dietary treatment. Body weights, totalfeed intake, feed conversion, carcass yield, fat pad, breast, wing,drum and thigh weight, percent moisture, protein and fat in thighand breast meat were similar across treatments. Adjusted feedconversion for birds on MON 89034 diet was significantly lower thanthe control diet, but similar to three of the commercial cornreference varieties. Further comparison of MON 89034 to control andcommercial corn varieties (as a group) showed no statisticallysignificant differences between the two groups. No adverse healtheffects were observed in the course of the trial.
The evidence provided by Monsanto supports the conclusion that thenutritional composition of MON 89034 corn is substantiallyequivalent to its non transgenic control varieties. 2. Potential Impact of Corn event MON89034 on Livestock andWorkers/Bystanders
Acute oral toxicity studies in mice demonstrated anon-observed-effect-level (NOEL) value of 2072 mg / kg - bw /day for Cry1A.105, which is much higher than the predicted worstcase exposure values of 19.38, 15.96, and 11.88 mg / kg - bw /day predicted for Cry1A.105 in broilers, swine, and cattle,respectively. Acute oral toxicity studies in mice demonstrated anon-observed-effect-level (NOEL) value of 2198 mg / kg - bw /day for Cry2Ab2, which is much higher than the predicted worstcase exposure values of 6.63, 5.46, and 4.06 mg / kg - bw /day predicted for Cry2Ab2 in broilers, swine, and cattle,respectively. A feeding trial in broiler chickens using realisticlevels of exposure of the two proteins in MON 89034 compared withconventional diets showed no effects on performance or health. Theproteins Cry1A.105 and Cry2Ab2 have been demonstrated to be heatlabile and rapidly digested under conditions present in thegastrointestinal tract. These proteins have shown no sequencesimilarities with known allergens or toxins which would indicatepotential allergenicity or toxicity issues. Maximum exposures tothe proteins are mitigated by their digestibility, processing, aswell as the blending of the MON 89034 corn with other varieties.
These findings indicate that MON 89034, which produces Cry1A.105and Cry2Ab2 proteins, is not expected to result in adversetoxicological effects in livestock fed MON 89034 corn or in humansexposed to the proteins. Based on the large disparity betweenpredicted exposure levels and no effect levels, and the lack ofinherent allergenic properties demonstrated by the proteins, nosignificant risk to livestock and workers/bystanders is expectedfrom exposure to the Cry1A.105 and Cry2Ab2 proteins produced by theMON 89034 corn.
Based on the detailed characterization provided for MON 89034(nutritional composition and agronomic data of the modified plantcompared to the unmodified comparator), it is unlikely that themodification has had any unintended effects on the modified plant.
The evidence provided by Monsanto Canada Inc. supports the conclusion that the potential impact on livestock andworkers/by-standers of corn event MON 89034 is equivalent to thatof currently commercialized corn lines. VI . New Information Requirements
If at any time Monsanto Canada Inc. becomes aware of any information regarding risk to theenvironment, including risk to human or animal health, which couldresult from release of corn event MON 89034 materials in Canada orelsewhere, Monsanto Canada Inc. will immediately provide such information to the CFIA . On the basis of such new information, the CFIA will re-evaluate the potential impact of event MON 89034 on theenvironment, livestock and human health, and may re-evaluate itsdecision with respect to the livestock feed use and environmentalrelease authorizations of corn event MON 89034. VII . Regulatory Decision
Based on the review of the data and information submitted byMonsanto Canada Inc. , and through comparisons of corn event MON 89034 with unmodifiedcorn counterparts, the Biotechnology Environmental ReleaseAssessment Unit, CFIA , has concluded that the novel genes and their corresponding traitsdo not confer to corn event MON 89034 any characteristic that wouldresult in unintended environmental effects following unconfinedrelease.
Based on the review of submitted data and information by MonsantoInc., including comparisons of corn event MON 89034 with itsunmodified corn counterparts, the Animal Feed Division of theAnimal Health Directorate has concluded that the introduced genesand their corresponding traits will not confer to corn event MON89034 any characteristic that would raise any concerns regardingthe safety or nutritional composition of corn event MON 89034.Grain corn, its byproducts and corn oil are currently listed inSchedule IV of the Feeds Regulations and are, therefore, approved for use inlivestock feeds in Canada. Corn event MON 89034 has been assessedand found to be as safe as and as nutritious as traditional cornvarieties. Corn event MON 89034 and its products are considered tomeet present ingredient definitions and are approved for use aslivestock feed ingredients in Canada.
Taking into account these evaluations, unconfined release into theenvironment and use as livestock feed of corn event MON 89034 aretherefore authorized by Plant Biosafety Office of the PlantProducts Directorate and the Animal Feed Division of the AnimalHealth Directorate as of June 19, 2008. Any corn lines derived fromevent MON 89034 may also be released into the environment and usedas livestock feed, provided that no inter-specific crosses areperformed; the intended uses are similar; it is known based oncharacterization, that these plants do not display any additionalnovel traits and are substantially equivalent to currently growncorn varieties in Canada, in terms of their potential environmentalimpact and livestock feed safety; the novel genes are expressed atlevels similar to that of the authorized line; and the insectresistance management requirements described in the presentdocument are applied.
Corn event MON 89034 is subject to the same phytosanitary importrequirements as its unmodified counterparts.
Please refer to Health Canada's Decisions on Novel Foods for adescription of the food safety assessment of corn event MON 89034.The food safety decisions are available at the following Health Canada web site:http://www.hc-sc.gc.ca/fn-an/gmf-agm/appro/index-eng.php
This bulletin is published by the Plant Health and ProductionDivision. For further information, please contact the PlantBiosafety Office or the Feed Section at:
Plant Biosafety Office
Plant Products Directorate
59 Camelot Drive, Nepean
Ontario, K1A 0Y9
613-225-2342
Feed Section
Animal Health Directorate
59 Camelot Drive, Nepean
Ontario, K1A 0Y9
613-225-2342