<p style="text-align: justify;"><b>Dietary Exposure</b></p>

<p style="text-align: justify;">MON802 maize was intended mainly for use in animal feed. The major human food uses for maize are extensively processed starch and oil fractions prepared by wet or dry milling procedures and products include corn syrup and corn oil, neither containing protein. Human exposure to the modified protein from whole grain corn in the diet was considered to be very low due both to its low abundance in the protein fraction of the grain and to the proportionately low percentage of protein in the kernal, compared with the major starch component. Overall, the dietary exposure of consumers in the United States and Canada to grain from MON802 maize was anticipated to be the same as for other lines of commercially available field corn.</p>

<p style="text-align: justify;"><b>Nutritional Data</b></p>

<p style="text-align: justify;">The major components of corn grain and forage for MON802 maize were analysed on plant material harvested from field trials. Compositional data for protein, fat, ash, carbohydrates, calories, moisture, amino acids, and fatty acids for line MON802 grain were comparable to the data from the control line and within published ranges for commercial hybrids. Similar promximate analysis, including acid detergent fibre and neutral detergent fibre analyses, were performed on forage for line MON802 and an appropriate control. Based on these compositional data, it was concluded that there were no significant differences between the forage from line MON802 and the control line.</p>

<p style="text-align: justify;"><b>Toxicity</b></p>

<p style="text-align: justify;">The low potential for toxicity of plant expressed Cry1Ab protein, and CP4 EPSPS and glyphosate oxidoreductase enzymes was demonstrated by studies on protein characterization, high oral dose feeding studies in laboratory animals, and a lack of amino acid sequence homologies with known protein toxins.</p>

<p style="text-align: justify;">Data were generated that demonstrated that the active Cry1Ab protein product of the inserted <i>cry1Ab</i> gene was equivalent to that produced in the naturally occurring bacterium <i>Bacillus thuringiensis</i> that have been in use for over 40 years as a commercial biopesticide. The inserted, plant expressed cry1Ab gene codes for a full length protein of 1156 amino acids which is processed by proteases into the 600 amino acid tryptic core fragment which is insecticidally active. After activation by trypsin to the insecticidal form, the resulting proteolytic fragments were compared to the bacterial proteins and shown to be of similar molecular weight, immunological reactivity and trypsin resistance. The protein was not glycosylated and showed similar bioactivity and host range specificity to the native protein.</p>

<p style="text-align: justify;">An acute oral toxicity study in mice was used to assess the potential mammalian toxicity of Cry1Ab, CP4 EPSPS and GOX proteins. There were no indications of toxicity as measured by the absence of treatment related adverse effects.<br />
A comparison of amino acid sequences showed that the Cry1Ab protein did not show meaningful homology with the sequences of known toxin or allergens contained in the public domain databases GenBank, EMBL, Pir and SwissProt.<br />
Similarly, the CP4 and GOX proteins did not show meaningful amino acid sequence homology when compared to known allergens or protein toxins. The Roundup Ready&reg; proteins showed no significant homology with any known toxins or allergens. EPSPS is an enzyme that is ubiquitous in nature, and therefore would not be expected to be toxic or allergenic. For the second novel enzyme, glyphosate oxidase, experiments demonstrated that the enzyme displayed a narrow substrate specificity, and appeared not to affect any plant specific pathways.</p>

<p style="text-align: justify;"><b>Allergenicity</b></p>

<p style="text-align: justify;">The likelihood of the Cry1Ab, CP4 EPSPS and GOX proteins being allergens was judged to be remote. No homologies were found when the deduced amino acid sequences of the introduced proteins were compared to the sequences of known allergens. In addition, the potential for allergenicity was assessed based upon the physiochemical properties of known food allergens, such as stability to acid and/or proteolytic digestion, heat stability, glycosylation, none which demonstrated characteristics normally associated with food allergens.</p>

<p style="text-align: justify;">Protein allergens are normally resistant to digestion unlike the Cry1Ab protein that was shown to degrade readily in simulated gastric fluid. The Cry1Ab protein was rapidly degraded and its insecticidal activity lost under conditions that simulate mammalian digestion. Similarly, the enzymatic activity of CP4 EPSPS and GOX protein disappeared rapidly as these proteins were rapidly degraded under these same conditions. Both CP4 EPSPS and GOX proteins were inactivated by heat.</p>

<p style="text-align: justify;">The Cry1Ab protein was shown to degrade readily in the environment. The plant expressed protein had DT50 and DT90 values (time to degrade to 50% and 90% of the original bioactivity) of 1.6 and 15 days respectively.<br />
Unlike many known allergens the insecticidal protein Cry1Ab is not glycosylated. A search for amino acid sequence similarity between the Cry1Ab protein and known allergens, using a database assembled from the public domain databases GenBank, EMBL, Pir and SwissProt, revealed no significant amino acid sequence homologies.<br />
Maize products are an important alternative to wheat flour for individuals afflicted with coeliac disease, an immune mediated food intolerance for which wheat gliadins have been implicated as the causal agent. In light of the importance of maize products to these individuals, a sequence similarity search was conducted and no amino acid sequence homologies between the Cry1Ab protein and gliadins were detected.</p>
