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

<p style="text-align: justify;">The genetic modification of COT102 cotton would not be expected to result in any change in the consumption pattern of cotton products. The seed and oil fractions from this cotton line did not exhibit any novel characteristics that would alter the consumption pattern. This was confirmed by comparison of the levels of proximates and fatty acids in COT102 compared to the parental line. Only refined cottonseed oil and cellulose from processed linters of cottonseed are consumed by humans. Processed linters are composed of pure (&gt;99%) cellulose and are treated with heat and solvent; these processes would be expected to remove and destroy any residual protein.</p>

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

<p style="text-align: justify;">The nutritional and anti-nutritional components of COT102 cottonseed were determined analytically and compared to cottonseed from the non-transgenic counterpart Coker312. Samples for analysis were obtained from field trials conducted at three locations in 2001, and two locations in 2002. Samples were obtained from each replicate of each trial in 2002, whereas only single samples were obtained from each location in 2001. The following components were determined: proximates (moisture, fat, crude protein, crude fibre, total dietary fibre, ash, carbohydrates, acid detergent fibre, and neutral detergent fibre); minerals (phosphorus, calcium, sodium, iron, magnesium, manganese, potassium, zinc, copper and chromium); fatty acids (myristic, palmitic, palmitoleic, stearic, oleic, linoleic, linolenic, arachidic, and behenic); amino acids (aspartate, threonine, serine, glutamate, praline, glycine, alanine, cysteine, valine, methionine, isoleucine, leucine, tyrosine, phenylalanine, histidine, lysine, arginine, and tryptophan), and natural toxicants (total gossypol, free gossypol, and the cyclopropenoid fatty acids: sterculic, malvalic and dihydrosterculic). Levels of total and free gossypol were also determined in oil, non-toasted meal, and toasted meal obtained from a 2001 cottonseed sample.</p>

<p style="text-align: justify;">Data from analyses of the 2001 samples demonstrated that, other than differences in the levels of the amino acids lysine and tyrosine, which were significantly higher in COT102, there were no statistically significant differences in the composition of COT102 compared to its non-transgenic parental line Coker 312. Analyses of the 2002 samples revealed significantly lower levels of iron, zinc, copper, as well as serine and lysine in COT102. However, none of the observed differences in nutritional composition, in both the 2001 and 2002 samples, were found to be biologically significant. No statistically significant differences were observed in the levels of gossypol (total and free) and cyclopropenoid fatty acids, in the seed, oil and meal. The levels of nutrients and anti-nutrients in both COT102 and Coker 312 were also compared to those in the published literature. Other than the levels of some of the amino acids from the 2001 samples, in both COT102 and Coker 312, the levels of all other components fell within the observed ranges for conventional, non-transgenic cotton cultivars. Based on these results, it was concluded that COT102 is equivalent to non-transgenic cotton, and that the genetic modification did not alter the nutritional and anti-nutritional composition.</p>

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

<p style="text-align: justify;">The potential for toxicity and allergenicity of the novel proteins VIP3A and APH4 was determined from results of acute feeding studies using mice, from <i>in-vitro</i> digestibility studies using simulated mammalian gastric fluids, and from the expression levels in COT102 tissues. No adverse effects were observed in the acute studies, in which mice were fed the VIP3A protein at levels far exceeding the estimated environmental exposure, based on the expression levels in tissues. The intestinal epithelial tissues of VIP3A-fed mice were studied, microscopically and immunologically, and no changes were observed, compared to mice fed a diet without VIP3A protein. No observed effects were observed in mice exposed to a very high oral dose of the APH4 protein; the exposure level to non-target organisms is also very low due to levels of APH4 in most tissues that were below quantifiable levels of detection. The VIP3A protein was completely hydrolyzed in simulated mammalian gut fluids, which further demonstrated the lack of toxicity and allergenicity of the protein. The results of these studies led to the conclusion that the novel proteins VIP3A and APH4 in event COT102 cotton are neither toxic or allergenic to non-target organisms, including mammalian species.</p>
