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

<p style="text-align: justify;">Refined cottonseed oil and cellulose from processed linters of cottonseed are the only cotton products consumed by humans. Processed linters are essentially pure cellulose (&gt;99%) and are subjected to heat and solvent treatment that would be expected to remove and destroy DNA and protein. Similarly, the refining process for cottonseed oil includes heat, solvent and alkali treatments that would remove and destroy DNA and protein. Typically, cottonseed oils are pooled and blended together and it is anticipated that the oil from 531, 757, and 1076 cottonseed will not be handled or treated any differently than other cottonseed oils. The genetic modification in these cotton lines will not result in any change in the consumption pattern for this product. As the introduced gene products are not detectable in the refined oil produced from transgenic cotton, there is no anticipated human exposure to these proteins based on normal consumption patterns.</p>

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

<p style="text-align: justify;">Compositional comparison of cottonseed from lines 531, 757 and 1076 was made to commercial non-transgenic cottonseed. The cottonseed used for compositional analyses was taken from four to six trial sites. The compositional analyses of cottonseed included proximates (crude protein, crude fat, crude fibre, ash and gross energy), amino acid composition, fatty acids profile, aflatoxins and levels of tocopherols.</p>

<p style="text-align: justify;">The concentrations of protein, oil, carbohydrate and ash were the same for transgenic cotton lines 531, 757 and 1076 and the control parent line Coker 312 line. Line 1076 contained slightly higher carbohydrate and slightly lower oil concentrations than the control line, but both were within the normal range for cottonseed. Fatty acid concentration was within the normal published range for cottonseed. Additional analyses of composite samples of cottonseed products (raw meal, toasted meal, kernel, refined oil) from each line showed that the products from the transgenic cotton lines were similar in composition to the control line. Feed studies of a four week rat feeding trial showed no difference in weight gain of animals fed diets containing 10% raw cottonseed meal from line 531 vs. Coker 312.<br />
The analysis of the fatty acid composition of refined oil from 531 and 757 cotton did not reveal any significant differences with the parent, non-transgenic variety and was within the normal range reported for cottonseed oils. In addition, the levels of alpha-tocopherol in refined oil from transgenic and control lines were similar. The consumption of refined oil from 531 and 757 cottonseed will have no significant impact on the nutritional quality of the food supply.</p>

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

<p style="text-align: justify;">Toxicity of cotton lines was assessed by analysis of naturally occurring toxins in cotton and an evaluation of the novel proteins Cry1Ac and NPTII. Toxicant analyses in cottonseed and refined oil evaluated levels of gossypol and cyclopropenoid fatty acids (sterculic and malvalic acid).</p>

<p style="text-align: justify;">Cotton is known for the production of anti-nutritional factors and untreated raw seed is unsuitable as livestock feed for monogastric animals. The transgenic and parental lines were assayed for the presence of potential toxins, including gossypol, dihydrosterculic acid, sterculic acid, malvalic acid and aflatoxins B1, B2, G1 and G2. At detection thresholds of 0.002% or 1 ppb, respectively, neither free gossypol nor any of the four aflatoxins were detected in the oil from transgenic cottonseed. Similarly, the respective levels of the cyclopropenoid fatty acids (dihydrosterculic, sterculic and malvalic) were statistically identical in cottonseed samples from transgenic and control lines.<br />
The deduced amino acid sequences of both Cry1Ac and NPTII were compared to the amino acid sequence of known protein toxins. No significant similarities were found.</p>

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

<p style="text-align: justify;">Refined cottonseed oil and cellulose from linters are devoid of detectable protein and their consumption is unlikely to result in an allergic reaction since most allergens are proteins. Therefore the potential for cottonseed oil or linters from transgenic cotton lines to constitute a source of allergens is extremely low.<br />
The Cry1Ac and NPTII proteins are not likely to be toxic or allergenic. The potential for allergenicity was assessed based on studies that included digestive degradation and sequence similarity to known allergens. The plant expressed proteins Cry1Ac and NPTII showed no significant protein sequence homology to a database of known toxins or allergens. Also, unlike known allergenic proteins, both Cry1Ac and NPTII were rapidly degraded when exposed to simulated gastric fluids (pH 1.2, pepsin digestion).</p>
