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

<p style="text-align: justify;">The genetic modification of 3006-210-23 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 line 3006-210-23 to null plants and conventional cotton cultivars. Only refined cottonseed oil and cellulose from processed linters of cottonseed are consumed by humans. There were no detectable proteins in the oil from line 3006-210-23. Processed linters are composed of pure (&gt;99%) cellulose and are treated with heat and solvent that 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 components of 3006-210-23 cottonseed, kernels, toasted meal, hulls and oil were determined analytically and compared to those of the non-transgenic control (a null segregant resulting from several backcrosses of 3006-210-23 with both parental lines). Cottonseed analysis included proximates (crude protein, crude fibre, acid detergent fibre, neutral detergent fibre, total fat, ash, and moisture), minerals, amino acids, and fatty acids, and anti-nutrients (cyclopropenoid fatty acids, gossypol [total and free], aflatoxins). Oil analysis included crude fat, crude protein, moisture, fatty acids, anti-nutrients and anti-oxidants. Kernels, toasted meal, hulls were analyzed as appropriate for the particular fraction. Other than small differences in the levels of some components in cotton seed, which were determined to be biologically insignificant, and most likely due to genotypic and environmental effects, the analyses demonstrated that the composition of 3006-210-23 cottonseed is comparable to that of the non-transgenic control, and seed of other commercial cotton varieties. The composition of cottonseed oil was also comparable to the non-transgenic control and commercial cottonseed oil. Anti-nutrient levels in 3006-210-23 were similar to those of the non-transgenic control and to values published in the literature.</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 Cry1Ac and PAT proteins in line 3006-210-23 was assessed from the following data and information: results of searches for amino acid sequence similarity between the Cry1Ac and PAT proteins and known toxins and allergens; analysis for possible glycosylation of Cry1Ac; in-vitro digestibility studies using simulated gastric fluid; assessment of heat stability; and acute oral toxicity studies with mice.</p>

<p style="text-align: justify;">Both the Cry1Ac and PAT proteins showed no amino acid sequence similarity to known allergens. Both of these proteins were degraded within one minute of exposure to simulated gastric fluid and were no longer detectable by either SDS-PAGE or Western blot analysis. The Cry1Ac protein was no longer biologically active after exposure to temperatures greater than 75&deg;C. No post-translational glycosylation was detected in cotton-expressed Cry1Ac protein. Levels of natural toxicants in cotton (<i>e.g.</i>, gossypol) were not significantly different in 3006-210-23 cotton compared to the non-transgenic control and conventional cotton varieties.</p>

<p style="text-align: justify;">Results of feeding studies on mammals, birds, soil invertebrates, aquatic organisms, and non-target insects showed no observable adverse effects of either the microbially expressed Cry1Ac or the protein expressed in cotton tissues. Generally, for most of the species tested, there was at least 5-fold safety margin between the lowest observed effect level and the highest estimated concentration to which these species might be exposed. A 2-fold safety margin was determined for green lacewing. An additional non-target insect feeding study was requested by the Environmental Protection Agency (EPA) of the United States as a condition of registration. This study was conducted with the minute pirate bug, an important predatory insect found in cotton fields.</p>

<p style="text-align: justify;">Cry proteins in <i>Bt</i> biopesticide formulations have a long history of safe use, with no evidence of adverse effects, other than insecticidal activity towards lepidopteran insect pests. The Cry1Ac protein also was expressed at very low levels in the various tissues of 3006-210-23 cotton. On the basis this information, and the results of the various safety studies and sequence homology investigations, a determination was made that 3006-210-23 cotton did not demonstrate potential for toxicity or allergenicity compared to conventional cotton varieties.</p>
