<p style="text-align: justify;"><b>Dietary Exposure:</b>&nbsp;&nbsp;The genetic modification of LLCotton25 would not be expected to result in any change in the consumption pattern of cotton products. LLCotton25 did not express any novel characteristics with regard to cotton seed, oil, and the lint fraction that would alter the consumption pattern. This was confirmed by comparison of the levels of these fractions, and those of the proximates, and fatty acids in LLCotton25 to the unmodified parent and conventional cotton cultivars.</p>

<p style="text-align: justify;">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 that would be expected to remove and destroy any residual protein. PAT protein was detected in linters from LLCotton25; this protein would also be expected to be removed and during heat and solvent treatment. Cottonseed oil for human consumption is highly refined and does not contain detectable levels of protein. The PAT protein was not detected in either crude oil, or the more refined, deodorized oil. The consumption of these cotton fractions would therefore not be expected to result in an increase in exposure to PAT protein.</p>

<p style="text-align: justify;"><b>Nutritional and Compositional Data:</b>&nbsp;&nbsp;The nutritional components of LLCotton25 cottonseed were determined analytically and compared to those of its parental line, &lsquo;Coker 312.&rsquo; These components included proximates [moisture, ash, fat (ether extract), crude protein, crude fibre, acid detergent fibre (ADF), neutral detergent fibre (NDF), and carbohydrates], amino acids, fatty acids, minerals (calcium, phosphorus, iron, magnesium, potassium, and zinc), and vitamin E. Lint samples were analyzed for crude protein, fat, ash, crude fibre, ADF and NDF. The nutritional composition of the products of cottonseed processing (linters, delinted seed, meal, toasted meal, hull, crude oil and deodorized oil) was also determined, as appropriate for the particular fraction. The data from the nutritional analyses demonstrated that the composition of LLCotton25 cottonseed is comparable to that of the nonmodified parental line &lsquo;Coker 312,&rsquo; and seed of other commercial cotton varieties. The composition of the refined (deodorized) oil fraction was also comparable to Coker 312 and commercial cottonseed oil.</p>

<p style="text-align: justify;">The levels of the anti-nutritional factors phytic acid, gossypol (total and free), and cyclopropenoid fatty acids, were determined in cottonseed of LLCotton25 and Coker 312. The levels of these anti-nutrients in LLCotton25 were comparable to those of Coker 312, and to values published in the literature, except for levels of free gossypol, which were higher in both LLCotton25 and its parental line Coker312. The higher level of gossypol was therefore attributed to genotype, and not to any unintended effect of the genetic modification in LLCotton25.</p>

<p style="text-align: justify;">The feeding performance of LLCotton25 was assessed in a 33-day broiler chicken study. Treatments also included the parental line, and a conventional cotton variety. Cottonseed meal comprised 10% of the total ration fed to the broilers. No differences in animal performance, (weight gain, feed efficiency), feed consumption, and chilled carcass weight were observed among the treatments.</p>

<p style="text-align: justify;"><b>Potential Allergenicity and Toxicity:</b>&nbsp;&nbsp;The potential for toxicity and allergenicity of LLCotton25 was investigated using the following data and information: comparison of the amino acid sequence of the PAT to known toxins and allergens and the assessment potential glycosylation sites using several public databases (<i>e.g.</i>, SwissProt, trEMBL, GeneSequ-Prot); citations from published and unpublished safety studies using the PAT protein (<i>i.e</i>., <i>ex vivo</i> digestibility using pig and cattle gastric fluids, <i>in vitro</i> digestibility using simulated gastric and intestinal fluid conditions, assessment of heat sensitivity, acute intravenous and subchronic oral toxicity studies in rodents); results from a feeding study of broiler chickens using with LLCotton25 cottonseed meal.</p>

<p style="text-align: justify;">The PAT protein in LLCotton25, as expressed by the bar gene, showed no amino acid sequence similarity with known toxins and allergens, and no potential glycosylation sites. The protein was rapidly digested under simulated gastric fluid conditions but demonstrated some resistance to heat degradation. The results of acute and subchronic rodent studies showed no adverse effects of the PAT protein, thereby addressing the concern with regard to the heat stability. The broiler chicken feeding study using cottonseed meal (10% of the total ration) also showed no adverse effects that could be attributed to the PAT protein in LLCotton25.</p>

<p style="text-align: justify;">The PAT protein was expressed at very low levels in LLCotton25 cottonseed and was virtually absent from the oil. This fact, along with negative results of the various safety studies and sequence homology investigations, led to the conclusion that LLCotton25 did not demonstrate any potential for toxicity and allergenicity compared to conventional cotton varieties.</p>
