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

<p style="text-align: justify;">The genetic modification of MON 88913 would not be expected to result in any change in the consumption pattern of cotton products. MON88913 did not express any novel characteristics with regard to the seed and oil that would alter the consumption pattern. This was confirmed by comparison of the levels of proximates, and fatty acids in MON 88913 to the non-transgenic counterpart and to 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. Cottonseed oil for human consumption is highly refined and does not contain detectable levels of protein. Cottonseed oil and linters are therefore not expected to contain any detectable levels of CP4 EPSPS protein.</p>

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

<p style="text-align: justify;">The nutritional components of MON 88913 whole delinted cottonseed, cottonseed meal, and refined cottonseed oil were determined analytically and compared to those of the non-transgenic control line, and several commercial cottonseed cultivars grown in replicated field trials at four locations in 2002. The non-transgenic control line consisted of a null segregant containing the same genetic background as MON 88913, with the exception of the transgenes.<br />
The analysis of whole delinted (acid-delinted) cottonseed included proximates (crude protein, crude fibre, total fat, ash, moisture, and carbohydrates), acid detergent fibre, neutral detergent fibre, total dietary fibre, amino acids, fatty acids (C8 to C22), minerals (calcium, copper, iron, magnesium, phosphorus, potassium, sodium and zinc), and antinutritional compounds (cyclopropenoid fatty acids, gossypol [total and free], and aflatoxins). A combined statistical analysis across all locations revealed significant differences among the cultivars in the levels of the amino acids phenylalanine and tryptophan, the fatty acids oleic acid and linoleic acid, moisture, and manganese. Phenylalanine, linoleic acid and manganese levels were higher, and tryptophan and oleic acid levels were lower in MON 88913 compared to the non-transgenic control. However, the levels of these nutrients were within the 99% tolerance interval established for the conventional cotton varieties. The moisture levels for MON 88913 were outside of the tolerance interval, but within the range of values in the literature for conventional cotton. Analyses on mechanically-delinted whole cottonseed also revealed significantly higher values of phenylalanine in MON 88913; however, these values were within the range of those in the literature, and within the 99% tolerance for the conventional cultivars.<br />
The composition of cottonseed meal was similar to that of whole cottonseed, with the exception of analyses for fatty acids, vitamin E and total dietary fibre. No significant differences were observed in the levels of nutrients between MON 88913 and the non-transgenic control.</p>

<p style="text-align: justify;">The analysis of refined cottonseed oil consisted of fatty acids (C8 to C22), vitamin E, cyclopropenoid fatty acids (malvalic acid, sterculic acid and dihyroterculic acid), and gossypol (free and total). Statistically significant differences were observed between MON 88913 and the non-transgenic control in the levels of the fatty acids myristic acid and behenic acid. The mean levels of these fatty acids were lower in MON 88913, but within the 99% tolerance interval established for the conventional cultivars, and the range of values in the literature. Other than for those specific fatty acids, refined cottonseed oil from MON 88913 was not significantly different in composition compared to oil from conventional cotton.</p>

<p style="text-align: justify;">No significant differences were observed in the levels of antinutritional compounds in whole cottonseed, cottonseed meal and oil, with the exception of gossypol in cottonseed meal. The level of total gossypol were significantly lower in MON 88913 compared to the non-transgenic control, but fell within the range of literature values, and the 99% tolerance interval for conventional cotton.</p>

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

<p style="text-align: justify;">The toxicity of MON 88913 was assessed by an analysis of the levels of naturally occurring toxins in cotton and, an evaluation of the novel protein CP4 EPSPS. The levels of the toxicants gossypol (free and total) and cyclopropenoid fatty acids (malvalic, sterculic, and dihydrosterculic acid) were quantified in whole delinted cottonseed, cottonseed meal, and cottonseed oil.</p>

<p style="text-align: justify;">Gossypol has numerous toxic effects on mammals, including heart damage and heart failure, such that whole cottonseed or cottonseed meal is not used in human foods, and its use as a monogastric livestock feed is limited. However, the removal or inactivation of gossypol during processing enables the use of some cottonseed meal as feed for poultry, pigs and fish.</p>

<p style="text-align: justify;">Statistically significant differences in the levels of gossypol were observed only in cottonseed meal from MON 88913 and the non-transgenic control. Levels of total gossypol were lower in MON 88913, compared to the non-transgenic control, and were within the range observed in the published literature and in the 99% tolerance interval calculated for several commercial varieties grown at various locations.</p>

<p style="text-align: justify;">The cyclopropenoid fatty acids, sterculic (C-17), dihydrosterculic (C-19) and malvalic (C-18), are naturally found in the seed oil of cotton and other species of the Malvales (<i>e.g.</i>, kapok). These compounds produce undesirable biological effects, including the inhibition of biodesaturation of stearic to oleic acid, which detrimentally affects phospholipid biosynthesis. There were no statistically significant differences observed in the levels of these fatty acids in cottonseed, cottonseed meal or cottonseed oil from MON 88913 and the non-transgenic control.</p>

<p style="text-align: justify;">The potential for toxicity of the CP4 EPSPS was previously tested by acute gavage exposure in mice, and by conducting searches for amino acid sequence homology of the novel protein with those of known toxins. No adverse effects were observed when CP4 EPSPS was administered at levels up to 572 mg/kg of body weight. This dose represented a level of CP4 EPSPS that was 1000-fold the anticipated level that would be consumed in food products. The sequence homology searches were conducted using public protein databases. The results of the searches revealed no biologically relevant sequence similarities between CP4 EPSPS and known toxins.</p>

<p style="text-align: justify;">The CP4 EPSPS was also evaluated for potential allergenicity. An amino acid sequence homology search revealed no similarity to known allergens, including gliadin. Results of previous in vitro digestibility studies on CP4 EPSPS demonstrated its rapid digestibility in simulated gastric and intestinal fluids, which is characteristic of non-allergenic proteins. These results, and those of other studies previously submitted to regulatory authorities in support of the safety of CP4 EPSPS, substantiate its low potential for allergenicity.</p>
