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

<p style="text-align: justify;">Cotton (<i>Gossypium hirsutum</i>) is mainly a self-pollinating plant, but pollen is also routinely transferred by insects, particularly bumblebees and honey bees. The pollen is heavy and sticky and the range of natural crossing is limited. Outcrossing rates of up to 28% to other cotton cultivars grown directly adjacent to the pollen source have been observed under field conditions when sufficient insect pollinators have been present. The rate of outcrossing declines rapidly with increased distance from the pollen source. When cotton line MON 15985 (Bollgard II) is grown in proximity with other cotton cultivars and sufficient insect pollinators are present and active, the possibility exists for hybridization to occur between line MON 15985 and other cotton cultivars.</p>

<p style="text-align: justify;">In the United States, species compatible with line MON 15985 and other transgenic cottons include <i>G. hirsutum</i> (wild or under cultivation), <i>G. barbadens</i>e (cultivated Pima cotton), and <i>G. tomentosum</i>. It was reported that gene movement from <i>G. hirsutum</i> to <i>G. barbadense</i> may be possible given suitable conditions, while gene transfer to <i>G. tomentosum</i> is less probable due to chromosomal incompatibility, non-synchronous flowering periods and a lack of common pollinators. Populations of wild Gossypium species occur in southern Florida (wild <i>G. hirsutum</i>), Hawaii (<i>G. tomentosum</i>), Puerto Rico and the U.S. Virgin Islands, however, to reduce to potential for gene flow to wild species, the conditions of registration for cotton line MON 15985 include a prohibition on planting of event MON 15985 south of Route 60 (Tampa) in Florida and prohibition of commercial cultivation of the line in Hawaii, Puerto Rico and the U.S. Virgin Islands. These prohibitions, along with the different breeding systems and genome incompatibility in <i>Gossypium</i> species contribute to a low likelihood of gene transfer in the field Hybrids resulting from artificial crosses between cotton and wild species are generally sterile, unstable and of poor fitness.</p>

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

<p style="text-align: justify;">Cultivated <i>Gossypium hirsutum</i> is not typically considered a weed species in the United States or other countries, although it is listed as a potential weed in southern Florida. Cotton does not tolerate cold conditions limiting its potential to survive the winter to southern Florida, Hawaii, and Puerto Rico. The transformation that resulted in event MON 15985 did not confer any ecological advantage to MON 15985 cotton or its offspring that would increase its potential for weediness beyond that of currently cultivated commercial varieties.</p>

<p style="text-align: justify;">Data collected from field trials conducted in the United States, Puerto Rico, Argentina, South Africa, Costa Rica, and Australia indicated that the agronomic performance of event MON 15985 was similar to the parental cotton line. There were no reports of increased weediness in the parent line DP50B (MON 531 hybrid), a <i>Bt</i>-cotton engineered to express Cry1Ac protein, during its five years of commercial cultivation history in the United States. These data were used to determine that event MON 15985 did not pose any increased risk of weediness that was substantially different from that of conventional cultivars.</p>

<p style="text-align: justify;"><b>Secondary and Non-target Effects</b></p>

<p style="text-align: justify;">Studies were conducted to determine whether beneficial insects (ladybird beetle, adult and larval honeybees, collembola, green lacewing, parasitic wasp and earthworm) were susceptible to the Cry2Ab2 protein. Due to problems with the control population in the parasitic wasp trial, Monsanto was requested to submit an additional study. However, as parasitic wasps are not likely to come into contact with the Cry2Ab2 protein expressed in cotton, it was determined that the study was not necessary for obtaining regulatory approval for environmental release of cotton line MON 15985.</p>

<p style="text-align: justify;">For larval honeybees, the No Observed Effects Concentration (NOEC) was found to be greater than 100 ppm, which was expected to exceed the maximum concentration encountered in the field. Adult honeybee NOEC was greater than 68 ppm. Green lacewing larval NOEC was greater than 1100 ppm. All these exceeded the maximum environmental concentration expected in cotton plant tissue (51 ppm).</p>

<p style="text-align: justify;">In the case of ladybird beetles, the reasonable route of exposure would be through contact with cotton pollen. In the test, the beetles exhibited lethargy and low motility, thus not allowing for the determination of a NOEC. However, the LC50 of Cry2Ab2 protein for ladybird beetles was &gt;4500 ppm, which greatly exceeded the level expected in the field (Cotton event MON 15985 contains both Cry1Ac and Cry2Ab2 proteins. Non-target testing of Cry1Ac protein revealed no hazards. Unexpected synergistic effects from MON 15985 line containing both proteins were not anticipated as no adverse effects were seen in non-target tests of avian, earthworm and collembola species feeding on tissue containing both Cry1Ac and Cry2Ab2 proteins.</p>

<p style="text-align: justify;"><b>Impact on Biodiversity</b></p>

<p style="text-align: justify;">The transgenic cotton line MON 15985 has no novel phenotypic characteristics that would extend its use beyond the current geographical range of cotton production. Since there is no occurrence of cotton species in Canada and the line is not intended for cultivation in Canada, there will be no transfer of novel traits to unmanaged environments in Canada. Similarly, the risk of gene transfer to wild relatives in the United States was determined to be very remote, thus resulting in an insignificant risk of transferring genetic traits from line MON 15985 to wild species in unmanaged environments.</p>

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

<p style="text-align: justify;">In the United States, registration of event MON 15985 plant incorporated protectant (PIP) by the Environmental Protection Agency (EPA) was contingent upon the implementation of an Insect Resistance Management (IRM) program. IRM programs are intended to prolong the effectiveness of plant expressed <i>Bt</i> toxins and microbial spray formulations. In the case of cotton line MON 15985, it was proposed by Monsanto that the pyramiding effect of the two <i>Bt</i> toxins, Cry1Ac and Cry2Ab2 would act to delay insect resistance development, which indicated that refuge size could be reduced for this cotton line. Nonetheless, the United States EPA required the same 20% minimum non-<i>Bt</i> cotton refuge for event 15985 as for other <i>Bt</i> cotton events. In addition, monitoring requirements similar to those required for other <i>Bt</i> cotton lines must be adhered to.</p>
