# Bayer CropScience Petition (08-340-01p) FOR Determination of Nonregulated Status of Insect Resistant and Glufosinate Ammonium-Tolerant (TwinLinkTM) Cotton, Gossypium hirsutum , Events T304-40 x GHB119

OECD Unique Identifier: BCS-GH004-7 x BCS-GH005-8



## Final Environmental Assessment

September 2011

Agency Contact Ms. Cynthia Eck, Document Cont rol Officer USDA, APHIS, BRS 4700 River Road, Unit 147 Riverdale, MD 20737-1237

Phone: (301) 734-0667 Fax: (301) 734-8669 Cynthia.A.Eck@aphis.usda.gov

The U.S. Department of Agriculture (U SDA) prohibits discrimination in all its programs and activities on the basis of race, color, national origin, sex, religion, age, disability, political beliefs, sexual or ientation, or marital or family statu s. (Not all prohibited bases apply to all programs.) Persons with disabilitie s who require alternative means for communication of program information (Braille, large print, audiotape, etc.) should contact USDA’S TARGET Center at (202) 720–2600 (voice and TDD).

To file a complaint of discrimination, write USDA, Dire ctor, Office of Civil Rights, Room 326–W, Whitten Building, 1400 Independence Avenue, SW, Washi ngton, DC 20250–9410 or call (202) 720–5964 (voice and TDD). USDA is an equal opportunity provider and employer.

Mention of companies or commercial products in this re port does not imply recommendation or endorsement by the U.S. Department of Agriculture over others not mentioned. USDA neither guarantees nor warrants the standard of any product mentioned. Product names are mentioned solely to report factually on available da ta and to provide specific information.

This publication reports research involving pesticides. All uses of pesticides must be regi stered by appropriate State and/or Federal agencies before they can be recommended.BAYER TWINLINKTM COTTON



### TABLE OF CONTENTS PAGE

i ACRONYMS ...................................................................................................................... .......... iv

1 PURPOSE AND NEED ..........................................................................................................1

1.1 REGULATORY AUTHORITY ................................................................................................... 1

1.2 REGULATED ORGANISMS ...................................................................................................... 2

1.3 PETITION FOR DETERMINATION OF NONREGULATED STATUS: BAYERCROPSCIENCE LEPIDOPTERAN-RESISTANT AND GLUFOSINATE AMMONIUMTOLERANT (TWINLINKTM) COTTON, EVENTS GHB119 AND T304-40 ........................... 2

1.4 PURPOSE OF PRODUCT ........................................................................................................... 3

1.5 APHIS RESPONSE TO PETITION FOR NONREGULATED STATUS ................................. 5

1.6 COORDINATED FRAMEWORK REVIEW .............................................................................. 6

1.7 PUBLIC INVOLVEMENT .......................................................................................................... 7

1.8 ISSUES CONSIDERED ............................................................................................................. .. 7
2 AFFECTED ENVIRONMENT .............................................................................................9

2.1 COTTON USE AND BIOLOGY ................................................................................................. 9

2.1.1 Cotton Taxonomy ............................................................................................................... .. 9

2.1.2 Cotton Use .................................................................................................................... ...... 10

2.2 AGRICULTURAL PRODUCTION OF COTTON .................................................................... 10

2.2.1 Acreage and Areas of Cotton Production ............................................................................ 10

2.2.2 Cropping Practices: Crop Rotation and Pesticide Use ....................................................... 11

2.2.3 Seed Production ............................................................................................................... ... 13

2.2.4 Organic Farming ............................................................................................................... .. 14

2.2.5 Specialty Cotton Production ............................................................................................... 15

2.3 PHYSICAL ENVIRONMENT ................................................................................................... 16

2.3.1 Water Resources ............................................................................................................... .. 16

2.3.2 Soil .......................................................................................................................... ............ 17

2.3.3 Air Quality ................................................................................................................... ....... 18

2.3.4 Climate Change ................................................................................................................ ... 19

2.4 ANIMAL AND PLANT COMMUNITIES ................................................................................ 21

2.4.1 Animals ....................................................................................................................... ........ 21

2.4.2 Plants ........................................................................................................................ ........... 22

2.4.3 Biological Diversity .......................................................................................................... .. 24

2.4.4 Gene Movement ................................................................................................................. . 25

2.5 PUBLIC HEALTH ................................................................................................................. .... 27

2.6 ANIMAL FEED ................................................................................................................... ....... 29

2.7 SOCIOECONOMIC ISSUES ..................................................................................................... 32
3 ALTERNATIVES .................................................................................................................3 4

3.1 NO ACTION: CONTINUATION AS A REGULATED ARTICLE ......................................... 34BAYER TWINLINKTM COTTON



### TABLE OF CONTENTS, CONTINUED PAGE

ii 3.2 PREFERRED ALTERNATIVE: DETERMINATION THAT TWINLINKTM COTTON IS NO LONGER A REGULATED ARTICLE ..................................................................................... 34

3.3 ALTERNATIVES CONSIDERED BUT REJECTED FROM FURTHER CONSIDERATION35

3.3.1 Prohibit Any TwinLinkTM Cotton from Being Released .................................................... 35

3.3.2 Approve the petition in part .............................................................................................. 35

3.3.3 Isolation distance between TwinLinkTM Cotton a nd non-GE cotton and geographical restrictions .................................................................................................................. ........ 36

3.3.4 Requirement of Testing For TwinLinkTM Cotton ............................................................... 36

3.4 COMPARISON OF AL TERNATIVES ..................................................................................... 37

4 ENVIRONMENTAL CONSEQUENCES ..........................................................................39

4.1 SCOPE OF THE ENVIRONMENTAL ANALYSIS ................................................................. 39

4.2 AGRICULTURAL PRODUCTION OF COTTON .................................................................... 40

4.2.1 Acreage and Areas of Cotton Production ............................................................................ 40

4.2.2 Cropping Practices: Crop Rotation and Pesticide Use ....................................................... 41

4.2.3 Seed Production ............................................................................................................... ... 44

4.2.4 Organic Farming ............................................................................................................... .. 44

4.2.5 Specialty Cotton Production ............................................................................................... 46

4.3 PHYSICAL ENVIRONMENT ................................................................................................... 47

4.3.1 Water Resources ............................................................................................................... .. 47

4.3.2 Soil .......................................................................................................................... ............ 48

4.3.3 Air Quality ................................................................................................................... ....... 50

4.3.4 Climate Change ................................................................................................................ ... 51

4.4 ANIMAL AND PLANT COMMUNITIES ................................................................................ 52

4.4.1 Animals ....................................................................................................................... ........ 52

4.4.2 Plants ........................................................................................................................ ........... 55

4.4.3 Biological Diversity .......................................................................................................... .. 56

4.4.4 Gene Movement ................................................................................................................. . 59

4.5 PUBLIC HEATH .................................................................................................................. ...... 62

4.5.1 Human Health .................................................................................................................. ... 62

4.5.2 Worker Safety ................................................................................................................. .... 65

4.6 ANIMAL FEED ................................................................................................................... ....... 66

4.7 SOCIOECONOMIC ISSUES ..................................................................................................... 68

4.7.1 Domestic Economic Environment at Risk .......................................................................... 68

4.7.2 Trade Economic Environment at Risk ................................................................................ 70

4.7.3 Social Environment at Risk ................................................................................................. 72

4.8 OTHER CUMULATIVE EFFECTS .......................................................................................... 73

4.9 THREATENED AND ENDANGERED SPECIES .................................................................... 73

4.9.1 Potential Effects of TwinLinkTM Cott on and Agricultural Practices on TES ..................... 75BAYER TWINLINKTM COTTON



### TABLE OF CONTENTS, CONTINUED PAGE

iii 4.9.2 Potential Effects of the Use of Glufosinate Ammonium ..................................................... 79

4.10 CONSIDERATION OF EXECUTIVE ORDERS, STANDARDS, AND TREATIES RELATING TO ENVIRONMENTAL IMPACTS .................................................................... 81

4.10.1 Executive Orders with Domestic Implications ................................................................... 81

4.10.2 International Implications ................................................................................................... 83

4.10.3 Compliance with Clean Water Act and Clean Air Act ....................................................... 84

4.10.4 Impacts on Unique Characteristics of Geographic Areas ................................................... 85

4.10.5 National Historic Preservation Act (NHPA) of 1966 as Amended ..................................... 85

5 BIBLIOGRAPHY .................................................................................................................8 7

6 LIST OF PREPARERS .....................................................................................................101

APPENDICES

Appendix A APHIS Threatened and Enda ngered Species Decision Tree for FWS Consultations

Appendix B Threatened and Endangered Species

LIST OF TABLES PAGE

Table 2-1: Cotton Fiber: World Production in Select Countries, 2006 to 2010/11 (estimated) (1,000 metric tons) .................................................................................................10 Table 2-2: Cotton Pests Ranked in Order of Bales Lost, Presen ting Percent U.S. Cotton Yield Reduction, Number of Cotton Acres Infest ed, Percent of U.S. Cotton Infested, Bales Lost, in 2009 ................................................................................................12 Table 3-1: Summary of Potential Impact s and Consequences of Alternatives .......................38 Table 4-1: Summary of Insecticide Use for Le pidopteran Control in Cotton Compared with Adoption of Bt Cotton ...........................................................................................43BAYER TWINLINKTM COTTON



### ACRONYMS

iv AIA advanced informed agreement AMS Agricultural Marketing Service ANZFA Australia and New Zealand Food Authority AOSCA Association of Official Seed Certifying Agencies APHIS Animal and Plant Health Inspection Service ARMS Agricultural Resource Management Survey BCS Bayer Crop Science BRAD Biopesticides Registration Action Document BRS Biotechnology Re gulatory Service BXN bromoxynil-tolerant CBD Convention on Biological Diversity CBW cotton bollworm CEQ Council of Environmental Quality CFIA Canadian Food Inspection Agency CFR Code of Federal Regulations DNA deoxyribonucleic acid EA Environmental Assessment ECOS Environmental Cons ervation Online System ELS extra-long staple EO Executive Order EPA Environmental Protection Agency ERA ecological risk assessment ESA Endangered Species Act EU European Union FAW fall armyworm FDA Food and Drug Administration FFDCA Federal Food, Drug, and Cosmetic Act FFP food, feed, or processing FIFRA Federal Insecticide, Fungicide, and Rodenticide Act FR Federal Register GE genetically engineered GHG greenhouse gases IPCC Intergovernmental Panel on Climate Change LMO living modified organism MFA Multifibre Arrangement MOU memorandum of understanding
N 2 O nitrous oxide NABI North American Biotechnology Initiative NAPPO North American Plant Protection Organization NEPA National Environmental Policy Act NHPA National Historic Preservation Act NMFS National Marine Fisheries Service NOP National Organic Program NRCS National Resources Conservation ServiceBAYER TWINLINKTM COTTON



### ACRONYMS, CONTINUED

v OECD Organization for Economic Cooperation and Development OSTP Office of Science and Technology Policy PAT phosphinothricin-acetyl-transferase PBO Plant Biosafety Office PGF pollen-mediated gene flow PPA Plant Protection Act RR Roundup Ready (registered trademark) RSPM Regional Standards for Phytosanitary Measures TBW tobacco budworm TES threatened and endangered species TSCA Toxic Substances Control Act U.S.C. United States Code USDA U.S. Department of Agriculture USFWS U.S. Fish and Wildlife Service WPS worker protection standardBAYER TWINLINKTM COTTON

1 

### 1 PURPOSE AND NEED

1.1 REGULATORY AUTHORITY

“Protecting American agriculture” is the basic charge of the U.S. Department of Agriculture's (USDA) Animal and Plant Health Inspection Serv ice (APHIS). APHIS provides leadership in ensuring the health and care of plants and animals. The agency improves agricultural productivity and competitiveness, and contributes to the national economy and the public health. USDA asserts that all methods of agricultural producti on (conventional, organi c, or the use of genetically engineered varieties) can provide benefits to the e nvironment, consumers, and farm income.

Since 1986, the United States government has regulat ed genetically engineered (GE) organisms pursuant to a regulatory framework known as the Coordinated Framework for the Regulation of Biotechnology (Coordinated Framework) (51 FR 23302, 57 FR 22984). The Coordinated Framework, published by the Office of Scie nce and Technology Policy, describes the comprehensive federal regulatory policy for ensu ring the safety of biotechnology research and products and explains how federal agencies will use existing Federal statutes in a manner to ensure public health and environmental safety wh ile maintaining regulatory flexibility to avoid impeding the growth of the biotechnology indus try. The Coordinated Framework is based on several important guiding principles: (1) agenci es should define those transgenic organisms subject to review to the extent permitted by their respective statutory authorities; (2) agencies are required to focus on the characteristics and risk s of the biotechnology pro duct, not the process by which it is created; (3) agencies are mandated to exercise oversight of GE organisms only when there is evidence of “unreasonable” risk.

The Coordinated Framework explains the regulato ry roles and authorities for the three major agencies involved in regulating GE organism s: USDA’s Animal and Plant Health Inspection Service (APHIS), the Food and Drug Administra tion (FDA), and the Environmental Protection Agency (EPA).

APHIS is responsible for regulating GE organism s and plants under the plant pest provisions in the Plant Protection Act of 2000, as amended (7 USC § 7701 et seq .) to ensure that they do not pose a plant pest risk to the environment.

The FDA regulates GE organisms under the author ity of the Federal Food, Drug, and Cosmetic Act (FFDCA). The FDA is responsible for ensu ring the safety and proper labeling of all plantderived foods and feeds, including those that are genetically engin eered. To help developers of food and feed derived from GE crops comply with their obligations under Federal food safety laws, FDA encourages them to participate in a voluntary consultation proce ss. All food and feed derived from GE crops currently on the market in the United States have successfully completed this consultation process. The FDA policy st atement concerning regulation of products derived from new plant varieties, including those gene tically engineered, was published in the Federal Register on May 29, 1992 (57 FR 22984-23005). Under this policy, FDA uses what is termed a consultation process to ensure th at human food and animal feed sa fety issues or other regulatory issues (e.g., labeling) are resolv ed prior to commercial dist ribution of bioengineered food.BAYER TWINLINKTM COTTON

2 The EPA regulates plant-incorpor ated protectants under the Federa l Insecticide, Fungicide, and Rodenticide Act (FIFRA). EPA also sets tolera nce limits for residues of pesticides on and in food and animal feed, or estab lishes an exemption from the re quirement for a tolerance, under the Federal Food, Drug and Cosmetic Act (FFDCA ) and regulates certain biological control organisms under the Toxic Substances Control Act (TSCA). The EPA is responsible for regulating the sale, distribution a nd use of pesticides, including pe sticides that are produced by an organism through techniques of modern biotechnology.

1.2 REGULATED ORGANISMS

The APHIS Biotechnology Regulatory Service’s (BRS) mission is to protect America’s agriculture and environment using a dynamic a nd science-based regulatory framework that allows for the safe development and use of GE organisms. APHIS regulations at 7 Code of Federal Regulations (CFR) Part 340 (hereafter Part 340), which were promulgated pursuant to authority granted by the PPA, regulate the intr oduction (importation, interstate movement, or release into the environment) of certain GE organisms and products. A GE organism is no longer subject to the plant pest provisions of the Plant Protection Act or to the regulatory requirements of Part 340 when APHIS determines th at it is unlikely to pos e a plant pest risk. A GE organism is considered a regulated article under Part 340 if the donor organism, recipient organism, vector, or vector agen t used in engineering the organi sm belongs to one of the taxa listed in the regulation (7 CFR 340.2) and is also c onsidered a plant pest. A GE organism is also regulated under Part 340 when APHIS has reason to believe that the GE organism may be a plant pest or APHIS does not have information to dete rmine if the GE organism is unlikely to pose a plant pest risk.

A person may petition the agency that a particular regulated article is unlikely to pose a plant pest risk, and therefore, is no longer regulated under the plant pest pr ovisions of the Plant Protection Act or the regulations at 7 CFR 340. The petitioner is re quired to provide information under §340.6(c)(4) related to plant pest risk that the agency may use to determine whether the regulated article is unlikely to present a greater pl ant pest risk than the unmodified organism. A GE organism is no longer subject to the regulator y requirements of 7 CFR Part 340 or the plant pest provisions of the Plant Protection Act when APHIS determines that it is unlikely to pose a plant pest risk.

1.3 PETITION FOR DETERMINATION OF NONREGULATED STATUS: BAYER CROPSCIENCE LEPIDOPTERAN-RE SISTANT AND GLUFOSINATE AMMONIUM-TOLERANT (TWINLINKTM ) COTTON, EVENTS GHB119 AND T304-40

Bayer Crop Science (BCS) has submitted a petition (APHIS Number 08-340-01p) seeking a determination that TwinLinkTM Cotton Events GH B119 and T304-40 are unlikely to pose a plant pest risk and, therefore, should no longer be a re gulated article under regulations at 7 CFR Part
340.

TwinLinkTM Cotton is a combined-trait cotton deve loped using conventional breeding techniques to link two deoxyribonucleic acid (DNA) transformation even ts; each developed using DNA recombinant techniques. By crossing BCS’ Cr y1Ab Cotton (event T304-40) with BCS’ Cry2AeBAYER TWINLINKTM COTTON

3 Cotton (event GHB119), BCS has developed a co tton resistant to lepidopteran pests. The TwinLinkTM Cotton also expresses a glufosinate ammonium herbicide to lerance trait based on LibertyLink ® technology.

TwinLinkTM Cotton is currently regulated under 7 CFR Part 340. Interstate movement, importation, and field testing of TwinLinkTM Co tton have been conducted under notifications acknowledged by APHIS.

1.4 PURPOSE OF PRODUCT

BCS has developed the TwinLinkTM Cotton as an alternative insect-res istant and herbicidetolerant cotton product.

BCS has developed upland or Mexican cotton ( Gossypium hirsutum ) plants that express two insecticidal crystalline protei ns, Cry1Ab and Cry2Ae, derived from the common soil bacterium, Bacillus thuringiensis ( Bt ). The Cry1Ab and Cry2Ae prot eins in TwinLinkTM Cotton are effective in controlling lepidopteran larvae such as bollworm (CBW, Helicoverpa zea ), tobacco budworm (TBW, Heliothis virenscens ), and fall armyworm (FAW, Spodoptera frugiperda ) which are common pests of cotton. The rationale for using two Bt genes is that target insects are much less likely to develop resi stance to both proteins simultaneous ly than to develop resistance to one toxic protein. In addition to the Cr y1Ab and Cry2Ae protei ns, the TwinLinkTM Cotton contains the modifying phosphi nothricin-acetyl-transferase (PAT) enzyme, encoded by the bar gene which confers tolerance to glufos inate ammonium-based herbicides. The bar gene, derived from Streptomyces hygroscopicus, encodes the PAT enzyme which acts to convert glufosinate ammonium into its inactive form, thus rendering the plant tolerant to the herbicide. This is the same enzyme that is expressed in BCS LibertyLink ® Cotton (LLCotton25) that also confers tolerance to glufosinate ammonium herbicides.

Cotton producers in the U.S. are among the most technically advanced in the world, annually harvesting about 17 million bales or 7.2 billi on pounds of cotton (US-EPA, 2009). In 2010, the USDA estimates that 18.9 million bales we re harvested (USDA-NASS, 2010a) with approximately 10.9 million acres planted (USDA-NA SS, 2010b). Cotton growers strive for high yields of high-value cotton fiber at the lowest production cost. To enhance successful cotton fiber production, the cotton industr y relies heavily on pest control measures to control both weeds and lepidopteran insects. Infestation of cotton by lepidopteran larvae represents the largest insect threat to cotton production in the U.S. (Bayer, 2010) . To combat these threats and enhance production, cotton growers in the U.S. have adopted genetically improved cotton, to reduce pest management costs, obtain greater yields, and decrease pesticide costs; thereby producing greater grower return on investment (Bayer, 2010). In 2010, 78% of all cotton grown in the U.S. contained herbicide-tolerant traits and 73% expressed insect -resistant traits (USDAERS, 2010a).

Cotton producers are currently limited to ins ect-resistant cotton vari eties containing three Bacillus thuringiensis Cry1 endotoxin protein-based plant inco rporated protectants. Each is no longer subject to the plant pestprovisions of the Plant Prot ection Act and 7 CFR part 340 and registered by the EPA under FIFRA. The thre e proteins are Cry1Ac (B ollgard)—a combination of CrylAc and Cry2Ab2 (Bollgard II), Cry1F, an d Cry1Ac (Widestrike) (USDA-APHIS, 2010a).BAYER TWINLINKTM COTTON

4 These transgenic varieties offer almost complete protection against tobacco budworm, but may require additional applications of insecticide for control of cott on bollworm. Hence, despite the wide adoption of these Bt cotton varieties to c ontrol lepidopteran insect s, these same species continue to be the most economically important pests of the crop. In 2009, it was estimated that the Heliothine complex of tobacco budworm and cotton bollworm infested 4.3 million acres of cotton and reduced yields across the Cotton Belt by 91,119 bales (Williams, 2010). Total 2009 cost and loss estimates for arthropod cotton pests were $502 million dollars (Williams, 2010).

TwinLinkTM Cotton has been developed by BCS as an alternative insect-res istant and herbicidetolerant cotton. The introduction of the Bt pr otein Cry2Ae provides an additional measure of insect protection not currently available in cott on. The tolerance to gl ufosinate ammonium, in addition to the insect resistance provided by th e two Cry proteins, will provide cotton growers with a new approach for managing lepidopteran pe sts of cotton, as well as herbicide-resistant weeds.

Current insect resistance manage ment strategies have delayed th e development of resistance to Bt toxins. However, in a review of GE crops, Lemaux (2009) reports that some cotton lepidopteran-insect resistance to Bt toxins ha s occurred. The availability of additional lepidopteran insect-resistant traits in cotton, such as that conferred by incorporation of the Cry2Ae protein, further reduces the likelihood of pest resistance to existing pest control methods (Gould, 2003). A recent study evaluated several cultivation practices which might delay the development of insect resistan ce, including the planting of refuge areas with crops not engineered to contain Bt (Tabashnik et al., 2008).

Glufosinate ammonium tolerance was in troduced by Bayer in its LibertyLink ® Cotton to provide cotton growers with an alternative herbicide choice. The LibertyLink ® Cotton product was no longer subject to the plant pest provisions of the Plant Protection Act and 7 CFR part 340 in 2003 (USDA-APHIS, 2003) and is widely availabl e. EPA has published a voluntary pesticide resistant management labeling guideline prov iding guidance on reducing or managing the potential for herbicide-resistant weeds or volunteers (Pesticide Registration Notice 2001-5) (USEPA, 2001a). Glufosinate ammonium tolerance provides growers with a nother herbicide option in cotton cultivation.

TwinLinkTM Cotton, either alone or when comb ined by traditional breeding with other genetically-modified insect-resistant cotton va rieties, by incorporat ing a new Cry protein, provides growers with an additiona l pest management option for le pidopteran-insect pest control and may contribute to a reduction in the likelihood of insect resist ance to Bt insect-resistant cotton varieties. Specificall y, BCS indicates that the use of TwinLinkTM Cotton will provide:

1. Improved efficacy in reduction of major insect pests of cotton, independent of weather conditions.
2. Combined Bt trait product for insect resistance management.
3. Additional weed control options.
4. Excellent human, animal, non-target organi sm, and environmental safety profile.
5. Potentially reduced pesticide use.
6. Potentially enhanced yield.
7. Potentially decreased pest management costs.BAYER TWINLINKTM COTTON

5 1.5 APHIS RESPONSE TO PETITION FOR NONREGULATED STATUS

Under the authority of the plant pest provisions of the Plant Protection Act and 7 CFR Part 340, APHIS has issued regulations for the safe development and use of GE organisms. As required by 7 CFR 340.6, APHIS must respond to petitioners who request a determination of the regulated status of GE organisms, including GE plants such as BCS TwinLinkTM Cotton. When a petition for nonregulated status is submitted, APHIS must make a determination if the GE organism is unlikely to pose a plant pest risk. If APHIS determines based on its Plant Pest Risk Assessment (PPRA) that the genetically engineered organism is unlikely to pose a plant pest risk, the genetically engineered organism is no longer subject the plant pest provisions of the Plant Protection Act and 7 CFR part 340.

TwinLinkTM Cotton has been field tested in the U. S. since 2005, as authorized by APHIS. Data were provided in the petition for field trials comple ted prior to the petition submission. Field test reports can be found in the BCS TwinLinkTM Cotton petition in Appendix 1, p.102 (Bayer, 2010).

BCS has conducted 66 field trials under field release authoriza tions granted by USDA APHIS in diverse regions of the U.S., including Arizona, California, Georgia, L ouisiana, Mississippi, North Carolina, South Carolina, and Texas. Field tests conduc ted under APHIS oversight allow for evaluation in agricultural settings under c onfinement measures designed to minimize the likelihood of persistence in the environment after completion of the field trial. Under confined field trial conditions, data are gathered on multiple parameters and used by applicants to evaluate agronomic characteristics and produ ct performance. These data ar e also valuable to APHIS for assessing the potential for a new variety to pose a plant pest risk. The data evaluated for TwinLinkTM Cotton may be found in the document Determination of Plant Pest Risk for InsectResistant and Glufosinate A mmonium-Tolerant (TwinLinkTM) Cotton, Gossypium hirsutum, Events T304-40 x GHB119 (USDA-APHIS, 2010b).

APHIS has prepared this environmental assessm ent to consider the potential environmental effects of an agency determ ination of nonregulated status consistent with Council of Environmental Quality’s (CEQ) regulations implementing NEPA (40 CFR Parts 1500-1508), and the USDA and APHIS NEPA implementing re gulations and procedures (7 CFR 1b, and 7 CFR Part 372) and the USDA and APHIS NEPA implementing regulations and procedures. This EA has been prepared in order to specifi cally evaluate the effects on the quality of the human environment 1 that may result from a determina tion of nonregulated status of BCS TwinLinkTM Cotton.

1 Under NEPA regulations, the “human environment” includes “the natural and physical environment and the relationship of people with that environment” (40 CFR §1508.14).BAYER TWINLINKTM COTTON

6 1.6 COORDINATED FRAMEWORK REVIEW

Food and Drug Administration

TwinLinkTM Cotton (events T304-40 x GHB119) is within the scope of the 1992 FDA policy statement concerning regulation of products deri ved from new plant vari eties, including those developed through biotec hnology (US-FDA, 1992).

Bayer has provided the FDA with information on th e identity, function, and characterization of the genes, for TwinLinkTM Cotton, including expression of th e gene products. The FDA is currently reviewing the informati on submitted by the applicant.

APHIS considers the FDA food and feed safety and nutritional assessment determination when assessing potential impacts that may result from a determination of nonregulated status of a GE organism. In the absence of a completed FDA determination, APHIS takes into consideration prior FDA reviews of comparable products to ma ke a preliminary assessment of the potential impacts.

Note that with regard to the e xpression of glufosinate ammonium to lerance, the gene construct in the Bayer TwinLinkTM Cotton is the same as that approved by the FDA in June 2003 for the LibertyLink ® Cotton product (US-FDA, 2003). In that approval, the F DA noted that the transformational event in LibertyLink ® Cotton was not materially different in composition, safety, or any other relevant parameter in co tton grown, marketed, and consumed at that time (US-FDA, 2003). This previous FDA review will be used by APHIS to analyze the food and safety impacts associated with the incorpor ation and expression of glufosinate ammonium tolerance in TwinLinkTM Cotton.

The FDA’s oversight of the food and safety im pacts associated with the incorporation and expression of pesticidal substances , in this case, the Cry proteins associated with Bt, are morelimited. As discussed below, the EPA is the primary authority for the review of plantincorporated protectants.

Environmental Protection Agency

The EPA has authority over the use of pesticidal substances and plant-in corporated protectants under the FIFRA as amended (7 USC §136, et seq. ) and the FFDCA (21 USC §301, et seq.). EPA is currently reviewing information submitted by the applicant on the efficacy and potential environmental concerns associated with the us e of this product. APHIS considers the EPA’s regulatory assessment when assessing potential imp acts that may result from a determination of nonregulated status of a GE organism. In th e absence of a completed EPA determination, APHIS takes into consideration prior EPA re views of comparable products to make a preliminary assessment of the potential impacts.

Note that EPA has issued a tolerance exemp tion for Cry1Ab protein in all crops (40 CFR §174.511; US-EPA, 2010d), as well as for the PA T protein (40 CFR §17 4.522; US-EPA, 2010g). A temporary exemption from the requirement of a tolerance has been issued for Cry2Ae (40 CFR §174.530; US-EPA, 2010e). These previous EPA reviews will be used by APHIS toBAYER TWINLINKTM COTTON

7 analyze the food and safety impact s associated with the incorporat ion and expression of the Cry proteins in TwinLinkTM Cotton.

In addition to review of the crop with plant- incorporated protectants, EPA has authority under FIFRA to establish pesticide use restrictions; these use restric tions are presented on pesticide labels which are prepared during the pesticide registration process. The development of a transformed cotton crop providing tolerance to glufosinate ammoni um may require a change in the EPA-approved label for this herbicide.

The current glufosinate ammonium label provides for its use on transformed crops expressing resistance to glufosinate ammonium, and specifi cally references products marketed under the trade name “LibertyLink” (BCS, 2010a). Gluf osinate ammonium-toler ant cotton was first available in the U.S. with the introduction of LibertyLink ® Cotton in 2004. Although the glufosinate ammonium toleran ce trait expressed in TwinLinkTM cotton is the same as that expressed in the LibertyLink ® Cotton, the EPA is expected to publish a new label for glufosinate ammonium that also references the TwinLinkTM product varieties. APHIS will use current glufosinate label as the basis for its evaluation of the potential impacts associated with the use of and exposure to glufos inate ammonium.

1.7 PUBLIC INVOLVEMENT

APHIS routinely seeks public comment on draft EA s prepared in response to petitions seeking a determination of nonregulated status of GE organisms. APHIS does this through a notice published in the Federal Register. The issues discussed in this EA were developed by considering public concerns, as well as issues raised in public comments submitted for other environmental assessments of GE organisms, concer ns raised in lawsuits, as well as those issues that have been raised by various stakeholders . These issues, includ ing those regarding the agricultural production of cotton using various production methods, and the environmental and food/feed safety of GE plants, were addressed to analyze the potential en vironmental impacts of TwinLinkTM Cotton.

This EA, the petition submitted by BCS, and APHIS’s Determination of Plant Pest Risk for Insect-Resistant and Glufosinate Ammonium -Tolerant (TwinLinkTM) Cotton, Gossypium hirsutum, Events T304-40 x GHB119 (USDA-APHIS, 2010b) will be available for public comment for a period of 60 days (7 CFR §340.6(d )(2)). Comments received by the end of the 60-day period will be reviewed and used to in form APHIS’s determination decision of the regulated status of TwinLinkTM Cotton and to assist APHIS in determining whether an Environmental Impact Statement is required prior to a determination of th e regulated status of TwinLinkTM Cotton.

1.8 ISSUES CONSIDERED

As stated above, the issues considered in this EA were developed based on APHIS’s determination that certain genetically engineered organisms are no longer subject to the plant pest provisions of the Plant Protection Act and 7 CFR part 340; and, for this particular EA, the specific petition seeking a determination of nonr egulated status of TwinLinkTM Cotton. These issues include:BAYER TWINLINKTM COTTON

8 Cotton Production:

 Acreage and Areas of Cotton Production

 Cropping Practices

 Seed Production

 Organic Farming

 Specialty Cotton Production

Environmental Considerations:

 Water Resources

 Soil

 Air Quality

 Climate Change

 Animals

 Plants

 Biological Diversity

 Gene Movement

Public Health Considerations:

 Human Health

 Worker Safety

Animal Feed

Socioeconomic Issues:

 Domestic Economic Environment at Risk

 Trade Economic Environment at Risk

 Social Environment at Risk

Other Cumulative Effects

Threatened and Endangered Species

Other U.S. Regulatory Approvals and Compliance with Other LawsBAYER TWINLINKTM COTTON

9 

### 2 AFFECTED ENVIRONMENT

The Affected Environment Section provides an overview of the use and biology of cotton, followed by a discussion of the current conditions of those aspects of the human environment potentially impacted by a determination of nonre gulated status of Tw inLinkTM Cotton. For the purposes of this EA, those aspects of the human environment are: cotton production practices, the physical environment, biological resources, public health, animal f eed, and socioeconomic issues.

2.1 COTTON USE AND BIOLOGY

2.1.1 Cotton Taxonomy

Cotton is a perennial plant that is cultivat ed as an annual crop (USDA-ERS, 2009). The Gossypium genus is made up of approximately 50 speci es, but only five species are cultivated worldwide (OECD, 2008). The cultivated species upland cotton ( G. hirsutum ) represents 97% of cotton crop grown in the U.S. (USDA-ERS, 2009). The balance of U.S.-grown cotton is American Pima, the generic name for extra-long staple (ELS) cotton, G. barbadense (Meyer et al., 2007; USDA-ERS, 2009). ELS is cultivated primarily in irrigated regions of California, New Mexico, Arizona, and Texas (Meyer et al., 2007; USDA-ERS, 2009). The three other cultivated species are G. arboretum, G. herbaceum, and G. lanceolatum (Bayer, 2010; OECD, 2008).

Seven genomes, designated A, B, C, D, E, F, and G, are found in the Gossypium genus (Endrizzi et al., 1984). Diploid (i.e., two sets of chromosomes) species (2n=26) are found on all continents, and a few are of some agricultural importance. The A genome is restricted in diploids to two species ( G. arboreum and G. herbaceum ) of the Old World. The D genome is restricted in diploids to some sp ecies of the New World, such as G. thurberi . The most important agricultural cottons, G. hirsutum and G. barbadense, are both allotetraploids (i.e., hybrids containing 4 complete haploid chromosome sets representing the complete chromosome sets from both parent organism s) (2n=4x=52). These cultivated varieties are of New World origin and are presumed to be the result of an ancient interspecific hybridization between Old World A genomes and New World D genomes (B ayer, 2010). How and when the original crosses occurred has been subject to much specu lation. Euploids (i.e., organisms containing equal numbers of the haploid chromosome sets) of these plants have 52 somatic chromosomes and are frequently designated as AADD (they be have as disomic polyploids, i.e., have two complete sets of chromosomes from both original ancestors). The New World allotetraploids are peculiar in the genus, because the species, at leas t in their wild forms, grow near the ocean as invaders in the constantly distur bed habitats of strand and associat ed environs. It is from these "weedy" or invader species that the cu ltivated cottons developed (Fryxell, 1984).

Due to the difference in ploidy level (i .e., number of sets of chromosomes), G. hirsutum cannot cross with wild diploid cottons. G. hirsutum is readily cross-compatible , i.e., can hybridize, only with other tetraploid members of the tribe Gossypium , which include G. tomentosum in Hawaii

, G. darwinii in the Galapagos Islands, G. mustelinum in northeastern Brazil, G. hirsutum and G. lanceolatum in tropical/subtropical America, and G. barbadense in South America, as well as cultivated forms of G. hirsutum and G. barbadense (Fryxell, 1984).BAYER TWINLINKTM COTTON

10 These distinctions in ge netic construct are impo rtant to recognize, as the genetic differences directly affect the ability of cultivated cotton to interbreed with wild cotton species as well as wild or feral cotton. There are only tw o wild cotton species found in the U.S.: G. thurberi (desert cotton), f ound in Arizona; and G. tomentosum (Hawaiian cotton), found in Hawaii (ITIS, 2010; USBG, 2010; USDA-ARS, 2010a). G. tomentosum has been crossed with G. hirsutum in breeding programs; however, no commercial cott on is produced in Hawaii (Bayer, 2010; USDANASS, 2010c). Feral populat ions of upland cotton ( G. hirsutum ) can be found in southern Florida (USDA-FS, 2010). These populations are a pparently self-sustaining because there is no commercial production of cotton in this region (Bayer, 2010; USDA-NASS, 2010c).

2.1.2 Cotton Use

Cotton ( Gossypium spp.) is a member of the Malvaceae family and is the world’s most widely grown fiber crop, accounting for over 40% of the total world fiber use (Meyer et al., 2007). Cotton has been cultivated for its fiber for se veral thousand years (Bayer, 2010). Other cotton products, such as cottonseed oil, cake, and co tton linters, are by-products of cotton fiber production (Bayer, 2010).

2.2 AGRICULTURAL PRODUCTION OF COTTON

2.2.1 Acreage and Areas of Cotton Production

Cotton is cultivated mostly in subtropical and warm temperate zones (OECD, 2008). Temperature is the main climatic factor determ ining the geographic range in which cotton can be grown, but cultivation is also dependent upon hi gh light intensity, good soil moisture, and soil fertility (Meyer et al., 2007; OECD, 2008).

Cotton is mainly produced in China, India, U.S ., Pakistan, Brazil, and Uzbekistan, with these five countries contributing nearly 83% of worl d production (Table 2-1) (USDA-FAS, 2010a). The total planted area throughout the world in 2007-2008 was 33.2 million hectares, for a production of 116 million bales (approximately 26 million metric tons) (USDA-FAS, 2010a). The U.S. is the third largest producer of cotton fiber.

Table 2-1: Cotton Fiber: World Produc tion in Select Countries, 2006 to 2010/11 (estimated) (1,000 metric tons)

Country Year 2006/2007 2007/2008 2008/2009 2009/2010 2010/2011 (est.) China 7,729 8,056 7,991 6,967 6,858 India 4,746 5,225 4,921 5,051 5,661 United States 4,700 4,182 2,790 2,654 4,109 Pakistan 2,090 1,872 1,894 2,090 2,025 Brazil 1,524 1,602 1,193 1,165 1,524 Uzbekistan 1,165 1,165 1,002 849 1,045
Australia 294 139 327 386 718 Other 4,258 3,814 3,210 2,907 3,463 Total 26,507 26,056 23,326 22,070 25,404 Source: (USDA-FAS, 2010a)BAYER TWINLINKTM COTTON

11 In the U.S. for the 2010 production year, cotton was grown on 4.4 million hectares (approximately 10.8 to 10.9 million acres) (USD A-NASS, 2010b). In th e U.S., cotton is produced in 17 southern states identified as the Cotton Belt: Alabama, Arizona, Arkansas, California, Florida, Ge orgia, Kansas, Louisiana, Mississi ppi, Missouri, New Mexico, North Carolina, Oklahoma, South Carolina, Tennessee, Texas, and Virginia (Meyer et al., 2007; USDA-NASS, 2010c). The major cotton producing st ates are Texas (5.4 million acres), Georgia (1.3 million acres), Arkansas (0.540 million acre s), Mississippi (0.420 million acres), North Carolina (0.545 million acres), and Tennessee (0.387 million acres) (USDA-NASS, 2010c).

Cotton acreage in the U.S. rose slightly duri ng the first half of the 2000s, continuing a multidecade trend. In the 1970s and 1980s, the area planted for cotton averaged about 12 million acres (USDA-ERS, 2009). The area planted ro se to about 14 million acres in the 1990s and averaged over 14.5 million acres during the first half of the 2000s, repres enting an increase of 20% over the 1970s and 1980s acreage (USDA-ER S, 2009). Since 2006, however, U.S. cotton planted acreage has been considerably lower, as prices have favored the planting of alternative crops such as corn and soybeans (USDA-ER S, 2009). All cotton growing regions have experienced significant d eclines compared with the first half of the 2000s (USDA-ERS, 2009).

GE varieties of cotton have been very rapi dly adopted. In 1996, when Bt cotton was first commercially introduced, initial plantings were estimated at 1.7 million acres (Benbrook, 2009). Of the approximate 11 million acres planted in cotton in 2010, about 93% (10.2 million acres) were GE cotton. Of this, 73% of the GE cotton acreage was GE insect-res istant (Bt) cotton, and 78% was herbicide-tolerant (USDA-ERS, 2010a).

2.2.2 Cropping Practices: Crop Rotation and Pesticide Use

Growers can choose from many cultivars of co tton marketed by companies that produce seed, including GE varieties (USDA-AMS, 2010a). Plan ting of cotton occurs after any danger of frost has passed and soil temperatures are at least 14 o C (57 o F) (OECD, 2008). Planting dates vary depending on the region, but generally range from early March in southern Texas to early May or June in the northern areas of the Cotton Be lt. Cotton requires 180 to 200 days from planting to maturity (OECD, 2008). Ther efore, cotton planted in March or April is ready for harvest in September.

Crop rotations (successive planting of different crops on the same la nd) are used to optimize soil nutrition and fertility and reduce pathogen loads (Gazaway et al., 2007; University of California IPM Online, 2008a). Cotton is ofte n rotated with other crops in order to control various cotton pests including nematodes, verticillium wilt, seed ling diseases, and pink bollworm (University of California IPM Online, 2008a). Rotation crops may include small grains, cowpea, corn, sorghum, alfalfa, onions or garlic, and nematode-re sistant tomatoes. Rota tions may last for two or three years. Winter cover crops are also u tilized in cotton. These cover crops are used to provide winter soil cover and protection, build soil nitrogen and organic matter, reduce nitrogen leaching, suppress weeds, and provide a habitat for beneficial predatory a nd parasitic insects and spiders (Guerena a nd Sullivan, 2003).

Cotton production in the U.S. is highly mechaniz ed and involves the exte nsive use of agronomic inputs and technology (USDA-APHIS, 2010a). Th e cost of production includes annual directBAYER TWINLINKTM COTTON

12 costs (seed, fertilizer, chemicals, irrigation, etc.), annual fixed co sts (depreciation on equipment), and annual land rent (Hogan et al., 2005). Th ese costs will vary depending on region and practices used. In 2009, average per acre costs fo r U.S. cotton growers for cotton seed, fertilizer (commercial fertilizers, soil conditioners, and ma nure), and chemicals (including pesticides and herbicides) accounted for 54% of the total dollars spent per planted acre or $232 per acre (USDA-ERS, 2010b).

Worldwide, cotton has proven vulnerable to the atta ck of many insect species. In the U.S., the cotton industry has consistently relied heavily on insecticide use strategies to manage arthropod pests (Gianessi and Carpenter, 1999). Resistan ce to commonly used insecticides (pyrethroids, organophosphates, and carbamates) has led to a need for new pesticide chem istries or other novel approaches, such as the use of sterile insects, pathogens, and transgen ic cotton (Gianessi and Carpenter, 1999). Pest management in agricultu ral crop production is ac hieved through various management methods, including chemical (pes ticides), cultural (mechanical cultivation, planting/harvesting dates, crop rotation, etc.), biological (antagonistic organisms), and bioengineering (primarily herbic ide-tolerant and insect-resist ant crops) (USDA-ERS, 2005a). GE insect-resistant cotton has been focused on incorporating Bacillus thuringiensis ( Bt ) Cry endotoxin protein based plant incorp orated protectants (Stewart, 2007).

The most damaging insect pests of cotton attack th e cotton square (the flow er bud) or the cotton boll (the ovary containing develo ping seeds and fibers) (Gianessi and Carpenter, 1999). There are over 20 economically important pests of cott on. The major insect pests of U.S. cotton in 2009 are listed in Table 2-2 (Williams, 2010). Thrips ( Frankliniella occidentalis ), lygus ( Lygus hesperus ), and the budworm/bollworm complex each infest ed more than half of the U.S. cotton crop. The budworm/bollworm complex, which infested 56% of the acreage, was the third leading cause of yield loss due to insects. It is referred to as a complex because the larvae of these moth insects are identical when observed in the field (Gianessi an d Carpenter, 1999). The complex consists of the cotton bollworm ( Helicoverpa zea ) and tobacco budworm ( Helicoverpa
virescens ) whose small larvae feed on smaller squares and terminal buds, but whose older, larger larvae devour buds, flowers, and bolls, consumi ng both lint and seed (Gia nessi and Carpenter, 1999). In 2009, the complex reduced yields by 0.486% or 91,119 bales of cotton (Williams, 2010). Bt cotton provides good control over bollw orm and tobacco budworm, as well as loopers, fall armyworm, and beet armyworm (Stewart, 2007).

Table 2-2: Cotton Pests Ranked in Order of Bales Lost, Presenting Percent U.S. Cotton Yield Reduction, Number of Cotton Acres Infested, Percent of U.S. Cotton Infested, Bales Lost, in 2009

Pest Percent U.S. Cotton Yield Reduction Attributed to Pest Acres Infested with Pest Percentage of Cotton Acreage Infested Bales Lost Thrips 0.713% 7,437,609 94.49% 138,207
Lygus 0.614% 4,209,086 53.47% 126,871
Bollworm/Budworm 0.486% 4,376,640 55.60% 91,119
Stink Bugs 0.371% 3,265,604 41.49% 79,327
Fall Armyworm 0.113% 1,835,299 23.32% 20,238BAYER TWINLINKTM COTTON

13 Pest Percent U.S. Cotton Yield Reduction Attributed to Pest Acres Infested with Pest Percentage of Cotton Acreage Infested Bales Lost Spider Mites 0.099% 2,022,705 25.70% 21,673
Cotton Fleahoppers 0.093% 1,490,760 18.94% 15,598
Clouded Plant Bugs 0.038% 452,752 5.75% 8,300
Aphids 0.030% 3,851,114 48.93% 6,456
Silverleaf Whitefly ( Bemisia ) 0.008% 289,731 3.68% 2,245
Beet Armyworm 0.004% 703,067 8.93% 643
Other Insects 0.003% 62,193 0.79% 668
Boll Weevil 0.002% 116,247 1.48% 400
Grasshoppers 0.001% 1,128,763 14.34% 266
Banded Wing Whitefly 0.001% 524,519 6.66% 249

Cutworms 0.001% 386,007 4.90% 200
Loopers 0.000% 808,131 10.27% 12
Saltmarsh Caterpillars 0.000% 487,934 6.20% 0
Cotton Leaf Perforator 0.000% 2,585 0.03% 1
Pink Bollworm 0.000% 81,211 1.03% 0
European Cornborer 0.000% 655 0.01% 0
Southern Armyworms 0.000% 68,798 0.87% 0
Source: (Williams, 2010)

2.2.3 Seed Production

On an annual basis, certified seed of all varietie s of cotton combined must be able to plant over 10 million acres in the U.S. alone (USDA- NASS, 2010b) This requires between 60,000 and 70,000 short tons of planting seed (National Cottonseed Products Association, 2011).

Public cotton breeding programs began in Califor nia in 1898 and private programs began in the early 1920s (Marra and Martin, 2007). Cotton was one of the earliest crops for which the Association of Official Seed Certifying Agen cies (AOSCA) develope d seed certification standards to maintain genetic purity of cotton varietal seed and specialty cotton crops by precluding gene flow and preventing cross pollin ation between species and varieties (AOSCA, 2010b; Wozniak, 2002).

In a seed certification program, classes of seed are identified to designate the seed generation from the original breeder source (Hartman and Kester, 1975). Foundation seed, Registered seed, and Certified seed production is controlled by public or private seed certification programs (see, e.g., AOSCA, 2010b). The original seed breeder seed stock is controlled by the developer of the variety (Adam, 2005; Hartman and Kester, 1975). The breeder stock is used to produce Foundation seed stock (Adam, 2005). The institution associated w ith the breeder controls the production of Foundation seed stock. Foundation seed stock, in turn, is used to produce Registered seed for distribution to licensees, su ch as seed companies (Adam, 2005). Registered seed is used by seed companies to produce la rge quantities of Certified seed (Adam, 2005;BAYER TWINLINKTM COTTON

14 Hartman and Kester, 1975). The Certified (or Se lect) seed is then sold to growers through commercial channels (Adam, 2005; Hartman and Kester, 1975).

Seed certification cultivation practices co mmonly include recommendations for minimum isolation distances between various seed lines a nd planting border or barrier rows to prevent pollen movement (Hartman and Kester, 1975; Wozniak, 2002). The isolation distance for Foundation, Registered, and Certified seeds, as dictated by the USDA Agricultural Marketing Service's (AMS) Federal Seed Act in 7 CFR Part 201.76, is 1,320, 1,320, and 660 feet, respectively (USDA-AMS, 2010c). During the grow ing season, seed certification agencies will monitor the fields for off-types, other crop s, weeds, and disease (Wozniak, 2002). These certifying agencies also establis h seed handling standards to redu ce the likelihood of seed source mixing during production stages, in cluding planting, harvesting, tran sporting, storage, cleaning, and ginning (Wozniak, 2002). Further discussi on of cross-pollination, gene transfer, and weediness is presented in Subsection 2.4.4, Gene Movement.

2.2.4 Organic Farming

In the U.S., only products produced using specific methods and certified under the USDA National Organic Program (NOP) definition of organic farming can be marketed and labeled as “organic” (Ronald and Fouche, 2006; USDA-AMS, 2010b). The NOP is administered by USDA’s AMS.

Organic certification is a process-based certifi cation, not a certification of the end product. The certification process specifies and audits the methods and procedures by which the product is produced (Ronald and Fouche, 2006). In accordance with NOP, an accredited organic certifying agent conducts an annual review of the certified operation’s organic system plan and makes on-site inspections of the certified operation and its records. Organic growers must maintain records to show that productio n and handling procedures comply with USDA organic standards.

The NOP regulations preclude the use of exclud ed methods. The NOP provides the following guidance under 7 CFR §205.105—

To be sold or labeled as “100 percent or ganic,” “organic,” or “made with organic (specified ingredients or group(s)),” th e product must be produced and handled without the use of: (a) Synthetic substanc es and ingredients,... (e) Excluded methods,...

Excluded methods are then defined at 7 CFR §205.2 as—

A variety of methods used to genetically modify organisms or influence their growth and development by means that ar e not possible under natural conditions or processes and are not considered co mpatible with organic production. Such methods include cell fusion, microencapsulation and macroencapsulation, and recombinant DNA technology (including gene deletion, gene doubling, introducing a foreign gene, and changing the positions of genes when achieved byBAYER TWINLINKTM COTTON

15 recombinant DNA technology). Such methods do not include the use of traditional breeding, conjugation, fermentation, hybridization, in vitro fertilization, or tissue culture.

The NOP has recognized the feasibility of protec ting organically-produced crops from accidental contamination by GE crops by requiring that orga nic production plans incl ude practical methods to protect organica lly-produced crops—

Organic crops must be protected from contamination by prohibited substances used on adjoining lands (for example, dr ifting pesticides, fertilizer-laden runoff water, and pollen drift from genetic ally engineered...) (NCAT, 2003).

Organic farming operations, as described by the NOP, require or ganic production operations to have distinct, defined boundaries and buffer zones to prevent uni ntended contact with excluded methods from adjoining land that is not unde r organic management. Organic production operations must also develop and maintain an or ganic production system plan approved by their accredited certifying agent. This plan enables the production operation to achieve and document compliance with the National Organic Standards, including the prohib ition on the use of excluded methods. In NOP organic systems, the use of GE crops, such as TwinLinkTM Cotton, is excluded (USDA-AMS, 2010b).

The organic plan used as the basis for organi c certification should incl ude a description of practices used to prevent or reduce the likelih ood of unwanted substances, like GE pollen or seed, at each step in the farming operation, such as planting, harvesting, storing, and transporting the crop (Krueger, 2007; Riddle, 2004). Organic pl ans should also include how the risk of GE pollen or co-mingling of seed will be monito red (Krueger, 2007; Riddle, 2004). Farmers using organic methods are requested to let neighbori ng farmers know that they are using organic production practices and request that the neighbors also help the organic farmer reduce contamination events (Krueger, 2007; NCAT, 2003).

Organic cotton provides price premiums for the grower (Guerena and Sullivan, 2003). Organic production begins with certified organically grown seed (Guerena and Sullivan, 2003). Certified organic cotton acreage is a re latively small percentage of overall cotton production in the U.S. The most recently av ailable data show 15,377 acres of certified organic cotton production in 2008 (USDA-ERS, 2010c ). This is 0.16% of the total 9.41 million acres of cotton planted in 2008 (USD A-ERS, 2009). Bayer CropScience FM 958, FM989, and ADF 2485 were the predominate va rieties planted in 2010 by organic cotton producers (USDA-AMS, 2010b).

2.2.5 Specialty Cotton Production

Several specialty cottons are cultiv ated in the U.S., including ELS ( G. barbadense ) Pima varieties, specialty l ong-staple upland cotton ( G. hirustum ) Acala varieties, organic cotton, and naturally colored cotton (Lee, 1996; USDA-ERS, 2009). Historically, ELS cotton acreage has ranged from a low of 63,000 acres in 1983 to a high of 377,000 acres in 1989 (National Cotton Council of America, 2010). This represented 0. 83% and 3.5% of the total cotton acreage planted for those years, respectively (National Cotton Council of America, 2010). Naturally coloredBAYER TWINLINKTM COTTON

16 cotton acreage was reported to be 4,000 acres in 1992 (Dickerson et al., 1999) and between 5,000 and 7,000 acres in 1995 (Lee, 1996).

Similar to the production of conventional seed, industry quality standards for specialty crop products have led these seed producers and grower s to employ a variety of techniques to ensure that their products are not pollinated by or co mmingled with conventional or genetically engineered crops (Bradford, 2006). Common practices include maintaining isolation distances to prevent pollen movement from other cotton sources , planting border or barri er rows to intercept pollen, and employing natural ba rriers to pollen (NCAT, 2003; Wozniak, 2002). The Federal Seed Act Regulations provide additional detail s on Certified seed pr oduction (see 7 CFR 201, et

seq ). Field monitoring for off-t ypes, other crops, weeds, disease etc. is also carried out by company staff and state crop improvement a ssociations (Bradford, 2006). Seed handling standards are established by AOSCA to reduce the likelihood of seed source mixing during planting, harvesting, transporting, storage, clean ing, and ginning (AOSCA, 2004). In general, the conventional management practices used for conventional seed production are sufficient to meet standards for the production of specialty crop seed (Bradford, 2006).

2.3 PHYSICAL ENVIRONMENT

The use of fertilizers, pesticides, and water may affect segments of th e environment including, but not limited to: waterways by increases in nu trient pollution; biodive rsity because pesticide inputs cause species changes; th e water table because of excessive irrigation practices; and productivity of cropland because irrigation incr eases salinity (Hoeft et al., 2000). Sediment, siltation, nutrients, pestic ides, salinity, and pathogens are pr imary agricultural pollutants (USDAERS, 2005b).

2.3.1 Water Resources

Cotton has been developed as a drought-tolerant crop; cotton production water use varies according to the growing environment (Cotton Incorporated, 2010b). Successful cultivation of dryland (non-irrigated) cotton requires at least 500 millimeters (mm) (40 inches) of rainfall during the growing season (OECD, 2008). Cotton cultivated in th e southwestern region of the U.S., including west Texas, southern New Mexic o, southern Arizona, and southern California, requires irrigation (USDA-APHIS, 2010a). Where irrigation wate r is needed (approximately 35% of the U.S. cotton grown), co tton yields are also much highe r (Cotton Incorporated, 2010a). The desert southwest requires a maximum of 40 in ches of irrigation per year, although the humid southeast may only require about 18 inches of ir rigation per year (Cotton Incorporated, 2010a). Carefully timing the application of irrigation wa ter optimizes the plant’s vegetative growth, flowering, and boll production (OECD, 2008). The l ack of affordable water has been noted as one factor in the reduction of acres of cotton grown in Californ ia, Arizona, and New Mexico in the past decade (Cotton Incorporated, 2010a). In these areas, cotton has been displaced by higher value crops and land uses (Cotton Incorporated, 2010a).

Conservation tillage and no-till practices have be en shown to minimize surface water runoff and soil erosion and to improve soil quality by increasing the soil organic matter that helps bind soil nutrients and prevent their loss to runoff, erosion, and leaching (L eep et al., 2003). This decrease in soil erosion carries a corresponding decrease in non-point source surface water pollution ofBAYER TWINLINKTM COTTON

17 fertilizer and pesticides (Co tton Incorporated, 2010a, 2010c). Precision agricu lture, a practice carefully managing nutrients and chemical applica tions, also ensures that inputs are used by the crop and are not entering ground or surface waters (Cotton Incorporated, 2010a). Precision agriculture practices may include computer mode ls to predict water use based on plant growth stage, weather, soil moisture probe data collect ion, and thermal infrared imagery of plant leaf temperature (Cotton Incorporated, 2010a). Each of these practices has the potential to reduce soil-laden water runoff (USDA-NRCS, 2006b).

2.3.2 Soil

Cotton is cultivated in a wide variety of soils, but develops best in deep, arable soils with good drainage, high organic content, and a hi gh moisture-retention capacity (OECD, 2008). Irrigation allows cultivation in poor-quality soils with necessary nutrients provided in the irrigation water (OECD, 2008).

Conventional tillage traditionally requires that the producer remove all plant residues and weeds from the soil surface prior to planting, a nd then continue to cultivate the soil while the crop is growing to control late emergi ng weeds (Cotton Incorporated, 2010c). This practice results in soil loss to wind and water erosion.

Conservation practices have been developed to reduce field tillage and thus reduce the corresponding soil loss (USDA-NRCS, 2006a). As conservation practices are adopted, there are increases in soil organic matter that helps bind soil nutrients and corresponding significant reductions over time in the lo ss of cropland soil from runoff, erosion, and leaching (Leep et al., 2003; USDA-NRCS, 2006a, 2006b). Total soil loss on highly erodible croplands and non-highly erodible croplands decr eased from 462 million tons per year to 281 million tons per year or by 39.2% from 1982 to 2003 (USDA-NRCS, 2006b). This decrease in soil erosion carri es a corresponding decrease in non-point source surface water pollution of fertilizer and pesticides (Cotton Incorporated, 2010a, 2010c). The reduction in soil erosion is also attributed to a decrease in the number of acres of highly erodible cropland being cultivated (USDA-NRCS, 2006b).

Over the last 10 years, cotton cultivation ha s contributed to this reduction in soil loss through a shift towards soil conservation practices (Cotton Incorporated, 2010c). Conservation practices adopted in cotton cu ltivation include use of seed treatment fungicides, herbicides, and herbicide-tolerant cotton varieties, each of which provide for the control of cotton disease and weeds with minimal tillage (Cotton Incorporated, 2010c; USDA-NRCS, 2006b). In the U.S., conservation til lage in cotton fields has increased from 0.5 million acres in 1990 to about 2.75 million acres in 2004; in 2008, approximately 67% of cotton growers reported adopting some for m of conservation tillage (Cotton Incorporated, 2010c). Increases in total acres dedicated to conservation tillage have been at tributed to an increased use of GE seed which eliminates th e need for mechanical w eed control (Alabama Cooperative Extension System, 1996; McClella nd et al., 2000; Towery and Werblow, 2010; USDA-NRCS, 2006b).

By definition, conservation tillage leaves at least 30% of the soil covered by crop residue (Peet, 2001). The new crop is planted into the plant residue or in narrow strips of tilled soil.BAYER TWINLINKTM COTTON

18 This is in comparison to conventional tillage where the seedbed is prepared through plowing (to turn the soil surface over), disking (to redu ce the size of soil clods created by plowing), and harrowing (to reduce the size of clods left by disking) (Peet, 2001). Benefits of reduced tillage practices include maintenance of soil organic matter and beneficial insects, increased soil water-holding capacity, less soil and nutri ent loss from the field, reduced soil compaction, and less time and labor required to prepare the field for planting (Peet, 2001).

Weed control in conservation tillage cotton is primarily through the use of herbicides (Alabama Cooperative Extension System, 1996). Preplant “burndown” herbicides such as paraquat and glyphosate and some pre-emerge nt herbicides make up the primary weed control methods. Winter and cover crops are also utilized in conservation tillage for the purpose of suppressing weeds (Alabama Coopera tive Extension System , 1996). Wheat and rye are commonly employed because of their ease in killing prior to cotton planting. The use of herbicide-resistant cotton has allowe d cotton growers to more readily adopt soil conservation practices because it provides an economical, effective means of controlling weeds in post-plant cotton (Alabama Cooperativ e Extension System, 1996; McClelland et al., 2000).

In some regions, conservation tillage is requ ired. Farmers, including cotton growers, producing crops on highly erodible land are required by law to maintain a soil conservation plan approved by the USDA National Res ources Conservation Service (NRCS) (USDAERS, 2010d). These soil conservation plans are prepared by the grower pursuant to the 1985 Food Security Act Conservation Compliance and Sodbuster programs to minimize soil erosion (USDA-ERS, 2010d).

The adoption of Bt cotton, whic h may require reduced quantities of pesticides, may benefit soil quality by reducing the risks associated with envi ronmental spills or misapplications of chemical insecticides to the soil, and reducing the fre quency of application. Reduced insecticide applications also mean a reduction in the number of trips across the field with heavy farm equipment which contributes to soil compaction, esp ecially when the soil is wet. The number of acre-treatments per year of budworm/bollworm insect icides declined from over 10 million to less than 3 million between the years 1996 and 2007 (US DA-NASS, 2008). This is attributed to the widespread adoption of Bt cotton, introduced commercially in 1996, which increased from 1.73 million acres of cotton planted to 7.6 million acre s in the U.S. during the same time period (Benbrook, 2009).

2.3.3 Air Quality

Agricultural air emission sources include sm oke from agricultural burning, vehicle exhaust associated with equipment used in tillage and harv est, suspended soil particulates associated with tillage, pesticide drift from spraying, and nitrou s oxide emissions from the use of nitrogen fertilizer (Aneja et al., 2009; Hoeft et al ., 2000; US-EPA, 2010b; USDA-NRCS, 2006c). These agricultural activities individually have the pote ntial to cause negative impacts to air quality.

Aerial application of pesticides may cause air quality impacts from drift and diffusion. Pesticides may volatilize after application to so il or plant surfaces and may also move as constituents of entrained materials in wind eroded soils (Vogel et al., 2008).BAYER TWINLINKTM COTTON

19 Many of the conservation plans and practices bei ng developed by cotton growers to comply with the Conservation Compliance and Sodbuster program s have an air quality focus which target reductions in air emissions from agricultural operations (Cotton In corporated, 2010c; USDANRCS, 2006a). Practices to improve air quality include c onservation tillage, residue management, wind breaks, road treatments, burn management, prunings shredding, feed management, manure management, integrated pe st management, chemical storage, nutrient management, fertilizer injection, chemigation and fertigation (inclusion in irrigation systems), conservation irrigation, sc rubbers, and equipment calibration (USDA-NRCS, 2006a). Conservation tillage practices re sulting in improved air quality in clude: fewer tractor passes across a field, thus decreasing dus t generation and tractor emissions ; and an increase in surface plant residues and untilled organic matter wh ich physically hold the soil in place and reduce wind erosion (Baker et al., 2005; Cotton Inco rporated, 2010d; USDA-NRCS, 2006a). The USDA has estimated that by 2006, the adoption of conservation management plans in the San Joaquin Valley of California had re duced air emissions by 34 tons da ily, or more than 20% of the total emissions attributed to agricultural pract ices (Baker et al., 2005; USDA-NRCS, 2006a).

2.3.4 Climate Change

Agriculture, including land-use ch anges for farming, is responsible for an estimated 6% of all human-induced greenhouse gases (GHG) in the U.S. (US-EPA, 2010b). Emissions of GHG released from agricultural equipment (e.g., irrigation pumps and tr actors) include carbon monoxide, nitrogen oxides, reactiv e organic gases, particulate ma tter, and sulfur oxides (USEPA, 2010b). Agricultural soil management pr actices, including nitrogen-based fertilizer application and cropping practices, are the largest source of U.S. nitrous oxide (N 2 O) emissions; croplands account for 69% of the total N 2 O emissions attributable to agricultural land uses (USEPA, 2010b). Agriculture source s of methane emissions are prim arily associated with enteric emissions of gas from cattle and manure manageme nt. Carbon dioxide is also a significant GHG gas associated with several agricultural practi ces, including land uses and energy consumption (US-EPA, 2010b).

Tillage contributes to the release of GHG because of the loss of carbon dioxide (CO 2 ) to the atmosphere, and the exposure and oxidation of soil organic matter (Baker et al., 2005). The carbon footprint for cotton production has been estimated to be approximately 300 pounds of carbon equivalent emission per acre (Nelson et al., 2009). The carbon f ootprint of cotton is directly affected by the associated cultivation pr actices. In a no-tillage production environment, if credit is taken for the sequest ration of carbon in soil, cotton act ually stores more carbon than is emitted from the production (Cotton Incorpor ated, 2010d). Reduced tillage increases sequestration rates of carbon emissions in soils, w ith the estimated increase to be as high as 400 pounds of carbon per acre per year (Causarano et al., 2006).

The contribution of agriculture to climate change is largely dependent on the production practices employed to grow various commoditi es, the region in which the commodities are grown, and the individual choices made by grower s. For example, emissi ons of nitrous oxide, produced naturally in soils through microbial nitrification and denitr ification, can be dramatically influenced by fertilization, introduction of grazi ng animals, cultivation of nitrogen-fixing crops and forage, retention of crop re sidues (i.e., no-till conservation) , irrigation, and fallowing of land (US-EPA, 2010b). These same agricultural pract ices can influence the decomposition of carbon-BAYER TWINLINKTM COTTON

20 containing organic matter sequestered in soil, re sulting in conversion to carbon dioxide loss to the atmosphere (US-EPA, 2010b). Conversion of crop land to pasture results in an increase in carbon and nitrogen sequestrati on in soils (US-EPA, 2010b).

The EPA has identified regional differences in GHG emissions associated with agricultural practices on different soil type s, noting that carbon emission ra tes differ between mineral soils and organic soils (US-EPA, 2010b) . Mineral soils contain from 1 to 6% organic carbon by weight in their natural state, whereas organi c soils may contain as much as 20% carbon by weight (US-EPA, 2010b). In mineral soils, up to 50% of the soil organic carbon can be released to the atmosphere on the initial conversion, however, over time, the soil establishes a new equilibrium that reflects a bala nce between carbon inputs from decaying plant matter and organic amendments and carbon losses from microbial de composition (US-EPA, 2010b). Organic soils, with their depth and richness in carbon content, continue to release carbon to the atmosphere for a longer period of time (US-EPA, 2010b). The EPA has estimated that mineral soil-based cropland areas sequestered over 45.7 Tg CO 2 Eq 1 in 2008, as compared with carbon emissions from organic soil-based croplands of 27.7 Tg CO 2 Eq (US-EPA, 2010b). The adoption of conservation tillage, particularly in the Midwest regions with mine ral soil shows the highest rates of carbon sequestration (US-EPA, 2010b).

Changes in agriculture-relate d GHG production will not be signi ficant unless large amounts of crop plantings produce changes in measureable concentrations (USDA-APHIS, 2010a). For example, the EPA has identified a net reduction in the sequestration of carbon in soil over an 18- year time scale, which it attributes to the de clining influence of th e Conservation Reserve Program which had encouraged gr owers to take marginal lands out of production (US-EPA, 2010b). To a certain extent, the EP A also noted that adoption of conservation tillage resulted in increases in carbon sequestration on those cropla nds (US-EPA, 2010b). The highest rates of carbon sequestration in mineral so ils occurred in the Midwest, which is the region with the largest area of cropland managed with conservation tillage (US-EPA, 2010b). This is in contrast to the highest emission rates from organic soils no ted in the southeastern coastal region, the areas around the Great Lakes, and the central and nor thern agricultural areas along the West Coast (US-EPA, 2010b).

One outcome of the potential effects of agriculture on climate change is the potential effect of the climate change on agriculture. In response to climate change, the current range of weeds and pests of agriculture is expected to increase. Current agricultural practi ces will be required to change in response to these changes in the ranges of weeds and pests of agriculture (IPCC, 2007).

Climate change may potentially provide a positiv e impact to agriculture in general. The Intergovernmental Panel on Climate Change (IPCC) predicts that potential climate change in

1 The global warming potential of greenhouse gases are measured against the reference gas CO 2 , and are reported as teragrams (or million metric tons) of CO 2 Equivalent, expressed as Tg CO 2 Eq.BAYER TWINLINKTM COTTON

21 North America may result in an increase in crop yield by 5-20% for this century (IPCC, 2007). However, this positive impact will not be observ ed across all growing regions. The IPCC report notes that certain regions of the U.S. will be negatively impacted because the available water resources may be substantially reduced. Note that the extent of c limate change effects on agriculture is highly sp eculative. Nevertheless, North Amer ican production is expected to adapt to climate change impacts wi th improved cultivars and res ponsive farm management (IPCC, 2007).

2.4 ANIMAL AND PLANT COMMUNITIES

Modern conservation practices incorporated in cotton cultivation have brought a positive impact to animal and plant communities, through reduced tillage, more carefully controlled and targeted chemical placement (fertilizers and pesticides), a nd better control of irriga tion systems. The GE crop systems provide opportunities to optimize the introduction and implementation of these practices, and have the potential to create more of these benefits. For example, herbicide tolerance allows cultivation with minimal till age required to control volunteers and weeds (Towery and Werblow, 2010). Th is subsection provides an overv iew of the relationships of these practices to the biotic community.

2.4.1 Animals

Intensive agricultura l lands, such as those used in crop pr oduction, usually have low levels of biodiversity compared with adjacent natural areas. Tillage, seed bed preparation, planting of a monoculture crop, pesticide use, fertilizer use, and harvest result in limited habitat and a correspondingly limited diversity of plants and animals (Lovett et al., 2003). However, the implementation of better cropland management strategi es can increase the value of crop fields to wildlife (Sharpe, 2010). Some of th ese strategies are noted below.

 Conservation tillage and no-till practices ha ve a positive impact on wildlife (Towery and Werblow, 2010). Benefits include improve d water quality, retention of cover, availability of waste grain on the soil surf ace for feed, and increased populations of invertebrates as a food source for turk ey, quail, and songbirds (Sharpe, 2010).

 Crop rotations reduce the likelihood of crop dis ease, insect pests, weed pests, and the need for pesticides (Universit y of California IPM Online, 2 008a). Reduced pesticide use has a direct positive effect on wildlife by reducing the direct exposure of birds, mammals, and fish to pesticides. Indirect benefits include less alteration of suitable wildlife habitat and an available food supply of insects fo r insectivores (Pal mer and Bromley, 2010; Sharpe, 2010). Crop rotations with legumes a nd small grains have been shown to provide excellent wildlife nesting cover, food, a nd brood-rearing habitat for quail in North Carolina (Sharpe, 2010).

 Field edges can be managed to promote wild life. These borders are often the least productive areas in a farm field and in some cases, the cost of producing crop areas along field edges exceeds the value of the crop pr oduced (Sharpe, 2010). Allowing field edges to return to volunteer vegetation does not contribute to ma jor pest problems in the crop field itself (Sharpe, 2010). Volunteer bo rder vegetation, such as ragweed, goldenrod,BAYER TWINLINKTM COTTON

22 asters, and forbs, quickly develops into nesting and brood habitat for quail and a multitude of songbirds (Sharpe, 2010). Re search conducted at North Carolina State University and the North Carolina Wild life Resources Commission found that quail populations doubled when field borders were used (Sharpe, 2010).

 Contour-strip cropping is another management practice that can be used to promote wildlife habitat. This practice alternates stri ps of row crops with strips of solid stand crops (i.e., grasses, legumes, or small grains ) with the strips follo wing the contour of the land (Sharpe, 2010). The primary purpose of contour-strip cropping is to reduce soil erosion and water runoff, but the solid sta nd crop also provides a nesting and roosting cover for wildlife (Sharpe, 2010).

 Drainage ditches, hedgerows, riparian areas, and adjacent woodlands to a cotton field also contribute to wildlife populations. D itch banks, for example, function as narrow wetlands that provide nesting sites and cove r, serve as wildlife corridors, and provide areas for the wildlife to occupy when crop fiel ds lack cover (Sharpe, 2010). Ditches have been shown to support birds, rodents, reptil es, furbearers, amphibians, fish, and aquatic organisms (Sharpe, 2010). Minimizing pestic ide exposure of ditches, aquatic habitats, border areas, strip-crop areas, a nd non-crop habitats may help protect fish and wildlife resources (Palmer and Bromley, 2010).

Although many of the invertebrate organisms found in cotton-producing areas are considered pests, such as the cotton bollwor m and tobacco budworm, most invertebrates are considered beneficial (University of Arka nsas Cooperative Extension Service, 2006). Beneficial insects include a wide variety of predators which catch and eat smaller insects and parasitic insects that live on or in the bod y of other insects during at least one stage of their life cycle. Other beneficial ins ects are the pollinators. Major pollinators of G. hirsutum are b umble bees ( Bombus spp .), black bees ( Melissodes spp .), and honey bees ( Apis mellifera ) (McGregor, 1976). Pollinators of G. tomentosum are principally moths (McGregor, 1976). Other beneficial organism s, including earthworms, termites, ants, beetles, millipedes, and others contribute to the decay of organic matter and th e cycling of soil nutrients (Ruiz et al., 2008).

2.4.2 Plants

The landscape surrounding a cotton field varies depending on the region. In certain areas, cotton fields may be bordered by other cotton (or any other crop) fields or may also be surrounded by woodland, rangelands, and/or pasture/grassland areas. These plant communities may be natural or managed plan t habitats for the control of soil and wind erosion and serve as wildlife ha bitats (USDA-AP HIS, 2010a).

Weed control programs are important aspects of cotton cultivation. In th is context, weeds are those plants which, when growing in the cotton field, compete with the cotton for space, water, nutrients, and sunlight, and may thus include native species (University of California IPM Online, 2009c). Weeds also interfere with harvesting equipment and can stain lint (USDAAPHIS, 2010a).BAYER TWINLINKTM COTTON

23 Cotton is susceptible to weeds because it is easily outgrown during its early season growth and total crop failure can occur if weeds are not properly controlled (Alabama Cooperative Extension System, 1996). Weed control typically involves an integrated approach that includes herbicides, crop rotation, weed surve illance, and weed monitoring (University of California IPM Online, 2009b). Crop rotation is an important component of a successful weed management program in cotton cultiva tion (University of California IPM Online, 2009b). If a crop is cultivated year after year in the same fields, with the same cultivation practices followed, there is a very good chance that one or more weed species will increase in these fields (University of California IPM Online, 2009b). It is important that cotton cultivation practices employ a variety of cultura l practices, as well as chemical controls (University of California IPM Online, 2009b).

The types of weeds in and around a cotton fiel d depend on the immediate area in which the cotton is planted. Those weed species also vary depending on the geographic region in which the cotton is planted (USDA-APHIS, 2010a). For example, California has over 50 common weeds found in cotton (University of California IPM Online, 2009b). Common weeds in cotton include annual and perennial grasses (monocots), broad-leaf weeds (dicots), and sedges ( Cyperus spp. ). To assist growers in ma naging weeds, individual states, typically through their state agri cultural extension service, will list the prevalent weeds of a crop and what is the most effective means for their control. For example, common weeds in California cotton and recommended he rbicidal control can be found at University of California IPM Online (2009b).

Herbicide-tolerant cotton provides several weed ma nagement advantages to the growers. Broad spectrum post-emergent herbicides, such as gl ufosinate ammonium, provi de control of weeds early in the cultivation cycle, minimizing co mpetition in the fields, and providing optimal conditions for cotton growth (University of Ca lifornia IPM Online, 2009b). Applications of over-the-top post-emergent broad spectrum herbicid es to an herbicide-tolerant crop allow the grower to decrease the overall use of herbicides before cultivation, reduce use of soil-applied herbicides, and streamline field cultivation activ ities for weed control (Marra and Martin, 2007; O'Sullivan and Sikkema, 2004). Production cost s decrease because growers apply fewer herbicides and field cultivation requirements ar e reduced (University of California IPM Online, 2009b).

Herbicide-tolerant cotton has the potential to imp act the cultivation of other crops in the same fields in later years. Cotton is periodically s een as a volunteer in soybeans, either when cotton seeds have successfully overwintered or in fiel ds where a failed cotton crop is replanted to soybeans (Stewart et al., 2003). Cotton volunte ers are limited by the geography in which they are initially planted; cotton does not survive as a perennial where freezing temperatures are reached in the winter (Bayer, 2010). These cott on volunteers typically do not reduce crop yield, but can act as reservoirs for in sect pests of cotton (Stewart et al., 2003). Successful control of cotton volunteers, including herb icide-resistant varie ties, is accomplished through using various combinations of herbicides (Mille r et al., 2004; Stewart et al., 2003).BAYER TWINLINKTM COTTON

24 2.4.3 Biological Diversity

Biodiversity refers to all plants, animals, a nd microorganisms interacting in an ecosystem (Wilson, 1988). Among other benefits, biodiversity provides valuable genetic resources for crop improvement (Harlan, 1975) and also provides other functions beyond food, fiber, fuel, and income. These include pollination, genetic introg ression, biological control, nutrient recycling, competition against natural enemies, soil structure, soil and water conservation, disease suppression, control of local microclimate, control of local hydrological processes, and detoxification of noxious chemicals (Altieri, 2000). The loss of biodiversity results in a need for costly external inputs in order to provid e these functions to th e crop (Altieri, 2000).

The degree of biodiversity in an agroecosystem depends on four primary characteristics: 1) diversity of vegetation within and around the agroecosystem; 2) permanence of various crops within the system; 3) intensity of management; a nd 4) extent of isolation of the agroecosystem from natural vegetation (Altieri, 1999). The re introduction of woodlots, fencerows, hedgerows, wetlands, etc. is one way to reintroduce biodive rsity into large scale monocultures. Some enhancement strategies include intercropping (the planting of tw o or more crops simultaneously to occupy the same field), agroforestry, crop ro tations, cover crops, no-til lage, composting, green manuring (growing a crop specifically for the purpose of incorporating it into the soil in order to provide nutrients and organic ma tter), addition of organic matter (compost, green manure, animal manure, etc.), and hedgerows a nd windbreaks (Altieri, 1999). To some de gree these practices are being utilized by cotton growers to increase biodiversity (Cotton In corporated, 2010e).

The use of genetically modified cotton affect s biological diversity by: 1) providing the opportunity to use ecological management pract ices that enhance habitat creation and 2) decreasing the use of pesticides which in tu rn, limits the impact on the floral and faunal communities that can potentially inhabit the fiel ds and immediate surroundings. The use of notill, cover crops, crop rotation, intercroppi ng, and good ditch/border/hedgerow management contributes to biodiversity in and around cotton fields (Pal mer and Bromley, 2010; Sharpe, 2010).

Habitat preservation and biodiversity, as well as cotton production, have be nefited from modern cotton technology. It is now possible to grow 50% more cotton on the same land required 40 years ago (Cotton Incorporated, 2010e). Incr eased productivity on the same amount of land allows growers to preserve habitat while maintaining food and fi ber security (Cotton Incorporated, 2010e). Various methods for prom oting animal and plant biodiversity in and around cotton fields have already been discu ssed above. Habitat preservation and plant biodiversity have a large impact on insect popu lations. Research cond ucted by Altieri (1999) and Altieri and Letourneau (Altieri and Letourneau, 1982, 1984) indicates:

1) Maintaining some weeds harbors and suppor ts beneficial arthropods that suppress herbivore insect pests; 2) Polycultures of plants support lower herbivore populations be cause they provide a more stable and continuous availability of food and habitat for benefi cial insects; and 3) Adjacent wild vegetation provides alternate fo od and habitat for natural enemies to pest herbivores.BAYER TWINLINKTM COTTON

25 The use of broad-spectrum insecticides is one of the most severe co nstraints for biological diversity in crops (Catta neo et al., 2006; Sisterson et al., 2007; USDA-APHIS, 2010a). One of the benefits of Bt cotton has been the reduction of broad-spectrum insecticide use during cotton production (Fernandez-Cornejo and Caswell, 2006).

However, there is concern over th e non-target effects of transgenic cotton, especially Bt cotton, on beneficial insects and non-target pests. Research in Arizona has shown that Bt cotton has a minimum effect on both the number and function of foliar-dwelling arthropod natural enemies (Naranjo, 2005a, 2005b). In contrast, broad-spec trum insecticide use caused a 48% reduction in 13 of the 22 taxa that were studied (Naranjo, 2005a). A co mparison of non-transgenic and transgenic cotton revealed no substantial impact on ant and beetle diversity in cotton fields, but broad-spectrum insecticide use considerably re duced this diversity (C attaneo et al., 2006).

Soil microorganisms play a key role in soil stru cture formation, decomposition of organic matter, toxin removal, nutrient cycling, and most biochemical soil pr ocesses (Garbeva et al., 2004). They also suppress soil-borne plant diseases and promote plant growth (Doran et al., 1996). The main factors affecting microbial population size and diversity incl ude plant type (providers of specific carbon and energy sources in to the soil), soil ty pe (texture, structure, organic matter, aggregate stability, pH, and nutrient content), and agricultural management practices (crop rotation, tillage, herbicide and fe rtilizer application, and irrigatio n) (Garbeva et al., 2004). Plant roots, including cotton, release a large variet y of compounds into the soil creating a unique environment for microorganisms in the rhizosphere. Microbial diversity in the rhizosphere is extensive and differs from the microbial community in the bulk soil (Garbeva et al., 2004).

2.4.4 Gene Movement

The possibility of gene movement from the host plant into native or feral populations of Gossypium species or wild or weedy relatives of cott on has been raised as an issue of concern (US-EPA, 2010a). This subsect ion provides a basis for evaluati ng the weediness potential of cotton as well as the potential for outcrossing and gene escape in cotton. The potential for outcrossing or gene escape is define d as the ability of the gene to es cape to wild cotton relatives.

G. hirsutum is a perennial plant that is cultivated as an annual in the U.S. (USDA-ERS, 2009). Cultivation as an annual means th at the entire cotton plant is harvested at the end of each growing year, and new seed is planted in following years. Because of the structure of the cotton boll, cotton seed dispersal in th e field is rarely successful (Bay er, 2010). Pollen dispersal is the most likely potential source of gene movement (Bayer, 2010).

The cotton flower is perfect, co ntaining both male and female pa rts, and can self- or crosspollinate (Meyer et al., 2007). However, the flowers of most cotton species, including G. hirsutum , are generally considered to be self-pollinating (Bay er, 2010; Meyer et al., 2007).

Cotton pollen is heavy and somewhat sticky and does not lend itself to wind dispersal and subsequent pollination (Bayer, 2010). Cotton ca n, however, be pollinated by insects. Bees — wild bees, honeybees ( Apis mellifera ), and bumblebees ( Bombus spp .) — are the primary insect pollinators (Pleasants and Wendel, 2010). Bees colle ct mainly the nectar from the cotton plants, and rarely the pollen. In addition, physical is olation from plants a ttractive to the beesBAYER TWINLINKTM COTTON

26 significantly reduces the potential for pollen move ment, as cotton flowers have a nectar high in glucose and low in sucrose, which probably make s it less attractive to bees. McGregor (1976) traced the movement of pollen from a cott on field surrounded by a large number of honeybee colonies using fluorescent partic les on source cotton plants. McGre gor illustrated a rapid decline in the transference of cotton pollen over distance , and found that at 150 to 200 feet away from the source plant, only 1.6% of th e bees showed the presence of the fluorescent particles. By comparison, the isolation distances for Foundation, Re gistered, and Certified seeds in 7 CFR Part 201 are 1320, 1320, and 660 feet, resp ectively (USDA-AMS, 2010c).

Despite these characteristics of cotton pollen which limit wind di spersal and bee pollination, it is common practice to maintain isolation distances to prevent pollen movement from other cotton sources and to plant border or barrier rows to in tercept pollen to maintain and protect cotton seed characteristics for seed certification programs (W ozniak, 2002). Field monitoring for off-types, other crops, weeds, disease etc. is carried out by both growers and state crop improvement associations (AOSCA, 2010a). Seed handling standards are established by AOSCA to reduce the likelihood of seed source mixing during plan ting, harvesting, transporting, storage, cleaning, and ginning (Bradford, 2006).

A majority of field-based research with cotton sh ows an outcrossing rate of 10% or less within 1 meter of the pollen source (USDA-APHIS, 2010a). In upland cotton, outcrossing studies suggest that pollen carryover decreases very rapidly as the distance to the closest marker pollen row increases, and that very little pollen is tr ansferred beyond 12 meters (Bayer, 2010; McGregor, 1976; Vaissiere and Vinson, 1994).

Reports of outcrossing of a brom oxynil-tolerant cotton are desc ribed in the USDA petition for CalGene’s GlyTol cotton 06-332-01p and in a st udy of pollen mediated gene flow by Van Deynze et al. (2005). Seed samples were collect ed in the border rows of Calgene's winter nursery sites in Catamarca, Argentina, and P ongola, Republic of South Africa, as well as in Stoneville, MS, USA. Sampling distance was one to 20 meters away from the bromoxyniltolerant cotton. The frequency of outcrossing wa s determined by the crop and the pollinator. It is interesting to note th at although the rate is higher for Ar gentina and South Africa (most likely due to the behavioral differences between Eu ropean and African honeybees), the pattern of decline with distance was the same. Van Deynze et al. (2005) measured pollen-mediated gene flow (PGF) in four dire ctions over two years from commercial seed fields of bromoxynil-tolerant (BXN) and Roundup Ready ® (RR) cotton in the California co tton growing region at various distances from non-transgenic cotton fields. Thei r results confirmed, as we ll as refined, those of Kareiva et al. (1994), as larger di stances were studied. In spite of variations due to the respective cardinal positions of the fields, the same declin e with distance was observed. Van Deynze et al. further reported that a 55-meter non-planted area was a sufficiently broad distance to maintain varietal purity (Van Deynze et al., 2005). This finding is consistent with the distance limitations identified by McGregor.

There are few wild relatives of cultivated cotton where cotton is cultivated. Two species of cotton are cultivated in the U.S.: upland cotton ( G

. hirsutum ) and Pima cotton ( G. barbadense ) (USDA-ERS, 2009). There are two wild rela tives of interest to U.S. growers: G. thurberi and G. tomentosum, found in the mountains of southern Arizona and in Hawaii, respectively. Outcrossing of genes from TwinLinkTM Cotton to G. thurberi is unlikely to occur because G.BAYER TWINLINKTM COTTON

27 thurberi contains only the D genome whereas G. hirsutum contains both the A and D genome. The difference in chromosome number s precludes sexual compatibility of G. hirsutum with G. thurberi (OECD, 2008).

G. tomentosum is native to the Hawaiian Islands, occurring primarily in arid, rock, or clay coastal plains (Pleasants and Wendel, 2010; USDA-APHIS, 2010a). G. tomentosum is not known to be weedy or to have invasive characteris tics, and is considered a rare plant in Hawaii (Pleasants and Wendel, 2010; Un iversity of Hawaii at Manoa, 2001). In laboratory and greenhouse breeding programs with hand pollination, G. tomentosum and G. hirsutum are sexually compatible and form viable progeny (Pleasants and Wendel, 2010). However, DNA marker analyses have not found evidence of genes from G hirsutum occurring in native populations of G. tomentosum (DeJoode and Wendel, 1992). It is possible that the lack of evidence of movement of G. hirsutum genes into G. tomentosum is the result of lack of opportunity because cotton has not been grown comm ercially in Hawaii for at least the last 45 years (USDA-APHIS, 2010a).

Historic literature suggests that there are barriers to cross pollination between G. tomentosum and either G. hirsutum or G. barbadense based on different flowering patterns and pollinators (Pleasants and Wendel, 2010). Field and laborato ry studies demonstrated that the historic literature is incorrect, and that these three species share comm on pollinators, and further that differences in flower structure and flowering habits do not serve as barriers to cross pollination (Pleasants and Wendel, 2010).

EPA recognizes a possibility of gene transfer from Bt cotton to feral cotton relatives in Hawaii, Florida, Puerto Rico, and the U.S. Virgin Isla nds (US-EPA, 2008a). Wher e feral populations of cotton species similar to cultivated cotton exist, EP A has prohibited the sale or distribution of Bt cotton in those areas, on the premise that th ese containment measures would prevent the movement of the registered Bt endotoxin from Bt cotton to wild or feral relatives (US-EPA, 2008a).

Feral populations of G. hirsutum have been listed as threatened and endangered by the State of Florida (Coile and Garland, 2003). However, in Florida, wild G. hirsutum is not present in the northwestern panhandle where co tton cultivation occurs (Co ile and Garland, 2003; USDANASS, 2007; Wunderlin and Hansen, 2010).

Horizontal gene transfer and c onsequent expression of DNA from a plant species to bacteria is unlikely to occur (Keese, 2008). Ma ny bacteria (or parts thereof) th at are closely associated with plants have been sequenced, including Agrobacterium and Rhizobium (Kaneko et al., 2000; Kaneko et al., 2002; Wood et al., 2001). There is no evidence that these organisms contain genes derived from plants. In cases where the review of sequence data implied that horizontal gene transfer occurred, these events were inferred to occur on an evolutionary time scale in the order of millions of years (Brown, 2003; Koonin et al., 2001).

2.5 PUBLIC HEALTH

Public health concerns surrounding GE cotton, such as TwinLinkTM C otton , focus primarily on human and animal consumption. Alt hough cotton is grown primarily for textileBAYER TWINLINKTM COTTON

28 production, valuable food oil is produced from cotton seed. The remaining seed residue (cottonseed meal and hulls) is used as live stock feed and as a hi gh-protein flour (OECD, 2009). Cotton seed meal animal feed provides prot ein, fat, and energy (National Cottonseed Products Association, 2002).

Non-GE cotton varieties, both those develope d for conventional use and for use in organic production systems, are not routinely required to be evaluated by any regulatory agency in the U.S. for human food safety prior to release in the market. Under the FFDCA, it is the responsibility of food manufacturers to ensure that the products they market are safe and properly labeled. Food derived from TwinLinkTM Cotton must be in compliance with all applicable legal and regulatory requirements. GE organisms for food and feed may undergo a voluntary consultation process with the FDA pri or to release onto the market. Although a voluntary process, thus far, all applicants who have wished to commercialize a GE variety that will be included in the food supply have completed a consultation with the FDA. In a consultation, a developer who intends to co mmercialize a bioengineer ed food meets with the agency to identify and discuss relevant sa fety, nutritional, or other regulatory issues regarding the bioengineered food and then submits to FDA a summary of its scientific and regulatory assessment of the food. FDA eval uates the submission and responds to the developer by letter (US-FDA, 2010).

Bayer has provided the FDA with information on th e identity, function, and characterization of the genes for TwinLinkTM Cotton, including expres sion of the gene products. The FDA is currently reviewing the information submitted by th e applicant. In the absence of a completed FDA determination, APHIS takes into considera tion prior FDA reviews of comparable products to make a preliminary assessment of the potential food and safety imp acts. Note that with regard to the expression of glufosin ate ammonium tolerance, the gene construct in the Bayer TwinLinkTM Cotton is the same as that approve d by the FDA in June 2003 for the LibertyLink ® Cotton product (US-FDA, 2003). In that approva l, the FDA noted that the transformational event in LibertyLink ® Cotton was not materially different in composition, safety, or any other relevant parameter than other cotton grown, ma rketed and consumed at that time (US-FDA, 2003). This previous FDA review will be used by APHIS to analyze the food and safety impacts associated with the incorporation and expre ssion of glufosinate ammonium tolerance in TwinLinkTM Cotton.

The FDA’s oversight of the food and safety im pacts associated with the incorporation and expression of pesticidal substances , in this case, the Cry proteins associated with Bt, are more limited. The EPA is the primary authority for th e review of plant-inco rporated protectants.

EPA Section 3 FIFRA registration applications have been submitted by Bayer to the EPA for Cry1Ab cotton, Cry2Ae cotton and TwinLinkTM Cotton. EPA is currently reviewing information submitted by the applicant on the efficacy and potential environmental concerns associated with the use of this product. APHIS has been advise d that the product will n eed three separate new active ingredient registrations under Section 3 of FIFRA:

 One for Cry1Ab from event 3T04-40.

 One for Cry2Ae from event GHB119.

 One for the stacked event combining both Cry proteins.BAYER TWINLINKTM COTTON

29 In the absence of a completed EPA determination, APHIS takes into consideration prior EPA reviews of comparable products to make a prelimin ary assessment of the pot ential impacts. The EPA, in 2008, published a Biopesticides Re gistration Action Document (BRAD) for Bacillus thuringiensis ( Bt ) incorporated protectant s (US-EPA, 2008a). Based on a comprehensive human health effects assessment, the Bt BRAD found no unreasonable adverse effe cts associated with exposure to the Bt products. The Bt protein is a registered micr obial pesticide wh ich is approved for use on food crops. The EPA concluded in th e BRAD that the Bt produc ts incorporated as plant protectants would be expected to behave as expected of a dietary protein (US-EPA, 2008a). A tolerance exemption is in place for Cry1Ab protein in all crops (40 CFR §174.511; US-EPA, 2010d), as well as for the PA T protein (40 CFR §174.522; US-EPA, 2010g). A temporary exemption from the requirement of a toleranc e has been issued for Cry2Ae (40 CFR §174.530; US-EPA, 2010e). This previous EPA review will be used by APHIS to analyze the food and safety impacts associated with the incorpora tion and expression of Cry proteins in TwinLinkTM
Cotton.

Glufosinate ammonium is currently registered un der various trade names for control of weeds in cotton and other crops; the Ignite ® /Liberty ® trade name is registered for use on LibertyLink ® corn, cotton, soy, rice and canola varieties (Bayer , 2010; BCS, 2010a). EPA’s core pesticide risk assessment and regulatory processes ensure that protections are in place for all populations of nontarget species. These assessme nts provide EPA with information needed to develop label use restrictions for the pesticide. Growers are required to use pe sticides, such as glufosinate ammonium, consistent with the application inst ructions provided on the EPA-approved pesticide label (see, e.g., BCS, 2010a). Thes e label restrictions carry the we ight of law and are enforced by EPA and the states (Federal Insecticide, Fungicide, and Rodent icide Act 7 USC 136j (a)(2)(G) Unlawful Acts). Therefore, it is ex pected that glufosinate ammonium use on the TwinLinkTM Cotton product would be cons istent with the EPA-approved label.

Glufosinate ammonium acts in pl ants by inhibiting the enzyme glutamine synthase causing a toxic buildup of ammonia within the treated pl ant (Bayer, 2010). Glufosinate ammonium is a nonselective herbicide for both non-crop and crop us es. It is highly biodegradable, has no residual activity, and has very low toxicity for hum ans. Glufosinate ammonium degrades rapidly in microbially active soils and also readily binds to soil particles, and its short-lived metabolites have not been found to accumulate in the enviro nment. Soil microorganisms, bees, earthworms, birds, and mammals are unaffected by glufosin ate ammonium (Bayer, 2010). A food tolerance for residues of glufosinate ammonium has been published (40 CFR §180.473; US-EPA, 2010c).

2.6 ANIMAL FEED

As noted in the discussion of Public Health (Subsection 2.5), seed residue remaining after fiber removal for textile production (cottonseed meal and hulls) is marketed as c ottonseed meal, cottonseed hulls, and whole cottonseed and uti lized in the animal f eed industry as sources of protein, fiber and energy (National Cottonseed Products Association, 2002; OECD, 2009). The value of cottonseed as animal feed represents a substantial portion of the grower’s income from cotton (Blasi and Drouillard, 2002).BAYER TWINLINKTM COTTON

30 Cottonseed meal is usually sold as a high protein animal feed supplement, providing as much as 41% protein by weight (National Cottonseed Products Association, 2002) . The meal can be used alone or in combination with ot her plant and animal protein sources (National Cottonseed Products Association, 2002) . Cottonseed hulls are the outer c overing of the cotton seed which are removed prior to oil extraction (National Cottonseed Products Association, 2002) . The hulls provide a source of fi ber to animal feed rations (National Cottonseed Products Association, 2002) . Whole cottonseed is the seed remain ing after the long fibers have been removed (National Cottonseed Products Association, 2002) . The whole cotton seed contains a small quantity of attached linters, and as animal feed provides protein, energy and fiber (National Cottonseed Products Association, 2002)

.
Although cottonseed meal is a widely used anim al feed (Blasi and Drouillard, 2002; Calhoun, 2011), gossypol, one of the chemical constituen ts of cottonseed meal, is potentially toxic (Gadberry, 2011). Ruminants (i.e. cattle) have the ability to detoxify gossypol during the fermentation process in the gut (Gadberry, 2011). Gossypol has be en identified as potentially causing reduced fertility in young developing bulls, resulting in recommendations to limit cottonseed meal in the diet for developing bu lls (Blasi and Drouillar d, 2002; University of Arkansas Cooperative Extension Service, 2011).

Non-GE cotton varieties, both those develope d for conventional use and for use in organic production systems, are not routinely required to be evaluated by any regulatory agency in the U.S. for animal feed safety prior to releas e in the market. Under the FFDCA, it is the responsibility of food and feed manufacturers to ensure that the products they market are safe and properly labeled. Feed derived from TwinLinkTM Cotton must be in compliance with all applicable legal and regulatory requi rements. GE organisms for feed may undergo a voluntary consultation process with the FDA pri or to release onto the market. Although a voluntary process, thus far, all applicants who have wished to commercialize a GE variety that will be included in the food supply have completed a consultation with the FDA. In a consultation, a developer who intends to co mmercialize a bioengineer ed food meets with the agency to identify and discuss relevant sa fety, nutritional, or other regulatory issues regarding the bioengineered food and then submits to FDA a summary of its scientific and regulatory assessment of the food. FDA eval uates the submission and responds to the developer by letter (US-FDA, 2010).

Bayer has provided the FDA with information on th e identity, function, and characterization of the genes for TwinLinkTM Cotton, including expres sion of the gene products. The FDA is currently reviewing the information submitted by th e applicant. In the absence of a completed FDA determination, APHIS takes into considera tion prior FDA reviews of comparable products to make a preliminary assessment of the potential food and safety imp acts. Note that with regard to the expression of glufosin ate ammonium tolerance, the gene construct in the Bayer TwinLinkTM Cotton is the same as that approve d by the FDA in June 2003 for the LibertyLink ® Cotton product (US-FDA, 2003). In that approva l, the FDA noted that the transformational event in LibertyLink ® Cotton was not materially different in composition, safety, or any other relevant parameter than other cotton grown, ma rketed and consumed at that time (US-FDA, 2003). This previous FDA review will be used by APHIS to analyze the food and safety impacts associated with the incorporation and expre ssion of glufosinate ammonium tolerance in TwinLinkTM Cotton.BAYER TWINLINKTM COTTON

31 The FDA’s oversight of the food and safety im pacts associated with the incorporation and expression of pesticidal substances , in this case, the Cry proteins associated with Bt, are more limited. The EPA is the primary authority for th e review of plant-inco rporated protectants.

EPA Section 3 FIFRA registration applications have been submitted by Bayer to the EPA for Cry1Ab cotton, Cry2Ae cotton and TwinLinkTM Cotton. EPA is currently reviewing information submitted by the applicant on the efficacy and potential environmental concerns associated with the use of this product. APHIS has been advise d that the product will n eed three separate new active ingredient registrations under Section 3 of FIFRA:

 One for Cry1Ab from event 3T04-40.

 One for Cry2Ae from event GHB119.

 One for the stacked event combining both Cry proteins.

In the absence of a completed EPA determination, APHIS takes into consideration prior EPA reviews of comparable products to make a prelimin ary assessment of the pot ential impacts. The EPA, in 2008, published a Biopesticides Re gistration Action Document (BRAD) for Bacillus thuringiensis ( Bt ) incorporated protectant s (US-EPA, 2008a). Based on a comprehensive human health effects assessment, the Bt BRAD found no unreasonable adverse effe cts associated with exposure to the Bt products. The Bt protein is a registered micr obial pesticide wh ich is approved for use on food crops. The EPA concluded in th e BRAD that the Bt produc ts incorporated as plant protectants would be expected to behave as expected of a dietary protein (US-EPA, 2008a). A tolerance exemption is in place for Cry1Ab protein in all crops (40 CFR §174.511; US-EPA, 2010d), as well as for the PA T protein (40 CFR §174.522; US-EPA, 2010g). A temporary exemption from the requirement of a toleranc e has been issued for Cry2Ae (40 CFR §174.530; US-EPA, 2010e). This previous EPA review will be used by APHIS to analyze the food and safety impacts associated with the incorpora tion and expression of Cry proteins in TwinLinkTM
Cotton.

Glufosinate ammonium is currently registered un der various trade names for control of weeds in cotton and other crops; the Ignite ® /Liberty ® trade name is registered for use on LibertyLink ® corn, cotton, soy, rice and canola varieties (Bayer , 2010; BCS, 2010a). EPA’s core pesticide risk assessment and regulatory processes ensure that protections are in place for all populations of nontarget species. These assessme nts provide EPA with information needed to develop label use restrictions for the pesticide. Growers are required to use pe sticides, such as glufosinate ammonium, consistent with the application inst ructions provided on the EPA-approved pesticide label (see, e.g., BCS, 2010a). Thes e label restrictions carry the we ight of law and are enforced by EPA and the states (Federal Insecticide, Fungicide, and Rodent icide Act 7 USC 136j (a)(2)(G) Unlawful Acts). Therefore, it is ex pected that glufosinate ammonium use on the TwinLinkTM Cotton product would be cons istent with the EPA-approved label.

Glufosinate ammonium acts in pl ants by inhibiting the enzyme glutamine synthase causing a toxic buildup of ammonia within the treated pl ant (Bayer, 2010). Glufosinate ammonium is a nonselective herbicide for both non-crop and crop us es. It is highly biodegradable, has no residual activity, and has very low toxicity for hum ans. Glufosinate ammonium degrades rapidly in microbially active soils and also readily binds to soil particles, and its short-lived metabolitesBAYER TWINLINKTM COTTON

32 have not been found to accumulate in the enviro nment. Soil microorganisms, bees, earthworms, birds, and mammals are unaffected by glufosin ate ammonium (Bayer, 2010). A food tolerance for residues of glufosinate ammonium has been published (40 CFR §180.473; US-EPA, 2010c).

2.7 SOCIOECONOMIC ISSUES

Cotton has been cultivated for fiber for over 5,000 years (OECD, 2008). Cotton is grown primarily for its lint fibers which are made in to textiles (OECD, 2 009). Cotton seed also produces valuable food oil and the remaining s eed residue (cottonseed meal and hulls) is used as livestock feed and as a high-protei n flour (OECD, 2009). The remainder of the plant is left in the field for deco mposition as fertilizer (OECD, 2009).

In the U.S. for the 2010 production year, cott on was grown on 4.4 million hectares (10.9 million acres (USDA-NASS, 2010b). Cotton is produced in the Cotton Belt, 17 southern states, from Virginia to California (Meyer et al., 2007; USDA-NASS, 2010c). The major cotton producing states are Texas (5.4 million acres), Georgia ( 1.3 million acres), Arkansas (0.540 million acres), Mississippi (0.420 million acres), North Caroli na (0.545 million acres) and Tennessee (0.387 million acres) (USDA-NASS, 2010c). Texas, Arkansas, Georgia, North Carolina, and Mississippi collectively produced over 58% of the cotton in 2010 (USDA-NASS, 2010e). In 2007, total cotton cultivated land in the U.S. wa s about 10,500,000 acres (sligh tly larger than the combined size of the states of Maryland and Connecticut) with a ma rket value of about $5 billion (USDA-NASS, 2007).

Cotton acreage in the U.S. rose slightly duri ng the first half of the 2000s, with a corresponding increase in production (USDA-ERS, 2009). The increase in producti on has been attributed to advances in technology (seed va rieties, fertilizers, pesticides, and machinery) and production practices (reduced tillage, irriga tion, crop rotations, and pest ma nagement systems) (USDA-ERS, 2009). The impact of these changes has been pa rticularly evident, with yields and production reaching new highs (USDA-ERS, 2009). Since 2006, however, U.S. cotton planted area has been considerably lower, as relative prices have favored the planting of a lternative crops, such as corn and soybeans (USDA-ERS, 2009). All regi ons of the Cotton Belt have experienced significant declines compared with the first half of the 2000s (USDA-ERS, 2009). Although U.S. cotton production decreased considerably fo llowing the area reductions of the late 2000s, consistently higher yields help ed limit the effect of these acr eage declines (USDA-ERS, 2009).

According to the Census of Agriculture, U. S. cotton farms numbered 18,605 in 2007, down from 24,805 in 2002 (USDA-ERS, 2009). Although the num ber has fallen, cotton acreage per farm has risen, averaging 564 acres per farm in 2007 compared with 502 acres in 2002. The percentage of large cotton farms (over 1,000 acres) has continued to incr ease, but the share of small cotton farms (under 100 acres) has declined (USDA-ERS, 2009).

The U.S. cotton industry generates about 200,000 j obs among the various sectors from farm to textile mill, and accounts for more than $25 bill ion in products and services annually (USDAERS, 2009).

Total 2010 cotton production is forecast at 18.9 million 480-pound bales, up 55% from 2009 production of 12.2 million bales (U SDA-NASS, 2010e). Yield is expected to average 841BAYER TWINLINKTM COTTON

33 pounds per harvested acre, up 64 pounds from 2009 (USDA-NASS, 2010e). Upland cotton production is forecast at 18.4 million 480-pound bales, and 56% above 2009 (USDA-NASS, 2010e). American Pima production is fo recast at 497,800 bales (USDA-NASS, 2010e).

Trade is particularly important for cotton. About 30% of the world's consumption of cotton fiber crosses international borders before processing, a larger share than for wheat, corn, soybeans, or rice (USDA-ERS, 2009). Through trad e in yarn, fabric, and clothi ng, much of the world's cotton again crosses international borders at least once more before reaching the final consumer (USDA-ERS, 2009).

Consumption of cotton by U.S. textile mills peak ed in 1997; since then, U.S. mill use of cotton has plummeted, dropping about 50% by 2005 and nearly 70% by 2009 (USDA-ERS, 2009). Although the end of the Multifibre Arrangement's (MFA in 2005 was a factor, much of the decline in U.S. textile production occurred befo re then. The MFA was a multilateral agreement signed in 1974, but it began in the 1930s as a mech anism to protect domestic textile markets, establishing strict limits on imports from Ja pan, Hong Kong, Pakistan and India, among many others (Meyer et al., 2007). U nder the MFA, Canada, the U.S., and the EU could set limits on the amount of foreign-made apparel and textiles th ey would allow into their countries (Meyer et al., 2007). The MFA ended after the 1995 Urugua y Round Agreements on trade liberalization (Meyer et al., 2007). The result of this change is increased global competition for import and export of raw cotton, as well as fi nished textiles (Mey er et al., 2007). U.S. consumer demand for cotton products remains strong, but imported cl othing now accounts for most purchases by U.S. consumers (USDA-ERS, 2006). The changes afte r the end of the MFA is reflected in the importance of exports for cotton growers. The USDA ERS reports that U.S. cotton mills consumed 60% of the domestic cotton through the 1990s, but not long af ter the ending of the MFA quotas, 70% of the U.S. cotton was exported (Meyer et al., 2007).

The cotton industry continues to face many of the supply and dema nd concerns confronting other field crops. However, because cotton is used primarily in manufactured products, such as clothing and home furnishings, the industry faces additional challenges associated with the economic well-being of downstream manufacturing industries, as well as the economic wellbeing of the final consumer (USDA-ERS, 2009).

The USDA ERS notes that U.S. cotton productivity ha s increased in recent years as a result of technological advancements, including biotech nology-derived varieties (Meyer et al., 2007). The introduction of cotton providing insect resi stance or herbicide tolerance also has global implications. Herbicide-tolerant cotton is one of the most widely and ra pidly adopted crops in the U.S., followed by insect-resistant cotton (USDA-ERS, 2010a). In 2010, GE varieties of cotton were cultivated on 93% of the acreage in the U.S., with herbicide-tolerant cotton cultivated on 78% of the acreage, and Bt cotton cultivated on 73% of the acreage (USDA-ERS, 2010a).

Outside the U.S., Bt cotton has been widely adopted in India and China, two major cotton producers (Meyer et al., 2007). The cost savings of Bt cotton brought milli ons of hectares back into cotton production in eastern China, and has also helped India’s co tton area rebound by more than 1 million hectares (Meyer et al., 2007). Bt cotton has also been adopted in smaller producing countries like Australi a, Argentina, Mexico, and S outh Africa (Meyer et al., 2007).BAYER TWINLINKTM COTTON

34 

### 3 ALTERNATIVES

This document analyzes the potential environmen tal consequences of a determination of nonregulated status of BCS TwinLinkTM Cotton. To respond favorably to a petition for nonregulated status, APHI S must determine that TwinLinkTM Cotton is unlikely to pose a plant pest risk. Based on its Plant Pest Risk Assessment (USDA-APHIS, 2010b) APHIS has concluded that TwinLinkTM Cotton is unlikely to pose a plant pest risk. Therefore, APHIS must determine that COT67B cotton is no longer s ubject to 7 CFR part 340 or the plant pest provisions of the Plant Protection Act.

Two alternatives will be evaluated in this EA: 1) no action; and 2) determination of nonregulated status of TwinLinkTM Cotton . APHIS has assessed the potential for environmental impacts for each alternative in Section 4 of this EA, Environmental Consequences.

3.1 NO ACTION: CONTINUATION AS A REGULATED ARTICLE

Under the No Action Alterna tive, APHIS would deny the pe tition. TwinLinkTM Cotton and progeny derived from TwinLinkTM Co tton would continue to be regulated articles under the regulations at 7 CFR Part 340. Permits issued or notifica tions acknowledged by APHIS would still be required for introducti ons of TwinLinkTM Cotton and measur es to ensure physical and reproductive confinement would continue to be implemented. APHIS might choose this alternative if there were insufficient evidence to de monstrate the lack of plant pest risk from the unconfined cultivation of TwinLinkTM Cotton.

This alternative is not the Preferred Alternat ive because APHIS has concluded through a Plant Pest Risk Assessment (USDA-APHIS, 2010b) th at TwinLinkTM Cotton is unlikely to pose a plant pest risk. Choosing this alternative would not satisfy the purpose and need of making a determination of plant pest risk status and resp onding to the petition for nonregulated status.

3.2 PREFERRED ALTERNATIVE: DETERMINATION THAT TWINLINKTM COTTON IS NO LONGER A REGULATED ARTICLE

Under this alternative, TwinLinkTM Cotton and progeny derived from them would no longer be regulated articles under the regulations at 7 CFR Part 340. TwinLinkTM Cotton is unlikely to pose a plant pest risk (USDA-APHIS, 2010b). Pe rmits issued or notific ations acknowledged by APHIS would no longer be required for intr oductions of TwinLinkTM Cotton and progeny derived from this event. This alternative best meets the agency’s purpose and need to respond appropriately to a petition fo r nonregulated status based on th e requirements in 7 CFR part 340 and the agency’s authority under the plant pest pr ovisions of the Plant Protection Act. Because the agency has concluded that TwinLinkTM Cott on is unlikely to pose a plant pest risk, a determination of nonregulated stat us of TwinLinkTM Cotton is a res ponse that is consistent with the plant pest provisions of the PPA, the regu lations codified in 7 CFR part 340, and the biotechnology regulatory policies in the Coordinated Framework.

Under this alternative, grow ers may have future access to TwinLinkTM Cotton and progeny derived from this event if the developer de cides to commercialize TwinLinkTM Cotton.BAYER TWINLINKTM COTTON

35 3.3 ALTERNATIVES CONSIDERED BU T REJECTED FROM FURTHER CONSIDERATION

APHIS assembled a list of alternatives that might be considered for TwinLinkTM Cotton. The agency evaluated these alternatives, in light of the agency's authority under the plant pest provisions of the Plant Protecti on Act, and the regulations at 7 CFR part 340, with respect to environmental safety, efficacy, a nd practicality to identify which alternatives would be further considered for TwinLinkTM Cotton. Based on th is evaluation, APHIS rejected several alternatives. These alternatives are discussed briefly below al ong with the specific reasons for rejecting each.

3.3.1 Prohibit Any TwinLinkTM Cotton from Being Released

In response to public comments that stated a preference that no GE organisms enter the marketplace, APHIS considered prohibiting th e release of TwinLinkTM Cotton, including denying any permits associated with the field testi ng. APHIS determined that this alternative is not appropriate given that AP HIS has concluded that TwinLinkTM Cotton is unlikely to pose a plant pest risk (USDA-APHIS, 2010b).

In enacting the Plant Protecti on Act, Congress found that

[D]ecisions affecting imports, exports, and interstate movement of products regulated under [the Plant Protection Act] shall be based on sound science... § 402(4).

On March 11, 2011, in a Memorandum for the H eads of Executive Depart ments and Agencies, the White House Emerging Technologies Interage ncy Policy Coordination Committee developed broad principles, consistent with Executiv e Order 13563, to guide the development and implementation of policies for oversight of emer ging technologies (such as genetic engineering) at the agency level. In accord ance with this memorandum, agencies should adhere to Executive Order 13563 and, consistent with that Executive Order, the followi ng principle, among others, to the extent permitted by law, when regulating emerging technologies:

“[D]ecisions should be based on the best re asonably obtainable scientific, technical, economic, and other information, within the boundaries of the authorities and mandates of each agency”

Based on our Plant Pest Risk Assessment (USDA-APHIS, 2010b) and the scientific data evaluated therein, APHIS has concluded that TwinLinkTM Cotton is unlikely to pose a plant pest risk . Accordingly, there is no basis in science for prohibiting the release of TwinLinkTM
Cotton.

3.3.2 Approve the peti tion in part

The regulations at 7 CFR 340.6(d)(3)(i) state that APHIS may "approve the petition in whole or in part." For example, a determination of nonregulated status in part may be appropriate if there is a plant pest risk asso ciated with some, but not all lines described in a petition. Because APHIS has concluded that TwinLinkTM Cotton is unlikely to pose a plant pest risk,BAYER TWINLINKTM COTTON

36 there is no regulatory basis unde r the plant pest provisions of the Plant Protection Act for considering approval of the petition only in part.

3.3.3 Isolation distance between TwinLinkTM Cotton a nd non-GE cotton and geographical restrictions

In response to public concerns of gene movement between GE and non-GE plants, APHIS considered requiring an isolation distance separating TwinLinkTM Cotton from non-GE cotton production. However, b ecause APHIS has concluded that TwinLinkTM Cotton is unlikely to pose a plant pest risk (USDA-APHIS, 2010b), an alternative based on requiring isolation distances would be inconsistent with the statutor y authority under the plant pest provisions of the Plant Protection Act and regul ations in 7 CFR part 340.

APHIS also considered geographically re stricting the producti on of TwinLinkTM Cotton based on the location of production of non-GE cotton in orga nic production systems in response to public concerns regarding possible gene movement be tween GE and non-GE plants. However, as presented in APHIS’ plant pest risk assessm ent for TwinLinkTM Cotton, there are no geographic differences associated with any identifia ble plant pest risks for TwinLinkTM Cotton(USDAAPHIS, 2010b). This alternative was rejected and not analyzed in detail because APHIS has concluded that TwinLinkTM Cotton does not pose a plant pest risk, and will not exhibit a greater plant pest risk in any geographica lly restricted area. Therefore, such an alternative would not be consistent with APHIS’ statutor y authority under the plant pest pr ovisions of the Plant Protection Act and regulations in Part 340 and the biotechnology regulat ory policies embodied in the Coordinated Framework.

Based on the foregoing, the impositi on of isolation distances or ge ographic restrictions would not meet APHIS’ purpose and need to respond appropriately to a petition for nonregulated status based on the requirements in 7 CFR part 340 and the agency’s au thority under the plant pest provisions of the Plant Protection Act. Nevertheless, APHIS is not expecting significant effects. However, individuals might choose on their own to geographi cally isolate their nonGE cotton productions systems from TwinLinkTM Cotton or to use isolation distances and other management practices to minimize gene movement between cotton fields.

3.3.4 Requirement of Testing For TwinLinkTM Cotton

During the comment periods for other petitions for nonregulated status, some commenters requested USDA to require and provide testi ng to identify GE produc ts in non-GE production systems. APHIS notes there are no nationa lly-established regula tions involving testing, criteria, or limits of GE mate rial in non-GE systems. Such a requirement would be extremely difficult to implement and maintain. Additionally, because TwinLinkTM Cotton does not pose a plant pest risk (USDA-APHIS, 2010b), the imposition of any type of testing requirements is inconsistent with the plant pest provisions of th e Plant Protection Act, th e regulations at 7 CFR part 340 and the biotechnology regulatory polic ies embodied in the Coordinated Framework. Therefore, imposing such a requirement fo r TwinLinkTM Cotton would not meet APHIS’ purpose and need to respond appropriately to the petition in accordance with its regulatory authorities.BAYER TWINLINKTM COTTON

37 3.4 COMPARISON OF ALTERNATIVES

Table 3-1 presents a summary of the potential impact s associated with selection of either of the alternatives evaluated in this EA. The impact assessment is presented in Section 4 of this EA.

Table 3-1: Summary of Potential Impact s and Consequences of Alternatives

Attribute/Measure Alternative A: No Action Alternative B: Determination of Nonregulated Status Meets Purpose and Need and Objectives No Yes Unlikely to pose a plant pest risk Satisfied through use of regulated field trials Satisfied – risk assessment (USDA-APHIS, 2010b) Management Practices

Acreage and Areas of Cotton Pr oduction Unchanged Unchanged Cropping Practices Unchanged Unchanged

Seed Production Un changed Unchanged Organic Farming Unchanged Unchanged Specialty Cotton Productio n Unchanged Unchanged Physical Environment Water Resources Unchanged Unchanged Soil Unchanged Unchanged Air Quality Unchanged Unchanged Climate Change Unchanged Unchanged

Animal and Plant Communication Animals Unchanged Unchanged

Plants Unchanged Minimal Biological Diversity Unchanged Minimal Gene Movement Unchanged Unchanged

Public Health Human Health Unchanged Unchanged Worker Safety Unchanged Unchanged

Animal Feed Unchanged Unchanged

Socioeconomic Issues Domestic Economic Environment Unchanged Unchanged

Trade Economic Environmen t Unchanged Unchanged Social Environment Unchanged Unchanged

Other Cumulative Effects Unchanged Unchanged
Threatened and Endangered Species Unchanged Unchanged Other U.S Regulatory Approvals Unchanged for existing nonregulated GE organisms FDA consultation pending, EPA tolerance exemptions and conditional pesticide registrations be ing reviewed

Compliance with Other Laws CWW, CAA, EOs Fully co mpliant Fully compliantBAYER TWINLINKTM COTTON

38 Notes:

1. Unchanged – the current conditions will not change as a result of the selection of this alternative. 2. Minimal – the current conditions may change slightly as a result of the selection of this alternative, but the changes, if any, are not deemed significant.BAYER TWINLINKTM COTTON

39 

### 4 ENVIRONMENTAL CONSEQUENCES

This analysis of potential environmental consequences addresses the potential impact to the human environment from the alternatives analy zed in this EA, namely taking no action and a determination by the agency that TwinLinkTM Cotton does not pose a plant pest risk and therefore should no longer be regulated under 7 CFR 340. Potential environmental impacts from the No Action Alternative and the Preferred Alternative for TwinLinkTM Cotton are described in detail throughout this section. A cumulative effects analysis is also included for each environmental issue and includes a discussion of the cumulative impacts that are associated with the Preferred Alternative, when combined with other recent past, present and reasonably foreseeable future actions within the affected envi ronment. Certain aspects of this product and its cultivation would be no different between the alternatives; those instances are described below.

4.1 SCOPE OF THE ENVI RONMENTAL ANALYSIS

For the discussion of environmental consequen ces, this section addresses the following principal areas of potential environmental concern:

 Agricultural Production of Cotton (Subsection 4.2);

 Physical Environment (Subsection 4.3);

 Animal and Plant Commun ities (Subsection 4.4);

 Public Health (Subsection 4.5);

 Animal Feed (Subsection 4.6);

 Socioeconomic Issues (Subsection 4.7);

 Other Cumulative Effects (Subsection 4.8);

 Threatened and Endangered Sp ecies (Subsection 4.9); and

 Consideration of Executive Orders, Standard s, and Treaties Relating to Environmental Impacts (Subsection 4.10).

Although the Preferred Alternative w ould allow for new plantings of TwinLinkTM Cotton to occur anywhere in the U.S., APHIS is limiting the environmental analysis to those areas that currently support cotton production, as identified by the National Agricultural Statistics Service (NASS) 2007 Census of Agriculture. The NASS found that cotton is currently produced in the following 17 states: Alabama, Arizona, Arka nsas, California, Florida, Georgia, Kansas, Louisiana, Mississippi, Missouri, New Mexico, North Carolina, Oklahom a, South Carolina, Tennessee, Texas, and Virgini a (USDA-NASS, 2010c).

The environmental consequences of the No Acti on and Preferred Alternatives are analyzed under the assumption that farmers who produ ce conventional cotton, TwinLinkTM Cotton

, or cotton using organic methods are using reasonable, commonly accepted best management practices specific to their agricultural co tton production. However, APHIS r ecognizes that not all farmers follow these best management practices for cott on. Thus, the analyses of the environmental effects will also include the assumption that some farmers do not follow these best management practices.BAYER TWINLINKTM COTTON

40 4.2 AGRICULTURAL PRODUCTION OF COTTON

One of APHIS’s missions is to improve American agricultural productivity. Best management practices are commonly accepted, pract ical ways to grow cotton, re gardless of whether the cotton farmer is using conventional prac tices with non-GE or GE varietie s, or organic practices. These management practices consider crop-specific pl anting dates, seeding rates, and harvest times, among others. Over the years, cotton productio n has resulted in we ll-established, widelypracticed management practices that are avai lable through local Cooperative Extension Service offices and their respective websites. The National Information System for the Regional Integrated Pest Management Centers publishes cr op profiles for major crops on a state-by-state basis. These crop profiles provide produc tion guidance for local growers, including recommended practices for specific pest cont rol. Crop profiles for many of the cotton production states can be re viewed at (www.ipmcenters .org/cropprofiles/index.cfm)

.
BCS’ studies demonstrate that agronomic characte ristics and cultivation practices required for TwinLinkTM Cotton are essentially i ndistinguishable from other co tton varieties, including other herbicide-tolerant and insect-resistant va rieties (Bayer, 2010; USDA-APHIS, 2010b). Although TwinLinkTM Cotton might be expected to replace other varieties of cotton currently cultivated, new acreage is not expected to be develope d to accommodate the cu ltivation of TwinLinkTM Cotton (Bayer, 2010). None of the best manage ment practices currently employed for cotton production is expected to change if TwinLinkTM Co tton is no longer subject to the plant pest provisions of the Plant Protecti on Act and 7 CFR part 340. Acco rdingly, the potential impacts on agricultural production of TwinLinkTM Cott on resulting from management practices associated with the No Action and Preferred Alternatives are the same.

4.2.1 Acreage and Areas of Cotton Production

GE and non-GE cotton varieties are continually under developmen t. Cotton acre age averaged over 14.5 million acres during the first half of the 2000s (USDA-APHIS, 2010a). Since 2006, however, U.S. cotton planted acreage has been cons iderably lower as market prices have favored the planting of alternativ e crops, such as corn and soybeans (USDA-APHIS, 2010a). Most of the U.S. cotton acreage is planted in GE cotton. Of the 11.3 million acres planted in cotton in 2010,93% (10.5 million acres) were GE cotton. Of this, 73% of the GE cotton acreage was GE insectresistant (Bt) cotton and 78% was herbicide-to lerant (USDA-ERS, 2010a, 2010c). Most cotton is planted on farms that have been in cotton production for at least five years (USDA-NASS, 2007).

No Action: Acreage and Areas of Cotton Production

Based on current acreage trends, conventional cott on production practices with GE varieties will likely continue to dominate or perhaps increase in acreage under the No Action Alternative. Cotton is currently produced co mmercially in 17 states (USDA- NASS, 2010c) and under the No Action Alternative, this range of pr oduction is not expected to change.

Preferred Alternative: Acreage and Areas of Cotton Production

BCS’ studies demonstrate that agronomic characte ristics and cultivation practices required for TwinLinkTM Cotton are essentially i ndistinguishable from other co tton varieties, including otherBAYER TWINLINKTM COTTON

41 herbicide-tolerant and insect-resistant va rieties (Bayer, 2010; US DA-APHIS, 2010b). BCS posits that although TwinLinkTM Cotton might be exp ected to replace othe r varieties of cotton currently cultivated, new acreage is not expected to be developed to accommodate the cultivation of TwinLinkTM Cotton (Bayer, 2010). The intr oduced lepidopteran resi stance trait and the herbicide tolerance trait do not c onfer any competitive advantage in terms of weediness or to extend the range of cultivation outside of existin g cultivation areas (Bayer, 2010). The Preferred Alternative, a determination of nonregulated stat us of TwinLinkTM Cotton, is thus not expected to increase cotton production, either by its availa bility alone or accompanie d by other factors, or cause an increase in overall GE cotton acreage. Impacts would be similar to the No Action Alternative.

Cumulative Effects: Acreage and Areas of Cotton Production

Cumulative effects of a determination of nonregulated status of TwinLinkTM Cotton are unlikely. Neither the No Action Alternative nor a determination of nonregulated status of TwinLinkTM
Cotton is expected to directly cause an increase in agricultural acreage devoted to cotton production or those cotton acres devot ed to GE cotton cultivatio n. The availability of TwinLinkTM Cotton will not change cultivation ar eas for cotton production in the U.S. and there are no anticipated change s to the availability of GE and non-GE cotton varieties on the market under either alternative.

4.2.2 Cropping Practices: Crop Rotation and Pesticide Use

Crop rotation in cotton is conduc ted to optimize soil nutrition and fertility, reduce pathogen loads, and control certain cotto n pests (Gazaway et al., 2007); (University of California IPM Online, 2008a). Crop rotation practices have b een described previously in Section 2. Crop rotation practices are not expect ed to change with the planti ng and production of TwinLinkTM
Cotton.

In the U.S., the cotton industry has consistently relied heavily on insectic ide use strategies to manage arthropod pests (Gianessi an d Carpenter, 1999). Insecticides were applied to 66% of the cotton acreage in 2007 (USDA-NASS, 2008). Of thir ty listed insecticides, Acephate was the most utilized insecticide, with 26% of the plan ted acreage being treated at an average rate of 0.900 pounds per acre per crop year (USDA-NASS, 2008). Dicrotophos was the second most commonly utilized insecticide, applied to 21% of th e acreage at an aver age rate of 0.565 pounds per year (USDA-NASS, 2008). Other insecticid es applied in the 2007 growing year included Acetamiprid (6% of acreage), Cyfluthrin (8 % of acreage), Cypermethrin (7% of acreage), Imadacloprid (6% of acreage), Lambda-cyhalothri n (5% of acreage), Malathion (5% of acreage), and Thiamethoxam (11% of acreage).

Resistance to commonly used insecticides (pyr ethroids, organophosphates, and carbamates) has led to a need for new insecticide chemistries or ot her novel approaches such as the use of sterile insects, pathogens, and transgenic cotton (Gia nessi and Carpenter, 1999). Cotton varieties expressing the Bt endotoxin have emerged as a viable product replacing many of these commonly used pesticides (Bayer, 2010).

One of the documented benefits of Bt cotton has been the reduction in the use of broad-spectrumBAYER TWINLINKTM COTTON

42 insecticides during cotto n production (Benbrook, 2009; Fernand ez-Cornejo and Caswell, 2006). Each acre planted in Bt cotton is presumed to displace the average pounds of insecticide previously sprayed on an acre of conventiona l cotton for lepidopteran control (Benbrook, 2009). In a comprehensive evaluation of 13 year s of USDA NASS data (1996 through 2008) on pesticide usage (including both herbicides and insecticides), Benbroo k found that pesticide application rates for managing lepi dopteran pests in cotton change d in direct proportion to the adoption of Bt Cotton. Table 4-1 provides an excerpt from Benbrook’s summary of insecticide

use.

Table 4-1: Summary of Insecticide Use for Lepidopteran Control in Cotton Compared with Adoption of Bt Cotton*

Cotton Crop Type % Acreage by Year
1996 2001 2005 2008 Planted with Bt Cotton 12% 41% 60% 73%
Treated with Insecticides 48% 31% 30% 25% *Benbrook’s study reports all 13 years of data. This tabl e presents an extracted summary to show use trends.

Source: Benbrook (2009) Supplemental Table 12. Insec ticides Applied to Control the Budworm/Bollworm Complex of Insects on Conventional Varieties of Cotton, 1996 – 2008.

Benbrook’s assessment presents strong evidence that there has been a dramatic reduction in the use of insecticides to control lepidopteran pests in cotton directly associat ed with the adoption of
Bt cotton.
Cotton cultivation also requires substantial use of herbicides for weed control during the growing season and for chemical defoliation prior to harvest (Bayer, 2010). In 2007, 35 different herbicides were ap plied to 97% of the cotton acreage (USDA-NASS, 2008). Glyphosate isopropylamine salt was the most commonly applied herbicide, applied to 85% of the planted acres at a rate of 1.900 pounds per acre per crop year (USDA- NASS, 2008). The next two most commonly applied herbicides on a per acre basi s were Trifluralin and Diuron, at 29 and 26% with average applicat ion rates of 0.921 and 0.499 pounds per acre per crop year, respectively (USDA-NASS, 2008). Other herbicides applied to cotton included 2,4-D (6% of the acreage), Carfentrazone-ethyl (10% of the acreage), Flumi oxazin (6% of the acreage ), Pendimethalin (17% of the acreage), Prometryn (7% of the acreag e), Pyraflufen-ethyl (8% of the acreage), pyrithiobac-sodium (10% of the acreage), and S-metolachlor (6% of th e acreage) (USDA-NASS, 2008).

The consequences of adoption of herbicide tolerance show a contrasting trend. Herbicidetolerant cotton was first grown commercially in the U.S. in 1997 and by 2008, herbicide-tolerant cotton was planted on 68% of total cotton plan tings (Brookes and Barfoot, 2010). Stacked varieties, providing both herbicide tolerance and insect resistance, comprised nearly 93% of thetotal plantings in 2008 (Benbrook, 2009). Benbrook has reported that the adoption of herbicidetolerant crops has resulted in an increase in th e volume of herbicides applied to crops (Benbrook, 2009). Benbrook notes that herbicide use declin ed between 1996 and 2001, apparently in direct response to the adoption of he rbicide-tolerant cotton (Benbr ook, 2009). However, since that time, herbicide use has increased. Reported increases in herbicide use reflect an increase in use of conventional herbicides, as well as an in crease in glyphosate applications (Benbrook, 2009).BAYER TWINLINKTM COTTON

43 Benbrook concludes that this reported increase in herbicide use is a direct consequence of the development of glyphosate-resistant weeds in cott on. As of 2008, nine weeds were identified as glyphosate-resistant, with three of these weed s, horseweed (marestail), Johnsongrass, and pigweed (Palmer amaranth) specifically identif ied as difficult to c ontrol weeds in cotton.

No Action: Cropping Practices: Crop Rotation and Pesticide Use

Under the No Action Alternative, cotton crop rotation practices and pesticide use is expected to remain as practiced today by the farming community. Growers will continue to have access to existing GE lepidopteran-resistant and herbicide- tolerant cotton varieties that are no longer subject to the plant pest provisi ons of the Plant Protection Act and 7 CFR part 340, as well as conventional cotton varieties. It is likely that gr owers would continue to experience the continued emergence of glyphosate-tolerant weeds, requiring modifications of crop management practices to address thes e weeds. These changes may involve all of the techniques identified by Benbrook (2009), including use of other herbicides for w eed control.

Preferred Alternative: Cropping Practi ces: Crop Rotation and Pesticide Use

BCS’ studies demonstrate TwinLinkTM Cotton is e ssentially indistinguishable from other cotton varieties used in terms of ag ronomic characteristics and cultiv ation practices (Bayer, 2010; USDA-APHIS, 2010b). Therefore, cotton crop rota tion practices are expected to be unchanged under the Preferred Alternative.

If TwinLinkTM Cotton is adopted, a continue d reduction in the us e of budworm/bollworm insecticides applications and the number of acre-treatments pe r year as repor ted in Benbrook’s trend analysis is expected to occur. A determination of nonr egulated status of TwinLinkTM Cotton will provide growers with another alternativ e Bt cotton variety to cultivate. Herbicide use patterns have the potential to change as well. The introduc tion of TwinLinkTM Cotton provides a stacked variety, expressi ng Bt-based lepidopteran resist ance combined with tolerance to glufosinate ammonium, an a lternative herbicide. TwinLi nkTM Cotton provides growers with an alternative to those cotton varieties resistant to glyphosate, thus expand ing options in the field for weed control.

In those fields where glyphosate-resistant weeds have emerged, glufosinate ammonium tolerance provides the grower with an option to transition away from glyphosate herbicides to a different post-emergent herbicide. The transition to gluf osinate ammonium could reduce applications of those other herbicides needed to manage glyphosate-resistant weeds.

Cumulative Effects: Cropping Practices : Crop Rotation and Pesticide Use

A determination of nonregulated status of TwinLinkTM Cotton is not expected to result in changes in the current cotton cropping practices. APHIS has determined that there are no past, present, or reasonably foreseea ble actions that would aggregat e with effects of the proposed action to affect changes in crop rotation. Studies demonstrate TwinLinkTM Cotton is essentially indistinguishable from other co tton varieties used in terms of agronomic characteristics and cultivation practices (Bayer, 2010).BAYER TWINLINKTM COTTON

44 It is anticipated that the trend of reduced br oad-spectrum insecticide use by cotton growers will continue due to the adoption of Bt cotton and other cultural practices. There is the potential that the introduction of glufosinate ammonium toleran ce may result in a reduction in total herbicide use as growers adopt different he rbicide treatment strategies i nvolving glufosinate ammonium as a post-emergent crop treatment, thereby reducing th e use of some of the other herbicides which have been required in res ponse to the emergence of gl yphosate-resistant weeds.

4.2.3 Seed Production

As discussed in Subsection 2.2.3, cotton seed production is managed through the AOSCA standard procedures to preclude gene flow between species and varieties (Wozniak, 2002). Common practices include: 1) maintaining isola tion distances to preven t pollen movement from other cotton sources; 2) planting bor der or barrier rows to intercept pollen; 3) employing natural barriers to pollen; and 4) field monitoring fo r off-types, other crops, weeds, and disease.

No Action: Seed Production

Under the No Action Alternative, current cotton seed production practices are not expected to change.

Preferred Alternative: Seed Production

The production of cotton seed for TwinLinkTM Cott on will be conducted consistent with these standard practices; consequently , no changes to seed production pr actices are required. Based on the data provided by BCS for TwinLinkTM Cotton (B ayer, 2010), as well as previous experience with other Bt cotton varieties th at have been widely adopted by growers since their introduction in 1996 (USDA-ERS, 2010a), APHI S has concluded that the ava ilability of TwinLinkTM Cotton would not alter the agronomic practices, locations, and seed production and quality characteristics of conventional and GE seed production (USDA-APHIS, 2010b). A determination of nonregulated status of TwinLinkTM Cotton will not require a change to seed production practices.

Cumulative Impacts: Seed Production

Based on current acreage trends, GE cotton varieties will likely continue to dominate. To the extent that growers see value in the traits offered by TwinLi nkTM Cotton, this new variety may replace existing cotton varieties, both conventional as well as GE. The availability of TwinLinkTM Cotton is not anticipated to change cultivation areas for cotton production in the U.S. Because changes in the agronom ic practices and locations for cotton seed production using TwinLinkTM Cotton are not expected, no cu mulative effects have been identified for seed production.

4.2.4 Organic Farming

Organic production plans prepared pursuant to the National Organic Program (NOP) include practical methods to protect organically-produced crops from accidental contamination with GE materials. Typically, organic growers use more than one method to prevent unwanted material from entering their fields including: isolati on of the farm, physical barriers or buffer zonesBAYER TWINLINKTM COTTON

45 between organic production and non-organic production, as well as formal communications between neighboring farms (Bai er, 2008; Guerena and Sulliva n, 2003; NCAT, 2003). These practices follow the same system utilized for the cultivation of certified seed under the AOSCA procedures.

APHIS recognizes that producers of non-GE cott on, particularly producers who sell their products to markets sensitive to genetically engineered traits (e.g., organic or some export markets), can be reasonably assumed to be usin g practices on their farm to protect their crop from unwanted substances and thus maintain their price premium. APHIS will assume that growers of organic cotton are already using, or have the ability to use, these common practices as APHIS’s baseline for the analysis of the alternatives.

No Action: Organic Farming

Current availability of seed fo r conventional (both GE and non-GE ) cotton varieties, and those cotton varieties that are devel oped for organic production, is expected to remain the same under the No Action Alternative. Commercial produc tion of conventional and organic cotton is not expected to change and will likely remain the same under the No Action Alternative. Planting and production of GE, non-GE, and organic cotton will continue to fluctuate with market demands, as it has over the last 10 years, and thes e markets are likely to continue to fluctuate under the No Action Alternative (USDA-ERS, 2010c, 2010e).

Preferred Alternative: Organic Farming

Transgenic cotton lines including those that are resist ant to lepidopteran insects and herbicidetolerant are already in widesp read use by farmers. TwinLin kTM Cotton should not present any new and different issues and impacts for or ganic and other specialty cotton producers and

consumers.

Organic producers employ a variety of measures to manage identity and preserve the integrity of organic production systems (NCAT, 2003)

. The trend in the cultivation of GE cotton, nonGE, and organic cotton va rieties, and the corresponding produc tion systems to mai ntain varietal integrity, are likely to remain the same as the No Action Alternative.

According to the petition, agronomic trials conducted between 2005 and 2008 in a variety of locations in the U.S. demonstrat ed that TwinLinkTM Cotton is not significantly different in plant growth, yield, and reproductive capacity from its nontransgenic coun terpart (USDA-APHIS, 2010b). No differences were observed in pollen diameter, weight, and vi ability. Therefore, TwinLinkTM Cotton is expected to present a simila r risk of cross-pollination as existing cotton cultivars. The practices curr ently employed to preserve a nd maintain purity of organic production systems would not be required to change to accommodate the production of TwinLinkTM Cotton.

Historically, organic cotton produc tion represents a small per centage (approximately, 0.1%) of total U.S. cotton acreage (USDA-ERS, 2010e). It will likely remain small regardless of whether new varieties of GE or non-GE cotton varieties, including TwinLinkTM Cotton, become available for commerc ial cotton production.BAYER TWINLINKTM COTTON

46 Accordingly, a determination of nonregulated st atus of TwinLinkTM Cotton is not expected to have a significant impact on organic cotton production.

Cumulative Effects: Organic Farming

A determination of nonregulated status of TwinLinkTM Cotton is not expected to change the market demands for GE cotton or cotton produced using organic methods. A determination of nonregulated status of TwinLinkTM Cotton could add another GE cotton variety to the conventional cotton market. Based upon recent trend information, adding GE varieties to the market is not related to the ability of organic production system s to maintain their market share. Although 12 GE cotton events or lines were no longer subject to the plant pest provisions of the Plant Protection Act and 7 CFR part 340, between 2000 and 2008, the total acreage associated with the organic production of co tton remained at slightly above 15,000 acres (USDA-ERS, 2010e).

4.2.5 Specialty Cotton Production

Specialty crop growers employ practices and st andards for seed production, cultivation, and product handling and processing to ensure that their products are not pollin ated by or commingled with conventional or genetically engineered crops (Bradford, 2006). These management practices include maintaining isolat ion distances to prevent pollen movement from other cotton sources, planting border or barrier rows to intercept poll en, and employing natural barriers to pollen (Bradford, 2006; NCAT, 2003; Wozniak, 2002). These management practices allow the grower to meet standards for the produc tion of specialty crop seed, maintain genetic purity, and protect the genetic dive rsity of cotton (Bradford, 2006).

No Action: Specialty Systems

Current availability of seed for specialty cotton varieties are expected to remain the same under the No Action Alternative.

Preferred Alternative: Specialty Systems

As noted in the discussion of Seed Producti on, no changes in the production of seed or cultivation of cotton are required to accommodate TwinLinkTM Cotton.

It is unlikely that spec ialty system farmers, or other farm ers who choose to plant nontransgenic varieties or sell nontransgenic seed, will be subs tantially impacted by the expected commercial use of TwinLinkTM Cotton. Transg enic cotton lines including t hose that are resistant to lepidopteran insects and express herbicide toleran ce are already in widespread use by farmers. TwinLinkTM Cotton should not present any new and different issues and impacts for specialty cotton producers and consumers.

According to the petition, agronomic trials conducted between 2005 and 2008 in a variety of locations in the U.S. demonstrated that TwinLi nkTM Cotton is not significantly different in plant growth, yield, and reproductive capacity from its nontransgenic counte rpart (USDA-APHIS, 2010b). No differences were observed in pollen diameter, weight, and vi ability. Therefore, TwinLinkTM Cotton is expected to present a simila r risk of cross-pollin ation as existing cottonBAYER TWINLINKTM COTTON

47 cultivars including other GE cotton varieties. Th e practices currently employed to preserve and maintain purity of specialty cotton production systems would not be required to change to accommodate the production of TwinLinkTM Cotton. A determination of nonregulated status of TwinLinkTM Cotton under the Preferred Alternative woul d not change the availability and genetic purity of seed for specialty cotton varieties. Conventional management practices and procedures, as described previously for cotton seed production, proper seed handling, protection of wild relatives of cotton, and organic cotton farming, are in place to maintain the genetic diversity of cotton. Cotton grower s have utilized these methods effectively to meet the standards for the production of specialty crop seed. Impacts would be similar to the No Action Alternative.

Cumulative Effects: Specialty Systems

A determination of nonregulated status of TwinLinkTM Cotton is not expected to change the market demands for GE cotton or cott on produced using specialty systems. A determination of nonregulated status of TwinLinkTM Cotton will add another GE cotton variety to the cotton market. Between 2000 and 2009, 12 GE cotton events or lines were no longer subject to the plant pest provisions of the Plant Protection Act and 7 CFR part 340. Based on demonstrated agronomic characteristics and cul tivation practices, and because the market share of specialty cotton vari eties is unlikely to change by th e introduction of TwinLinkTM Cotton, APHIS has determined that there are no past, pr esent, or reasonably foreseeable changes that would impact specialty cotton producers and consumers.

4.3 PHYSICAL ENVIRONMENT

4.3.1 Water Resources

Although cotton has been developed as a droug ht-tolerant crop (Cotton Incorporated, 2010b), irrigation (used on approximately 35% of the U.S. cotton acreage) produces much higher cotton yields (Cotton Incorporated, 2010a). Despite high production levels in irrigated areas, the lack of affordable water has been noted as one fact or in the reduction of ac res of cotton grown in California, Arizona, and New Mexico in the past decade (Cotton Incorporated, 2010b). In these areas, cotton has been displaced by higher valu e crops and land uses (Cotton Incorporated, 2010b).

Water quality is also preserved in modern co tton productions systems. The increase in conservation tillage practices has resulted in a reduction of runoff from agricultural lands, decreasing non-point source pollution of suspended se diment, fertilizer, and pesticides. Intensive monitoring of surface water and gr oundwater proximate to agricultu ral fields has demonstrated the benefits of no-till cotton in protec ting both ground and surface water resources (University of Tennessee Agricultural Extension Service, 2010) . Better nutrient management and precision technologies are ensuring inputs are used by th e crop and are not ente ring ground or surface waters (Cotton Incorporated, 2010a).

No Action: Water Use

Under the No Action Alternative, current land acreage and agronomic practices, including irrigation, tillage, and nutrient management as sociated with cotton production would not beBAYER TWINLINKTM COTTON

48 expected to change. Therefor e, no expected change s to water use associ ated with cotton production is expected for this alternative.

Preferred Alternative: Water Use

In 2010, GE cotton occupied 93% of the cott on acreage (USDA-ERS, 2 010c). Conventional and GE cotton production occurs on land that is dedicated to crop production and most cotton is planted in fields that have been in crop production for years (USDA-APHIS, 2010a). The introduction of GE cotton has resulted in th e adoption of conserva tion practices (Cotton Incorporated, 2010a). These conservation prac tices, including reduced tillage and precision agriculture play a significant role in wate r conservation and maintaining water quality by minimizing soil erosion (USDA-NRCS, 2006b).

TwinLinkTM Cotton would not change cultivation practices fo r cotton production , nor would it increase the total acres and rang e of U.S. cotton production areas. Therefore, a determination of nonregulated status of TwinLinkTM Cotton will not change the current use of irrigation practices in commercial cotton production. Because the TwinLinkTM Cotton is expected to simply replace GE and non-GE cotton varieties already in use, the consequences of the Preferred Action Alternative on water use are the same as the No Action Alternative.

Cumulative Effect: Water Use

No cumulative effects on water use have been identified for a determin ation of nonregulated status of TwinLinkTM Cotton. A determination of nonregulated status of TwinLinkTM Cotton would not change the water use and irrigation practices used in commercial cotton production

.

4.3.2 Soil

This section discusses the potential consequences of the No Action and the Preferred Alternatives on soil. The USDA Natural Resources Con servation Service (NRCS) has identified significant reductions in the loss of so il from croplands in the U.S., finding that total soil loss on highly erodible croplands and non- highly erodible cropland decreased by 39.2% from 1982 to 2003 (USDA-NRCS, 2006b). Over the last 10 years, cultivation of herbicidetolerant cotton has contributed to this reduction in soil loss through a shift towards conservation tillage and the use of cover crops (Cotton Incorporated, 2010c).

Increases in total acres dedicated to conservation tillage have been attributed to an increased use of genetically modified crops which eliminates the need for mechanical weed control (Alabama Cooperative Extension System, 1996; McClella nd et al., 2000; Towery and Werblow, 2010; USDA-NRCS, 2010).

Additional soil quality benefits from adoption of GE co tton may also be realized by reducing the risks associated with environmental spills or misapp lications of chemical insecticides to the soil, and reductions in the frequency with wh ich insecticides must be applied.BAYER TWINLINKTM COTTON

49 No Action: Soil

Land acreage and agronomic practi ces associated with traditional and existing non-regulated GE cotton production would not be expected to change in response to the No Action Alternative.

Continued adoption of GE cotton varieties no l onger subject to the plant pest provisions of the Plant Protection Act and 7 CF R part 340 would be expected. In 2010, GE cotton occupied 93% of the cotton acreage (USDA- ERS, 2010c). Conventional and GE cotton production occurs on land that is dedicated to crop production and most cotton is planted in fields that have been in crop produc tion for years (USDA-APHIS, 2010a).

Current agronomic practices asso ciated with cotton production including tillage, cultivation, applications of pesticides and fertilizer, and the use of agricultural equipment would not be expected to change if TwinLinkTM Co tton is still a regulated article. To the extent that currently available GE cotton varieties can result in the grower’s adoption of co nservation tillage, soils would be positively affected by that adoption.

Preferred Alternative: Soil

No changes to agronomic practices typically applied in the management of conventional cotton, including other commercially ava ilable GE cotton varieties, are re quired for TwinLinkTM Cotton.

BCS’ field trial and laboratory analyses demonstrated that the agronomic performance of TwinLinkTM Cotton was functionally identical to its non-transgenic counterpart, the Coker 315 cultivar (Bayer, 2010). No increases in fertiliz ers and pesticides were required, nor were any changes in cultivation, planting, harvesting, and vol unteer control required (Bayer, 2010). It is expected that similar agronomic practices that are currently used for commercially available GE cotton will also be used by growers of TwinLinkTM Cotton. If TwinLinkTM Cotton is adopted and replaces non-GE cotton varieties, soils currently under non-GE cotton production may benefit from the use of TwinLinkTM Cotton due to a reduction in applications and the number of acre-treatments per year required for both insecticide and herbicide applications. Soils would also benefit from the reduction in soil disturban ce because of fewer pa sses by heavy farm equipment.
It is expected that only glufosinate ammonium would be applied as a post-emergent herbicide on TwinLinkTM Cotton. Glufosinate ammonium ha s been shown to rapidly dissipate from most agricultural ecosystems across a wide range of soil and climatic co nditions, with a median soil half-life (the time it takes for half of the glufos inate ammonium to dissipate in the soil) of 7.5 days (PPDB, 2010). When used in accordan ce with the label, glufosinate ammonium accumulation in soil has not been shown to be significant.

The bar gene which has been introdu ced into TwinLinkTM Cotton was originally isolated from Streptomyces hygroscopius , a common soil bacteria (Bayer, 2010 ). The PAT enzyme produced by the TwinLinkTM Cotton is naturally occurring in the soil (CFIA, 2004). The cultivation of TwinLinkTM Cotton and the attendan t production of the PAT enzyme will therefore not cause an impact to the physicochemical characteristics of the soil.BAYER TWINLINKTM COTTON

50 As explained above, if TwinLinkTM Cotton is adopted and cultivated, there may be positive impacts to soil when compared with traditional cotton cultivation practices.

Cumulative Effects: Soil

APHIS has not identified any cumulative effect s of the use of TwinLinkTM Cotton to soils. Bayer has compared phenotypic, agronomic, and cultivation characteris tics between TwinLinkTM Cotton and control cottons (Bayer, 2010). TwinLinkTM Cotton required the same soil, fertilizer, water and pest management practices (except fo r lepidopteran pest control) as non-GE cotton (Bayer, 2010). Consequently, the phenotypic, ag ronomic, and ecological data presented by BCS (Bayer, 2010) support the conclusion by APHIS that TwinLinkTM Cotton will not result in any significant impact to the soil that is not already found in conventional cotton production practices.

Based on these findings, and because the amount of cotton grown in the U.S. is unlikely to change by the introduction of TwinLinkTM Cotton, APHIS has determined that there are no cumulative impacts to soil.

4.3.3 Air Quality

Traditional agricultural practices have the potential to cause negative impacts to air quality. Agricultural emission sources in clude smoke from agricultural bur ning, tillage, heavy equipment emissions, pesticide drift from spraying, and nitr ous oxide emissions from the use of nitrogen fertilizer (Aneja et al., 2009; Hoef t et al., 2000; USDA-NRCS, 2006c).

The adoption of GE cotton has reduced air em issions from several of these sources. Conservation practices, including conservation tillage associated with GE cotton production requires fewer tractor passes across a field, thus decreasing dust ge neration and tractor emissions; surface residues and untilled organic matter physically hold the soil in place thus decreasing wind erosion of soils and pesticide dr ift in wind-eroded soils (Baker et al., 2005; Cotton Incorporated, 2010d; USDA-NRCS, 2006c). Reduced tillage also in creases sequestration rates of carbon emissions in soils (Causarano et al., 2006). This section discusses the potential consequences of the No Action and the Pr eferred Alternatives on air quality.

No Action: Air Quality

Under the No Action Alternative, c urrent impacts to air quality associated with land acreage and cultivation practices associated with cott on production would not be affected.

Adoption of GE cotton varieties would be expect ed to continue. To the extent that the adoption and cultivation of GE cotton varieties allows the grower to implement conservation practices, air quality improvement associated w ith these practices would be expected to follow. Air quality would continue to be af fected by agronomic prac tices associated with conventional methods of cotton pro duction such as tillage, cultivation, pesticide and fertilizer applications, and the use of agricultural equipment.BAYER TWINLINKTM COTTON

51 Preferred Alternative: Air Quality

TwinLinkTM Cotton production does not change land acreage or any cultivation practices for conventional, transgenic, or non-transgenic cotton production. There are no expected increases in cultivation, planting, pesticide use, fertilizer use, harvesting, or volunteer control compared to currently available GE and non-GE co tton cultivars. It is expected that similar agronomic practices that are currently used for commercially available Bt cotton or herbicidetolerant cotton will also be used by growers of TwinLinkTM Cotton.

If TwinLinkTM Cotton is adopted and replaces non-GE co tton varieties, air quality issues associated with pesticide application and us e in non-GE cotton production may benefit from the use of TwinLinkTM Cotton due to a reduction in the applic ation of lepidopteran targeted insecticides and similar reductions in herbic ide applications required as growers take advantage of the post-emergent herbicide gluf osinate ammonium. The collective impact is a reduction in the number of acre-treatments per year requiring the use of heavy farm equipment, with a positive impact on air quality associated with the decrease in equipmentrelated emissions, as well as a decr ease in dusts and pesticide drifts.

Based on this information, APHIS conclude s that the production of TwinLinkTM Cotton is not expected to adversely affect air qualit y and, in fact, may provide some benefit. However, considering the wide adoption of similar GE cotton lines, overall impacts are likely to be similar to the No Action Alternative.

Cumulative Effects: Air Quality

Based on the findings described abov e, APHIS has determined that there are no past, present, or reasonably foreseeable actions th at would aggregate with effect s of the proposed action that would have a negative impact on air quality. The consequences of the Preferred Action Alternative on commercial cotton production and the resulting air quality are similar to those expected for the No Action Alternative.

4.3.4 Climate Change

Agriculture, including land-use ch anges for farming, is responsible for an estimated 6% of all human-induced GHG emissions in the U.S. Moreover, nitrous oxide (N 2 O) emissions from agricultural soil management (primarily fertilizer use) represent 68% of all U.S. N 2 O emissions (US-EPA, 2010b). A more comprehensive discus sion of the contribution of agricultural practices to GHGs is provided in Subsection 2.3.4, Affected Environment – Climate Change. Agriculture-related GHG emissions include carbon dioxide (CO 2 ), N 2 O, and methane (CH 4 )

, produced through the combustion of fo ssil fuels to run farm equipment; the use of fertilizers; or the decomposition of agricultural waste products including crop residues, animal wastes, and enteric emissions from livestock. This section discusses the potential c onsequences of the No Action and the Preferred Alternatives on climate change.

No Action: Climate Change

Under the No Action Alternative, envi ronmental releases of TwinLinkTM Cotton would be under APHIS regulation. Due to the limited size of thes e field trials, there would be no measurableBAYER TWINLINKTM COTTON

52 effect on climate change from these confin ed environmental releases. Land acreage and cultivation practices associat ed with cotton production would not be affected. Current agronomic practices associated with convent ional cotton production and current GE cotton varieties which contribute to GHG emissions, in cluding tillage, cultivati on, irrigation, pesticide application, fertilizer ap plications and use of agriculture e quipment, would not be expected to change if TwinLinkTM Cotton remains a regulated ar ticle. To the extent that the adoption and cultivation of GE cotton varieties allows the grower to implement conservation practices, GHG emissions would be expected to be reduced commensurate with the ai r quality improvements anticipated from adoption of conservation tillage practices.

Preferred Alternative: Climate Change

A determination of nonregulated status of TwinLinkTM Cotton would not change the cultivation or agronomic practices , or agricultural land acr eage associated with growing cotton, and thus is expected to have the sa me effect on climate change as the No Action Alternative.

Cumulative Effects: Climate Change

APHIS has not identified any cumulative effects for this issue. The use of TwinLinkTM Cotton in commercial cotton production is not expected to cause any cu mulative effect on climate change because APHIS does not anticipate any changes in cotton production practices or an expansion of cotton acreage as a result of a determination of nonregulated status of TwinLinkTM Cotton . T he consequences of the Preferre d Action Alternative on commercial cotton production and acreage are the same as for the No Action Alternative.

4.4 ANIMAL AND PLANT COMMUNITIES

4.4.1 Animals

Cotton production systems in agriculture are host to many animal species. A variety of insects feed on cotton plants or prey upon other insect s inhabiting cotton fields . In addition, mammal and bird species use cotton fiel ds and adjacent vegetation for food and habitat throughout the year. This section discusses the potential consequences of the No Action and the Preferred Alternatives on animal populations associated with cotton production. The cumulative effects analysis for this issue is found discussed below under “Cumulative Effects: Animals, Plants, and Biodiversity” in Subsection 4.4.3.

No Action: Animals

Under the No Action Alternativ e, conventional and GE transg enic cotton production will continue while TwinLinkTM Cotton remains a regulat ed article. Potential impacts of GE and non-GE cotton production practices on non- target species would be unchanged.

Preferred Alternative: Animals

BCS data indicates that the agronomic practices used to produce TwinLinkTM Cotton will be the same as those used to p roduce other c onventionally grown GE and non-GE cotton. TwinLinkTMBAYER TWINLINKTM COTTON

53 Cotton production will not change land acreag e or any cultivation practices for cotton production.

Since the mid-1990s, transgenic cotton lines have been commerc ialized without substantiated reports of significant deleterious impacts on non- target organisms (Mendelsohn et al., 2003; OECD, 2007; US-EPA, 2008a; USDA-APHIS, 2010a). Evidence submitted by BCS from laboratory and field studies as well as other peer reviewed studies demonstr ate a lack of toxicity of the TwinLinkTM Cotton on non-target repres entative species (Bayer, 2010).

BCS has presented laboratory test data with indica tor test species to determine potential toxicity of Cry enzyme toxin at doses higher than woul d be anticipated under field conditions (Rose et al., 2007). BCS’ selection of repr esentative indicator test speci es was based upon their potential for exposure to the Cry1Ab and Cry2Ae proteins . BCS submitted non-target data for two above-ground arthropods (Ladybug, Coleomegilla maculate; and Lacewing , Chrysoperla

carnea ); a soil-dwelling insect (springtail, Folsomia candida ); a pollinator (honeybee, Apis mellifera ); a mammal (house mouse, Mus musculus ); and an aquatic inve rtebrate (water flea, Daphnia magna ) (Bayer, 2010). The data submitted in th e petition indicate that no significant adverse effects were observed at the maximum te st dose for either of the two Cry proteins for any of the tested species. Other research has also shown no direct adverse effects on insectivorous insects in field and laboratory st udies with transgenic plants expressing Cry proteins (Marvier et al., 2007; Naranjo, 2009b; Romeis et al ., 2006; Shelton et al., 2009).

The non-target risks associated with exposure of aquatic organisms is likely very low because cotton pollen is large, sticky, and is not tran sported long distances by the wind (USDA-APHIS, 2010a). Seed and plant debris are not expected to be readily transported via overland runoff or wind to aquatic habitats (USDA-APHIS, 2010a). Moreover, because the specific intestinal and midgut receptors for the Bt endotox in are not present in vertebrate s, these insecticidal proteins are not expected to adversely affect these or ganisms (Hofmann et al., 1988a; Hofmann et al., 1988b; Shimada et al., 2006b; US-EPA, 2008a; Van Ri e et al., 1989; Van Rie et al., 1990).

An additional advantage of the use of transg enic cotton producing the Bt proteins is the corresponding reduction in use of broad spectrum insecticides 1 and the consequent reduction of potential impact on diversity of non-target insects (Cattaneo et al., 2006; Dively, 2005; Marvier et al., 2007; Naranjo, 2005a; Romeis et al., 2006; Torres and Ruberson, 2005; Torres et al., 2006; Whitehouse et al., 2005). Several authors note that insect-resistant crops had a higher diversity and abundance of non-target animals than conve ntionally grown crops (Hardee et al., 2001; Moar et al., 2003; Sanvido et al., 20 06; Sisterson et al., 2007).

1 Broad spectrum insecticides are chemical insecticides whic h kill insects that are causing injury to plants and also kill other insects that are not causing injury to the plant. Insects that are inadvertently killed by the application of insecticide are called “non-target” ins ects. Because the Cry proteins are speci fic for a narrow range of lepidopteran insects, use of Cry proteins to control plant pests is recognized as being beneficial to the survival of non-target insects (US-EPA, 2008a).BAYER TWINLINKTM COTTON

54 BCS has filed applications with the FDA and the EPA for the TwinLinkTM Cotton variety. The FDA and EPA are currently reviewing the information submitted by the applicant. Both agencies have previously reviewed and approv ed comparable products, including several crop varieties expressing the Bt-based Cry proteins as plant-incorpor ated protectants, as well as the bar gene, resulting in the expr ession of the PAT enzyme provi ding tolerance to glufosinate ammonium. In the absence of a completed FDA and EPA determinations, APHIS takes into consideration prior agency revi ews of comparable products to conduct its assessment of the potential impacts.

EPA has issued a tolerance exemption for Cry1Ab protein in all crops (40 CFR §174.511; USEPA, 2010d), as well as for the PAT protein (40 CFR §174.522; US-EPA, 2010g). A temporary exemption from the requirement of a toleranc e has been issued for Cry2Ae (40 CFR §174.530; US-EPA, 2010e).

The PAT enzyme bar gene construct was no longer subject to the plant pest provisions of the Plant Protection Act and 7 CFR part 340 in 2003 in the BCS LibertyLink ® Cotton (USDAAPHIS, 2003). In 2003, the FDA completed its food safety consultation on LibertyLink ® Cotton and granted tolerance exemptions for both the pl ant-incorporated protectant, as well as the genetic material necessary for its production (U S-FDA, 2003). The EPA granted exemptions from the requirement of a tolerance for residues of glufosinate ammonium, as well as the plantpesticide PAT in or on food and feed comm odities of cotton (see: 62 FR 17719; 40 CFR §180.473; US-EPA, 2010c).

The conclusions presented by the EPA and the FDA were based upon their independent reviews of human health and environmen tal impacts associated with e xposure to and consumption of plant material incorporating the bar gene and expressing the PAT enzyme. In both cases, these agencies determined that the cottonseed, refine d oil, and cotton seed meal derived from the LibertyLink ® variety was not materially different in composition, safety and nutritional value from other cotton varieties then on the market.

Similar international reviews and approvals have been published for the LibertyLink ® Cotton. The Australia and New Zealand Food Authority (ANZFA) completed a Toxicological Review and Risk Assessment, and a Technical Report was published in 2005 in which ANZFA concluded that LibertyLink ® Cotton was comparable to non- genetically engineered (non-GE) cotton in terms of their safety and nutritiona l adequacy (ANZFA, 2005). In 2004, the Plant Biosafety Office (PBO) of the Canadian Food In spection Agency (CFIA) also concluded that BCS’ LibertyLink ® Cotton was substantially equivalent to currently grown cotton in terms of their potential environmental impact and livestock feed safety, and the novel traits would not have any substantial negative effect on the e nvironment (CFIA, 2004). The CFIA specifically noted that the PAT enzyme responsible for gluf osinate ammonium tolerance has very specific enzymatic activity associated with the acetyla tion of glufosinate ammonium, and is rapidly inactivated in mammalian stomach and intestinal fluids by enzymatic degradation and pH mediated proteolysis (CFIA, 2004).

To the extent that TwinLinkTM Cotton displace s conventional, non-GE cotton varieties, the expression of the stacked Cry proteins providing lepidopteran resistance in conjunction with the tolerance to glufosinate ammonium may have an overall positive impact on animals. AsBAYER TWINLINKTM COTTON

55 previously discussed in the anal ysis of water, soil, in those fields where TwinLinkTM Cotton is cultivated, growers would be expected to re duce their use of pestic ides for control of lepidopteran pests and take advantage of the weed control offered by glufosinate ammonium, thus allowing for a shift to in creased conservation practices.

The use of insecticides other than Bt crops may affect non- target organisms including honey bees, soil invertebrates, or culturable microbial flora (D ively and Rose, 2003; US-EPA, 2010a). The cultivation of insect-resistant crops allows a reduction and replaceme nt of broad-spectrum insecticides, with a correspondi ng positive effect on biodiversit y, including non-target insects (Cantrell, 2006). A notable advantage of GE insecticidal (Bt) crops over conventional insecticides is the high specificity of the Bt t oxins, which minimize the pot ential toxic effects on non-target insects (Bigler et al ., 2008; Sanvido et al., 2006).

The incorporation of glufosinate ammonium tolera nce provides several pot ential advantages over conventional weed management programs, includi ng increased flexibility to manage problem weeds, reduced use of soil-app lied herbicides, reduced total use of herbicides, and more widespread adoption of conservation tillage (O 'Sullivan and Sikkema, 2004). The associated adoption of conservation tillage an d reduced use of soil-applied herb icides have the potential to positively impact animal populations in fields pl anted with TwinLinkTM Cotton (see, e.g., Eggert et al., 2004).

Based on the above information, APHIS concludes that a determination of nonregulated status and cultivation of TwinLinkTM Cotton is expected to have no adverse effects on non-target animals. Overall impacts would be similar to the No Action Alternative.

4.4.2 Plants

The landscape surrounding a cotton field may be bordered by other cott on (or any other crop) fields or may also be surrou nded by woodland, rangel ands, and/or pasture/grassland areas. These plant communities represent natural or managed plant buffers for the control of soil and wind erosion and also serve as ha bitats for a variety of transi ent and non-transient wildlife species. The potential impacts to off-site plan t communities is discussed in Subsection 4.4.3, Biological Diversity.

Weed control programs are important aspects of cotton cultivation. In th is context, weeds are those plants which, when growing in the cotton field, compete with the cotton for space, water, nutrients, and sunlight, and may thus include native species (University of California IPM Online, 2008b). The types of weeds in and around a co tton field will vary depending on the geographic region where the cotton is grown. Common weeds in cotton include grasses, broadleaf weeds, and sedges ( Cyperus spp.). Some of thes e have been discussed in Subsection 4.2.2, Cropping Practices. This section discusses the potential consequences of the No Action and the Preferred Alternatives on plants. The cumulative effects analysis for this issue is discussed below under “Cumulative Effects: Animals, Pl ants, and Biodiversity” in Subsection 4.4.3.

No Action: Plants

Under the No Action Alternative, environmental releases of TwinLinkTM Cotton would be under APHIS regulation. Plant species (i.e., weeds) that typically inhabit GE and non-GEBAYER TWINLINKTM COTTON

56 cotton production systems will continue to be managed through the use of mechanical, cultural, and chemical control methods.

Preferred Alternative: Plants

I n the event of a determination of nonregulated status of TwinLinkTM Cotton , the risks to wild plants and agricultural productivity from weedy cotton popul ations are low; volunteer cotton populations are easily managed an d feral populations occur rarely in the U.S. Cotton Belt (Wozniak, 2002).

Agronomic studies conducted by BCS tested the hypothesis that the weediness potential of TwinLinkTM Cotton is unchanged w ith respect to conv entional cotton (Bayer, 2010). No differences were detected between TwinLi nkTM Cotton and nontransgenic cotton in growth, reproduction, or interactions with pests and diseas es, other than the intended effect of protection from lepidopteran pests. The main natural cont rols on feral populations of cotton are poor seed dispersal, poor seed germination, competition from other plants, and a high requirement for moisture (US-EPA, 2001b). Therefore, the incorpor ation of either the inse ct resistance trait or the herbicide tolerance trait is unlikely to appreciably improve seedling establishment and increase weediness potential. Any TwinLinkTM Cott on volunteers could be r eadily controlled by other readily available herbicides , including glyphosate (Miller et al., 2004; Stewart et al., 2003).

The incorporation of glufosinate ammonium tolera nce provides several pot ential advantages over conventional weed management programs, includi ng increased flexibility to manage problem weeds, reduced use of soil-app lied herbicides, reduced total use of herbicides, and more widespread adoption of conservation tillage (O 'Sullivan and Sikkema, 2004). The associated adoption of conservation tillage and reduced use of herbicides have the potential to positively impact plant populations in fields and fallow ar eas adjacent to those planted with TwinLinkTM Cotton (see, e.g., Eggert et al., 2004).

The effect of cotton production on plant communities is likely to be unchanged by the introduction of TwinLinkTM Cotton. Overall im pacts would be simila r to the No Action Alternative.

4.4.3 Biological Diversity

Biodiversity in an agroecosystem depends on four primary charac teristics: 1) diversity of vegetation within and around the agroecosystem; 2) permanence of various crops within the system; 3) intensity of management, including sele ction and use of insecticides and herbicides; and 4) extent of isolation of the agroecosyst em from natural vegetation (Altieri, 1999). The introduction of woodlots, fencerows, hedgero ws, wetlands, etc. is one way to enhance biodiversity in large scale monocultures. Additional enhancement strategies include intercropping (the planting of two or more crops simultaneously to occupy the same field), agroforestry, crop rotations, cover crops, no-till age, composting, green manuring (growing a crop specifically for the purpose of incorporating it into the soil in order to provide nutrients and organic matter), addition of organic matter (compost, green manure, animal manure, etc.), as well as the introduction of hedgerows and wi ndbreaks (Altieri, 1999). To some degree these practices currently are being utilized by co tton growers to increase biodiversity (CottonBAYER TWINLINKTM COTTON

57 Incorporated, 2010a). The adopti on of GE crops, with the concom itant reduction in insecticide use and enhanced soil conservati on practices, has also contributed to the increase in biodiversity of soil microorganisms, beneficial organisms, and plants (Dively and Rose, 2003; Naranjo, 2009a; Sanvido et al., 2006).

No Action: Biological Diversity

Under the No Action Alternative, TwinLinkTM Cotton would continue to be a regulated article. Growers and other parties who are involved in production, handling, processing, or consumption of cotton would continue to have access to GE le pidopteran-resistant (Bt) and herbicide-tolerant cotton products that are no longer subject to the plan t pest provisions of the Plant Protection Act and 7 CFR part 340 , stacked varieties pres enting both, and conventi onal cotton varieties. The implications of a gronomic practices associated with cotton production, whether traditional or GE varieties, would not change.

Preferred Alternative: Biological Diversity

The use of genetically modified cotton varieties currently availa ble affects biological diversity by: 1) providing the opportunity to use conservation practices th at enhance habitat creation; and 2) decreasing the use of pesticides which in tu rn, limits the impact on the floral and faunal communities that can potentially inhabit the fields and immediate surroundings (Cotton Incorporated, 2010a; Naranjo, 2009a; Sanvido et al., 2006). Incorporation of herbicide tolerance in the crop allows the gr ower to adopt the use of no-till, cove r crops, crop rotati on, intercropping, and good ditch/border/hedgerow management, each of which contributes to biodiversity in and around cotton fields (Palmer and Bromley, 2010; Sharpe, 2010). The use of broad-spectrum insecticides is one of the most severe cons traints for biological di versity in crops (USDAAPHIS, 2010a). One of the benefits of Bt co tton has been the reducti on of broad-spectrum insecticide use during cott on production (Fernandez-Corn ejo and Caswell, 2006).

A comparison of non-transgenic an d transgenic cotton production re vealed no substantial impact on ant and beetle diversity in cotton fields; by contrast, br oad-spectrum insecticide use demonstrated considerably reduced diversity in these populations (Cattaneo et al., 2006). Cotton expressing the Bt Cry pr oteins allows for the reduction in use of these broad spectrum insecticides, and is not expected to present a ri sk to beneficial insect s and non-target pests. Research in Arizona has shown that Bt cott on has a minimum effect on both the number and function of foliar-dwelling arthropod natural enemies; on the other hand, broad-spectrum insecticide use caused a 48% reduc tion in 13 of the 22 taxa that were studied (Naranjo, 2005a, 2005b).

Bt toxin has been found to be present in every ma jor part of Bt cotton plants (leaves, stems, and roots) (Vadakattu and Watson, 2004). Bt cotton roots were also found to release the Bt protein into the soil (Vadakattu and Watson, 2004). The effects of Bt on non-target soil microorganisms in Bt maize and Bt cotton cultivation found th at microbial biodiversity and activity was not different from that of their non-Bt counterparts (EFSA, 2008; Höne mann et al., 2008; Icoz et al., 2008; Shen et al., 2006).BAYER TWINLINKTM COTTON

58 Plant biodiversity associated with cotton cultivation is also an issue of co ncern. Although cotton fields are cultivated as monocultures to optim ize cotton yield, areas surrounding a cotton field may harbor a wide variety of plan ts. Broad spectrum herbicides have the potential to impact offsite plant communities. Glufosinate ammonium provides opportunities to the grower to control weeds in the field with fewer applications of herb icides than in traditional cultivation systems or those systems where glyphosat e resistance has emerged (University of California IPM Online, 2008b) . Accordingly, adoption of a glufosinate am monium protocol could minimize off-site impacts. The application proce sses for glufosinate ammonium a llow the grower to apply fewer herbicides and reduce field cult ivation. Reducing the number of applications of herbicides during the cotton production cycle minimizes instan ces of unintentional drif t of these chemicals to adjacent fields, buffer zones, ditches, and fallow areas.

Although TwinLinkTM Cotton does provide the grower with a new choi ce for a herbicide-tolerant and insect-resistant variety, cultivation of TwinLinkTM Cotton does not otherwise require a change in the rates of fertilizer application, tillage, planting, or harvesting from existing commercial cotton varieties, including other GM cotton varieties providing either insect resistance, herbicide tolerance, or stacked with both (Bayer, 2010) . Therefore, the effect of cotton production on plant commun ities is likely to be unch anged by the introduction of TwinLinkTM Cotton.

A determination of nonregulated status of Tw inLinkTM Cotton will not change the cultivation or agronomic practices, or agricultural land acreag e associated with growing cotton. TwinLinkTM Cotton would be an additional Bt cotton variety providing a different option for herbicide use (glufosinate ammonium) for growers to use and ther efore is expected to have the same effect on biological diversity as th e No Action Alternative.

Therefore, a determination of nonregulated st atus of TwinLinkTM Cotton for the control of lepidopteran pests and tolerance of glufosinate ammonium is highl y unlikely to have any direct toxic effects on non-target organisms and is likely to be neutral or beneficial to animal and plant biodiversity compared to non-transgenic cotto n managed with conventional broad-spectrum insecticides.

Cumulative Effects: Animals, Plants, and Biodiversity

APHIS has determined that ther e are no impacts from past, pres ent, or reasonably foreseeable actions that would aggregate with effects of th e proposed action to create cumulative impacts or reduce the long-term productivity or sustainability of any of the resources associated with the ecosystem in which TwinLinkTM Cotton is planted. Based on scientific ev idence, the diversity and abundance of non-target organisms in lepidopter an-resistant cott on is at least as high as in conventional cotton; and in many st udies, higher biodiversity is asso ciated with tr ansgenic cotton (Cattaneo et al., 2006; Head et al., 2005; Naranjo, 2005a, 2005b; Torres and Ruberson, 2007; Torres et al., 2006; Whitehouse et al., 2005).

Cultivation of TwinLinkTM Cotton, with the attendant expression of the two stacked Cry proteins and the PAT enzyme providing herbicide tolerance, is highly unlikely to have direct toxic effects on non-target organisms and is likely to be neutral or beneficial to biodiversity compared with conventionally managed cotton. Therefore, the like lihood of adverse cumulative effects on non-BAYER TWINLINKTM COTTON

59 target organisms and biodiversity following the introduction of TwinLi nkTM Cotton is minimal.

4.4.4 Gene Movement

Gene flow is the movement of genes from one or ganism to another. Vertical gene flow, or introgression, is the movement of genes to sexua lly compatible relatives (Ellstrand, 2003; Quist, 2010). In assessing the risk of gene introgression from Tw inLinkTM Cotton to its sexually compatible relatives, APHIS considered two prim ary issues: 1) the potential for gene flow and introgression to feral upland cotton and cotton relatives and 2) the potential impact of introgression. This is di scussed below under the “Prefe rred Alternative” heading.

Horizontal gene flow is the st able movement of genes from one organism to another without reproduction or human intervention (Keese, 2 008; Quist, 2010). There is no evidence of naturally occurring transgene move ment from transgenic crops to sexually incompatible species (Bayer, 2010; Stewart, 2008). Ho rizontal gene transfer and c onsequent expres sion of DNA from one plant to another plant or other phyla (e.g., sp ecies of bacteria) is unlikely to occur (Keese, 2008). This event would require ph ysical relocation of the comple te genetic material from the transgenic plant to the new location, including not only the genes which code for the production of specific proteins, but also those portions of the genome whic h regulate the activity of those genes (Bayer, 2010; Keese, 2008). There are no known naturally occurring vectors (such as plasmids, phages or transposable elements) that could be responsible for inter-domain gene transfer, and there is little ev idence that eukaryotic cells ar e naturally capable of stably incorporating genes from the environment into their genome (Brown, 2003). Although viruses do move genetic material, all viruses that infect higher plants have small RNA or DNA genomes, usually with fewer than 20 encoded proteins (Keese, 2008). These viruses are therefore constrained as to the type and size of novel gene tic material which can be acquired by horizontal gene transfer (Stewart, 2008).

Two bacteria species commonl y associated with plants, Agrobacterium and Rhizobium, have been evaluated to determine the probability of horizontal gene transfer between the bacterium and their host plants. Agrobacterium moves its genes from its bacterial plasmid to the plant causing the plant to produce the gall (abnormal outgrowth) (Unive rsity of Illinois, 2010). Rhizobium aids in nitrogen fixation in legume nodules (Wilkinson and Elevitch, 2011). The genomes of both bacteria have been sequenc ed, and the sequenced genes evaluated for

exogenous genes (Kaneko et al., 2000; Ka neko et al., 2002; Wood et al., 2001). Despite what would appear to be millennia of symbiotic relationships between these bacteria and their host plants, there is no evidence that these organi sms contain genes derived from plants (USDAAPHIS, 2010a). In cases where review of sequence data implied that horizontal gene transfer occurred, these events are inferred to occur on an evolutionary time scale in the order of millions of years (Brown, 2003; Koonin et al., 2001). Transgene DNA promoters and coding sequences are optimized for plant expression, not bacteria l expression (USDA-APHIS, 2010a). Horizontal gene flow, resulting in the relo cation of entire transgenes includi ng the regulatory portions of the DNA (those parts of the DNA which code for th e production of the specific proteins in that relocated transgene) has never been shown to o ccur in nature (Clarke, 2007; Stewart, 2008). Thus, even if horizontal gene tran sfer occurred, proteins associated with these transgenes are not likely to be produced in the new host organism (Stewart, 2008; USDA-AP HIS, 2010a). Based on this information, APHIS cons iders the horizontal gene flow from TwinLinkTM Cotton toBAYER TWINLINKTM COTTON

60 unrelated species to be highly unl ikely, and the same as potentia l horizontal gene flow from existing GE cotton varieties.

No Action: Gene Movement

Under the No Action Alternativ e, conventional and GE transg enic cotton production will continue while TwinLinkTM Cotton will re main a regulated article. Cotton land acreage and cultivation practices for both conventional and GE transgen ic cotton varieties will remain the same. In 2010, Bt cotton varieties occupi ed 93% of the cotton acr eage (USDA-ERS, 2010c). Gene flow from current commercially available GE cultivars to non-GE cotton cultivars, feral cotton populations, and wild sexually compatible relatives will remain unchanged from the current condition.

Preferred Alternative: Gene Movement

Under this alternative, TwinLinkTM Cotton would be available to growers, raising the potential for gene introgression of TwinLinkTM Cotton with ot her sexually compatible, related species.

The passage of the transgene to a cross-compatible wild relative could allo w the wild relative to become more abundant in the habitat or to in vade new habitats because of its lepidopteran resistance (Wilkinson and Ford, 2007). This in creased resistance of the wild relative to one group of insects could alter the balance of othe r insect herbivores, pr edators, and parasites (Wilkinson and Ford, 2007). The introgressed wild crop relative could also impact other plants through interspecific (between species) competiti on and indirectly affect the insect pollinators, herbivores, predators, and paras ites of those species (Wilkinson and Ford, 2007). Vertical gene flow, or introgression, is influe nced by many factors, including the subtle differences between flowers and pollinators in related species, the mode of seed dispersal, and habitat where the two species grow (Messeguer, 2003; Quist, 2010). Introgression can be managed by understanding these biological and physical ba rriers to pollen movement (Dan iell, 2002; Eastham and Sweet, 2002; Messeguer, 2003; Quist, 2010).

Two species of cotton are cultivated in the U.S.: upland cotton ( G. hirsutum ) and Pima cotton ( G. barbadense ) (USDA-ERS, 2009). There are two wild re latives of interest to U.S. growers, G. thurberi and G. tomentosum, found in the mountains of southern Arizona and in Hawaii, respectively. APHIS has considered the potenti al for gene flow from TwinLinkTM Cotton, a GE variety of G. hirsutum , to other upland cotton va rieties, as well as these other three species.

With regard to gene flow to other upland cotton varieties ( G. hirsutum ), such gene flow is controlled by careful adherence to isolation distances, border and barrier rows, and similar practices developed following AOSCA standard s for identity protection of seeds. These measures have been developed based on research on the pollination of the upland cotton flower.

Although upland cotton is considered self-polli nating (Bayer, 2010; Hartman and Kester, 1975), cotton can be pollinated by bees. McGregor (1976) illustrated a rapid decline in the transfer of cotton pollen by bees over distance and found that at 150 to 200 feet away from the source plant, only 1.6% of the bees showed the presence of the source plant polle n particles. Ve ry little pollen is transferred beyond 12 meters (Bayer, 2010; McGregor, 1976; Vaissi ere and Vinson, 1994). By comparison, the isolation distances for Founda tion, Registered, and Cer tified seeds specifiedBAYER TWINLINKTM COTTON

61 in 7 CFR Part 201 are 1320, 1320, and 660 feet, resp ectively. If these id entity practices are adhered to by growers, it is unlikely that Twin LinkTM Cotton will outcross to other upland cotton varieties.

Upland cotton and Pima cotton are frequently cultiv ated in the same regions (Hutmacher et al., 2006). Although cross pollination has been dem onstrated between these species, Van Deynze and others have found that the isolation distance s currently recommended to ensure seed quality for Foundation and Registered seed production ar e adequate to maintain varietal purity (Hutmacher et al., 2006; Van Deynze et al., 2005).

Outcrossing of genes from TwinLinkTM Cotton to G. thurberi is unlikely to occur because G. thurberi contains only the D genome whereas G. hirsutum contains both the A and D genome. The difference in chromosome number s precludes sexual compatibility of G. hirsutum with G. thurberi (OECD, 2008).

G. tomentosum is native to the Hawaiian Islands (P leasants and Wendel, 2010). Historic literature suggests that differences in flowerin g patterns and pollinators presented barriers to cross pollination between G. tomentosum and either G. hirsutum or G. barbadense (Pleasants and Wendel, 2010). Recent field and laboratory studi es contradict this historic literature, demonstrating that these three species share comm on pollinators, and further that differences in flower structure and flowering habits do not serve as barriers to cross pollination (Pleasants and Wendel, 2010). Although laboratory and greenhouse breeding programs with hand pollination have produced viable progeny, DNA marker anal yses have not found evidence of natural hybridization genes between G. hirsutum and G. tomentosum (DeJoode and Wendel, 1992).

The EPA has established cultivation restrictions on Bt cotton varieties to reduce the possibility of gene transfer to feral cotton relatives in Hawaii, Florida, Puerto Rico, and the U.S. Virgin Islands (US-EPA, 2008a). The terms and conditions of EPA’s conditional registration for Cry1Ab in cotton prohibits commercial cultivation sout h of Route 60 near Tampa (US-EPA, 2008a). TwinLinkTM Cotton is neither expected to be plante d commercially in the areas of Florida where wild populations of G. hirsutum occur nor would they likely be impacted by TwinLinkTM Cotton planted north of Route 60. In Hawaii, the EPA ( 2001b) has restricted the sa le and distribution of Bt cotton for commercial planting where it could outcross with the wild cotton species G.

tomentosum. These restrictions in Hawaii also appl y to Bt cotton test plots and breeding nurseries (US-EPA, 2001b). Because of these restri ctions, gene flow from Bt cotton to wild or feral populations of G. hirsutum or to G. tomentosum is extremely unlikely.

APHIS evaluated the potential for gene intr ogression to occur fr om TwinLinkTM Cotton to sexually compatible wild relatives and consider ed whether such introgression would result in increased weediness. Based on the plant pest risk assessment, APHIS has concluded that TwinLinkTM Cotton is not a plant pest and that gene flow between this product and wild or feral relatives will not occur in th e U.S. (USDA-APHIS, 2010b).

Based on the above information, APHIS has concluded that that a determination of nonregulated status of TwinLinkTM Cotton will not impact sexually compatible relatives, nor would it increase the weedy or invasive characteristic s of weedy or wild relatives if gene flow or introgression were to o ccur (USDA-APHIS, 2010b). TwinLinkTM Cotton would provideBAYER TWINLINKTM COTTON

62 growers with an additional cotton variety combin ing a Bt-based lepidopteran protection, as well as herbicide tolerance, and is expected to ha ve the same effect on ge ne movement as the No Action Alternative.

Cumulative Effect: Gene Movement

Based on available scientific evidence, APHIS ha s not identified any cumulative effects on gene movement that would occur from a determinatio n of nonregulated status of TwinLinkTM Cotton.

Gene movement between sexually compatible cotton species is no greater for TwinLinkTM Cotton than it is for other non-GE or GE cultiva rs. Many factors limit the likelihood of gene movement between cotton species. These include:

 Cotton is considered to be a self-pollinating crop with a very low frequency of outcrossing from insect pollinators or wind;

 The Cry1Ab

, Cry2Ae , and bar genes in TwinLinkTM Cotton and other GE cottons impart no agronomic advantages whereby a greater potential for weediness or invasiveness would result should in trogression occur;

 Neither GE or non-GE cotton cultivars form self-sustaining popul ations outside of cultivation because of limitati ons in seed dispersal, germination, and high moisture requirements (US-EPA, 2001b, 2008a);

 The cotton industry has measures in place to restrict pollen movement and gene flow between cotton fields through th e use of isolation distances, border and barrier rows, the staggering of planting dates and various seed handling, transportation and ginning procedures (Sundstrom et al., 2002); and

 Where a potential for GE cotton to introgress into wild or feral populations of cotton exists, the EPA has placed restrictions on GE cotton breeding, testing, and production (US-EPA, 2001b, 2008a).

In addition, there is no eviden ce that horizontal gene transf er and expression of DNA occurs between cotton and soil bacteria or unrelated plant species under natural field conditions, and even if this were to occur, prot eins corresponding to the transgenes are not likely to be produced.

4.5 PUBLIC HEATH

4.5.1 Human Health

Under FFDCA, it is the responsib ility of food and feed manufacturers to ensure that the products they market are safe an d properly labeled. Food and feed derived from TwinLinkTM Cotton must be in compliance with all applicable lega l and regulatory requirements. GE organisms for food and feed may undergo a voluntary consulta tion process with the FDA prior to release onto the market. The EPA also has authority over GE crops such as TwinLinkTM Cotton which present pesticidal proper ties such as Bt toxin.BAYER TWINLINKTM COTTON

63 With regard to human health effects, the FDA and the EPA assess the relative toxicity of the incorporated proteins on humans from the perspective of both consumption, as well as exposure. BCS has submitted a food and feed safety and nutritional assessment to the FDA, as well as a registration review to the EPA for this product. The FDA and EPA are currently reviewing the information submitted by the applicant.

No Action: Human Health

Under the No Action Alternative, TwinLinkTM Co tton would continue as a regulated article. Grower and consumer exposure to this product would be limited to those individuals involved in the cultivation under regulated conditions. Human exposure to existing traditional and GE cotton would not change under this a lternative.

Preferred Alternative: Human Health

APHIS considers the FDA and the EPA regulatory assessment in making its determination of the potential impacts of a determination of nonregulated status of the new agricultural product. BCS has submitted a food and feed safe ty and nutritional assessment to the FDA, as well as a registration review to the EPA for this product. The FDA and EPA are currently reviewing the information submitted by the applicant. In the absence of these completed agency reviews, APHIS takes into consid eration prior reviews of comparab le products as a starting point for its human health impact determination.

Similar products were no longer subject to the plant pest provisions of the Plant Protection Act and 7 CFR part 340 and approved for general cultivation beginning in 1996 with the introduction of Bt products and followed shor tly after by the introduc tion of the various “Liberty” products which provided tolerance to glufosinate ammonium. In each case, FDA and EPA reviews and approvals determined that the products met the agency’s review criteria for approval. The cultivation of these existing crop products would not change under either alternative. Both characteristics have been successfully cultivated in multiple crops in the ensuing years with no evidence of human health impacts.

Note that with regard to the e xpression of glufosinate ammonium to lerance, the gene construct in the Bayer TwinLinkTM Cotton is the same as that approved by the FDA in June, 2003 for the LibertyLink ® Cotton product (US-FDA, 2003). In that approval, the F DA noted that the transformational event in LibertyLink ® Cotton was not materially different in composition, safety, or any other relevant parameter in cott on then grown, marketed, and consumed (US-FDA, 2003).

The EPA, in 2008, published a Biopesticides Registration Action Document (BRAD) for Bacillus thuringiensis ( Bt ) incorporated protectants (US-EPA, 2008a). Based on a comprehensive human health effects assessmen t, the Bt BRAD found no unreasonable adverse effects associated from exposure to the Bt products. The Bt pr otein is a registered microbial pesticide which is approved for use on food crops . The EPA concluded in the BRAD that the Bt products incorporated as plant prot ectants would be expected to be have as expected of a dietary protein (US-EPA, 2008a). A tolerance exemption is in place for Cry1Ab protein in all crops (40 CFR §174.511; US-EPA, 2010d), as well as for the PAT protein (40 CFR §174.522; US-EPA,BAYER TWINLINKTM COTTON

64 2010g). A temporary exemption from the requirement of a tolerance has b een issued for Cry2Ae (40 CFR §174.530; US-EPA, 2010e).

For the majority of Bt proteins currently registered, the so urce bacterium has been a registered microbial pesticide, which has b een approved for use on food crops wi thout specific restrictions. Because of their use as microbial pesticides, a long history of safe use is associated with many Bt products (US-EPA, 2001b). Results from numerous animal and human ep idemiological studies together, over the years, indica te that biopesticides pose mini mal risks to humans and animals (Betz et al., 2000; US-EPA, 2001b, 2008a).

The Cry proteins presen t in the TwinLinkTM Cotton plant have been reviewed and registered by the EPA since 1996 and re-registered in 2001 and 2006 (US-EPA, 2001b, 2008a). The EPA has determined that the use of these Cry proteins as plant-incorporated protectants do not pose a risk to human health (Betz et al ., 2000; US-EPA, 2001b, 2008a).

The PAT protein in TwinLinkTM Cotton is derived from Streptomyces hygroscopius , a common soil bacteria. Humans are frequently exposed to th is bacteria when consuming vegetables or root crops, and there is no evidence of harm from such exposure (Delaney et al., 2008; Hérouet et al., 2005). The PAT enzyme bar gene construct in cotton was no longer subject to the plant pest provisions of the Plant Protection Act and 7 CFR part 340 in 2003 with the BCS LibertyLink ® Cotton (USDA-APHIS, 2003). In 2003, the FDA co mpleted its food safety consultation on LibertyLink ® Cotton and granted tolerance exemptions for both the plant-incorporated protectant, as well as the genetic material n ecessary for its production (US-FDA, 2003). The EPA granted exemptions from the requirement of a tolerance for re sidues of glufosinate ammonium, as well as the plan t-pesticide PAT in or on food and feed commodities of cotton (see: 62 FR 17719; 40 CFR §180.473; US-EPA, 2010c).

BCS conducted safety evaluations based on Codex Alimentari us Commission procedures to assess any potential adverse effects to humans or animals resulting from environmental releases of TwinLinkTM Cotton containing the Cry1Ab, Cry2Ae, and PAT proteins see (Bayer, 2010; FAO, 2001). These safety studies included eval uating protein structure and function, including homology searches of the amino acid sequences with comparison to all known allergens and toxins, an in vitro digestibility assay of the proteins, and an acute toxicity test in the mouse. The TwinLinkTM Cotton proteins were determined to have no amino acid sequence similar to known allergens and lacked toxic potential to mammals (Bayer, 2010). The results of studies conducted by BCS confirm that the crops containing these pr oteins can be safely used as food or feed (Bayer, 2010).

The Australia and New Zealand Food Authorit y (ANZFA) also completed a Toxicological Review and Risk Assessment, and a Techni cal Report was published in 2005 (ANZFA, 2005). ANZFA concluded that LibertyLink ® Cotton was comparable to non-genetically engineered (non-GE) cotton in terms of their safety and nutritional adeq uacy. In 2004, the Plant Biosafety Office (PBO) of the Canadian Food Inspecti on Agency (CFIA) concluded that BCS’s LibertyLink ® Cotton was substantially equivalent to currently grown cotton, in terms of their potential environmental impact a nd livestock feed safety, and the novel traits would not have any substantial negative effect on the environment (C FIA, 2004). CFIA further noted that the PAT enzyme responsible for glufosinate ammonium to lerance has very specif ic enzymatic activityBAYER TWINLINKTM COTTON

65 associated with the acetylation of glufosin ate ammonium, and is ra pidly inactivated in mammalian stomach and intestinal fluids by enzymatic degradation and pH mediated proteolysis (CFIA, 2004).

As with other Bt cotton and he rbicide-tolerant cotton products no longer subject to the plant pest provisions of the Plant Protection Act and 7 CFR part 340 and commercialized, including the LibertyLink ® variety in 2003, TwinLinkTM Cotton is expected to be used throughout cotton producing areas of the country. Based on the analysis of field and laboratory data and scientific literature provided by BCS (Bayer, 2010), and safety data available on other GE cotton, APHIS has concluded that a determin ation of nonregulated status of TwinLinkTM
Cotton would have no adverse impacts on human health. Overall impacts are similar to the No Action Alternative.

Cumulative Effects: Human Health

There are no significant impacts on human or animal health related to the No Action Alternative or a determination of nonregulated status of TwinLinkTM Cotton , and no cumulative effects have been identified.

4.5.2 Worker Safety

EPA’s Worker Protection Standa rd (WPS) (40 CFR Part 170) wa s published in 1992 to require actions to reduce the risk of pesticide poisoni ngs and injuries among agricultural workers and pesticide handlers. The WPS offers protec tions to more than two and a half million agricultural workers who work with pesticid es at more than 560,000 workplaces on farms, forests, nurseries, and greenhouses. The WPS contains requirements for pesticide safety training, notification of pesticide applications, use of personal prot ective equipment, restricted entry intervals following pesticide application, decontamination supplies, and emergency medical assistance.

No Action: Worker Safety

During agricultural production of cotton, agricu ltural workers and pesticide applicators may be exposed to a variety of EP A registered pesticides (US-EP A, 2001b). Under the No Action Alternative, agricultural work ers and pesticide a pplicators may be exposed to these agricultural chemicals during co tton production. Such chemicals would be expected to include those products currently used for insect pest and pl ant pest management in both GE and non-GE cotton cultivation.

Preferred Alternative: Worker Safety

Before the introduction of Bt cott on varieties in 1996, chemical pes ticides were widely used to control lepidopteran pests of cotton. Orga nophosphate, carbamate, a nd pyrethroid products accounted for a substantial percen tage of the insecticides used (USDA-NASS, 2008). However, these three classes of pesticides require numerous safety warnings and extensive use restrictions, which raise concerns for worker safety (US-EP A, 2001b). Bt cotton varieties have offered a very effective and environmentally benign alternat ive to chemical insecticides for lepidopteran insect pest control. As a result, Bt cotton va rieties have been extens ively adopted by cottonBAYER TWINLINKTM COTTON

66 farmers and now represent 73% of the comm ercial cotton planted (USDA-ERS, 2010a). Herbicide-tolerant cotto n varieties have seen similar adopt ion within the i ndustry, with a 78% adoption rate in 2010 (USDA-ERS, 2010a). The adoption of herbicide tolerance has allowed growers to shift their herbicide use to produc ts like glufosinate ammo nium which has a low toxicity to humans. Avoidance of other he rbicides provides corre sponding benefits.

Similar to the No Action Alternative, it is exp ected that EPA registered pesticides that are currently used for cotton production will contin ue to be used by grower s. If TwinLinkTM Cotton is adopted, agricultural workers and pesticide applicators may benefit from the use of TwinLinkTM Cotton due to a reductio n in the use of budworm/bollwor m insecticide applications and the use of the low-toxicity herbicide gluf osinate ammonium. The adoption of glufosinate ammonium-tolerant cotton allows growers to change cultivation practices in those fields where glyphosate resistance has emerged, with the resulti ng potential reduction in herbicide treatments required in those current glyphosate-based systems. A reduction in insec ticide and herbicide use would benefit worker safety.

Cumulative Effects: Worker Safety

A determination of nonregulated status of TwinLinkTM Cotton is not expected to increase the total acreage of cott on production or the use of Bt cotton, he rbicide-tolerant cotton, or stacked varieties presenting both traits. BCS anticipat es that TwinLinkTM Cotton will primarily replace some of the insect-resistant and herbicide-tolera nt cotton cultivars alread y on the market today. As a result, worker safety issues related to the use of EPA registered pesticides during conventional and GM cotton production should remain the same. However, if a grower replaces a non-Bt cotton or non-herbicide-tolerant cotton variety with TwinLinkTM Cotton, then it would be expected that there would be a decrease in the use of lepidopteran pest in secticides. Similarly, replacement of herbicide-tolerant cotton curren tly based on expressing glyphosate tolerance with TwinLinkTM Cotton has the potential to reduce herbicide use to ma nage weedy species which are glyphosate-resistant. This has been the case with the adoption of ot her GM cotton cultivars (Benbrook, 2009). A reduction in pe sticide use and exposure should positively benefit worker safety.

4.6 ANIMAL FEED

Cotton is grown primarily for its lint, but the seed residue, called cottonseed meal, provides a valuable animal feed (Meyer et al., 2007). Cott on seed meal animal feed provides protein, fat, and energy (National Cottonseed Products Asso ciation, 2002). Under FFDCA, it is the responsibility of feed manufacture rs to ensure that the produc ts they market are safe and properly labeled. Feed derived from TwinLinkTM Cotton must be in compliance with all applicable legal and regulatory requir ements. GE organisms for feed may undergo a voluntary consultation process with the FDA pri or to release onto the market. BCS has submitted a food and feed safety and nutritiona l assessment to the FDA, as well as a registration review to the EPA for this product. The FDA and EPA are currently reviewing the information submitted by the applicant. In the absence of these completed agency reviews, APHIS takes into consideration prior reviews of comparable products as a starting point for its animal health impact determination.BAYER TWINLINKTM COTTON

67 No Action: Animal Feed

Under the No Action Alternative, TwinLinkTM Cotton will remain a regulated product and will not be available as an animal feed.

Cotton-based animal feed will still be availabl e from currently cultivated cotton crops, both conventional varieties as well as GE cotton no longer subject to the plant pest provisions of the Plant Protection Act and 7 CFR part 340 expressing either insect resistance or herbicide tolerance. No change in the availability of the residues of these crops as animal feed is expected under this alternative.

Preferred Alternative: Animal Feed

BCS has submitted compositional and nutritio nal characteristics of TwinLinkTM Cotton seed to APHIS (Bayer, 2010). Th e nutritional components of TwinLinkTM Cotton were compared with the non-transgenic cotton cult ivar, Coker 315. The compositio nal comparison was based on analysis of products collected from seven field trials (Bayer, 2010). Chemical analysis of the cotton samples included the nutritio nal factors proximate (moisture, crude protein, lipids, ash, carbohydrates, and energy); minerals, vitamins, ami no acids, and fatty acids (cottonseed oil); and the antinutrient factors of cotton including the goss ypols, phytic acid, and the cyclic fatty acids (Bayer, 2010). The field trials were conducted to simulate modern cotton cultivation techniques, so that the replicates evaluated included fields treated with glufosinate ammonium in accordance with the label provisions and unt reated fields (Bayer, 2010).
The results provided by BCS show no differences between TwinLinkTM Cotton and the nontransgenic Coker 315 cultivars; reported result s for each parameter were also within the reference ranges for cotton composition (Bayer, 2010). All the analytes meas ured fell within the normal range of natural variation in cotton (ILSI, 2008; OECD, 2004).

The EPA has established a permanent exemption from the requirement of a tolerance for the PAT enzyme and the genetic mate rial necessary for its producti on in plants (40 CFR 174.522; US-EPA, 2010g). A similar perm anent exemption has been publi shed for the Cr y1Ab protein in all plants (40 CFR 174.511; US-EPA 2010d). A temporary exemption for the requirement of a tolerance for the Cry2Ae protein in cott on has been published at 40 CFR 174.530, in accordance with Experimental Use Permit 264-EU-143 (US-EPA, 2010e, 2010f).

APHIS’s assessment of the safety of this product for animals fo cuses on plant pest risk (USDAAPHIS, 2010b) and effects on wildlife and thr eatened and endangered species and those analyses are based on the comparison of the GE TwinLinkTM Cotton to its non-GE counterpart. No new issues appear to be associated with the Cry and the PAT proteins in TwinLinkTM
Cotton

.
Based on this information APHIS has concluded that a determination of nonregulated status of TwinLinkTM Cotton would have no significant impacts on animal feed or animal health. Overall impacts are similar to the No Action Alternative.BAYER TWINLINKTM COTTON

68 Cumulative Effects: Animal Feed

There are no significant impacts on animal health related to the No Action Alternative or a determination of nonregulated status of TwinLinkTM Cotton , and no cumulative effects have been identified.

4.7 SOCIOECONOMIC ISSUES

4.7.1 Domestic Economic Environment at Risk

Cotton producers in the U.S. are among the most technically advanced in the world, annually harvesting about 17 million bales or 7.2 billi on pounds of cotton (US-EPA, 2009). The USDA ERS notes that U.S. cotton productivity has increase d in recent years as a result of technological advancements, including biot echnology-derived varieties (Mey er et al., 2007). Herbicidetolerant cotton is one of the most widely and rapidly adopt ed crops in the U.S., followed by insect-resistant cotton (Fernandez-Cornejo an d Caswell, 2006; USDA-ERS, 2010a). By the adoption of new technologies, including agrono mic traits delivered through biotechnology, the yields of cotton lint per acre in the U.S. ranks among the highest in the world (USDA-APHIS, 2010a). Bt varieties of cotton offer excellent prot ection from the damage incurred by insect pests and an economical alternative to broad-spectrum insecticides. In addition, the adoption of postemergent herbicide-tolerant va rieties provide opportunities to apply fewer herbicides and to reduce field cultivation (University of California IP M Online, 2009c; USDA-APHIS, 2010a)

.

No Action: Domestic Economic Environment

Under the No Action Alte rnative, TwinLinkTM Cotton would continue to be regulated articles under the regulations at 7 CFR Part 340. Grow ers and other parties who are involved in production, handling, processi ng, or consumption of cott on would not have access to TwinLinkTM Cotton, but would continue to ha ve access to GE lepidopteran-resistant or herbicide-tolerant (or stacked ) cotton varieties that are no longer subject to the plant pest provisions of the Plant Protection Act and 7 CFR part 340, as well as other GE cotton verities no longer subject to the plant pest provisions of the Plant Protection Act and 7 CFR part 340 and conventional cotton varieties. Domestic growers will continue to utilize currently available traditional and GE cotton varieties based upon ava ilability and market demand. Consequently, the economic profile of cotton production a nd marketing is not expected to change.

Preferred Alternative: Do mestic Economic Environment

BCS anticipates that TwinLinkTM Cotton would re place currently available Bt cotton varieties and may be selected by growers of conventi onal cotton varieties making a change to a GE variety; however, availability of TwinLinkTM Cotton is not expected to result in an increase in cotton acreage (Bayer, 2010). BCS considers the major benefits resulting from the introduction of TwinLinkTM Cotton and its progeny to be additional grower choice, increased competition, and extended useful life of Bt cotton technology.

In its review and analysis of the public intere st documents submitted to support current Bt cotton products, EPA determined that economic value would result from the sale and use of these products (US-EPA, 2001b). Other studies have reached the same conclusions. For example,BAYER TWINLINKTM COTTON

69 based on an analysis of biot echnology-derived crops planted in 2004, the National Center for Food and Agricultural Policy estimated that products providing prot ection against cotton bollworm, tobacco budworm, and pink bollworm increased cotton production by almost 600 million pounds, improved farm income by almost $300 million, and reduced chemical pesticide use by more than 1.6 million pounds (Sankula et al., 2005). In 2006, Fernandez-Cornejo and Caswell provided a summary of other studies that characterize effects on yi eld, pesticide use, and grower returns, suggesting that in addition to the economic benefits to the GE developers and seed firms associated with technology fees and seed premiums, both U.S. and foreign consumers indirectly benefit thr ough lower commodity prices resulti ng from increased supplies (FernandezCornejo and Caswell, 2006). Market benefits were estimated at $230 million for herbicidetolerant cotton and $210 million for insect-resistan t cotton (Bt cotton) after only two years of availability (Fernandez-Corn ejo and Caswell, 2006).

BCS’ field trials demonstrated that TwinLinkTM Co tton was effective in c ontrolling lepidopteran larvae, such as cotton bollworm (CBW, Helicoverpa zea ), tobacco budworm (TBW, Heliothis
virenscens ) larvae, and fall armyworm (FAW, Spodoptera frugiperda ) which are common pests of cotton (Bayer, 2010). These pests cause seve re economic damage to the cotton crop if not controlled. If controlled by chemi cal pesticides, there is the n eed for large insecticide input annually to control these pests (Bayer, 2010). The incorporation of the two Bt-based Cry proteins obviates the need for the input of these broad spectrum insecticides, with a commensurate economic benefit (red uced cost) to the grower.

Despite the wide adoption of Bt cotton varieties to control lepidopteran insects, these same species continue to be the most economically important pests of the crop. In 2009, it was estimated that the Heliothine complex of H. virescens and H. zea infested 4.4 million acres of cotton and reduced yields across the Cotton Belt by 91,119 bales (Williams, 2010). Total 2009 cost and loss estimates for arthropod cotton pests were $502 million (Williams, 2010).

BCS anticipates that TwinLinkTM Cotton will repl ace or displace existing GE cotton varieties currently in the market. Specific economic projec tions are not provided. To the extent that the planting of TwinLinkTM Cotton results in a decrease in insecticide applications for lepidopteran pest control and a potential decrease in herbicide applications where the ut ilization of glufosinate ammonium allows growers to reduce or elimin ate multiple herbicides to control glyphosateresistant weeds, farms adopting TwinLinkTM Cotton mi ght experience an increase in net income. However, net income differentials cannot be proj ected. The net income of cotton producers just growing Bt cotton varieties or he rbicide-tolerant vari eties is not availabl e (USDA-NASS, 2007). However, as BCS anticipates that TwinLinkTM Cotton would only replace existing GE cotton varieties and not result in increased cotton acreage, resu lting socioeconomic impacts are expected to be similar to the No Action Alternative.

Cumulative Effects: Dome stic Economic Environment

Based on the information described above, APHI S concludes that a de termination of nonregulated status of TwinLinkTM Cotton in itself will have no foreseeable adverse cumulative domestic economic effects. TwinLinkTM Cotton w ould simply replace or displace existing cotton varieties for those growers seeking a new comb ination of insect resistance and herbicide tolerance. TwinLinkTM Cotton is not expected to l ead to an expansion of U.S. cotton acreage.BAYER TWINLINKTM COTTON

70 No net cumulative effects have been identified a ssociated with a determination of non-regulated status and cultivation of TwinLinkTM Cotton.

4.7.2 Trade Economic Envi ronment at Risk

Cotton is the single most importa nt textile fiber in the world, accounting for about 40% of all fibers produced. On average, the U.S. produces 20% of the global cotton production and is the leading supplier in the international market (M eyer et al., 2007). However, the U.S. cotton sector has faced a number of challenges as it sh ifts from a domestic-oriented market to one focused largely on the global marketplace. Domest ic mill demand has declined significantly (in the U.S.) from only a decade ago as competition from imported textile and apparel products has risen dramatically. Meanwhile, export demand ha s increased rapidly with the recent expansion of global textile production (Meyer et al., 2007). The USDA ERS reports that U.S. cotton mills consumed 60% of the domestic cotton through the 1990s, but by the mid-2000s 70% of the U.S. cotton was exported (Meyer et al., 2007). As a result, U.S. cotton prices are no longer determined solely by domestic supplies and stocks (Isengildina-Massa and MacDonald, 2009; Meyer et al., 2007).

Research using a multi-region computable genera l equilibrium model assessed the impacts of international Bt cotton adoption on cotton and re lated sectors of regional economies (Frisvold and Reeves, 2007). Productivity gain estimates were based on 2005 adoption rates for Bt cotton in seven countries. Global economic benefits we re nearly $1.4 billion, with U.S. benefits being over $200 million. Increased production from Bt cotton adoption led to a 3% reduction in the world cotton price. Employment and trade balances in the textile and apparel sectors increased for China and India, but generally declined el sewhere. Individual countries obtained greater economic welfare gains upon adoption of Bt cott on; non-adopting regions lost cotton market share to adopting regions (Frisvold and Reeves, 2007).

U.S. cotton production reached consecutive records in 2004 and 2005 seasons, with rising global cotton demand providing a market for much of th e increased output (M eyer et al., 2007). However, the growing use of be tter crop production t echnologies overseas may narrow the gap between foreign production and mill use, constraining growth in foreign import demand and U.S. cotton exports (Meyer et al., 2007 ). Beginning in the 2006 growing year, U.S. cotton acreage was considerably lower, as prices favored the planting of alternative cr ops such as corn and soybeans (USDA-ERS, 2009). Increased acreage in cotton in 2010 was attributed to an export ban imposed by India early that same year (U SDA-FAS, 2010a). Meanwhile, debate over trade policy and the sustainability of current farm programs are a so urce of uncertainty for U.S. agricultural commodities in general and the cotton sector in partic ular (Meyer et al., 2007).

Around the world, new technology has made cotton more attractive to farmers in many countries, and policy reforms in other countries have increased farmers’ willingness to plant cotton (USDA-ERS, 2009). Outside the U.S., the spread of Bt cotton has recently revolutionized India’s cotton sector as had occurred in China (Meyer et al., 2007). The cost savings of Bt cotton brought millions of hectares back into cotton production in eastern China and has also helped India’s cotton area rebound by more than 1 million hectares (Meyer et al., 2007). Bt cotton has also been adopted in smaller produci ng countries like Australia , Argentina, Mexico, and South Africa (Meyer et al., 2007; USDA-ERS, 2009).BAYER TWINLINKTM COTTON

71 No Action: Trade Economic Environment

The cropping and marketing decisions made by cott on growers are unlikely to be influenced by the selection of this alternative and it is expect ed that approximately 93% of the cotton produced will continue to be planted with the currently available GE cotton (USDA-ERS, 2010c). U.S. cotton will continue to play a role in global co tton production, and will continue to be a supplier in the international market. How and where that cotton will be used will be subject to global market conditions.

Global economic benefits from the adoption of Bt cotton were nearly $1.4 billion for the U.S., China, India, Australia, Mexico, Argentina, a nd South Africa (Frisvold and Reeves, 2007). U.S. benefits of the use of Bt cotton were over $200 million (Frisvold and Reeves, 2007). The adoption of Bt cotton technology has brought milli ons of hectares of cotton land back into production in China and has revolutionized I ndia’s cotton industry (Meyer et al., 2007). Economic evidence shows that international trad e and productivity is enhanced by the adoption of Bt cotton and that these products can be cha nneled into suitable export markets. Increased global production of Bt cotton a doption led to a 3% reduction in the world cotton price (Frisvold and Reeves, 2007). This research also showed that individual countries obtained greater economic gains by adopting Bt cotton technology th an if they did not a dopt it, and further, found non-adopting regions to lose cotton market share to the adopting regions (Frisvold and Reeves, 2007). These conditions are not expect ed to change if TwinLinkTM Cotton remains a regulated article.

Preferred Alternative: Trade Economic Environment

BCS intends to submit dossiers to the proper re gulatory authorities of foreign governments to allow the importation of U.S. cottonseed and to ha ve regulatory processes in place so as to allow for the international movement of this product (Bayer, 2010). These may include submissions to the relevant regulatory authorities in Canada , Mexico, the European Union (EU), and Japan, among others. TwinLinkTM Cotton has been, or is currently, in field trials in cotton growing regions around the world (Bayer, 2010).

A determination of nonregulated status of TwinLinkTM Cotton is not expected to adversely impact the trade economic environment and may potentially enhance it through the subsequent development and global adoption of the Twin LinkTM Cotton line which c ould provide another lepidopteran insect resistance and herbicide-to lerant management choice for growers. A reduction in costs for domestic growers associated with the reduction in insecticide and herbicide use may make U.S. producers more competitive in the global market. The trade economic impacts associated with a determination of nonregulated status of TwinLinkTM Cotton are anticipated to be very similar to the No Action Alternative.

Cumulative Effects: Trade Economic Environment

Current and historic economic evidence indicates that Bt cotton technology enhances international cotton trade and production. Based on the information described above, APHIS has determined that there are no past, present, or reasonable foreseeable actions that in aggregate with effects of the proposed action would negati vely impact the trade economic environment.BAYER TWINLINKTM COTTON

72 4.7.3 Social Environment at Risk

U.S. cotton farms and their operators are similar in many respects to those of other crops, but are very different in some key areas. Accordin g to data from the 2003 Agricultural Resource Management Survey (ARMS), farms growing cotton tend to be larger than those growing other crops, with above average gross farm income s, government payments, farm expenses, net incomes, farm asset values, and debt -to-asset ratios (Meyer et al., 2007).

Large farm operations are more likely to be orga nized into partnerships , and cotton farms are no exception. Partnerships allow operators to pool their resources to achieve economies of scale and to combine their talents in managing the fa rm operation (Meyer et al., 2007). Cotton farm operators are also more likely to list farming as their occupation and to have completed high school and college compared with other farm operators (Meyer et al., 2007).

Cotton farms in 2003 generated an average net ca sh income of $127,354 per farm, far more than the average of $11,568 for non-cotton farms in the cotton production regions (USDA-ERS, 2009). The higher average income generated on cott on farms is mainly due to their larger farm operations. Cotton farms averaged 1,199 acres per farm, compared with 376 acres for noncotton farms (USDA-ERS, 2009). Cotton farms’ av erage ratio of cash expenses to gross cash income was 71%, compared with 91% for non- cotton farms (USDA-ERS, 2009). This means that cotton farms could generate $100 of gross income with less expenditures (i.e., cotton farms can earn $100 of gross income for each $71 of cash expense compared to $91 of cash expense for a non-cotton farm). Larger farms can achie ve economies of scale by spreading management, labor, and machinery costs over more units of output, thus gaining an advantage over smaller farms (USDA-ERS, 2009).

The household income for cotton producers av eraged $142,463 in 2003. In comparison, the household income for non-cotton farms in co tton-producing states averaged $71,447 (USDAERS, 2009). For most farm households, income fr om off-farm sources exceeds income from the farm operation. However, cotton producers derive th e majority of their fa mily income from the farm. Eleven percent of cotton producers list ed nonfarm jobs or busi nesses as their main occupation, compared with 48% of non- cotton farm operators (USDA-ERS, 2009).

No Action: Social Environment

Under the No Action Alternative, no direct im pact on the social envi ronment surrounding cotton farming is expected. The cropping and mark eting decisions made by cotton growers are unlikely to be influenced by the selection of this alternative.

Preferred Alternative: Social Environment

Most of the cotton acreage in the U.S. is pl anted with GE cotton. Of the 11.3 million acres planted in cotton in 2010, 93% were GE cotton: 73% of that GE cotton acreage was GE insectresistant (Bt) cotton and 78% was herbicide-to lerant (USDA-ERS, 2010a, 2010c). Most cotton is planted on farms that have been in cotton production for at least five years (USDA-NASS, 2007).BAYER TWINLINKTM COTTON

73 The introduced stacked lepidopteran-resistant and herbicide-tolerant trait in TwinLinkTM Cotton is not intended to confer any competitive advantage in terms of weediness or to extend the range of cultivation outside of existing cultivation areas. A determination of nonregulated status of TwinLinkTM Cotton by APHIS is not expected to signif icantly expand the number of cotton acres, and cotton acreage is expected to remain re latively stable. Overall impacts are similar to the No Action Alternative.

Cumulative Effects: Social Environment

Based on the information described above, APHIS ha s determined that there are no past, present, or reasonably foreseeable actions that in aggr egate with effects of the proposed action would impact the social environment surrounding cotton farming.

4.8 OTHER CUMULATIVE EFFECTS

The potential cumulative effects regarding specifi c issues have been analyzed and addressed above. No further potential cumulative effects have been identified. To date, none of the GE cotton varieties that are no longer subject to the plant pest provisions of the Plant Protection Act and 7 CFR part 340 and used for commercial cotton prod uction or cotton breeding programs have been subseq uently found to pose a plant pest risk.

Cultivation of stacked varieties, those crop varieties that may contain more than one trait, are currently found in the marketplace and in agricultural production. In the event APHIS reaches a determination of nonregulated status, TwinLinkTM Cotton may be combined with non-GE and GE cotton varieties by traditional breeding techniques , resulting in a plant that, for example, may also be resistant to other herb icides, or may present a different combination of insect pest-resistant, but may also have progeny with no transgenes at all. APHIS regulations at 7 CFR part 340 do not provide for Agency oversight of GE cotton varieties that are no longer subject to the plant pest provision s of the Plant Protection Act and 7 CFR part 340, nor over stacked varieties combining GE varieties that are no longer subject to the plant pest provisions of the Plant Protection Act and 7 CFR part 340 unless it can be positively shown that such stacked varieties were to pose a likely plant pest risk.

There is no guarantee that TwinLinkTM Cotton will be stacked with any particular non-GE or GE cotton varieties that are no longer subject to th e plant pest provisions of the Plant Protection Act and 7 CFR part 340 , as company plans and market demand s play a significant role in those business decisions. Predicting all potential combinations of stack ed varieties that could be created using both non-GE and GE cotton varieties that are no longer subject to the plant pest provisions of the Plant Prot ection Act and 7 CFR part 340 is hypothetical and purely speculative. No further an alysis is required.

4.9 THREATENED AND ENDANGERED SPECIES

The Endangered Species Act (ESA) of 1973, as amended, is one of the most far-reaching wildlife conservation laws ever enacted by any nation. Congress, on behalf of the American people, passed the ESA to prevent extinctio ns facing many species of fish, wildlife and plants. The purpose of the ESA is to conserve endangered and threatened species and the ecosystems on which they depend as key components of America’s heritage. To implementBAYER TWINLINKTM COTTON

74 the ESA, the U.S. Fish and Wildlife Service (USFWS) works in cooperation with the National Marine Fisheries Service (NMFS), other Federal, State, and local agencies, Tribes, non-governmental organizations, and private citizens. Before a plant or animal species can receive the protection provided by the ESA, it must first be added to the Federal list of threatened and endangered wildlife and plants.

A species is added to the list when it is determined by the USFWS/NMFS to be endangered or threatened because of any of the following factors:

 The present or threatened dest ruction, modification, or curtailment of its habitat or range;

 Overutilization for commercial, recreational, scientific, or educational purposes;

 Disease or predation;

 The inadequacy of existing regulatory mechanisms; and

 The natural or manmade fact ors affecting its survival.

Once an animal or plant is added to the li st, in accordance with the ESA, protective measures apply to the species and its hab itat. These measures include protection from adverse effects of Fede ral activities.

Section 7 (a)(2) of the ESA requires that Federal agencies, in consultation with USFWS and/or the NMFS, ensure that any action they authorize, fund, or carry out is not likely to jeopardize the continued existence of a listed sp ecies or result in the destruction or adverse modification of designated critical habitat. It is the responsibility of the Federal agency taking the action to assess the effects of thei r action and to consult with the USFWS and NMFS if it is determined that the action “may affect” listed species or critical habitat. To facilitate APHIS’ ESA consultation process, AP HIS met with the USFWS to discuss factors relevant to their effects analysis for petitio ns for nonregulated status and developed the following process for conducting an effects determination for biotechnology regulatory actions. Appendix A provides a copy of th e interagency consultation protocol.

As part the environmental review process, APHIS thoroughly reviews GE product information and data to inform the ESA effects analysis and, if ne cessary, the biological assessment. For each transgene(s)/transgenic plant the following information, data, and questions are consid ered by APHIS:

 A review of the biology, taxonomy, and weediness potential of the crop plant and its sexually compatible relatives;

 Characterization of each transgene with respect to its structure and function and the nature of the organism from which it was obtained;

 A determination of where the new transgen e and its products (if any) are produced in the plant and their quantity;

 A review of the agronomic performance of the plant including disease and pest susceptibilities, weediness potential, a nd agronomic and environmental impact;

 Determination of the concentrations of know n plant toxicants (i f any are known in the plant); andBAYER TWINLINKTM COTTON

75  Analysis to determine if the transgenic plant is sexually compatible with any threatened or endangered plant sp ecies (TES) or a host of any TES.

In following this process, APHIS evaluated the potential effects that a determination of nonregulated status of TwinLinkTM Cotton would have on Fe derally listed threatened and endangered species (TES) and species proposed for listing, as well as designated critical habitat and habitat proposed for designation. Based upon the scope of the EA and production areas identified in the Affected Environment section of the EA, APHIS obtained a list of TES species (listed and proposed) for each state where cotton is commercially produced from the USFWS Environmental Cons ervation Online System (ECOS; see Appendix B).

4.9.1 Potential Effects of TwinLinkTM Cotton and Agricultural Practices on TES

BCS anticipates that TwinLinkTM Cotton w ould replace currently available Bt cotton varieties and may be selected by growers of c onventional cotton varieties making a change to a GE variety; however, availability of Twin LinkTM Cotton is not exp ected to result in an increase in cotton acreage (Bayer, 2010). Accordi ngly, the issues discussed herein focus on the potential environmental consequences of a determination of nonregulated status of TwinLinkTM Cotton on TES species in the areas where cotton is grown.

APHIS therefore focuses its TES review on th e implications of exposure to the PAT and Cry proteins in cotton. This analysis also addresses the potential impacts of the use of glufosinate ammonium, and impacts to non-targe t organisms and the natural environment.

BCS has presented data evaluating the agronomic and morphological characteristics of TwinLinkTM Cotton, including compositional and nutritional characteristics, safety evaluations and toxicity tests, comparing th e product to a conventional cotton variety (i.e., Coker 315) (Bayer, 2010). The results presented by BCS show no differences between TwinLinkTM Cotton and the non-transgenic Coker 315 variety, other than the introduced PAT and Cry proteins. (See previ ous discussions on Human Hea lth and Animal Feed for an expanded discussion of BCS’ comparison.)

The agronomic and morphologic characteristics da ta provided by BCS were used in APHIS analysis of the weediness potential for TwinLi nkTM Cotton, and evaluated for the potential to impact TES. Agronomic studies conducted by BCS tested the hypothesis that the weediness potential of TwinLinkTM Cotton is unc hanged with respect to conventional cotton (Bayer, 2010). No differences were de tected between TwinLinkTM Cotton and nontransgenic cotton in growth, reproduction, or inte ractions with pests and diseases, other than the intended effect of protection from lepidopt eran pests. The main natural controls on feral populations of cotton are poor seed disper sal, poor seed germination, competition from other plants, and a high requi rement for moisture (US-EP A, 2001b). Therefore, the incorporation of either the insect resistance trait or the herbicide tolerance trait is unlikely to appreciably improve seedling establishment and increase weediness potential. Any TwinLinkTM Cotton volunteers could be readily controlled by other readily available herbicides, including glyphosate (Miller et al., 2004; Stewart et al., 2003). Based on thisBAYER TWINLINKTM COTTON

76 analysis, APHIS has concluded that a dete rmination of nonregulated status of TwinLinkTM Cotton has no potential to affect TES, as a result of weediness.

In addition to evaluating BCS’ comparisons of TwinLinkTM Cotton with the non-transgenic Coker 315 variety, for potential differences in agronomic and morphology APHIS also considers the EPA and FDA regulatory asse ssment in making its determination of the potential impacts of a determination of nonregula ted status of the new agricultural product. In that regard, BCS has submitted food and feed safety and nutritional assessments to the FDA as well as comparable exemptions from to lerance and registration reviews to the EPA for TwinLinkTM Cotton. The FDA and EPA are currently reviewing the information submitted by the applicant. In the absence of a completed FDA and EPA determinations, APHIS takes into consideration prior agency reviews of comparable products to conduct its assessment of the potential impacts. As discus sed above in Animal and Plant Communities (Subsection 4.4) and Public Health (Subsec tion 4.5), both agencies have previously reviewed and approved comparable products, including several crop varieties expressing the Bt-based Cry proteins as plant-incorpora ted protectants, as well as the bar gene, resulting in the expression of the PAT enzyme providing to lerance to glufosinate ammonium.

Data regarding the environmental safety of PAT proteins have been extensively reviewed in international scientific, peer-reviewed journals (Hérouet et al., 2005). In summary, the PAT protein is expressed in a number of GM crops that have been approved by regulatory agencies. These GM crops have been c onsumed by humans and animals since 1995 and have exhibited no significant adverse effects. The PAT protein mode of action is specific to the breakdown of ammonia in plant tissues. Because animals lack this enzymatic pathway (Hérouet et al., 2005), and there is no mechanis m to transfer this gene to unrelated plant species, APHIS limits its analysis of the impli cations of exposure to the PAT protein to the potential for gene movement to related TES cotton species.

None of the relatives of cotton are listed (or proposed) as endangered or threatened species under Federal (US-FWS, 2011a) or state listi ngs (Arizona, 2009; Ha waii, 2001) with the exception of G. hirsutum in the State of Florida, where wild populations of upland cotton, G. hirsutum have been listed as endangered by the state (Coile and Garland, 2003; Institute for Regional Conservation, 2010). However, in Florida, wild G. hirsutum is not present in the northwestern panhandle where cotton cultivation occurs, and cultivation of cotton is prohibited by the EPA in those areas of s outhern Florida where it is found (Coile and Garland, 2003; US-EPA, 2008a; USDA-NA SS, 2007; Wunderlin and Hansen, 2010). Accordingly, a determination of nonregulated st atus of TwinLinkTM Cotton is not expected to impact state endangered feral cotton populations.

Evaluation of the potential exposure of threaten ed and endangered species to the Bt-based Cry proteins focuses on the route of exposure. Cry1Ab and Cry2Ae proteins are highly specific to Lepidoptera. The specificity of Bt crystalline proteins to lepidopteran insect larvae, but not for other insects, birds, an d mammals results from the highly specific receptors for these proteins in the larvae midg ut (Lemaux, 2009). Once activated by insectspecific proteases in the insect midgut, Cry protei ns bind to receptors in the midgut. Such binding leads to the formation of pores in the mi dgut membranes and ultima tely to cell lysis and death. The specific binding of Bt-based Cry proteins to midgut membrane receptors is aBAYER TWINLINKTM COTTON

77 key determinant of pest specificity (Showalter et al., 2009). Accordingly, the APHIS review of the potential impacts is focused on Fede rally listed threatened and endangered lepidopterans in the cotton growing regions of the U.S.

APHIS obtained and reviewed the USFWS Federal List of listed (or proposed) TES and considered the potential impacts of a determination of nonr egulated status of TwinLinkTM Cotton on lepidopterans (Appendix B). There are currently 21 species of butterflies, moths, and skippers on this list (US-FWS, 2011b). Because it is not possible to use TES species to quantify sensitivity to the Cry and PAT proteins, APHIS’ evaluation focused on the likelihood of exposure of TES species to the Bt-endotoxin expressed in TwinLinkTM Cotton. Exposure of TES species to the Bt-based Cry protei ns in TwinLinkTM Cotton is only likely if the species occur in the areas where cotton is grown, as cotton plant pa rts (seeds, pollen, and crop debris) are not readily transported long distances without hum an involvement.

APHIS has thoroughly examined all listed a nd proposed threatened and endangered lepidopterans and compared their habitats to t hose habitats present in counties where cotton is grown. APHIS has determined that the breeding habitats of listed Lepidoptera do not overlap cotton growing areas. Therefore, it is highly unlikely that these species can be exposed to TwinLinkTM Cotton. Habitats in which threatened and endangered lepidopterans in the U.S. are likely to occur are very limite d in geographical distribution. Moreover, the larvae of these species typically feed on specific host plants, none of which include cotton, or plants likely to be found in cotton fields. In those states where cotton is cultivated, lepidopteran species on the Federal TES list only occur in California, Florida, and North Carolina (US-FWS, 2011b) (see Appendix B: T hreatened and Endangered Species). Of these species, only the Quino Checkerspot butterfly ( Euphydryas editha quino ), the Kern primrose sphinx moth ( Euproserpinus euterpe ), and the Saint Francis’ satyr ( Neonympha mitchellii francisci ) are known to occur in countie s were cotton is grown.

The Quino Checkerspot butterfly is found in co astal sage scrub, open chaparral, juniper woodland, and native grassland (US-FWS, 2003). Its primary host plants include: the dwarf plantain ( Plantago erecta ) that occurs in southern California within annual scrub, grassland, and open chaparral plant communities; the wooly plantain ( Plantago patagonica ) that overlaps in distribution with P. erecta at lower elevations ; the white snapdragon ( Antirrhinum coulterianum ) that appears to be a facultativ e fire-follower in non-desert areas and in desert plant communiti es often growing between shrubs; and the thread-leaved bird’s beak ( Cordylanthus rigidus ), a partially parasitic plant often found at high densities in disturbed areas, open slopes, and flats of foothill woodlands, chaparral margins, and coniferous forests (US-FWS, 2003). Areas wher e cotton is grown are likely to have been used in agricultural production for many years and thus would not change the habitat that is already unsuitable for the Quino Checkerspot butterfly.

The Kern primrose sphinx moth is also found in California. This moth may occupy habitat near cotton cultivation, known to occur in Kern, Santa Barbara, San Luis Obispo, and Ventura Counties, CA (Jump et al., 2006; US -FWS, 2007a). The larv ae feed primarily on their essential host plant Camissonia sp . (certain primrose and sun cup species). The host plant evening primrose is found along sandy washes with young alluvial sandy soils that are prone to regular flooding (Jump et al., 2006). Si nce listing, the primary threats to the KernBAYER TWINLINKTM COTTON

78 primrose sphinx moth are agricultural land use practices that degrade the moth habitat, particularly cattle grazing, disking, pesticide and herbicide use, and development. Many agricultural pesticides are specifically designed to target insect larvae (caterpillars) as well as adult moths (US-FWS, 2007a). All Kern primrose sphinx moth populations are potentially at risk from this effect. Kern primrose sphinx moth exposure to agricultural pesticides could occur when cotton is grown in and around sandy washes with sandy alluvial soil where moths perform morning basking and where the food plant Camissonia campestris

or C. contorta (field primrose and evening primros e, respectively) is supported. The potential route of Kern primrose sphinx moth exposure to the Bt protein contained in TwinLinkTM Cotton would be in the form of pollen deposited on the evening primrose, where the threatened caterpillars feed. Beca use cotton pollen is heavy and will not be dispersed much from the field (Thies, 1953), it is not likely that the larvae of the Kern primrose sphinx moth will be exposed to the Cry protein. Any pollen produced by TwinLinkTM Cotton plants is not expected to drift onto larval host plants.

The Saint Francis’ satyr is known only in the sand hills of North Carolina, in Cumberland and Hoke Counties (US-FWS, 2010a). The species is known only to be located on government land (Fort Bragg U.S. Army installati on). The habitat occupied by this satyr consists primarily of wide, wet meadows dominated by a high diversity of sedges ( Carex ) and other wetland graminoids. Sain t Francis’ satyr has also been observed in pitcher plant ( Sarracenia flava ) swales, with cane ( Arundinaria tecta ), rough-leaved loosestrife ( Lysimachia as perulaefolia ), and pocosin lily ( Lilium iridollai ). It is unknown whether satyr uses such habitat for reproduction or simply as a dispersal corridor. Adult food habits are not really known; larva probably feed on Carex , but this has not been established. Oviposition has been observed on small Panicu m grass. Areas in H oke and Cumberland County where cotton is grown are likely to have been used in agricultural production for many years and would be unsuitable habita t for the Saint Francis’ satyr.

As noted above, the EP A conducts a comprehensive asse ssment of the implications of pesticide registrations on TES. EPA’s eval uations include exposure assessments and ecological effects data for each registered pesticide. The EPA (2001b, 2008a) has published the following specific statements regardi ng potential effects of Bt-based Cry proteins incorporated in cotton—

...The larvae of endangered Lepidoptera species in cotton growing counties (Quino Checkerspot butterfly, Saint Fran cis' satyr butterfly and Kern primrose sphinx moth) are not going to be e xposed to Cry1Ac protein [the Bt-based Cry protein that was the focus of this report] because their habitats do not overlap with cotton fields. The Quino Checkerspot butterfly is found only in the coastal sage scrub habitat in southern California, the Kern primrose sphinx moth (threatened, not endangered) is found only on a privately owned ranch in the Walker Basin, Kern County Ca lifornia. Finally, the only known population of Saint Francis’ satyr butte rfly is found in wetlands dominated by sedges and grasses on Department of De fense property in North Carolina. None of the larvae of these insects feed on cotton, and will not be exposed to Cry protein in pollen. The amount of pollen that would drift from these cotton plants onto plants fed upon by endangered/threatened species, wouldBAYER TWINLINKTM COTTON

79 be very small (if measurable) compared to the levels fed to the test species. Therefore, EPA does not expect that any endangered/threatened species will be affected by pollen contai ning the Cry1Ac protein.

In addition, because EPA is imposing conditi ons for geographic areas (Hawaii and Florida) that have sexually compatible wild or weed y relatives of cotton, the Cry1Ac protein gene cannot escape into related wild plants which could serve as a source of Bt pollen for plants on which endangered/threatened species may feed on in these areas.

Because EPA expects that no listed endangered species of Lepidoptera will be exposed to the Bt Cry protein expressed in cotton plan ts, and because the most probable exposure scenario does not appear to affect listed species, EPA believes that this action will have no effect on listed species (US-EPA, 2001b; see also US-EPA 2008a).

Based on the information above, APHIS has determ ined there would be no effects to the three listed lepidopteran species as a result of a determination of nonregulated status of TwinLinkTM Cotton.

4.9.2 Potential Effects of the Use of Glufosinate Ammonium

As the action agency for pesticide registration s, EPA has the responsibility to conduct an assessment of effects of a registration action on endangered species. The EPA Endangered Species Protection Program web site, http ://www.epa.gov/espp/, describes the EPA assessment process for endangered species. Some of the elements of that process, generally taken from the web site, are summarized below.

When registering a pesticide or reassessing the potential ecological risks from use of a currently registered pesticide, EPA evaluates extensive exposure and ecological effects data to determine how a pesticide will move throu gh and break down in the environment. Risks to birds, fish, invertebrates, mammals, and plants are routinely assessed and used in EPA’s determinations of whether a pesticide may be licensed for use in the U.S.

EPA’s core pesticide risk assessment and regul atory processes ensure that protections are in place for all populations of nontarget species, including TES. These assessments provide EPA with information needed to develop label use restrictions for the pesticide. These label restrictions carry the weight of law and ar e enforced by EPA and the states (Federal Insecticide, Fungicide, and Rodenticide Act 7 USC 136j (a)(2)(G) Unlawful acts). Because TES may need specific protection, EPA has de veloped risk assessment procedures described in the Overview of the Ecological Risk Asse ssment Process (US-EP A, 2004) to determine whether individuals of a listed spec ies have the potential to be harmed by a pesticide, and if so, what specific protections may be appropriate. EPA’s conclusion regarding the potential risks a pesticide may pose to a listed species and any designated critical habitat for the species, after conducting a thorough ecological risk assessment, results in an "effects determination" in accordance with Section 7 (a)(2) of the ESA.

The EPA first registered glufosinate ammoni um in 2000 as a non-selective foliar herbicide for use on a wide range of crops, including cotton (US-EPA, 2008b). In 2008, the EPA announced that it was undertaking a registrati on review of this product, and has published aBAYER TWINLINKTM COTTON

80 preliminary work plan for this process (US-EP A, 2008b). The EPS’ preliminary work plan provides for an ecological risk assessment (ERA) to allow the EPA to determine whether glufosinate ammonium use has “no effect” or “m ay affect” Federally listed threatened or endangered species (listed species) or their de signated critical hab itat (US-EPA, 2008b). If the ERA indicates that glufosinate ammonium “m ay affect” a listed species or its designated critical habitat, the assessment will be refine d to allow the EPA to determine whether the use of glufosinate ammonium is “likely to adversely affect” the species or critical habitat or “not likely to adversely affect” the species or critical habitat (US-EPA, 2008b). When an ERA concludes that a pesticide’s use “may a ffect” a listed species or its designated critical habitat, the Agency will consult with the U.S. Fish and Wildlife Service and National Marine Fisheries Servi ce, as appropriate.

To mitigate potential adverse effects to TES, EP A has imposed specific label use restrictions for glufosinate ammonium use when applied with aerial equipment including “The product should only be applied when the potential for drift to adjacent sensitive areas (e.g.,residential areas, bodies of water, known habitat for threatened or endangered species, nontarget crops) is minimal (e.g., when wind is blowing away from the sensitive areas)” (BCS, 2010a).

To facilitate pesticide applicators adheren ce to EPA label use restrictions for glufosinate ammonium, BCS’ label for Ignite, the commercial form of glufosinate ammonium, provides specific instructions for managing spray drift and cautions pertaining to applications around water so as to minimize potential effect s to non-target organisms (BCS, 2010a). Label requirements for BCS’ Ignite formulations prohib it application in conditions or locations where adverse impact on Federally listed endange red/threatened plants or aquatic species is likely (BCS, 2010a). BCS has also provided a t echnical and trade manual with additional guidance on the proper storage, handling and use of the product (BCS, 2010b).

In conclusion, there are legal precautions in place (EPA label use restrictions) and “best practice” guidance to reduce the possibility of exposure and adverse impacts to TES from glufosinate ammonium application to TwinLi nkTM Cotton; EPA has considered potential impacts to TES as part of their registrati on and labeling process for glufosinate ammonium; and adherence to EPA label use restrictions by the pesticide applicator will ensure that the use of glufosinate ammonium will not adversel y affect TES or critical habitat. Based on these factors and the legal requirements for pe sticide applicators to follow EPA label use restrictions, APHIS has determined that the use of EPA registered glufosinate ammonium for TwinLinkTM Cotton production will not adversely impact listed species or species proposed for listing and would not adversely im pact designated critical habitat or habitat proposed for designation.

Based on the above information, APHIS has de termined that the Preferred Alternative, a determination of nonr egulated status of TwinLinkTM Cotton, woul d have no effect on Federally listed TES and species proposed for lis ting, or on designated critical habitat or habitat proposed for designation. Conse quently, a written concurrence or formal consultation with the USFWS is not required for this action.BAYER TWINLINKTM COTTON

81 4.10 CONSIDERATION OF EXECUTIVE ORDE RS, STANDARDS, AND TREATIES RELATING TO ENVIRONMENTAL IMPACTS

4.10.1 Executive Orders with Domestic Implications

The following two executive orders require cons ideration of the potential impacts to minority and low income populations and children:

 Executive Order (EO) 12898 (US-NARA, 2 010), "Federal Actions to Address Environmental Justice in Minority P opulations and Low-Income Populations," requires Federal agencies to conduct thei r programs, policies, and activities that substantially affect human health or the e nvironment in a manner so as not to exclude persons and populations from participation in or benefiting from such programs. It also enforces existing statutes to preven t minority and low-income communities from being subjected to disproportionately high and adverse human health or environmental effects.

 EO 13045, “Protection of Children from En vironmental Health Risks and Safety Risks,” acknowledges that children may suffer disproportionately from environmental health and safety risks because of their developmental stage, greater metabolic activity levels, and behavior patter ns, as compared to adults. The EO (to the extent permitted by law and consistent with the agency’s mission) requires each Federal agency to identify, assess, and addr ess environmental hea lth risks and safety risks that may disproporti onately affect children.

Each alternative was analyzed with respect to EO 12898 and EO 13045. Neither alternative is expected to have a disproporti onate adverse effect on minoriti es, low-income populations, or children.

Available mammalian toxicity, along with the history of safe us e of microbial Bt products and other cotton varieties expressing Bt and PAT pr oteins, establishes the safety of TwinLinkTM Cotton and its products to humans, including minor ities, low income populations, and children who might be exposed to them through agricultu ral production and/or processing. No additional safety precautions would need to be taken.

None of the impacts on agricultural practices expect ed to be associated with a determination of nonregulated status of TwinLinkTM Cotton are expect ed to have a disproportionate adverse effect on minorities, low income populations, or children. As noted above, the cultivation of cotton varieties that are no longer subject to the plant pest provisions of the Plant Protection Act and 7 CFR part 340 with similar insect-resistant traits has been associated with a decrease and/or shift in pesticide applications for those who adopt thes e varieties that is either favorable or neutral with respect to environmental and human toxicit y. If pesticide applications are reduced, there may be a beneficial effect on children and low in come populations that might be exposed to the chemicals. These populations might include migr ant farm workers and their families, and other rural dwelling individuals who are exposed to pe sticides through groundwater contamination or other means of exposure. It is expected that EPA and USDA Economic Research Service wouldBAYER TWINLINKTM COTTON

82 monitor the use of this product to determine impacts on agricultura l practices, such as chemical use, as they have done prev iously for other products.

The following executive order addresses Federal re sponsibilities regardi ng the introduction and effects of invasive species:

EO 1311 (US-NARA, 2010), “Invasive Species,” states that Federal agencies take action to prevent the introductio n of invasive species, to provide for th eir control, and to minimize the economic, ecological, and human health impacts that invasive species cause.

Non-engineered cotton as well as other Bt and herbic ide-tolerant cotton varieties are widely grown in the U.S. Based on historical experience with these varieties and the data submitted by the applicant and revi ewed by APHIS, TwinLinkTM Cotton plan ts are sufficiently similar in fitness characteristics to other cotton varieties currently grown and are not expected to become weedy or invasive (USDA-APHIS, 2010b).

The following executive order requires the pr otection of migratory bird populations:

EO 13186 (US-NARA, 2010), “Responsibilities of Federal Agen cies to Protect Migratory Birds,” states that Federal agencies taking actions that have, or are likely to have, a measurable negative e ffect on migratory bird populations are directed to develop and implement, within two years, a Memorandum of Understanding (MOU) with the Fish and Wi ldlife Service that shall promote the conservation of migrator y bird populations.

Data submitted by the applicant has shown no difference in compositional and nutritional quality of TwinLinkTM Cotton compared with other GE cotton or non-GE cotton, apart from the presence of the two Cry proteins and the PAT protein . The migratory bird s that occasionally forage in cotton fields are unlikely to ingest high amounts of TwinLinkTM Cotton seed as cotton seed is limited by harvest.

The introduction of Bt cotton has had a positive effect on bird co unts in North American cotton fields. Comparing bird populations from 19 91 to 1995 and 1996 to 2000, EPA (2001b) showed that bird counts increa sed and were positively correlated with Bt cotton adoption, the reduction in insectic ide use, and the relative presence of th e species in cotton fields prior to the introduction of Bt cott on (US-EPA, 2001b). B road-spectrum insecticides, which are considered to be one of the largest impe diments to agroecosystem biodive rsity, have been reduced by the commercial use of Bt cotton (Benbrook, 2009; US DA-NASS, 2008). Since the introduction of Bt cotton there has been a two thirds decrease in use for the most toxic insecticides products to birds and fish, and a comparable one third decrease in use for the most toxic products to humans (US-EPA, 2001b). Additionally, there have been no reported adverse e ffects of Bt cotton on non-target insects which can serve as a food source for birds.

Collectively, the weight of considerable field studies with Bt crops, including Bt cotton, have consistently demonstrated that Bt crops have only minor effects, if any, on a large number of non-target insect taxa which is very small in comparison to the use of broad-spectrum insecticides (N aranjo, 2009a).BAYER TWINLINKTM COTTON

83 The history of safe use of cr ops expressing the PAT enzyme strongly suggests that TwinLinkTM Cotton will not present a risk to birds. The PAT enzyme bar gene construct in cotton was no longer subject to the plant pest provisions of the Plant Protection Act and 7 CFR part 340 in 2003 with the BCS LibertyLink ® Cotton (USDA-APHIS, 2003). In 2003, the FDA completed its food safety consultation on LibertyLink ® Cotton, and granted tolerance exemptions for both the plantincorporated protectant as well as the genetic material nece ssary for its production (US-FDA, 2003). The EPA granted exemptions from the requirement of a tole rance for residues of glufosinate ammonium as well as the plant-pes ticide PAT in or on food and feed commodities of cotton (see: 62 FR 17719; 40 CFR §180.473; US-EPA , 2010c). When used in accordance with label restrictions, glufosinate ammonium doe s not present a risk to migratory birds.

Based on APHIS’ assessment of TwinLinkTM Cotton, it is unlikely that a determination of nonregulated status of this cotton variety will have a negative effect on migratory bird populations.

4.10.2 International Implications

EO 12114 (US-NARA, 2010), “Environmental E ffects Abroad of Major Federal Actions” requires Federal officials to take into consider ation any potential environmental effects outside the U.S., its territories, and possessions th at result from actions being taken.

APHIS has given this EO carefulconsideration and does not expect a significant environmental impact outside the U.S. in the event of a de termination of nonregulat ed status of TwinLinkTM
Cotton.

It should be noted that all the existing national and international regulatory authorities, and phytosanitary regimes that currently apply to introductions of new cotton cultivars internationally apply equally to those covered by an APHIS determ ination of nonregulated status under 7 CFR Part 340.

Any international trade of TwinLinkTM Cotton subsequent to a determination of nonregulated status of the product would be fully subject to national phytosanitary requirements and be in accordance with phytosanitary standards devel oped under the Internati onal Plant Protection Convention (IPPC, 2010). The purpose of the IPPC “is to secure a common and effective action to prevent the spread and introduction of pe sts of plants and plant products and to promote appropriate measures for their control” (IPPC, 2 010). The protection it affords extends to natural flora and plant products and includes both direct and indirect damage by pests, including weeds.

The IPPC establishes a standard for the reci procal acceptance of phytosanitary certification among the nations that have sign ed or acceded to the Convention (172 countries as of March 2010). In April 2004, a standard for PRA of living modified organisms (LMOs) was adopted at a meeting of the governing body of the IPPC as a supplement to an existing standard, International Standard for Phytosanitary Meas ure No. 11 (ISPM-11, Pest Risk Analysis for Quarantine Pests). The standard acknowledges that all LMOs will not present a pest risk and that a determination needs to be made early in the PRA for importation as to whether the LMO poses a potential pest risk resulting from the genetic modification. APHIS pest risk assessmentBAYER TWINLINKTM COTTON

84 procedures for genetically engineered organisms are consistent with the guidance developed under the IPPC. In addition, issues that may relate to commercialization and transboundary movement of particular agricu ltural commodities produced through biotechnology are being addressed in other international forums and through national regulations.

The Cartagena Protocol on Biosafety is a treaty under the Un ited Nations Convention on Biological Diversity (CBD) that established a framework for the safe transboundary movement, with respect to the environment a nd biodiversity, of LMOs, which include those modified through biotechnology. The Prot ocol came into force on September 11, 2003, and 160 countries are Parties to it as of December 2010 (CBD, 2010). Although the U.S. is not a party to the CBD, and thus not a party to the Cartagena Protocol on Biosafety, U.S. exporters will still need to comp ly with those regulations that importing countries which are Parties to the Protocol have promulgated to comply with their obligations. The first intentional transboundary movement of LMOs intended for environmental release (field trials or commercial planting) will requi re consent from the importing country under an advanced informed agreement (AIA) provision, which in cludes a requirement for a risk assessment consistent with Annex III of the Protocol and the required documentation.

LMOs imported for food, feed, or processing (FFP) are exempt from the AIA procedure, and are covered under Article 11 and Annex II of the Protocol. Under Article 11, Parties must post decisions to the Biosafety Clearinghouse databa se on domestic use of LMOs for FFP that may be subject to transbou ndary movement. To f acilitate compliance with obligations to this protocol, the U.S. Government has developed a website that provides the status of all regulatory reviews completed for different uses of bioengineered products (see the NBII listings posted at http ://usbiotechreg.nbii.gov/). These data will be ava ilable to the Biosafety Clearinghouse.

APHIS continues to work toward harmonizati on of biosafety and biotechnology consensus documents, guidelines, and regulations, incl uding within the North American Plant Protection Organization (NAPPO ), which includes Mexico, Canada, and the U.S., and within the Organization for Economic Cooperati on and Development (OECD). NAPPO has completed three modules of the Regional Standards for Phytosanitary Measures (RSPM) No. 14, Importation and Release into the Envir onment of Transgeni c Plants in NAPPO Member Countries (NAPPO, 2003).

APHIS also participates in the North American Biotechnology Initiative (NABI) , a forum for information exchange and cooperation on agri cultural biotechnology issues for the U.S., Mexico, and Canada. In addition, bilateral discussions on biotechnology regulatory issues are held regularly with other c ountries including Argentina, Braz il, Japan, China, and Korea.

4.10.3 Compliance with Clean Water Act and Clean Air Act

This EA evaluated the changes in cotto n production due to the unrestricted use of TwinLinkTM Cotton . Cultivation of TwinLinkTM Cotton is not expected to lead to the increased production of cotton in U.S. agriculture.BAYER TWINLINKTM COTTON

85 There is no expected change in water use and quality due to the cultivation of TwinLinkTM
Cotton compared with current cotton seed and co tton production. Also, there is no expected change in air quality associated with the cultivation of TwinLinkTM Cotton.

Based on this review, APHIS concludes that the cultiva tion of TwinLinkTM Cotton would comply with the Clean Water Act and the Clean Air Act.

4.10.4 Impacts on Unique Characte ristics of Geographic Areas

A determination of non-regulated status of TwinLinkTM Cotton is not expected to impact unique characteristics of geographic areas such as pa rk lands, prime farm lands, wetlands, wild and scenic areas, or ecologi cally critical areas.

The common agricultural practices that would be carried out in the cultivation of TwinLinkTM Cotton are not expected to deviate from current practices. The produc t is expected to be deployed on agricultural land cu rrently suitable fo r production of cotton and replace existing varieties, and is not expe cted to increase the acr eage of cotton production.

There are no proposed major ground disturbances; no new physical destru ction or damage to property; no alterations of prope rty, wildlife habitat, or landscapes ; and no prescribed sale, lease, or transfer of ownership of any property. Th is action is limited to a determination of nonregulated status of TwinLinkTM Cotton. Th is action would not convert land use to nonagricultural use and therefore would have no adverse impact on prime farm land. Standard agricultural practices for land prep aration, planting, irrigation, and harvesting of plants would be used on agricultural lands planted to TwinLi nkTM Cotton, including the use of EPA registered pesticides. The Applicant’s adherence to EPA label use restrictions for all pesticides is expected to mitigate potential impacts to the human environment.

4.10.5 National Historic Preservation Ac t (NHPA) of 1966 as Amended

The NHPA of 1966 and its implementing regulations (36 CFR 800) require Federal agencies to: 1) determine whether activities they propose cons titute "undertakings" that have the potential to cause effects on historic properties and 2) if so, to evaluate the effects of such undertakings on such historic resources and consult with the A dvisory Council on Hist oric Preservation (i.e., State Historic Preservation Offi ce, Tribal Historic Preservati on Officers), as appropriate.

APHIS’ proposed action, a determ ination of nonregulated stat us of TwinLinkTM Cotton, is not expected to adversely impact cultural resources on tribal properties. A ny farming activity that may be taken by farmers on tribal lands would only be conducted at the tribe’s request; thus, the tribes would have control over any potential conflic t with cultural resource s on tribal properties.

APHIS’ proposed action would have no impact on districts, sites, highways, structures, or objects listed in or eligible for listing in the Na tional Register of Historic Places, nor would it likely cause any loss or destruction of significant scientific, cultural, or historical resources.

APHIS’ proposed action is not an undertaking that may directly or indirectly cause alteration in the character or use of historic properties pr otected under the NHPA. In general, common agricultural activ ities conducted under this action do not have the poten tial to introduce visual,BAYER TWINLINKTM COTTON

86 atmospheric, or noise elements to areas in which they are used that could result in effects on the character or use of historic properties. For exam ple, there is potential for increased noise on the use and enjoyment of a historic property during the operation of tractors and other mechanical equipment close to such sites. Nevertheless, it is expected that this noise would only be temporary and short-term. The cultivation of TwinLinkTM Cotton is not e xpected to change any of these agronomic practices that would result in an adverse impact under the NHPA.BAYER TWINLINKTM COTTON

87 

### 5 BIBLIOGRAPHY

Adam KL (2005). Seed Production and Variety Development for Organic Systems National Center for Appropriate Technology. Retrieved November 3, 2010 from http://www.attra.ncat.org/ attra-pub/PDF/seed_variety.pdf Alabama Cooperative Extension System (1996). C onservation Tillage: Cotton Production Guide, Circular ANR-952 Retrieved November 9, 2010 from http://hubcap.clemson.edu/~blpprt/constill.html Altieri MA. (1999) The ecological role of biodiversity in agroecosystems. Agriculture, Ecosystems and Environment, 74 , 19-31. Altieri MA. (2000) The ecologi cal impacts of transgenic cr ops on agroecosystem health. Ecosystem Health, 6 (1), 13-23. Altieri MA and Letourneau DK. (1982) Vegeta tion management and biological control in agroecosystems. Crop Protection, 1 (4), 405-430. Altieri MA and Letourneau DK. (1984) Vegeta tion diversity and insect pest outbreaks. Critical Reviews in Plant Sciences, 2 , 131-169. Aneja VP, Schlesinger WH, and Erisman JW. (2009) Effects of agricult ure upon the air quality and climate: Research, policy, and regulations. Environmental Science & Technology,
43 (12), 4234–4240. ANZFA (2005) Food Derived from Glyphos ate-Tolerant Cotton Line MON88913 (Final Assessment Report) (Report: Australia and New Zealand Food Authority. Retrieved from AOSCA (2004). Quality Assurance (QA) Program Th e Association of Offi cial Seed Certifying Agencies. Retrieved November 8, 2010 from http://www.certifiedseed .org/PDF/UGAHosted/QA.pdf AOSCA (2010a). General IP Protocol Standards The Association of Official Seed Certifying Agencies. Retrieved November 8, 2010 from http://www.identitypreserved.c om/handbook/aosca-general.htm AOSCA (2010b). Cotton Retrieved November 3, 2010 from http://www.aosca.org/Cotton.html Arizona (2009). Species Lists Arizona Game a nd Fish Department. Retrieved January 24, 2011 from http://www.azgfd.gov/w_c/edits/hdms_species_lists.shtml Baier AH (2008). Organic Standards for Crop Production National Cent er for Appropriate Technology. Retrieved December 7, 2010 from http://attra.ncat.org/attrapub/PDF/nopstandard_crops.pdf Baker J, Southard R, and Mitchell J. (2005) Agricultural dust production in standard and conservation tillage systems in the San Joaquin Valley. Journal of Environmental Quality, 34 , 1260-1269. Bayer (2010). Petition for Determination of N onregulated Status for Insect-Resistant and Glufosinate Ammonium-Tolerant Cott on: TwinLinkTM Cotton (Events T304-40 x GHB119). Submitted by AL Scott, Regulatory Affairs Manager. Bayer CropScience LP (See Table http://www.aphis.usda.gov/ biotechnology/not_reg.html)

. BCS (2010a). Ignite® 280 SL Herbicide Label Ba yer Crop Science. Retrieved January 2, 2011 from http://fs1.agrian.com/pdfs/Ignite_280_SL_Herbicide_Label3.pdf BCS (2010b). 2010 Trait and Technology Use Ma nual. Submitted by Registration Manager. Bayer Crop Science (See TableBAYER TWINLINKTM COTTON

88 Benbrook C (2009) Impacts of Genetically Engine ered Crops on Pesticide Use in the United States: The First Thirteen Years. Critical Issue Report Number 3 (Report: The Organic Center. Retrieved from Betz FS, Hammond BG, and Fuchs RL. (2000) Safety and advantages of Bacillus thuringiensis

protected plants to control insect pests. Regulatory Toxicology and Pharmacology, 32 (2),
156-173. Bigler F, Sanvido O, and Romeis J (2008) Can ge netically modified plan ts play a role in sustainable crop protection? (Report: Zürich, Switzerla nd: Agroscope ReckenholzTänikon Research Station ART. Retrieved from Blasi DA and Drouillard J (2002) Composition and Feeding Value of Cottonseed Feed Products for Beef Cattle (Report: Kansas State Univer sity Agricultural Experiment Station and Cooperative Extension Service. Retrieved May 2002, from Bradford KJ (2006). Methods to Maintain Gene tic Purity of Seed Stocks University of California, Division of Agricu lture and Natural Resources. Retrieved November 8, 2010 from http://ucanr.org/freepubs/docs/8189.pdf Brookes G and Barfoot P (2010) GM Crops: Global Soci o-Economic and Environmental Impacts 1996-2008 (Report: United Kingdom: PG Ec onomics Ltd. Retrieved from Brown JR. (2003) Ancient horizontal gene transfer. Nature Reviews: Genetics, 4 , 121-132. Calhoun MC (2011) Cottonseed Meal and Whole cottonseed: Optimizing Their Use in Dairy Cattle Rations (Report: San Angelo, TX: Te xas Agricultural Experiment Station. Retrieved from www.cottonseed.com/publications/default.asp Cantrell RG. (2006). The Role of Biotechnology in Impr oving the Sustainability of Cotton. Presented at 65th Plen ary Meeting, Brazil. Cattaneo MG, Yafuso C, Schmidt C, Huang C, Rahman M, Olson C, Ellers-Kirk C, Orr BJ, Marsh SE, Antilla L, Dutilleul P, and Carri ère Y. (2006) Farm-scale evaluation of the impacts of transgenic cotton on biodi versity, pesticide use, and yield. Proceedings of the National Academy of Sciences, 103 (20), 7571–7576. Causarano HJ, Franzluebbers AJ, Reeves DW , and Shaw JN. (2006) Soil organic carbon sequestration in cotton production systems of the Southeastern United States: A review. Journal of Environmental Quality, 35 (4), 1374-1383. CBD (2010). The Cartegena Protocol on Bios afety Convention on Bi ological Diversity. Retrieved January 31, 2011 from http://www.cbd.int/biosafety/ CFIA (2004). Decision Docume nt DD2004-49 Determination of the Safety of Bayer CropScience's Herbicide Tolerant Li bertyLink® Cotton Event LLCotton25 ( Gossypium hirsutum L. ) Canadian Food Inspection Agency. Retrieved December 11, 2010 from http://www.inspection.gc.ca/english/plaveg/bio/dd/dd0449e.shtml Clarke C (2007). Gene Flow between Genetically Modified and Non-GM Plants Retrieved January 9, 2011 from http://cosmos.ucdavis.edu/archiv es/2007/cluster1/c larke_cornelia.pdf Coile NC and Garland MA. (2003) Notes on Flor ida's Endangered and Threatened Plants. Bureau of Entomology, Nematology, and Plant Pathology ~ Botany Section Contribution,
38 (4), 127. Cotton Incorporated (2010a). Co tton and Water Cotton Incorporat ed. Retrieved November 8, 2010 from http://cottontoday.cottoninc.c om/natural-resources/water/BAYER TWINLINKTM COTTON

89 Cotton Incorporated (2010b). Cotton Production Water Requireme nts Cotton Incorporated. Retrieved November 8, 2010 from http://www.cottoninc.com/WaterManagement/Cotton-Water-Requirements/ Cotton Incorporated (2010c). Soil Cotton Inco rporated. Retrieved November 8, 2010 from http://cottontoday.cottoninc.c om/natural-resources/soil/ Cotton Incorporated (2010d). Air Quality Cott on Incorporated. Retrieved November 8, 2010 from http://cottontoday.cottoninc.com /natural-resources/air-quality/ Cotton Incorporated (2010e). Im pact on Habitat & Biodiversity Co tton Incorporated. Retrieved November 8, 2010 from http://cottontoday.cottoninc.com/natural-resources/habitatbiodiversity/ Daniell H. (2002) Molecular strategies fo r gene containment in transgeneic crops. Nature Biotechnology, 20 , 581-586. DeJoode DR and Wendel JF. (1992) Genetic diversity and origin of the Hawaiian Islands cotton, Gossypium tomentosum

. American Journal of Botany, 79 (11), 1311-1319. Delaney B, Astwood J, Cunny H, Conn R, Herouetgui cheney C, Macintosh S, Meyer L, Privalle L, Gao Y, and Mattsson J. (2008) Evaluati on of protein safety in the context of agricultural biotechnology. Food and Chemical Toxicology, 46

. Dickerson DK, Lane EF, and Rodriguez DF ( 1999). Naturally Colored Cotton: Resistance to Changes in Color and Durability when Refurbished with Selected Laundry Aids, CATI #990901. Submitted by Registration Manager. California Agricultural Technology Institute (See Table http://cati.csufres no.edu/ip/rese/99/990901.pdf)

. Dively GP. (2005) Impact of transgenic VIP3A x Cry1Ab lepidopteran-resistant field corn on the nontarget arthropod community. Environmental Entomology, 34 (5), 1267-1291. Dively GP and Rose R. (2003). Effects of Bt Transgenic and C onventional Insecticide Control on the Non-Target Natural Enemy Communtiy in Sweet Corn. Presented at 1st International Symposium on Biological C ontrol of Arthropods, Doran J, Sarrantonio M, and Liebig M. (1996) Soil health and sustainability. Advances in Agronomy, 56 , 1-54. Eastham K and Sweet J (2002). Genetically Modi fied Organisms (GMOs): The Significance of Gene Flow through Pollen Transfer. Subm itted by Registration Manager. European Environment Agency (See Table EFSA. (2008) Request from the European Comm ission related to the safeguard clause invoked by Hungary on maize MON810 according to Article 23 of Directive 2001/18/EC. The EFSA Journal, 756 , 1-18. Eggert D, Frederick JR, Robins on SJ, and Bowerman W. (2004). Impact of Soybean Conservation Systems on Bobwh ite Quail Habitat and Mortality . Paper presented at the 26th Southern Conservation Tillage Conf erence, Raleigh, NC. Retrieved from Ellstrand NC. (2003) Current knowledge of gene flow in plants: Implications for transgene flow. Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences,
358 (1434), 1163-1170. Endrizzi JE, Turcotte EL, and Kohel RJ . (1984). Qualitative Genetics, Cytology, and Cytogenetics Cotton (Vol. No. 24). Madison, WI: Agronomy Monograph. FAO (2001). Evaluation of Allergenicity of Gene tically Modified Foods Retrieved December 15, 2010 from http://www.fao.org/docrep/007/y0820e/y0820e00.htm Fernandez-Cornejo J and Caswell M (2006). The Fi rst Decade of Genetically Engineered Crops in the United States Submitted by Registration Manager. U.S. Department of Agriculture,BAYER TWINLINKTM COTTON

90 Economic Research Service (See Table http://www.ers.usda.gov/publ ications/eib11/eib11.pdf)

. Frisvold GB and Reeves JM. (2007). Economy-wide Impacts of Bt Cotton. Presented at Proceedings of the Beltwide Cotton Conferences, New Orleans. Fryxell PA. (1984). Taxonomy and Germplasm Resources Cotton (Vol. No. 24). Madison, WI: Agronomy Monograph. Gadberry S (2011) Alternative Feeds for Beef Ca ttle (Report: University of Arkansas, Division of Agriculture, Cooperative Extens ion Service. Retrieved from www.uaex.edu/other_areas/ publications/pdf/FSA-3047.pdf

, Garbeva P, van Veen JA, and van Elsas JD. (2004) Microbial diversity in soil: Selection of microbial populations by plan t and soil type and implications for disease suppressiveness. Annual Review of Phytopathology, 42 (1), 243-270. Gazaway WS, Lawrence KS, Akridge JR, and Monks CD. (2007). Crop rotation for the control of reniform nematodes . Department of Entomol ogy & Plant Pathology: Auburn University. Auburn, AL. Gianessi LP and Carpenter JE. (1999) Agricult ural Biotechnology: Ins ect Control Benefits. (pp. 98). Washington, DC: National Center for Food and Agricultural Policy. Gould FW. (2003) Bt-resistance management--theory meets data. Nature Biotechnology, 21 (12),
1450-1451. Guerena M and Sullivan P (2003). Organic Cotton Production National Center for Appropriate Technology. Retrieved November 3, 2010 from http://attra.ncat.org/attrapub/PDF/cotton.pdf Hardee DD, Van Duyn J, Layton MB, and Bagwell RD (2001) Bt Cotton and Management of the Tobacco Budworm-Bollworm Complex (Report: ARS-154). United States Department of Agriculture, Agricultural Resear ch Service. Retrieved from Harlan JR. (1975) Our vani shing genetic resources. Science, 188 (4188), 618-621. Hartman HT and Kester DE. (1975) Plant Propagation: Principles and Practices . Englewood Cliffs, NJ: Prentice-Hall, Inc. Hawaii (2001). Threatened and Endangered Plants of Hawaii State of Hawaii. Retrieved January 24, 2011 from http://www.state.hi.us/dlnr/dofaw/pubs/TEplant.html Head G, Moar W, Eubanks M, Freeman B, Ruberson J, Hagerty A, and Turnipseed S. (2005) A multiyear, large-scale compar ison of arthropod populations on commercially managed Bt and non- Bt cotton fields. Environmental Entomology, 34 (5), 1257-1266. Hérouet C, Esdaile DJ, Mallyon BA, Debruyne E, Schulz A, Currier T, Hendrickx K, van der Klis R-J, and Rouan D. (2005) Safe ty evaluation of the phosphinothricin acetyltransferase proteins encoded by the pat and bar sequences that confer tolerance to glufosinate-ammonium herbicide in transgenic plants. Regulatory Toxicology and Pharmacology, 41 (2), 134-149. Hoeft RG, Nafziger ED, Johns on RR, and Aldrich SR. (2000) Modern Corn and Soybean Production . Champaign, IL: MCSP Publications. Hofmann C, Luthy P, Hutter R, and Pliska V. (1988a) Binding of the delta endotoxins from Bacillus thuringiensis to brush-border membrane vesicles of the cabbage butterfly (Pieris brassicae). European Journal of Biochemistry, 173 , 85-91. Hofmann C, Vanderbruggen H, Höfte H, VanRie J, Jansens S, and VanMellaert H. (1988b) Specificity of Bacillus thuringiensis δ -endotoxins is correlated with the presence of high-BAYER TWINLINKTM COTTON

91 affinity binding sites in the brush borde r membrane of target insect midguts. Proceedings of the National Academy of Sciences of the United States of America, 85 , 7844-7848. Hogan R, Stiles S, Bryant KJ, and Cochran MJ (2005). Reducing the co st of cotton production University of Arkansas Cooperative Extensi on Service Printing Services. Retrieved December 3, 2010 from http://deltafarmpress.com/markets/reducing-cost-cottonproduction Hönemann L, Zurbrügg C, and Nentwig W. ( 2008) Effects of Bt-corn decomposition on the composition of the soil meso- and macrofauna. Applied Soil Ecology, 40 (2), 203-209. Hutmacher RB, Vargas RN, and Wright SD (20 06). Methods to Enable the Coexistence of Diverse Cotton Production Systems. Submitted by Registration Manager. University of California, Division of Agricultur e and Natural Resources (See Table http://anrcatalog.ucdavis.edu/pdf/8191.pdf)

. Icoz I, Saxena D, Andow A, Zwahlen C, a nd Stotzky G. (2008) Microbial populations and enzyme activities in soil in situ under tr ansgenic corn expressi ng cry proteins from Bacillus thuringiensis

. Journal of Environmental Quality, 37 (2), 647-662. ILSI (2008). International Life Sciences Inst itue Crop Composition Database, 3.0 International Life Sciences Institute. Re trieved March 30, 2010 from http://www.cropcomposition.org/ Institute for Regional Conservation (2010). W ild cotton, Upland cotton The Institute for Regional Conservation. Retrie ved November 3, 2010 from http://www.regionalconservation.org/beta/nfyn IPCC (2007). Subsection 14.4.4: Agriculture, Forest ry and Fisheries. Submitted by Registration Manager. Cambridge University Press (See Table http://www.ipcc.ch/publications_and _data/ar4/wg2/en/ch14s14-4-4.html)

. IPPC (2010). Official Web Site for the Internat ional Plant Protection C onvention: International Phytosanitary Portal, International Plant Pr otection Convention. Re trieved November 30, 2010 from https:// www.ippc.int/IPP/En/default.jsp Isengildina-Massa O and MacDonald S (2009) U.S. Cotton Prices and the World Cotton Market: Forecasting and Structural Change (Report: Economic Research Report Number 80). US Department of Agriculture, Economic Re search Service. Retrieved from ITIS (2010). Gossypium thurberi Todaro Integrated Taxonomic Information System. Retrieved November 3, 2010 from http://www.itis.gov/servlet/SingleRpt/Singl eRpt?search_topic=TSN&search_value=5028
19 Jump PM, Longcore T, and Rich C. (2006) Ecol ogy and distribution of a newly discovered population of the federally threatened Euproserpinus euterpe (Sphingidae). Journal of the Lepidopterist’s Society, 60 (1), 41-50. Kaneko T, Nakamura Y, Sato S, Asamizu E, Kato T, Sasamoto S, Watanabe A, Idesawa K, Ishikawa A, Kawashima K, Kimura T, Kish ida Y, Kiyokawa C, Kohara M, Matsumoto M, Matsuno A, Mochizuki Y, Nakayama S, Nakazaki N, Shimpo S, Sugimoto M, Takeuchi C, Yamada M, and Tabata S. (2000) Complete genome structure of the nitrogen-fixing symbiotic bacterium Mesorhizobium loti (supplement). DNA Research, 7

, 381-406. Kaneko T, Nakamura Y, Sato S, Minamisawa K, Uchiumi T, Sasamoto S, Watanabe A, Idesawa K, Iriguchi M, Kawashima K, Kohara M, Matsumoto M, Shimpo S, Tsuruoka H, WadaBAYER TWINLINKTM COTTON

92 T, Yamada M, and Tabata S. (2002) Comp lete genomic sequence of nitrogen-fixing symbiotic bacterium Bradyrhizobium japonicum USDA110. DNA Research, 9 , 189-197. Kareiva P, Morris W, and Jacobi CM. (1994) Studying and managing the risk of crossfertilization between transgenic crops and wild relatives. Molecular Ecology, 3 , 15-21. Keese P. (2008) Risks from GMOs due to horizontal gene transfer. Environmental Biosafety Research, 7 (3), 123-149. Koonin EV, Makarova KS, and Aravind L. (2001) Horizontal gene transfer in prokaryotes: Quantification and classification. Annual Review of Microbiology, 55 , 709-742. Krueger JE (2007). If Your Farm Is Organic, Must It Be GMO Free? Organic Farmers, Genetically Modified Organisms, and the Law. Submitted by Registration Manager. Farmers’ Legal Action Group, Inc. (See Table http://www.flaginc.org/topics/ pubs/arts/OrganicsAndGMOs2007.pdf)

. Lee J (1996). A New Spin on Naturally Colored Cottons Agricultural Research Magazine. Retrieved November 8, 2010 from http://www.ars.usda.gov/is/a r/archive/apr96/cotton0496.htm Leep R, Undersander D, Min D, Harrigan T, a nd Grigar J (2003). Step s to Successful No-till Establishment of Forages. Submitted by Registration Manager. Michigan State University Extension (See Table http://www.fieldcrop.msu.edu/documents/E2880.pdf)

. Lemaux PG. (2009) Genetically engineered plants a nd foods: A scientist's analysis of the issues (part II). Annual Review of Plant Biology, 60 , 511-559. Lovett S, Price P, and Lovett J (2003). Mana ging Riparian Lands in the Cotton Industry. Submitted by Registration Manager. Cotton Research and Development Corporation (See Table http://live.greeningaustralia .org.au/nativevegetation/pa ges/pdf/Authors%20L/18_Lovett_ Price.pdf)

. Marra MC and Martin S (2007). Important Innov ations in Cotton Production: An Assessment by U.S. Cotton Growers and Other Experts. Retrieved from http://cipm.ncsu.edu/cipmpubs/marra_cotton07.pdf Marvier M, McCreedy C, Regetz J, and Karieva P. (2007) A meta-ana lysis of effects of Bt cotton and maize on nontarget invertebrates. Science, 316 , 1475-1477. McClelland MR, Barrentine JL, Brya nt KJ, and Webster EP. (2000). Roundup Ready® Cotton Programs in Conservation Tillage . Paper presented at the Arkansas Cotton Research Meeting, Fayettesville, AR. Retrieved from http://arkansasagnews.uark.edu/198su32.pdf McGregor SE. (1976). Insect Pollination of Cultivated Crop Plants. USDA Agriculture Handbook: AH496 . Washington, DC: U.S. Gove rnment Printing Office. Mendelsohn M, Kough J, Vaituzis Z, and Ma tthews K. (2003) Are Bt crops safe? Nature Biotechnology, 21 (9), 1003-1009. Messeguer J. (2003) Gene flow asse ssment in transgenic plants. Plant Cell, Tissue and Organ Culture, 73 , 201-212. Meyer L, MacDonald S, and Foreman L (2007) Cotton Backgrounder No. CWS-07B-01 (Report: U.S. Department of Agriculture, Econom ic Research Service. Retrieved from http://www.ers.usda.gov/publicatio ns/cws/2007/03Mar/CWS07B01/CWS07B01.pdf Miller DK, Culpepper AS, Kelly ST, Lanclos DY, Stewart AM, Griffin JL, Webster EP, Strahan RE, Vidrine PR, Williams BJ, and York AC. (2004). When Crops Become Weeds: Control Strategies. Louisiana Agriculture Magazine

.BAYER TWINLINKTM COTTON

93 Moar WJ, Eubanks M, Freeman B, Turnipseed S, Ruberson J, and Head G. (2003). Effects of Bt cotton on biological control agents in the southeastern United States. Presented at 1st International Symposium on Biol ogical Control of Arthropods, NAPPO (2003). Regional Standards for Phytos anitary Measures (RSPM) 14: Importation and Release (into the Environment) of Transgenic Plants in NAPPO Member Countries Retrieved December 6, 2010 from http://www.nappo.org/St andards/Std-e.html Naranjo SE. (2005a) Long-term assessment of th e effects of transgenic Bt cotton on the abundance of nontarget arthropod natural enemies. Environmental Entomology, 34 (5),
1193-1210. Naranjo SE. (2005b) Long-term assessment of the e ffects of transgenic Bt cotton on the function of the natural enemy community. Environmental Entomology, 34 (5), 1211-1223. Naranjo SE. (2009a) Impacts of Bt crops on non-targ et invertebrates and insecticide use patterns. CAB Review: Perspectives in Agriculture, Ve terinary Science, Nutrition and Natural Resources, 4 (11), 1-23. Naranjo SE (2009b). Bt Crops and Invertebrate Non-target Effects – Revisited. Submitted by Registration Manager. U.S. Department of Agriculture, Agricultural Research Service, Arid-Land Agricultural Res earch Center (See Table National Cotton Council of America (2010). U.S. Cotton Planted Acres National Cotton Council of America. Retrieved January 11, 2011 from http://www.cotton.org/econ/cropinfo/pr oduction/us-cotton-pl anted-acres.cfm National Cottonseed Products A ssociation (2002). Cottonseed Feed Products Guide Retrieved December 15, 2010 from http://www.cottonseed.com/publi cations/feedproductsguide.asp National Cottonseed Products Association ( 2011). Supply and Demand Estimates for U.S. Cotton and Cottonseed Products Re trieved January 17, 2011 from http://www.cottonseed.com/Members/Statistics/cs&ip_sd.pdf NBII (2010). United States Regulatory Agencies Unified Biotechnology Website United States National Biological Information Infrastruc ture, Center for Biological Informatics Geological Survey. Retrieve d November 12, 2010 from http://usbiotechreg.nbii.gov/ NCAT (2003). NCAT's Organi c Crops Workbook: A Guide to Sustainable and Allowed Practices National Center for Appropriate Technology. Retrieved November 8, 2010 from http://attra.ncat.org/att ra-pub/PDF/cropsworkbook.pdf Nelson RG, Hellwinckel CM, Brandt CC, West TO , De La Torre Ugarte DG, and Marland G. (2009) Energy use and carbon dioxide emissions from cropland production in the United States, 1990–2004. Journal of Environmental Quality, 38 (2), 418-425. NSFC-IPM (2010). Crop Profiles and Timelin es National Science Foundation National Information System of the Regional Integrated Pest Management Ce nters. Retrieved January 13, 2010 from http://www.ipmcenters.org/cropprofiles/index.cfm O'Sullivan J and Sikkema PH. (2004) Response of glufosinate-resistant Bt sweet corn ( Zea

mays ) cultivars to glufosinat e and residual herbicides. Canadian Journal of Plant Science, 85 , 285-289. OECD (2004) Consensus Docume nt of Compositional Consider ations for New Varieties of Cotton ( Gossypium hirsutum and Gossypium barbadense ): Key food and Feed Nutrients and Anti-Nutrients. ENV/JM/MONO(2004)16 (R eport: Organization for Economic Cooperation and Development. Retrieved fromBAYER TWINLINKTM COTTON

94 OECD (2007) Consensus Document on Safety Information on Transgenic Plants Expressing Bacillus thuringiensis - Derived Insect Contro l Proteins. ENV/JM/MONO(2007)14 (Report: Organization for Economic Co-opera tion and Development. Retrieved from OECD (2008) Consensus documen t of the biology of cotton ( Gossypium spp.). ENV/JM/MONO(2008)33 (Report: Organization for Economic Co-operation and Development. Retrieved from OECD (2009) Consensus Document on Compositi onal Considerations for New Varieties of Coton ( Gossypium hirsutum and Gossypium barbadense ): Key Food and Feed Nutrients and Anti-Nutrients. ENV/JM/MONO(2004)16 (R eport: Organization for Economic Cooperation and Development. Retrieved from Palmer WE and Bromley PT (2010). Pesticides and Wildlife - Cotton Retrieved November 9, 2010 from http://ipm.ncsu.edu/wildlife/cotton_wildlife.html Peet M (2001). Conservation Tillage Retrieved December 5, 2010 from http://www.ncsu.edu/sustai nable/tillage/tillage.html Pleasants JM and Wendel JF. (2010) Reproduc tive and pollination bi ology of the endemic Hawaiian cotton, Gossypium tomentosum (Malvaceae). Pacific Science, 64 (1), 45-55. PPDB (2010). Glufosinate-ammonium Retrieved December 8, 2010 from http://sitem.herts.ac.uk/aeru /footprint/en/Reports/372.htm Quist D. (2010). Vertical (Trans)gene Flow: Imp lications for Crop Diversity and Wild Relatives. Biotechnology & Biosafety Series 11 . Penang, Malaysia: Third World Network. Riddle J (2004) Best Management Practi ces for Producers of GMO and non-GMO Crops (Report: The School of Agriculture, Univ ersity of Minnesota. Retrieved from http://swroc.cfans.umn.edu/ prod/groups/cfans/@pub/@cfans /@swroc/documents/asset/cf ans_asset_229637.pdf Romeis J, Meissle M, and Bigler F. (2006) Transgenic crops expressing Bacillus thuringensis toxins and biological control. Nature Biotechnology, 24 (1), 63-71. Ronald P and Fouche B (2006) Genetic Engineeri ng and Organic Production Systems. (Report: ). Oakland, CA: University of California, Divisi on of Agriculture and Natural Resources. Retrieved November 8, 2010, from http://anrcatalog.ucdavis.edu/pdf/8188.pdf Rose R, Dively GP, and Pettis J. (2007) Effects of Bt corn pollen on honey bees: Emphasis on protocol development. Apidologie, 38 (4), 368-377. Ruiz N, Lavelle P, and Jimenez J (2008). Soil Macrofauna Field Manua l: Technical Level Food and Agriculture Organization of the United Nations. Retrieved January 17, 2011 from ftp://ftp.fao.org/docrep/fao/011/i0211e/i0211e.pdf Sankula S, Marmon G, and Blumenthal E (2005 ) Biotechnology-Derived Crops Planted in 2004 - Impacts on US Agriculture (Report: Wash ington: National Center for Food and Agricultural Policy. Retrieved from http://www.whybiotech.com/resources/tp s/BiotechnologyDerivedCropsPlantedin2004.pd f Sanvido O, Stark M, Romeis J, and Bigler F (20 06) Ecological Impacts of Genetically Modified Crops: Experiences from Ten Years of Expe rimental Field Research and Commercial Cultivation (Report: Zürich, Switzerland: Agroscope Reckenholz-Tänikon Research Station ART. Retrieved from Sharpe T. (2010). Cropland Management ( Chapter 4 ). In MD Jones and JS Braden (Eds.), Tarheel Wildlife: A Guide for Managing Wildlife on Private Lands in North Carolina (pp. 26-29). Raleigh: North Carolin a Wildlife Resources Commission.BAYER TWINLINKTM COTTON

95 Shelton AM, Naranjo SE, Romeis J, Hellmich RL, Wolt JD, Federici BA, Albajes R, Bigler F, Burgess EP, Dively GP, Gatehouse AM, Malone LA, Roush R, Sears M, and Sehnal F. (2009) Setting the record stra ight: a rebuttal to an errone ous analysis on transgenic insecticidal crops and natural enemies. [Letter]. Transgenic research, 18 (3), 317-322. Shen RF, Cai H, and Gong WH. (2006) Transg enic Bt cotton has no apparent effect on enzymatic activities or functional diversity of microbial communities in rhizosphere soil. Plant Soil, 285 , 149-159. Shimada NK, Miyamoto K, Kanda K, and Murata H. (2006b) Bacillus thuringiensis insecticidal Cry1Ab toxin does not affect the membrane integrity of the mammalian intestinal epithelial cells: an in vitro study. In Vitro Cellular & Developmental Biology - Animal,
42 , 45-49. Showalter AM, Heuberger S, Tabashnik BE, and Carrière Y. (2009) A primer for using transgenic insecticidal cotton in developing countries. Journal of Insect Science, 9 (22),
39. Sisterson MS, Biggs RW, Manhardt NM, Carrièr e Y, Dennehy TJ, and Tabashnik BE. (2007) Effects of transgenic Bt cotton on insecticide use and abundance of two generalist predators. Entomologia Experiment alis et Applicata, 124 (3), 305-311. Stewart CN (2008). Gene Flow and the Risk of Tr ansgene Spread University of Tennessee, Plant Molecular Genetics

Retrieved November 30, 2010 from http://agribiotech.info/details/StewartGeneFlow%20Mar%208%20-%2003.pdf Stewart S, York A, and Vidrine PR. (2003) Volunteer cotton problem in soybeans. Delta Farm Press, 2010 (November 10). Stewart SD. (2007) Bt Cotton The University of Tennessee Extension Publication W129 , 3. Sundstrom FJ, Williams J, Van Deynze A, and Bradford KJ (2002). Identity Preservation of Agricultural Commodities. University of Calif ornia, Division of Agriculture and Natural Resources. Retrieved Publication 8077, from http://anrcatalog.ucdavis.edu/pdf/8077.pdf Tabashnik BE, Gassmann AJ, Crowder DW, and Carriere Y. (2008) Insect resistance to Bt crops: Evidence versus theory. Nature Biotechnology, 26 (2), 199-202. Thies SA. (1953) Agents concerned with natural crossing of cotton in Oklahoma. Agronomy Journal, 45 , 481-484. Torres JB and Ruberson JR. (2005) Canopy- a nd ground-dwelling predatory arthropods in commerical Bt and non-Bt cotton fi elds: patterns and mechanisms. Environmental Entomology, 34 (5), 1242-1256. Torres JB and Ruberson JR. (2007) Abundance a nd diversity of ground-dwelling arthropods of pest management importance in commercial Bt and non-Bt cotton fields. Annals of Applied Biology, 150 (1), 27-39. Torres JB, Ruberson JR, and Ad ang MJ. (2006) Expression of Bacillus thuringiensis Cry1Ac protein in cotton plants, ac quisition by pests and predator s: A tritrophic analysis. Agricultural and Forest Entomology, 8 , 191-202. Towery D and Werblow S (2010) Facilitating Co nservation Farming Practices and Enhancing Environmental Sustainability with Agricultu ral Biotechnology (Re port: West Lafayette, IN: Conservation Technology Information Center,. Retrieved from http://www.utextension.utk.e du/publications/wfiles/W129.pdf University of Arkansas Cooperative Extensi on Service (2006). Cotton Insect Management: Beneficial Insects and Diseases of Cott on University of Arkansas, Division ofBAYER TWINLINKTM COTTON

96 Agriculture. Retrieve d November 9, 2010 from http://www.aragriculture.org/ins ects/cotton/beneficial.htm#Spiders University of Arkansas Cooperative Extensi on Service (2011). Beef Cattle Nutrition and Feeding: Frequently Asked Questions, Feed ing Cottonseed Meal to Developing Bulls Retrieved March 28, 2011 from www.aragriculture.org/livest ock/beef/nutrition/frequent _questions/cottonseed.htm University of California IPM Online (2008a). Cotton: Crop Rotation Agriculture and Natural Resources, University of California. Retrieved November 9, 2010 from http://www.ipm.ucdavis.edu/PMG/r114900611.html University of California IPM Online (2008b). Cott on: Transgenic Cotton Agriculture and Natural Resources, University of California. Retrieved November 9, 2010 from http://www.ipm.ucdavis.edu/PMG/r114901011.html University of California IPM Online (2009b) UC IPM Pest Management Guidelines: Cotton (Report: Publication 3444). University of Ca lifornia Agriculture and Natural Resources. Retrieved from http://www.ipm.ucdavis.edu/PMG/selectnewpest.cotton.html University of California IPM Online (2009c). Cott on: Integrated Weed Ma nagement Agriculture and Natural Resources, University of Calif ornia. Retrieved November 9, 2010 from http://www.ipm.ucdavis.edu/PMG/r114700111.html University of Hawaii at Manoa (2001). Hawaiian Native Plant Propagation Database: Gossypium

tomentosum Retrieved January 17, 2010 from http://www2.hawaii.edu/~eherring/hawnprop/gos-tome.htm University of Illinois (2010). Bacterial Disease, Crown Gall , Agrobacterium tumefaciens Retrieved December 19, 2010 from http://urbanext.illinois.edu/hortans wers/detailproblem.cfm?PathogenID=23 University of Tennessee Agricultural Extens ion Service (2010). Beaver Creek Study Final Report: Making a Splash AE03-63 Retrieved January 20, 2011 from http://economics.ag.utk.edu/bcstudy.html US-EPA (2001a). Pesticide Registration (P R) Notice 2001-5: Noti ce to Manufacturers, Formulators, Producers and Registrants of Pesticide Products. Submitted by Registration Manager. U.S. Environmental Protection Ag ency, Office of Pesticide Programs (See Table www.epa.gov/PR_notices/pr2001-5.pdf)

. US-EPA (2001b). Bt Plant-Incorporated Protect ants October 15, 2001 Biope sticides Registration Action Document Retrieved January 22, 2010 from http://www.epa.gov/opp00001/biopesticid es/pips/bt_brad2/5-benefits.pdf US-EPA (2004). Overview of the Ecological Risk Assessment Process in the Office of Pesticide Programs, U.S. Environmental Protection Agency. Submitted by Registration Manager. U.S. Environmental Protection Agency, O ffice of Prevention, Pesticides and Toxic Substances, Office of Pestic ide Programs (See Table US-EPA (2008a) Biopesticides Registration Action Document: Bacillus thuringiensis modified Cry1Ab (SYN-IR67B-1) and Vip3Aa19 (SYN-IR102- 7) insecticidal proteins and the genetic material necessary for their produc tion in COT102 X COT67B cotton (Report: Retrieved from http://www.epa.gov/oppbppd1/piopesticid es/ingredients/t ech_docs/brad_006529.pdf US-EPA (2008b). Glufosinate Final Work Pl an Registration Review August 2008. Submitted by S Bradbury, Registration Manager. U.S. Envi ronmental Protection Agency (See TableBAYER TWINLINKTM COTTON

97 US-EPA (2009). Major crops grown in the Unite d States Retrieved November 17, 2010, from http://www.epa.gov/agricu lture/ag101/cropmajor.html US-EPA (2010a). Introduction to Biotechnology Regulation for Pes ticides Retrieved November 3, 2010 from http://www.epa.gov/oppbppd1/biopesticide s/regtools/biotec h-reg-prod.htm US-EPA (2010b). Inventory of U.S. Gree nhouse Gas Emissions and Sinks: 1990-2008 Submitted by Registration Manager. United States E nvironmental Protection Agency (See Table http://www.epa.gov/climatechange/emissions/)

. US-EPA. (2010c) Glufosinate Ammonium; To lerances for Residues, 40 CFR §180.473 US-EPA. (2010d) Bacillus thuringiensis Cry1Ab Protein in all Pl ants; Exemption from the Requirement of a Tolerance, 40 CFR §174.511. US-EPA. (2010e) Bacillus thuringiensis Cry2Ae Protein in Cotton; Temporary Exemption from the Requirement of a Tolerance, 40 CFR §174.530 US-EPA. (2010f) Issuance of an Experimental Use Permit. Federal Register, 73 (215), 65848. US-EPA. (2010g) Phosphinothricin Acetyltransferase (PAT): Exemption from the Requirement of Tolerance. 40 CFR §174.522, first published at 40 CFR §180.1151. US-FDA (1992). Statement of Policy - Foods Derive d from New Plant Varieties. Retrieved January 18, 2011 from http://www.fda.gov/Food/GuidanceComplianc eRegulatoryInformation/GuidanceDocume nts/Biotechnology/ucm096095.htm US-FDA (2003). Biotechnology Consultation Note to the File BNF No. 000086 Retrieved October 14, 2010 from http://www.fda.gov/Food/Biotec hnology/Submissions/ucm155782.htm US-FDA (2010). The FDA List of Completed Consultations on Bioengineered Foods United States Food and Drug Administratio n. Retrieved May 17, 2010 from http://www.fda.gov/Food/Biotechno logy/Submissions/default.htm US-FWS (2003). Recovery Plan for the Quino Checkerspot Butterfly ( Euphydryas editha quino ). Submitted by Registration Manager. U.S. Fish and Wildlife Service (See Table http://ecos.fws.gov/docs/recovery_plan/030917.pdf)

. US-FWS (2007a). Kern Primrose Sphinx Moth ( Euproserpinus euterpe ) 5-Year Review: Summary and Evaluation. Submitted by Regist ration Manager. U.S. Fish and Wildlife Service (See Table http://ecos.fws.gov/docs/five_year_review/doc1157.pdf)

. US-FWS (2010a). Saint Fran cis' Satyr Butterfly ( Neonympha mitchellii francisci ) Retrieved January 25, 2011 from http://ecos.fws.gov/speciesProfile/profile/s peciesProfile.action? spcode=I0MZ#crithab US-FWS (2011a). Endangered Species Progr am Retrieved January 31, 2011 from http://www.fws.gov/endangered/index.html US-FWS (2011b). Species Reports Retrieved January 25, 2011 from http://ecos.fws.gov/tess_public/SpeciesRe port.do?groups=I&listingType=L&mapstatus=
1 US-NARA (2010). Executive Orders Disposition Tables Index Un ited States National Archives and Records Administration. Re trieved January 31, 2011 from http://www.archives.gov/f ederal-register/executiv e-orders/dis position.html USBG. (2010). Plant Collections: Rare and Enda ngered Plant. Retrieved November 3, 2010, from United States Botanic Garden [Online Database] http://www.usbg.gov/plantcollections/conservation/Gossypium-tomentosumBAYER TWINLINKTM COTTON

98 USDA-AMS (2010a) Cotton Varieties Planted 2010 Crop (Report: Memphis, TN: U. S. Department of Agriculture, Agricultural Marketing Service. Retrieved from http://www.ams.usda.gov/mnreports/cnavar.pdf USDA-AMS (2010b). National Organi c Program Agricultural Marke ting Service, United States Department of Agriculture. Retrieved November 23, 2010 from http://www.ams.usda.gov/AMSv1.0/nop USDA-AMS. (2010c) Minimum Land, Isolation, Field, and Seed Standards, 7 CFR Part 201.76 USDA-APHIS (2003) Approval of Aventis Crop Science USA LP Petition (02-042-01p) Seeking a Determentation of Non-regulated Status fo r Glufosinate-ammonium Herbicide-tolerant Cotton Transformation Event LLCotton25 (Env ironmental Assessment) (Report: U.S. Department of Agriculture, Animal and Plan t Health Inspection Se rvice, Biotechnology Regulatory Services. Retrieved from USDA-APHIS (2010a) Draft Envi ronmental Assessment, Syngenta Biotechnology, Inc. Petition (07-108-01p) for Determination of Nonregulated Status for Lepidopteran-Resistant Event COT67B Cotton (Report: Riverd ale, MD: U.S. Department of Agriculture, Animal and Plant Health Inspection Serv ice. Retrieved from USDA-APHIS (2010b) Determination of Plant Pest Risk for Insect-Resistant and Glufosinate Ammonium-Tolerant (TwinLinkTM) Cotton, Gossypium hirsutum , events T304-40 x GHB119 (Report: U.S. Department of Agricu lture, Animal and Plan t Health Inspection Service. Retrieved from USDA-ARS. (2010a). Taxon: Gossypium thurberi Tod. Retrieved November 3, 2010, from National Germplasm Resources Laboratory [Online Database] http://www.arsgrin.gov/cgi-bin/npgs/html/taxon.pl?17947 USDA-ERS (2005a). Agricultura l Chemicals and Production Tec hnology: Pest Management Retrieved November 3, 2010 from http://www.ers.usda.gov/Briefing/ AgChemicals/sustainability.htm USDA-ERS (2005b). Agricultural Chemicals and Production Technology: Sustainability and Production Systems Economic Research Servic e. Retrieved November 3, 2010 from http://www.ers.usda.gov/Briefing/ AgChemicals/sustainability.htm USDA-ERS (2006). U.S. Textile and Apparel In dustries and Rural America Retrieved December 5, 2010 from http://www.ers.usda.gov/briefi ng/cotton/ustextileapparel.htm USDA-ERS (2009). Cotton: Background Retrieved November 2, 2010 from http://www.ers.usda.gov/Bri efing/Cotton/background.htm USDA-ERS (2010a). Adoption of Ge netically Engineered Crops in the U.S. Economic Research Service. Retrieved November 2, 2010 from http://www.ers.usda.gov/data/BiotechCrops/ USDA-ERS (2010b). Cotton production costs and returns per planted acre, excluding Government payments, 2008-2009 [Data table]. Retrieved December 3, 2010 from http://www.ers.usda.gov/data/cost sandreturns/testpick.htm#recent USDA-ERS (2010c). Adoption of Genetically En gineered Crops in the U.S.: Upland Cotton Varieties Economic Research Service. Retrieved December 3, 2010 from http://www.ers.usda.gov/Data/Biotec hCrops/ExtentofAdoptionTable2.htm USDA-ERS (2010d). Conservation Policy: Co mpliance Provisions for Soil and Wetland Conservation Retrieved December 3, 2010 from http://www.ers.usda.gov/Briefing/Co nservationPolicy/compliance.htmBAYER TWINLINKTM COTTON

99 USDA-ERS (2010e). Table 3--Certified Organic and Total U.S. Acreage, Selected Crops and Livestock, 1995-2008 Economic Research Servi ce. Retrieved November 5, 2010 from http://www.ers.usda.gov/Data/Organic/ USDA-FAS (2010a). Cotton: World Markets and Trad e Foreign Agricultural Service, Office of Global Analysis. Retrieved November 2, 2010 from http://www.fas.usda.gov/cotton/circul ar/2010/october/Co tton_full10-08.pdf USDA-FS (2010) Gossypium hirsutum L. (Report: San Juan, PR: Fore st Service, International Institute of Tropical Forestry. Retrieved from http://www.fs.fed.us/global/iitf/pdf /shrubs/Gossypium%20hirsutum.pdf USDA-NASS (2007). 2007 Census of Agriculture : Cotton Industry Retrieved November 5, 2010 from http://www.agcensus.usda.gov/Publications/ 2007/Online_Highlights/Fact_Sheets/cotton. pdf USDA-NASS (2008). Agricultural Chemical Usage 2007 Field Crops Summary National Agricultural Statistics Service. Retrieved November 6, 2010 from http://usda.mannlib.cornell.edu/usda/cur rent/AgriChemUsFC/AgriChemUsFC-05-21- 2008.pdf USDA-NASS (2010a). U.S. All Cotton Production National Agricultural Statistics Service (NASS). Retrieved October 29, 2010 from http://www.nass.usda.gov/Charts _and_Maps/graphics/cotnprod.pdf USDA-NASS (2010b). U.S. All Cotton Acres Nationa l Agricultural Statisti cs Service (NASS). Retrieved October 29, 2010 from http://www.nass.usda.gov/Charts _and_Maps/graphics/cotnac.pdf USDA-NASS (2010c). 2010 Upland Cotton Harveste d Areas National Agriculture Statistics Service Board. Retrieved November 2, 2010 from http://www.nass.usda.gov/Charts _and_Maps/graphics/cotnacm.pdf USDA-NASS (2010e). Crop Production National Agricultural Statisti cs Service. Retrieved November 5, 2010 from http://usda.mannlib.cornell.edu/usda/n ass/CropProd//2010s/ 2010/CropProd-10-08- 2010.pdf USDA-NRCS (2006a). Conservation Resource Brief: Soil Quality, Number 0601 National Resources Conservation Service. Retrieved November 9, 2010 from http://www.nrcs.usda.gov/feat ure/outlook/Soil%20Quality.pdf USDA-NRCS (2006b). Conservation Resource Brief: Soil Erosion, Number 0602 National Resources Conservation Service. Retrieved November 9, 2010 from http://www.nrcs.usda.gov/feat ure/outlook/Soil%20Erosion.pdf USDA-NRCS (2006c). Conservation Resource Brief: Air Quality, Number 0605 National Resources Conservation Service. Retrieved November 9, 2010 from http://www.nrcs.usda.gov/feat ure/outlook/Air%20Quality.pdf USDA-NRCS (2010) 2007 National Resources Inve ntory: Soil Erosion on Cropland (Report: National Resources Conservation Service. Retrieved from Vadakattu G and Watson S (2004) Ecological Impacts of GM Cotton on Soil Biodiversity: Below Ground Production of Bt by GM Cotton a nd Bt Cotton Impacts on Soil Biological Processes (Report: CSIRO Land an d Water. Retrieved fromBAYER TWINLINKTM COTTON

100 Vaissiere BE and Vinson B. (1994) Pollen mo rphology and its effect on pollen collection by honey bees, Apis mellifera L. (Hymenoptera: Apidae), with special reference to upland cotton, Gossypium hirsutum L. (Malavaceae). Grana, 33 , 128-138. Van Deynze A, Sundstrom FJ, and Bradford KJ. ( 2005) Pollen-mediated gene flow in California cotton depends on pollinator activity. Crop Science, 45 (4), 1565-1570. Van Rie J, Jansens S, Höfte H, Degheele D, and Van Mellaert H. (1989) Specificity of Bacillus thuringiensis δ -endotoxins: Importance of specific receptors on the brush border membrane of the mid-gut of target insects. European Journal of Biochemistry, 186 , 239- 247. Van Rie J, Jansens S, Höfte H, Degheele D, an d Van Mellaert H. (1990) Receptors on the brush border membrane of the insect midgut as determinants of the specificity of Bacillus thuringiensis delta-endotoxins. Applied and Environmental Microbiology, 56 (5), 1378-
1385. Vogel JR, Majewski MS, and Capel PD. (2008) Pestic ides in rain in four agricultural watersheds in the United States. Journal of Environmental Quality, 37 (3), 1101-1115. Whitehouse MEA, Wilson LJ, and Fitt GP. (2005) A Comparison of Arthropod Communities in Transgenic Bt and Conventional Cotton in Australia. Environmental Entomology, 34 (5),
1224-1241. Wilkinson KM and Elevitch CR (2011). The Ov erstory #65: Biological Nitrogen Fixation Retrieved February 24, 2011 from http://www.agroforestry.net/ overstory/overstory65.html Wilkinson MJ and Ford CS. (2007) Estimating the pot ential for ecological ha rm from gene flow to crop wild relatives. Collection of Biosafety Reviews, 3 , 42-63. Williams MR. (2010). Cotton Insect Loss Estimates - 2009. Presented at 2010 Beltwide Cotton Conferences, New Orleans, LA. Wilson EO. (1988) Biodiversity . Washington, DC: National Academy Press. Wood D, Setubal J, Kaul R, Monks D, Kitajima J, Okura V, Zhou Y, Chen L, Wood G, Almeida N, Woo L, Chen Y, Paulsen I, Eisen J, Ka rp P, Bovee S, Chapman P, Clendenning J, Deatherage G, Gillet W, Grant C, Kutyavin T, Levy R, Li M, McClelland E, Palmieri A, Raymond C, Rouse G, Saenphimmachak C, Wu Z, Romero P, Gordon D, Zhang S, Yoo H, Tao Y, Biddle P, Jung M, Krespan W, Perry M, Gordon-Kamm B, Liao L, Kim S, Hendrick C, Zhao Z, Dolan M, Chumley F, Ti ngey S, Tomb J, Gordon M, Olson M, and Nester E. (2001) The genome of the natural genetic engineer Agrobacterium tumefaciens
C58. Science, 294 (5550), 2317-2323. Wozniak CA. (2002). Gene Flow Assessment for Plant-I ncorporated Protectants by the Biopesticide and Pollution Prevention Division, U.S. EPA. Presented at Scientific Methods Workshop: Ecological and Agronomi c Consequences of Gene Flow from Transgenic Crops to Wild Relatives, Columbus, Ohio. Wunderlin R and Hansen B (2010). Atlas of Florid a Vascular Plants Institute for Systematic Botany, University of South Florida. Retrieved December 5, 2010 from http://www.florida.plantatlas.usf.edu/BAYER TWINLINKTM COTTON

101 

### 6 LIST OF PREPARERS

Biotechnology Regulatory Services

Michael Blanchette

Senior Environmental Protection Specialist

David Reinhold

Assistant Director, Environmental Risk Analysis Program

Kham Vongpaseuth

APHIS Fellow

Patricia Beetham, Ph. D.

Biotechnologist

Avatar Environmental LLC

Michael T. Werner, J.D.

Project Manager

Charles Dobroski

Technical Director/PrincipalBAYER TWINLINKTM COTTON

1 

### APPENDIX A

APHIS THREATENED AND ENDANGE RED SPECIES DECISION TREE FOR FWS CONSULTATIONSBAYER TWINLINKTM COTTON

9/9/2011 A-1 

### APPENDIX A

APHIS THREATENED AND ENDANGE RED SPECIES DECISION TREE FOR FWS CONSULTATIONS

DECISION TREE ON WHETHER SECTI ON 7 CONSULTATION WITH FWS IS TRIGGERED FOR PETITIONS OF TRANSGENIC PLANTS

This decision tree document is based on the phenotypes (traits) that have been permitted for environmental releases under APHIS oversight (for a list of approved notifications and environmental releases, visit Inform ation Systems for Biotechnology, at http://isb.vt.edu .) APHIS will re-evaluate and update this decision document as it receives new applications for environmental releases of new traits that are genetically engineered into plants.

BACKGROUND

For each transgene(s)/transgenic plant the follo wing information, data, and questions will be addressed by APHIS, and the EAs on each petition will be publicly available. APHIS review will encompass:

 A review of the biology, taxonomy, and weed iness potential of th e crop plant and its sexually compatible relatives;

 Characterization of each transgene with re spect to its structure and function and the nature of the organism from which it was obtained;

 A determination of where the new transgene and its products (if any) are produced in the plant and their quantity;

 A review of the agronomic performance of the plant including disease and pest susceptibilities, weediness potential, a nd agronomic and environmental impact;

 Determination of the concentrations of know n plant toxicants (if any are known in the plant),

 Analysis to determine if the transgenic plan t is sexually compatible with any threatened or endangered plant species (TES) or a host of any TES.

FDA published a policy in 1992 on foods derived from new plant varieties, including those derived from transgenic plants ( http://vm.cfsan.fda.gov/~lrd/fr92529b.htm l and http://vm.cfsan.fda.gov/~lrd/consulpr.html ). The FDA’s policy requires that genetically engineered foods meet the same rigorous safety st andards as is required of all other foods. Many of the food crops curr ently being developed using biotechno logy do not contain substances that are significantly different from those already consumed by huma n and thus do not require premarket approval. Consistent with its 1992 polic y, FDA expects developers to consult with the agency on safety and regulatory questions. A list of consultations is available at http://vm.cfsan.fda .gov/~lrd/biocon.html . APHIS considers the stat us and conclusion of the FDA consultations in its EAs.BAYER TWINLINKTM COTTON

2 Below is a description of our review process to whether a c onsultation with U.S. Fish and Wildlife Service is necessary.

If the answer to any of the questions 1-4 below is yes, APHIS will contact FWS to determine if a consultation is required:

Is the transgenic plant sexually compatible with a TE plant 1 without human intervention?

1. Are naturally occurring plant toxins (toxica nts) or allelochemicals increased over the normal concentration range in parental plant species?

2. Does the transgene product or its metabolites have any significant si milarities to known
toxins 2 ?

3. Will the new phenotype(s) imparted to the transg enic plant allow the plant to be grown or employed in new habitats (e.g., outside agro-ecosystem) 3

. 4. Does the pest resistance 4 gene act by one of the mechanis ms listed below? If the answer is YES then a consultation with U.S. Fish and Wildlife Service is NOT necessary.

A. The transgene acts only in one or more of the following ways:

i. As a structural barrier to either the attachment of the pest to the host, to penetration of the host by the pest, to the spread of the pest in the host plant (e.g., the production of lignin, callose, thickened cuticles);
ii. In the plant by inactivating or resisting toxins or other disease causing substances produced by the pest;
iii. By creating a deficiency in the host of a component required for growth of the pest (such as with fungi and bacteria);
iv. By initiating, enhancing, or potentiating the endogenous host hypersensitive disease resistance response found in the plant;

v. In an indirect manner that does not result in killing or interfering with normal growth, development, or behavior of the pest;

1 APHIS will provide FWS a draft EA that will address the impacts, if any, of gene movement to the TES plant

2 Via a comparison of the amino acid sequ ence of the transgene’s protein with those found in the protein databases like PIR, Swiss-Prot and HIV amino acid data bases.

3 Such phenotypes might include tolerance to environmental stresses such as drought, salt, frost, aluminum or heavy metals.

4 Pest resistance would include any toxin or allelochemical that prevents, destroys, repels or mitigates a pest or effects any vertebrate or invertebrate animal, plant, or microorganism.BAYER TWINLINKTM COTTON

3 B. A pest derived transgene is expressed in the plant to confer resistance to that pest (such as with coat protein, replicase, and pathogen virulence genes).

For the biotechnologist:

Depending on the outcome of the decision tree , initial the appropriate decision below and incorporate its language into the EA. Retain a hard copy of this decision document in the petition’s file.

________ BRS has reviewed the data in accordance with a proces s mutually agreed upon with the U.S. Fish and Wildlife Service to determin e when a consultation, as required under Section 7 of the Endangered Species Act, is needed. APHIS has reached a determination that the release following a determination of non-regulated status would have no effects on listed threatened or endangered species and consequently, a written conc urrence or formal consultation with the Fish and Wildlife Service is not required for this EA.

________ BRS has reviewed the data in accordance with a proces s mutually agreed upon with the U.S. Fish and Wildlife Service to determin e when a consultation, as required under Section 7 of the Endangered Species Act, is needed. AP HIS reached a determination that the release following a determination of non-regulated status is not likely to adversely affect any listed threatened or endangered species and consequen tly obtained written concurrence from the Fish and Wildlife Service.

________ BRS has reviewed the data in accordance with a proces s mutually agreed upon with the U.S. Fish and Wildlife Service to determin e when a consultation, as required under Section 7 of the Endangered Species Act, is needed. AP HIS reached a determination that the release following a determination of non-regu lated status is likely to aff ect adversely one or more listed threatened or endangered specie s and has initiated a formal c onsultation with the Fish and Wildlife Service.BAYER TWINLINKTM COTTON



### APPENDIX B

THREATENED AND ENDANGERED SPECIES STATE LISTSBAYER TWINLINKTM COTTON

B-1 APPENDIX B

THREATENED AND ENDANGERED SPECIES

Distribution, Habitat, and Food Sources ofThreatened and Endangered Lepidoptera (USFWS, 2011b)

Given the specificity of activity of the Cry prot eins, species outside the insect order Lepidoptera should not be affected (Naranjo, 2009a; Sanvido et al., 2006). APHIS has thoroughly examined all FWS threatened and endangere d lepidopteran species to determine if there could be possible effects from the proteins found in Twin LinkTM Cotton. Threatened and endangered lepidopterans in the U.S. have restrictive habitats; and their larvae typically feed on specific host plants, none of which includes cotton or its sexually compatible rela tives, or plants expected to be found in cultivated agricultura l land. Furthermore, an examin ation of county distribution of endangered lepidopterans shows that most are not known to occur in co unties where cotton is grown, with the exception of the Kern primrose sphinx moth, the Quino Checkerspot butterfly and the Saint Francis’ satyr butte rfly. The EPA (2001b) has alrea dy concluded that the habitats of these species do not overlap with cotton fiel ds and their larvae do not feed on cotton. Pollen dispersal due to wind is considered insignificant because cotton pollen is sticky and heavy (Khan and Afzal, 1950; Thies, 1953). The EPA (2001b) has concluded that the amounts of cotton pollen that might be deposited on host plants of these species would be negligible, or perhaps none at all, and have no impact . If pollen were to deposit on host plants, the amount of pollen that would drift from these cotton plants onto pl ants fed upon by endangered/threatened species, would be very small compared to the levels fed to test species (Bayer, 2010; US-EPA, 2008; USDA-APHIS, 2010b).

Species Description

Butterfly, bay checkerspot Euphydryas editha bayensis The Bay checkerspot butterfly ( Euphydryas editha bayensis ) is restricted to serpentine outcrops near San Francisco Bay, in Santa Clara and San Matao, CA. The primary constituent elements of the habitat for the bay checkerspot are one or more of the following: stands of Plantago erecta

, Castilleja

exserta , or Castilleja densiflora ; spring flowers providing nectar; pollinators of the bay checkerspot’s food and nectar plants; soils derived from serpentinic rock; and space for dispersal between habitable areas (NatureServe, 2010a; US-FWS, 2001a).

Butterfly, Behren's silverspot Speyeria zerene behrensii Currently inhabits one site in southern Mendocino County, CA. The Behren’s silverspot butterfly inhabits coastal prairie habitat that contains Viola adunca (early blue violet), the larval host plant, adult nectar sources, and adult courtship (NatureServe, 2010b; US-FWS, 2004).

Butterfly, callippe silverspot Speyeria callippe callippe Is found in native grassland and associated habitats in the San Francisco Bay area, CA. The females lay their eggs on the dry remains of the larvae foodplant, Johnny jump-up ( Viola pedunculata ), or on the surrounding debris (Butterfly Conservation Initiative, 2006c; US-FWS, 2010b).BAYER TWINLINKTM COTTON

B-2 Species Description

Butterfly, El Segundo blue Euphilotes battoides allyni Now limited to one 302-acr e parcel (owned by the Los Angeles Airport) and one 2-acre parcel (owned by the Standa rd Oil Company of California) in El Segundo Dunes, Los Angeles County, CA in sand dunes with its larval and adult host plant Eriogonum parvifolium (NatureServe, 2010c; US-FWS, 2007b).

Butterfly, Fender's blue Icaricia icarioides fenderi Restricted primarily to the Willamette Valley of Oregon, in upland prairies in Douglas, Benton, Lane, Linn, Polk , and Yamhill Counties, OR and Lewis County, WA. Dry, fescue prairies make up the majority of habitat. Larvae feed exclusively on certain lupine, mainly Lupinus sulphereus var. kincaid occasionally L. laxiflorus and albicauliss (NatureServe, 2010d; US-FWS, 2000a).

Butterfly, Karner blue Lycaeides melissa samuelis Requires the wild lupine ( Lupinus perennis ) that occurs in Illinois, Indiana, Michigan, Minnesota, New Hampshire, New York, Ohio, and Wisconsin to deposit eggs for larval food. In eastern New York and New Hampshire, habitat typically is in sandplain communities, such as grassy openings within very dry, sandy pitch pine/scrub oak barrens. In the Midwest, the habitat is also dry and sandy, including oak savanna and jack pine barrens, and less often dune communities. Within the overall community remnantinhabited by a metapopulation, any patch of foodplant in open to semishaded setting is likely to be used. Females lay eggs on or near wild lupine plants, and main requirement seems to be thousands of stems of lupine in the short term (Nat ureServe, 2010e).

Butterfly, Lange's metalmark Apodemia mormo langei Is currently found only at Antioch Sand Dunes in Contra Costa County, CA, as part of the Antioch Dunes National Wildlife Refuge. The larvae are known to feed only on buckwheat ( Eriogonum nudum ssp. auriculatum ) (US-FWS, 2008).

Butterfly, lotis blue Lycaeides argyrognomon lotis Is found only in Mendocino County, CA in a sphagnum-willow bog of about five acres in size. Larvae probably feed on Lotus formosissimus ; if not then presumably some other legume such as Lathyrus vestitus bolanderi

. If not a legume, some species of Ericaceae would seem most likely. (NatureServe, 2010f).

Butterfly, mission blue Icaricia icarioides missionensis Restricted to a few sites in about three populations, including San Bruno Mountain in San Mateo County, Twin Peaks in San Francisco, and the vicinity of Skyline College in San Mateo County, CA. Limited to grasslands, with its larval hosts, Lupinus albifrons, L. varicolor, and L. formosus (NatureServe, 2010g)

.

Butterfly, Mitchell's satyr Neonympha mitchellii mitchellii Known to occur in Indiana, Michigan, and Ohio in northern limestone wetlands. Females oviposit on a variety of small forbs and sedges (e.g., Thelypeteris palustris, and Carex sp.) seedlings and individuals do not move great distances (Barton and Bach, 20 05; Szymanski et al., 2004; US-FWS, 1998a, 2010c).BAYER TWINLINKTM COTTON

B-3 Species Description

Butterfly, Myrtle's silverspot Speyeria zerene myrtleae Marin, San Mateo, and Sonoma, CA. Myrtle’s silverspot inhabits coastal dunes, coastal prairie, and coastal scr ub at elevations ranging from sea level to 300 meters (1,000 feet), and as far as 5 kilometers (3 miles) inland. The larval food plant is Viola adunca and possibly other Viola species (USFWS, 1998b).

Butterfly, Oregon silverspot Speyeria zerene hippolyta Known to occur in Del Norte County, CA; in Clatsop, Lane, Lincoln, Tillamook and Counties, OR and P acific County, WA in salt-spray meadows with its host Viola adunca (US-FWS, 2001b)

.

Butterfly, Palos Verdes blue Glaucopsyche lygdamus palosverdesensis Occurs only in Los Angeles County, CA, confined to coastal sage scrub community and is dependent on two known host plants, locoweed ( Astragalus trichopodus var. lonchus , also known as Santa Barbara milkvetch) and common deerweed ( Lotus scoparius ) (Butterfly Conservation Initiative, 2006a; US-FWS, 1980).

Butterfly, Quino checkerspot Euphydryas editha quino (=E. e. wrighti) Is known to occur in the San Diego National Wildlife Refuge, in Riverside and San Diego Counties, CA, in Chaparra l, coastal sage scrub, with primary host plants: dwarf plantain ( Plantago erecta ), wooly plantain ( Plantago patagonica ), the white snapdragon ( Antirrhinum coulterianum ), threadleaved bird’s beak ( Cordylanthus rigidus ) (US-FWS, 2003, 2010d).

Butterfly, Saint Francis' satyr Neonympha mitchellii francisci Located in one site in NC sandhills, may be restricted to artillery impact areas at Fort Bragg, military installation in North Carolina. Known only from a few sedge wetlands (NatureServe, 2010h).

Butterfly, San Bruno elfin Callophrys mossii bayensis Restricted to Contra Costa, Marin and San Mateo Counties, CA. The San Bruno elfin is found in coastal mountains near San Francisco Bay, in the fog-belt of steep north facing slopes th at receive little direct sunlight. It lives near prolific growths of th e larval food plant, stonecrop ( Sedum spathulifolium ), which is a low-growing succul ent. Stonecrop is associated with rocky outcrops that occur at 900-1075 feet elev ation (Butterfly Conservation Initiative, 2006b; NatureServe, 2010i; US-FWS, 2010e).

Butterfly, Schaus swallowtail Heraclides aristodemus ponceanus Restricted to Dade and Monroe Counties, FL. Habitat is tropical hardwood hammocks and their edges with the larval foodplant which is torchwood Amyris elemifera . Adults do stray into othe r nearby areas. Larvae feed mainly on Amyris elemifera and occasionally on other Rutaceae (NatureServe, 2010j; US-FWS, 1999).

Butterfly, Smith's blue Euphilotes enoptes smithi Restricted to Monterey, Santa Cruz, and San Mateo counties, CA. Coastal and inland sand dunes and steep slopes along the coast where coastal sanddune strand vegetation dominates. One population found in chaparralwoodland dominated area. Also has been found in serpentine grassland area. Area must contain seacliff and coastal buckwheat. An undescribed ecotype of E. latifolium (coastal buckwheat) is used by the females for oviposition as well as providing food for the larvae (NatureServe, 2010k).BAYER TWINLINKTM COTTON

B-4 Species Description

Butterfly, Uncompahgre fritillary Boloria acrocnema Restricted to: Gunnison, Hinsdale, and Chaffee counties in CO; isolated alpine habitats in the San Juan Mountains of southwestern Colorado. Habitat is moist alpine slopes above 12,000 feet with extensive snow willow ( Salix nivalis) (NatureServe, 2010l; US-FWS, 1994)

.

Moth, Blackburn's sphinx Manduca blackburni Today it occurs on Hawaii, Kalo ̀ owahe, Maui and is more or less restricted to tracts of dry forest. Inhabits mostly lowland dry forests and shrub lands where the larvae feed on native a nd a few introduced Solanaceae (tobacco family). However, adults do wander and larvae have turned up on cultivated Solanaceae. Larvae of Blackburn's sp hinx moth feed on plants in the nightshade family (Solanaceae). The natural host plants are native shrubs in the genus Solanum (popolo), and the native tree, Nothocestrum latifolium (`aiea), on which the larvae consume leaves, stems, flowers, and buds. However, many of the host plants reco rded for this species are not native to the Hawaiian Islands, and include Nicotiana tabacum (commercial tobacco), Nicotiana glauca (tree tobacco), Solanum melongena (eggplant), Lycopersicon esculentum (tomato), and possibly Datura stramonium (Jimson weed) (NatureServe, 2010m; US-FWS, 2000b, 2010f).

Moth, Kern primrose sphinx Euproserpinus euterpe Found in Kern, San Luis Obispo, Santa Barbara, and Ventura Counties, CA. The most important habitat factor is presence of the larval foodplant, which is Camissonia sp. Some of the habitat has been disked, and some roads and development are within the population areas. Sheep grazing has contributed to habitat destruction (US-FWS, 2007a).

BIBLIOGRAPHY

Barton BJ and Bach CE. (2005) Habitat use by the federally endangered Mitchell's Satyr butterfly ( Neonympha mitchellii mitchellii ) in a Michigan prairie fen. The American Midland Naturalist, 153 (1), 41-51. Bayer (2010). Petition for Determination of N onregulated Status for Insect-Resistant and Glufosinate Ammonium-Tolerant Cott on: TwinLinkTM Cotton (Events T304-40 x GHB119). Submitted by AL Scott, Regulatory Affairs Manager. Bayer CropScience LP (See Table http://www.aphis.usda.gov/ biotechnology/not_reg.html)

. Butterfly Conservation Initiative (2006a). Palos Ve rdes Blue Butterfly Retrieved January 31, 2011 from http://www.butterflyreco very.org/species_profil es/profiles.php?id=13 Butterfly Conservation Initiative (2006b). San Br uno Elfin Butterfly Retrieved January 31, 2011 from http://www.butterflyreco very.org/species_profil es/profiles.php?id=16 Butterfly Conservation Initiative (2006c). Callippe Silverspot Butterfly Retrieved March 8, 2011 from http://www.butterflyreco very.org/species_profil es/profiles.php?id=2 Khan AH and Afzal M. (1950) Natural crossing in cotton in Western Punjab IV. Agents of natural crossing. Agronomy Journal, 42 (5), 236-238. Naranjo SE. (2009a) Impacts of Bt crops on non-targ et invertebrates and insecticide use patterns. CAB Review: Perspectives in Agriculture, Veterinary Science, Nutrition and Natural Resources, 4 (11), 1-23.BAYER TWINLINKTM COTTON

B-5 NatureServe (2010a). Euphydryas editha bayensis : Bay Region Checkerspot Retrieved January 31, 2011 from http://www.natureserve.org/explorer/servl et/NatureServe?searchName=Euphydryas+edit ha+bay NatureServe (2010b). Speyeria zerene behrensii : Behren's Fritillary Retrieved January 31, 2011 from http://www.natureserve.org/explorer/servlet /NatureServe?searchName=Speyeria+zerene +behren NatureServe (2010c). Euphilotes battoides allyni : El Segundo Blue Retrieved January 31, 2011 from http://www.natureserve.org/explorer/servlet /NatureServe?searchN ame=Euphilotes+battoi des+all NatureServe (2010d). Plebejus icarioides fenderi : Fender's Blue Retrieved January 31, 2011 from http://www.natureserve.org/explorer/servlet /NatureServe?searchNam e=Icaricia+icarioide s+fend NatureServe (2010e). Plebejus melissa samuelis : Karner Blue Retrieved January 31, 2011 from http://www.natureserve.org/explorer/servlet /NatureServe?searchName=Lycaeides+meliss

a+sam NatureServe (2010f). Plebejus anna lotis : Lotis Blue Retrieved January 31, 2011 from http://www.fws.gov/sacramento/es/anim al_spp_acct/langes_metalmark_butterfly.pdf NatureServe (2010g). Plebejus icarioides missionensis : Mission Blue Retr ieved January 31, 2011 from http://www.natureserve.org/explorer/servlet /NatureServe?searchNam e=Icaricia+icarioide s+missi NatureServe (2010h). Neonympha mitchellii francisci : Mitchell's Satyr Retrieved January 31, 2011 from http://www.natureserve.org/explorer/servl et/NatureServe?searchName=Neonympha+mit chellii+francisci NatureServe (2010i). Callophrys mossii bayensis : San Bruno Elfin Retrieved January 31, 2011 from http://www.natureserve.org/explorer/servl et/NatureServe?searc hName=Callophrys+moss ii+bayensis NatureServe (2010j). Papilio aristodemus ponceanus : Schaus' Swallowtail Retrieved January 31, 2011 from http://www.natureserve.org/explorer/servlet /NatureServe?searchName=Heraclides+aristo demus+ponceanus NatureServe (2010k). Euphilotes enoptes smithi : Smith's Blue NatureServe. Retrieved January 31, 2011 from http://www.natureserve.org/explorer/servl et/NatureServe?searc hName=Euphilotes+enopt es+smithi NatureServe (2010l). Boloria improba acrocnema : Uncompahgre Fritillary NatureServe. Retrieved January 31, 2011 from http://www.natureserve.org/explorer/servlet /NatureServe?searchName=Boloria+acrocne

maBAYER TWINLINKTM COTTON

B-6 NatureServe (2010m). Manduca blackburni : Blackburn's Sphinx Moth Na tureServe. Retrieved January 31, 2011 from http://www.natureserve.org/explorer/servl et/NatureServe?searchName=Manduca+blackb urni Sanvido O, Stark M, Romeis J, a nd Bigler F (2006) Ecological Im pacts of Genetically Modified Crops: Experiences from Ten Years of Expe rimental Field Research and Commercial Cultivation (Report: Zürich, Switzerland: Agroscope Reckenholz-Tänikon Research Station ART. Retrieved from Szymanski J, Shuey JA, and Oberhauser K. (2004) Population structure of the endangered Mitchell's Satyr, Neonympha mitchellii mitchellii (French): Implications for conservation. The American Midland Naturalist, 152 (2), 304-322. Thies SA. (1953) Agents concerned with na tural crossing of cotton in Oklahoma. Agronomy Journal, 45 , 481-484. US-EPA (2001b). Bt Plant-Incorporated Protec tants October 15, 2001 Biope sticides Registration Action Document Retrieve d January 22, 2010 from http://www.epa.gov/opp00001/biopesticid es/pips/bt_brad2/5-benefits.pdf US-EPA (2008) Biopesticides Re gistration Action Document ( Bacillus thuringiensis Cry2Ab2 Protein and its Genetic Material Necessary for its Production in Cotton) (Chemical PC Code 006487) Amended (Report: U.S. Envi ronmental Protection Agency, Office of Pesticide Programs. Retrieved from http://www.epa.gov/oppbppd1/biopesticid es/ingredients/t ech_docs/brad_006487.pdf US-FWS. (1980) 50 CFR Part 17 Li sting the Palos Verdes Blue Butterfly as an Endangered Species with Critical Habitat. Federal Register, 45 (129), 44939-44942. US-FWS (1994). Uncompahgre Fritillary Butterf ly Recovery Plan. Submitted by Registration Manager. U.S. Fish and W ildlife Service (See Table http://ecos.fws.gov/docs/recovery_plan/940317.pdf)

. US-FWS (1998a). Recovery Plan for Mitchell's Satyr Butterfly Neonympha mitchellii mitchellii French. Submitted by Registration Manager. U.S. Department of the Interior, Fish and Wildlife Service (See Table http://ecos.fws.gov/docs/ recovery_plan/980402.pdf)

. US-FWS (1998b). Seven Coastal Plants and the My rtle's Silverspot Butte rfly Recovery Plan. Submitted by Registration Manager. U.S. Fish and Wildlife Service (See Table http://ecos.fws.gov/docs/recovery_plan/980930d.pdf)

. US-FWS (1999). Schaus Swallowtail Butterfly Heraclides aristodemus ponceanus . Submitted by Registration Manager. U.S. Fish and Wildlife Service (See Table US-FWS. (2000a) 50 CFR Part 17 Endangered and Threatened Wildlife and Plants; Endangered Status for " Erigeron decumbens " var. " decumbens " (Williamette Daisy) and Fender's Blue Butterfly (" Icaricia icarioides fenderi ") and Threatened Status for " Lupinus sulphureus " ssp. " kincaidii " (Kincaid's Lupine). Federal Register, 65 (16), 3875-3890. US-FWS. (2000b) 50 CFR Part 17 Endangered and Threatened Wildlife and Plants; Determination of Endangered Status for Blackburn's Sphinx Moth from the Hawaiian Islands. Federal Register, 65 (21), 4770-4779. US-FWS. (2001a) 50 CFR Part 17 Endangered a nd Threatened Wildlife and Plants; Final Determination of Critical Habitat fo r the Bay Checkerspot Butterfly ( Euphydryas editha bayensis ); Final Rule. Federal Register, 66 (83), 21450-21489.BAYER TWINLINKTM COTTON

B-7 US-FWS (2001b). Oregon S ilverspot Butterfly ( Speyeria zerene hippolyta ) Revised Recovery Plan. Submitted by Registration Manager. U. S. Fish and Wildlife Service (See Table http://ecos.fws.gov/docs/recovery_plan/010822.pdf)

. US-FWS (2003). Recovery Plan for the Quino Checkerspot Butterfly ( Euphydryas editha quino ). Submitted by Registration Manager. U.S. Fish and Wildlife Service (See Table http://ecos.fws.gov/docs/recovery_plan/030917.pdf)

. US-FWS. (2004) Draft Recovery Plan for Behren's Silverspot Butterfly ( Speyeria zerene behrensii ). Federal Register, 69 (12), 2725-2726. US-FWS (2007a). Kern Primrose Sphinx Moth ( Euproserpinus euterpe ) 5-Year Review: Summary and Evaluation. Submitted by Registration Manager. U.S. Fish and Wildlife Service (See Table http://ecos.fws.gov/docs/five_year_review/doc1157.pdf)

. US-FWS. (2007b) Endangered and Threatened Wildlif e and Plants; Initiation of 5-Year Reviews of 58 Species in California and Nevada; Avai lability of Completed 5-Year Reviews in California and Nevada. Federal Register, 72 (30), 7064-7068. US-FWS (2008). Lange's Metalmark Butterfly: Apodemia mormo langei Retrieved January 31, 2011 from http://www.fws.gov/sacramento/es/anim al_spp_acct/langes_metalmark_butterfly.pdf US-FWS (2010b). Callippe S ilverspot Butterfly ( Speyeria callippe callippe ) Retrieved January 31, 2011 from http://ecos.fws.gov/speciesProfile/prof ile/speciesProfile.a ction?spcode=I019 US-FWS (2010c). Mitchell's Satyr Butterfly ( Neonympha mitchellii mitchellii ) Retrieved January 31, 2011 from http://ecos.fws.gov/speciesProfile/prof ile/speciesProfile.a ction?spcode=I00K US-FWS (2010d). Quino Checkerspot Butterfly ( Euphydryas editha quino ) Retrieved January 31, 2011 from http://ecos.fws.gov/speciesProfile/prof ile/speciesProfile.a ction?spcode=I00P US-FWS (2010e). San Bruno Elfin Butterfly: Callophrys mossii bayensis U.S. Fish and Wildlife Service. Retrieved March 8, 2011 from http://www.fws.gov/sacramento/es/anim al_spp_acct/san_bruno_elfin_butterfly.pdf US-FWS (2010f). Endangered Species in the Paci fic Islands US-FWS Pacific Islands Fish and Wildlife Office. Retrieved January 31, 2011 from http://www.fws.gov/pacificislands/fauna/bsmoth.html US-FWS (2011b). Species Reports Retrieved January 25, 2011 from http://ecos.fws.gov/tess_public/SpeciesRe port.do?groups=I&listingType=L&mapstatus=
1 USDA-APHIS (2010b) Determination of Plant Pest Risk for Insect-Resistant and Glufosinate Ammonium-Tolerant (T winLinkTM) Cotton, Gossypium hirsutum , events T304-40 x GHB119 (Report: U.S. Department of Agricu lture, Animal and Plant Health Inspection Service. Retrieved from