# 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



## Draft Environmental Assessment

April 2011 Agency Contact Ms. Cynthia Eck, Document Control 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 (USDA) prohibits discrimination in all its programs and activities on the basis of race, color, national origin, sex, religion, age, disability, political beliefs, sexual orientation, or marital or family status

. (Not all prohibited bases apply to all programs.) Persons with disabilities who require alternative means for communication of program in formation (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, Director, Office of Civil Rights, Room 326

– W, Whitten Building, 1400 Independence Avenue, SW, Washington, 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 report does not imply recommendation or endorsement by the U.S. Departm ent 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 data and to provide specific information.
This publication rep orts research involving pesticides

. All uses of pesticides must be registered 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: BAYER CROPSCIENCE LEPIDOPTERAN

- RESISTANT AND GLUFOSINATE AMMONIUM

TOLERANT (TWINLINKTM) COTTON, EVENTS GHB119 AND T304

- 40

........................... 2 1.4 PURPOSE OF PRODUCT

................................ ................................ ................................ ........... 3 1.5 APHIS RESPONSE TO PETITION FOR NO NREGULATED 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 Wa ter Resources

................................ ................................ ................................ ................. 16 2.3.2 Soil

................................ ................................ ................................ ................................ ...... 17 2.3.3 Air Quality

................................ ................................ ................................ .......................... 18 2.3.4 Climate Change

................................ ................................ ................................ ................... 19 2.4 ANIMAL A ND 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 H EALTH

................................ ................................ ................................ ..................... 27 2.6 ANIMAL FEED

................................ ................................ ................................ .......................... 29 2.7 SOCIOECONOMIC ISSUES

................................ ................................ ................................ ..... 32 3 ALTERNATIVES

................................ ................................ ................................ ................. 34 3.1 NO ACTION: CONTINUATION AS A REGULATED ARTICLE

................................ ......... 34BAYER TWINLINKTM COTTON



### TABLE OF CONTENTS , CONTINUED PAGE

ii 3.2 PREF ERRED ALTERNATIVE: DETERMINATION THAT TWINLINKTM COTTON IS NO LONGER A REGULATED ARTICLE

................................ ................................ ..................... 34 3.3 ALTERNATIVES CONSIDERED BUT REJECTED FROM FURTHER CONSIDERATION 35 3.3.1 Prohibit Any TwinLinkTM Cotton from Being Released

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

................................ ................................ .............................. 35 3.3.3 Isol ation 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 ALTERNATIVES

................................ ................................ ..................... 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

................................ ................................ ................................ .................. 58 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 Do mestic Economic Environment at Risk

................................ ................................ .......... 68 4.7.2 Trade Economic Environment at Risk

................................ ................................ ................ 69 4.7.3 Social Environment at Risk

................................ ................................ ................................ . 71 4.8 OTHER CUMULATIVE EFFECTS

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

................................ ................................ .... 73 4.9.1 Potential Effects of TwinLinkTM Cotton 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

................................ ................................ .... 80 4.10.1 Executive Orders with Domestic Implications

................................ ................................ ... 80 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

................................ ................... 84 4.10.5 National Historic Preservation Act (NHPA) of 1966 as Amended

................................ ..... 85 5 BIBLIOGRAPHY

................................ ................................ ................................ ................. 86 6 LIST OF PREPARERS

................................ ................................ ................................ ........ 99

APPENDICES Appendix A A PHIS Threatened a nd Endangered Species Decision Tree f or F WS 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, Presenting Percent U.S. Cotton Yield Reduction, Number of Cotton Acres Infested, Percent of U.S. Cotton Infested, Bales Lost, in 2009

................................ ................................ ................................ 12 Table 3

- 1: Summary of Potential Impacts and Consequences of Alternatives

....................... 38 Table 4

- 1: Summary of Insecticide Use for Lepidopteran Control in Cotton Comp ared with Adoption of Bt Cotton

................................ ................................ ........................... 43BAYER TWINLINKTM COTTON



### ACRONYMS

iv AIA advanced informed agreement AMS Agricultural Ma rketing Service ANZFA Australia a nd New Zealand Food Authority AOSCA Association of Offi cial Seed Certifying Agencies APHIS Animal and P lant Health Inspection Service ARMS Agricultural Resource Management Survey BCS Bayer Crop Science BRAD Biopesticides Registration Action Document BRS Bi otechnology Regulatory Service BXN bromoxynil

- tolerant CBD Convention on Biological Diversity CBW cotton bollworm CEQ Co uncil of Environmental Quality CFIA Ca nadian Food Inspection Agency C FR Co de of Federal Regulations DNA deoxyribonucleic acid EA Environmental Assessment ECOS Environmental Conservation Online System ELS extra

- long staple EO Executive Order EPA E nvironmental Protection Agency ERA ecological risk assessment ESA Endangered Species Act EU European Union FAW fall armyworm FD A Food and Drug Administration FFDCA Federa l 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 Intergovernm ental Panel on Climate Change LMO living modified organism MFA Multifibre Arrangement MOU memorandum of understanding N 2 NABI North American Biotechnology Initiative O nitrous oxide

NAPPO North American Plant Protection Organization NEPA Nat ional Environmental Policy Act NHPA Natio nal 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 Deve lopment OSTP Office of Science and Technology Policy PAT phosp hinothricin

- acetyl

- transferase PBO Plant Biosafety Office PGF pollen

- mediated gene flow PPA Plant Prot ection 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. Departmen t 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 a griculture” is the basic charge of the U.S. Department of Agriculture's (USDA ) Animal and Plant Health Inspection Service (APHIS )

. APHIS provides leadership in ensuring the health and care of plants and animals

. The agency improves agri cultural productivity and competitiveness, and contributes to the national economy and the public health

. USDA asserts that all methods of agricultural production (conventional, organic, or the use of genetically engineered varieties) can provide benefits to the environment, consumers, and farm income. Since 1986, the United States government has regulated genetically engineered (GE) organisms pursuant to a regulatory framework known as the Coordinated Framework for the Regulation of Biotechnology (Coordi nated Framework) (51 FR 23302, 57 FR 22984). The Coordinated Framework, published by the Office of Science and Technology Policy, describes the comprehensive federal regulatory policy for ensuring the safety of biotechnology research and products and expla ins how federal agencies will use existing Federal statutes in a manner to ensure public health and environmental safety while maintaining regulatory flexibility to avoid impeding the growth of the biotechnology industry. The Coordinated Framework is base d on several important guiding principles: (1) agencies 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 risks of the biotechnology product, 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 explain s the regulatory roles and authorities fo r the three major agencies involved in regulating GE organisms: USDA’s Animal and Plant Health Inspection Service (APHIS), the Food and Drug Administration (FDA), and the Environmental Protection Agency (EPA). APHIS is responsible for regulating GE organ isms and plants under the plant pest authorities 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 authority of the Federal Food, Drug, and Cosmetic Act (FFDCA) . The FDA is responsible for ensuring the safety and proper labeling of all plant

derived foods and feeds, including those that are genetically engineered. To help developers of food and feed derived from GE crops com ply with their obligations under Federal food safety laws, FDA encourages them to participate in a voluntary consultation process. All food and feed derived from GE crops currently on the market in the United States have successfully completed this consul tation process. The FDA policy statement concerning regulation of products derived from new plant varieties, including those genetically engineered, was published in the Federal Register on May 29, 1992 (57 FR 22984

- 23005). Under this policy, FDA uses w hat is termed a consultation process to ensure that human food and animal feed safety issues or other regulatory issues (e.g., labeling) are resolved prior to commercial distribution of bioengineered food.BAYER TWINLINKTM COTTON

2 The EPA regulates plant

- incorporated protectants under the Federal Insecticide, Fungicide, and Rodenticide Act (FIFRA) and certain biological control organisms under the Toxic Substances Control Act (TSCA). The EPA is responsible for regulating the sale, distribution and use of pesticides, including pes ticides 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 agricu lture and environment using a dynamic and science

- based regulatory framework that allows for the safe development and use of GE organisms

. APHIS regulations at 7 Code of Federal Regulations (CFR ) Pa rt 340 (hereafter Part 340), which were promulgated pursuant to authority granted by the PPA, regulate the introduction (importation, interstate movement, or release into the environment) of certain GE organisms and products

. A GE organism is no longer su bject to the plant pest provisions of the Plant Protection Act or to the regulatory requirements of P art 340 when APHIS determines that it is unlikely to pose a plant pest risk. A GE organism is considered a regulated article under Part 340 if the donor organism, recipient organism, vector, or vector agent used in engineering the organism belongs to one of the taxa listed in the regulation (7 CFR 340.2) and is also considered 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 determine if the GE organism is unlikely to pose a plant pest risk. A person may petition the agency that a particular regulated article is un likely to pose a plant pest risk, and therefore, is no longer regulated under the plant pest provisions of the Plant Protection Act or the regulations at 7 CFR 340. The petitioner is required to provide information under § 340.6(c)(4) related to plant pes t risk that the agency may use to determine whether the regulated article is unlikely to present a greater plant pest risk than the unmodified organism

. A GE organism is no longer subject to the regulatory 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

- RESISTANT AND GLUFOSINATE AMMONIUM

- TOLERANT (TWINLINKTM) COTTON, EVENT S GHB119 AND T304

- 40 Bayer Crop Science (BCS ) has submitted a petition ( APHIS Number 08

- 340

- 01p) seeking a determination that TwinLinkTM C otton Events GHB119 and T 304

- 40 are unlikely to pose a plant pest risk and, therefore, should no longer be a regulated article under regulations at 7 CFR Part 340.
TwinLinkTM Cotton is a combined

- trait cotton developed using convention al breeding techniques to link two deoxyribonucleic aci d ( DNA ) transformation events; each developed using DNA recombinant techniques. By crossing BCS’ Cry1Ab C otton ( e vent T304

- 40) with BCS’ C ry 2Ae Cotton (event GHB119), BCS has developed a cotton resistant to lepidopteran pests. TheBAYER TWINLINKTM COTTON

3 TwinLinkTM Cotton also expresses a g lufosinate ammonium herbicide tolerance trait based on LibertyLink ®

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

1.4 PURPOSE OF PRODUCT BCS has developed the TwinLinkTM Cotton as an alternative insect

- resistant and herbicide

tolerant cot ton product. BCS has developed u pland or Mexican cotton ( Gossypium hirsutum ) plants that express two insecticidal crystalline proteins, Cry1Ab and Cry2Ae, derived from the common soil bacterium, Bacillus thuringiensis ( Bt ). The Cry1Ab and Cry2Ae protein s 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 resistance to both proteins simultaneously than to develop resistance to one toxic protein

. In addition to the Cry1Ab and Cry2Ae proteins , the TwinLink TM Cotton contains the modifying phosphinoth ricin

- acetyl

- transferase (PAT ) enzyme, encoded by the bar gene which confers tolerance to glufosinate ammonium

- based herbicides

. The bar gene, derived from Streptomyces hygroscopicus, encode s 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 producers in the U.S. are among the most technically advanced in the world, annually harvesting about 17 million bales or 7.2 billion pounds of cotton ( Cotton (LLCotton25) that also confers tolerance to glufos inate ammonium herbicides.

US

- EPA, 2009 )

. In 2010, the USDA estimates that 18. 9 million bales were harvested ( USDA

- NASS, 2010a ) with approximately 10.9 million acres planted ( USDA

- NASS, 2010b )

. Cotton g rowers striv e for high yield s of high

- value cotton fiber at the lowest production cost

. To enhance successful cotton fiber production, t he cotton industry relies heavily on pest control measures to control both weeds and lepidopteran insects

. Infestation of cotton b y lepidopteran larvae represents the largest insect threat to cotton production in the U.S. ( Scott, 2008 )

. To combat these threats and enhance production, cotton growers in the U .S. have adopted genetically improved cotton, to reduc e pest management costs, obtain greater yield s , and decrease pesticide costs ; thereby producing greater grower return on investment ( Scott, 2008 )

. In 20 10 , 78 % of all cotton grown in the U .S. contained herbicide

- tolerant traits and 73 % expresse d insect

- resistant traits ( USDA

ERS, 2010a )

. Cotton producers are currently limited to insect

- resistant cotton varieties containing three Bacillus thuringiensis Cry1 endotoxin protein

- based plant incorporated protectants

. Each is deregulated by the USDA and registered by the EPA under FIFRA

. The three proteins are Cry1Ac (Bollgard)

— a combination of CrylAc and Cry2Ab2 (Bollgard II), Cry1F , and Cry1Ac (Widestrike ) ( USDA

- APHIS, 2010a )

. These transgenic varieties offer almost complete protection against tobacco budworm, but may require additional applications of insecticide forBAYER TWINLINKTM COTTON

4 control of cotton bollworm

. Hence, despite the wide adoption of these Bt cotton varieties to control lepidopteran insects, these same species continue to be the most economically important pests of the crop

. In 200 9 , it was estimated that the Helioth ine 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 200 9 cost and loss estimates for arthropod cotton pests were $ 502 million dollars ( Williams, 2010 )

. TwinLi nk TM Cotton has been developed by BCS as an alternative insect

- resistant and herbicide

tolerant cotton

. The introduction of the Bt protein Cry2Ae provides an additional measure of insect protection not currently available in cotton. The tolerance to glufosinate ammonium, in addition to the insect resistance provided by the two Cry proteins, will provide cotton growers with a new approach for managing lepidopteran pests of cotton , as well as herbicide

- resistant weeds. Current insect resistance manageme nt strategies have delayed the 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 has occu rred

. The availability of additional lepidopte r an 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 resistance, including the planting of refuge areas with crops not engineered to contain Bt ( Tabashnik et al., 2008 )

. Glufosinate ammonium t olerance was introduced by Bayer in its LibertyLink ® Cotton to provide cotton growers with an alternative herbicide choice. The Lib ertyLink ® USDA

- APHIS, 2003 Cotton product was deregulated by APHIS in 2003 ( ) and is widely available. EPA has published a vol untary pesticide resistant management labeling guideline providing guidance on reducing or managing the potential for herbicide

- resistant weeds or volunteers (Pesticide Registration Notice 2001

- 5 ) ( US

- EPA, 2001a )

. Glufosinate a mmoni um tolerance provides growers with another herbicide option in cotton cultivation.
TwinLinkTM Cotton , either alone or when combined by traditional breeding with other genetically

- modified insect

- resistant cotton varieties, by incorporating a new Cry protein, provide s growers with an additional pest management option for lepidopteran

- insect pest control and may contribute to a reduction in the likelihood of insect resistance to Bt insect

- resistant cotton varieties. Specifically, BCS indicates that the use of TwinLinkTM Cotton will provide:

1. Improved efficacy in reduction of major insect pests of cotton, independen t of weather conditions. 2. Combined Bt trait product for insect resistance management. 3. A dditional weed control options. 4. Excellent human, animal, non

- target organism , and environmental safety profile. 5. Potentially r educed pesticide use. 6. Potentially e nhanced yield. 7. Potentially d ecreased 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 a nd 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 submit ted, 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 geneticall y 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 completed prior to the petition submission. Field test reports can be found in the BCS TwinLinkTM Cotton petition in Appendix 1, p.102 ( Scott, 2008 )

. BCS has conducted 66 field trials under field release authorizations granted by USDA APHIS in diverse regions of the U.S., including Arizona, California, Georgia, Louisiana, Mississippi, North Carolina, South Carolina , and Texas

. Field tests conducted under APHIS oversight allow for evaluation in agricultural settings under confinement 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 pr oduct performance

. These data are also valuable to APHIS for assessing the potential for a new variety to pose a plant pest risk

. The data evaluated for TwinLink TM Cotton may be found in the document Determination of Plant Pest Risk for Insect

Resistant and Glufosinate Ammonium

- Tolerant (TwinLinkTM) Cotton, Gossypium hirsutum, E

vents T304

- 40 x GHB119 ( USDA

- APHIS, 2010b )

. APHIS has prepared this environmental assessment to consider the potential environmental effects of an agency determination of nonregulated status consistent with Council of Environmental Quality’s (CEQ ) regulations implementing NEPA (40 CFR Part s 1500

- 1508 ) , and the USDA and APHIS NEPA implementing regulations 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 specifically evaluate the effects on the quality of the human environment 1

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). that may result from the deregulation of B CS TwinLinkTM Cotton

.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 derived from new plant varieties, including th ose developed through biotechnology ( US

- FDA, 1992 )

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

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

. Note that with regard to the expre ssion of glufosinate ammonium tolerance, the gene construct in the Bayer TwinLinkTM Cotton is the same as that approved by the FDA in June 2003 for the LibertyLink ® US

- FDA, 2003 Cotton product ( ) . In that approval, the FDA noted that the transformational event in LibertyLink ®

US

- FDA, 2003 Cotton was not materially different in composition, safety, or any other relevant parameter in cotton grown, marketed , and consumed at that time ( )

. This previous FDA review will be used by APHIS to analyze the food and safety impacts associated with the incorporation and expression of glufosinate ammonium tolerance in TwinLinkTM Cotton. The FDA’s oversight of the food and safety impacts associated with the incorporation and expression of pesticidal substances, in this case, the Cry proteins associated with Bt, are more limited. As discussed below, the EPA is the primary authority for the review of plant

incorporated protectants. Environmental Protection Agency The EPA has authority over the use of pesticidal substances and plant

- incorporated protectants under the FIFRA as amended (7 US C § 136 , et s eq. ) 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 use of this product

. APHIS considers the EPA’s regulatory assessment when assessing potential impacts that may result from the deregulation of a GE organism. In the absence of a completed EPA determination, APHIS take s into consideration prior EPA reviews of comparable products to make a preliminary assessment of the potential impacts.
Note that EPA has issued a tolerance exemption 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, 2010g )

. A temporary exemption from the requirement of a tolerance has been issued for Cry2Ae ( 40 CFR §174.530; US

- EPA, 2010e )

. Th ese previous EPA review s will be used by APHIS to analyze the food and safety impacts associated with the incorporation and expression of the Cry proteins in TwinL inkTM Cotton.BAYER TWINLINKTM COTTON

7 In addition to review of the crop with plant

- incorporated protectants, EPA has authority under FIFRA to establish pesticide use restrictions; these use restrictions are presented on pesticide labels which are prepared during the pesticide regi stration process

. The development of a transformed cotton crop providing tolerance to glufosinate ammonium may require a change in the EPA

- approved label for this herbicide.
The current glufosinate ammonium label provides for its use on transformed crop s expressing resistance to glufosinate ammonium , and specifically references products marketed under the trade name “LibertyLink” ( BCS, 2010a )

. Glufosinate ammonium

- tolerant cotton was first available in the U.S. with the introduction of LibertyLink ® C otton in 2004. Although the glufosinate ammonium tolerance trait expressed in TwinLink TM cotton is the same as that expressed in the LibertyLink ®

1.7 PUBLIC INVOLVEMENT C otton, the EPA is expected to publish a new lab e l for glufosinate ammonium that also references the TwinLink TM 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 glufosinate ammonium.

APHIS routinely seeks public comment on draft EAs prepared in response to petitions to deregulate 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, concerns raised in lawsuits, as well as those issues that have been raised by various stakeholders

. These issues, including those regarding the agricultural pr oduction of cotton using various production methods , and the environmental and food/feed safety of GE plants , were addressed to analyze the potential environmental 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, E vents 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))

. Com ments received by the end of the 60

- day period will be reviewed and used to inform APHIS’ s determination decision of the regulated status of TwinLinkTM Cotton an d to assist APHIS in determining whether an Environmental Impact Statement is required prior to a determination of the regulat ed status of TwinLink TM Cotton

. 1.8 ISSUES CONSIDERED As stated above, the issues considered in this EA were developed based on APHIS’ s determination to deregulate certain GE organisms ; and , for this particular EA, the specific deregulation of TwinLinkTM Cotton

. The se issues include:

Cotton Production :

**•** Acreage and Areas of Cotton Production

**•** Cropping Practices

**•** Seed ProductionBAYER TWINLINKTM COTTON

8 **•** Organic Farming

**•** Specialty Cotton Production
Environmental Considerat ions :

**•** 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 the deregulation of TwinLinkTM Cotton. For the purposes of this draft EA, those aspects of the human environment are : cotton production practices, the physical environment, biological resources, public health, animal feed, and socioeconomic issues.
2.1 COTTON USE AND BIOLOGY
2.1.1 Cotton Taxonomy Cotton is a perennial plant that is cultivated as an annual crop ( USDA

- ERS, 2009 )

. The Gossypium genus is made up of approximately 50 species, but only five sp ecies are cultivated worldwide ( OECD, 2008 )

. The cultivated species u pland cotton ( G. hirsutum ) represents 97% of cotton crop g rown in the U.S. ( USDA

- ERS, 2009 ) . The balance of U.S.

- grown cotton i s 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 ( OECD, 2008 ; Scott, 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 g enome is restricted in diploids to some species 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 representi ng the complete chromosome sets from both parent organisms) (2n=4x=52) . These cultivated varieties are of New World origin and are presum ed to be the result of an ancient interspecific hybridization between Old World A g enomes and New World D genomes ( Scott, 2008 ) . How and when the original crosses occurred has been subject to much speculation. Euploids (i.e., organisms containing equal numbers of the haploid chromosome sets) of these plants have 52 somatic chromosomes and are frequently des ignated as AADD (they behave 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 least in their wild forms, grow near the ocean as invaders in the constantly disturbed habitats of strand and associated environs. It is from these "weedy" or invader species that the cultivated 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 genetic construct are important to recognize, as the genetic differences direc tly affect the ability of cultivated cotton to interbreed with wild cotton species as well as wild or feral cotton. There are only two wild cotton species found in the U.S.: G. thurberi (desert cotton), found in Arizona ; and G. tomentosum (Hawaiian cotto n), found in Hawaii ( ITIS, 2010 ; USBG, 2010 ; USDA

- ARS, 2010a )

. G. tomentosum has been crossed with G. hirsutum in breeding programs; however, no commercial cotton is produced in Hawaii ( Scott, 2008 ; USDA

NASS, 2010c )

. F eral populations of u pland cotton ( G. hirsutum ) can be found in southern Florida ( USDA

- FS, 2010 )

. These populations are apparently self

- sustaining becaus e there is no commercial production of cotton in this region ( Scott, 2008 ; 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 fo r over 40% of the total world fiber use ( Meyer et al., 2007 )

. Cotton has been cultivated for its fi ber for several thousand years ( Scott, 2008 ) . Other cotton products, such as cottonseed oil, cake, and cotton linters, are by

- produ cts of cotton fiber production ( Scott, 2008 )

. 2.2 AGRICULTURAL PRODUCTION OF COTTON
2.2.1 Acreage and Areas of Cotton Production Cotton is cultivated mostly in subtr opical and warm temperate zones ( OECD, 2008 )

. Temperature is the main climatic factor determining the geographic range in which cotton can be grown, but cultivation is a lso dependent upon high light intensity, good soil moisture, and soil fertility ( Meyer et al., 2007 ; OECD, 2008 )

. Cotton is mainly produced i n China, India, U.S., Pakistan, Brazil , and Uzbekistan, with these five countries contributing nearly 83% of world 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 Production in Select Countries, 2006 to 2010/11 (est imated ) (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 acre s) ( USDA

- NASS, 2010b )

. In the U.S., c otton is produced in 17 southern states identified as the Cotton Belt: Alabama, Arizona, Arkansas, California, Florida, Georgia , Kansas, Louisiana, Mississippi, Missouri, New Mexico, North Carolina, Oklahoma, South Carolina, Tennessee, Texas, and Virginia ( 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 Carolina (0.545 million acres), and T ennessee (0.387 million acr es) ( USDA

- NASS, 2010c )

. Cotton acreage in the U.S. rose slightly during the first half of the 2000s, continuing a multi

decade trend. In the 1970s and 1980s, the area planted for cotton averaged about 12 million
acres ( USDA

- ERS, 2009 )

. The area planted rose to about 14 million acres in the 1990s and averaged over 14.5 million acres during the fir st half of the 2000s , representing an increase of 20 % over the 1970s and 1980s acreage ( USDA

- ERS, 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

- ERS, 2009 )

. All cotton growing regions have experienced significant declines compared with the first half of the 2000s ( USDA

- ERS, 2009 )

. GE varieties of cotton have been very rapidly 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

- resistant (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 cotton marketed by companies that produce seed , including GE varieties ( USDA

- AMS, 2010a ) . Planting of cotton occurs after any danger of frost has passed and soil temperatures are at least 14 o C (57 o OECD, 2008 F) ( ) . Planting dates vary depending on the region , but generally range from early March in southern Texas to early May or June in the northe rn areas of the Cotton Belt. Cotton requires 180 to 200 days from planting to maturity ( OECD, 2008 )

. Therefore, cotton planted in March or April is ready for harvest in Sept ember. Crop rotations (successive planting of different crops on the same land) are used to optimize soil nutrition and fert ility and reduce pathogen loads ( Gazaway et al., 2007 ; University of California IPM Online, 2008a )

. Cotton is often 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. Rotations may last for two or three years. Winter cover crops are also utilized in cotton. These cover crops are used to provide winter soil cover and protection, build soil nitrogen and organic matter, reduce nitrogen leaching, suppre ss weeds, and provide a habitat for beneficial predatory and parasitic insects and spiders ( Guerena and Sullivan, 2003 )

. Cotton production in the U.S. is highly mechanized and involves the extensive use of agronomic inputs and technology ( USDA

- A PHIS, 2010a )

. The c ost of production includes annual directBAYER TWINLINKTM COTTON

12 costs (seed, fertilizer, chemicals, irrigation, etc.), annual fixed costs (depreciation on equipment), and annual land rent ( Hogan et al., 2005 )

. These costs will vary depending on region and practices used. In 2009, average per acre costs for U.S. cotton growers for cotton seed, fertilizer (commercial fertilizers, soil conditioners, and manure) , and chemicals (including pesticides and herbicides) accounted fo r 54% of the total dollars spent per planted acre or $232 per acre ( USDA

- ERS, 2010b )

. Worldwide, cotton has proven vulnerable to the attack 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 )

. Resistance to commonly used insecticides (pyrethroids, organophosphates, and carbamates) has led to a need for new pesticide chemistries or other novel approaches , such as the use of sterile insects, pathogens, and transgenic cotton ( Gianessi and Carpenter, 1999 )

. Pest management in agricultural crop production is achieved through various management methods , in cluding chemical (pesticides), cultural (mechanical cultivation, planting /harvesting dates, crop rotation , etc.), biological (antagonistic organisms), and bioengineer ing (primarily herbicide

- tolerant and insect

- resistant crops) ( USDA

- ERS, 2005a )

. GE insect

- resistant cotton has been focused on incorporating Bacillus thuringiensis ( Bt ) Cry endotoxin protein based plant incorporated protectants ( Stewart, 2007 )

. The most damaging insect pests of cotton attack the cotton square (the flower bud) or the cotton boll ( the ovary containi ng developing seeds and fibers) ( Gianessi and Carpenter, 1999 )

. There are over 20 economically important pests of cotton. 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 infested more than half of the U.S. cotton crop

. The bu dworm/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 and Carpenter, 1 999 )

. 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 , consuming both lint and seed ( Gianessi and Carpenter, 1999 ) . In 2009, the complex reduced yields by 0.486% or 91,119 bales of cotton ( Will iams, 2010 ) . Bt cotton provides good control over bollworm 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
w ith 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
w ith 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 varieties of cotton combined must be able to plant over 10 millio n 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 breed ing programs began in California 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 Agencies (AOSCA ) developed seed certification standards to m aintain genetic purity of cotton varietal seed and specialty cotton crop s by precluding gene flow and prevent ing cross pollination between species and varieties ( AOSCA, 2010b ; Wozniak, 2002 )

. In a seed ce rtification program, classes of seed are identified to designate the seed generation fro m the original breeder source ( Hartman and Kester, 1975 ) . Foundation seed, R egistered seed, and C ertified seed production is controlled by public or private seed certification pro grams ( see, e.g., AOSCA, 2010b ) . The original seed breeder seed stock is controlled b y the developer of the variety ( Adam, 2005 ; Hartman and Kest er, 1975 )

. The breeder stock is used to produce F oundation seed stock ( Adam, 2005 ) . The institution associated with the breeder controls the production of F oundation seed stock. Foundation seed stock, in turn, is used to produce R egistered seed for distribution to lic ensees, such as seed companies ( Adam, 2005 ) . Registered seed is used by seed companies to produce larg e quantities of C ertified seed ( Adam, 2005 ;BAYER TWINLINKTM COTTON

14 Hartman and Kester, 1975 )

. The C ertified (or S elect) seed is then sold to growe rs through commercial channels ( Adam, 2005 ; Hartman and Kester, 1975 )

. Seed certification cultivation practices commonly include recommendations for minimum isolation distances between various seed lines and planting border or barrier rows to pr event 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 A ct in 7 CFR Part 201 .76, is 1,320 , 1,320 , and 660 feet, respectively ( USDA

- AMS, 2010c )

. During the growing season, seed certification agencies will monitor the fields for off

- types, other crops, weeds, and disease ( Wozniak, 2002 ) . These certifying agencies also establish seed handling standards to reduce the likelihood of seed source mixi ng during production stages, including planting, harvesting, transporting, storage, cleaning, and ginning ( Wozniak, 2002 )

. Further discussion of cross

- pollination, gene transfer, and weediness is pr esented 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 certification, 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 certif ied operation and its records. Organic growers must maintain records to show that production and handling procedures comply with USDA organic standards. The NOP regulations preclude the use of excluded methods. The NOP provides the following guidance un der 7 CFR § 205.105

—
T o be sold or labeled as “100 percent organic , ” “organic , ” or “made with organic (specified ingredients or group(s)),” the product must be produced and handled without the use of: (a) Synthetic substances 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 are not possible under natural conditions or processes and are not considered compatible 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 protecting organically

- pro duced crops from accidental contamination by GE crops by requiring that organic production plans include practical methods to protect organically

- produced crops

— Organic crops must be protected from contamination by prohibited substances used on adjoining lands (for example, drifting pesticides, fertilizer

- laden runoff water, and pollen drift from genetically engineered...) ( NCAT, 2003 )

. Organic farming operations , as described by the NOP , require organic production operations to have distinct, defined boundaries and buffer zones to prevent unintended contact with excluded methods from adjoining land that is not under organic management. Organic production operations must also develop and maintain an organic 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 prohibition on the use of excluded m ethods. 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 organic certification should include a descr iption of practices used to prevent or reduce the likelihood of unwanted substances, like GE pollen or seed, at each step in the farming operation, such as planting, harvesting, sto ring, and transporting the crop ( Krueger, 2007 ; Riddle, 2004 )

. Organic plans should also include how the risk of GE pollen or co

- mingling of seed will be monitored ( Krueger, 2007 ; Riddle, 2004 )

. Farmers using organic methods are requested to let neighboring 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 relatively small percentage of overall cotton production in the U.S. The most recently available dat a show 15,377 acres of certified
or ganic cotton production in 2008 ( USDA

- ERS, 2010c )

. This is 0.16% of the total 9.41 million acres of cotton planted in 2008 ( USDA

- ERS, 2009 )

. Bayer CropScience FM 958, FM989, and ADF 2485 were the predominate varieties planted in 2010 by organic cotton producers ( USDA

- AMS, 2010b )

. 2.2.5 Specialty Cotton Production
Several specialty cottons are cultivated in the U.S. , including ELS ( G. barbadense ) Pima varieties , specialty long

- 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 q uality standards for specialty crop products have led these seed producers and growers to employ a variety of techniques to ensure that their products are not pollinated by or commingled 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 barrier rows to intercept pollen, and employing natural barriers to pollen ( NCAT, 2003 ; Wozniak, 2002 ) . The Federal Seed Act Regulations provide additional details on C ertified seed production ( s ee 7 CFR 201, et seq ) . Field monitoring for off

- types, other crops, weeds, disease etc. is also carried out by company staff and state crop improvement associations ( Bradford, 2006 ) . Seed handling standards are established by AOSCA t o reduce the likelihood of seed source mixing during planting, harvesting, transporting, storage, cleaning, and ginning ( AOSCA, 2004 )

. In general, the conventional management practices used for conventional seed production are sufficient to meet standard s for the production of specialty crop seed ( Bradford, 2006 )

. 2.3 PHYSICAL ENVIRONMENT The use of fertilizers, pesticides, and water may affect segments of the environment including, but not limited to : waterways by increases in nutrient pollution; biodiversity because pesticide inputs cause species changes; the water table because of excessive irrigation practices; and productiv ity of cropland because irrigation increases salinity ( Hoeft et al., 2000 )

. Sediment , siltation, nutrients, pes ticides, salinity, and pathogens are primary agricultural pollutants ( USDA

ERS, 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

- irri gated) cotton requires at least 500 m illi m eters ( mm ) (40 inches) of rai nfall during the growing season ( OECD, 2008 )

. Cotton cultivated in the southwestern region of the U.S., including west Texas, southern New Mexico, southern Arizona, and southern California, requires irrigation ( USDA

- APHIS, 2010a )

. Where irrigation water is needed (a pproximately 35% of the U.S. cotton grown), cotton yields are also much higher ( Cotton Incorporated, 2010a )

. The desert southwest requires a maximum of 40 inches of irrigation per year, although the humid southeast may only require ab out 18 inches of irrigation per year ( Cotton Incorporated, 2010a )

. Carefully timing the application of irrigation water optimizes the plant’s vegetative growth, flowering, and boll production ( OECD, 2008 ) . The lack of affordable water has been noted as one factor in the reduction of acres of cotton grown in California, 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 practice s ha ve been shown to minimize surface water runoff and soil erosion and t o improve soil quality by increasing the soil organic matter that helps bind soil nutrients and prevent their loss to runoff, erosion , and leaching ( Leep et al., 2003 )

. This decrease in soil erosion carries a corresponding decrease in non

- point source surface water pollutio n ofBAYER TWINLINKTM COTTON

17 fertilizer and pesticides ( Cotton Incorporated, 20 10a , 2010c )

. Precision agriculture, a practice carefully managing nutrients and chemical applications, also ensure s that inputs are used by the crop and are not en tering ground or surfac e waters ( Cotton Incorporated, 2010a ) . Precision agriculture practices may include computer models to predict water use based on plant growth stage , weather, soil moisture probe data collection, and thermal infrared imagery of plant l eaf 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 high 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 requi res that the producer remove all plant residues and weeds from the soil surface prior to planting, and then continue to cultivate the soil while the crop is growing to control late emerging weeds ( Cotton Incorporated, 2010c ) . This practice resu lts 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 loss 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 decreased 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 carries a corresponding decreas e in non

- point source surface water pollution of fertilizer and pesticides ( Cotton Incorporated, 2010a , 2010c )

. T he 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 has contributed to this reduction in soil loss through a shift towards soil co nservation practices ( Cotton Incorporated, 2010c )

. Conservation practices adopted in cotton cultivation include use of seed treatment fungicides, herbicides, and herbicide

- tolerant cotton varieties, each of whi ch provide for the control of cotton disease and weeds with minimal tillage ( Cotton Incorporated, 2010c ; USDA

- NRCS, 2006b )

. In the U.S., conservation tillage 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 form of conservation tillage ( Cotton Incorporated, 2010c )

. I ncreases in total acres dedicated to conservation tillage have been attributed to an increased use of GE seed which eliminates the need for mechanical weed control ( Alabama Cooperative Extension System, 1996 ; McClelland et al., 2000 ; Towery and Werblow, 2010 ; USDA

- NRCS, 2006b )

. By definition, conservation tillage lea ves 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 reduce 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 c apacity, less soil and nutrient 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

- emergent herbicides make up the primary weed control methods. Winter and co ver crops are also utilized in conservation tillage for the purpose of suppressin g weeds ( Alabama Cooperative Extension System, 1996 )

. Wheat and rye are commonly employed be cause of their ease in killing prior to cotton planting. The use of herbicide

- resistant cotton has allowed cotton growers to more readily adopt soil conservation practices because it provides an economical, effective means of controlling weeds in post

- pla nt cotton ( Alabama Cooperative Extension System, 1996 ; McClelland et al., 2000 )

. In some regions, conservation tillage is required. F armers, including cotton growers, producing crops on highly erodible land are required by law to maintain a soil conservati on p lan approved by the USDA National Resources Conservation Service (NRCS ) ( USDA

ERS, 2010d )

. These soil conservation plans are prepared by the grower pursuant to t he 1985 Food Security Act Conservation Compliance and Sodbuster programs to minimize soil erosion ( USDA

- ERS, 2010d )

. The adoption of Bt cotton, which may require redu ced quantities of pesticides, may benefit s oil quality by reducing the risks associated with environmental spills or misapplications of chemical insecticides to the soil, and reducing the frequency of application

. Reduced insecticide applications also mea n a reduction in the number of trips across the field with heavy farm equipment which contributes to soil compaction, especially when the soil is wet. The number of
acre

- treatments per year of budworm/bollworm insecticides declined from over 10 million to less than 3 million between the years 1996 and 2007 ( USDA

- 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 acres in the U.S. during the same time period ( Benbrook, 2009 )

. 2.3.3 Air Quality Agricultural air emission sources include smoke from agricultural burning, vehicle exhaust associated with equipment used in tillage and harvest, suspended soil particulates associated with tillage, pesticide drift from spraying, and nitrous 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 potential to cause negative impacts to air quality. Aerial application of pesticides may cause air quality impacts from drift and diffusion. Pe sticides may volatilize after application to soil or plant surfaces and may also move as constituents of entrained materials in wind ero ded soils ( Vogel et al., 2008 )

.BAYER TWINLINKTM COTTON

19 Many of the conservation plans and practices being developed by cotton growers to comply with the Conservation Compliance and Sodbuster programs have an air quality focus which target reductions in air emissions from agricultural operations ( Cotton Incorporated, 2010c ; USDA

NRCS, 2006a ) . Practices to improve air quality include conservation tillage, residue
man agement, wind breaks, road treatments, burn management, prunings shredding, feed management, manure management, integrated pest management, chemical storage, nutrient management, fertilizer injection, chemigation and fertigation (inclusion in irrigation sy stems), conservation irrigation, scrubbers, and equipment calibration ( USDA

- NRCS, 2006a )

. Conservation tillage practices resulting in improve d air quality include: fewer tractor passes across a field , thus decreas ing dust generation and tractor emission s; and an increase in surface plant residues and untilled organic matter which physically hold the soil in place and reduce wind erosion ( Baker et al., 2005 ; Cotton Incorporated, 2010d ; USDA

- NRCS, 2006a ) . The USDA has e stimated that by 2006, the adoption of conservation management plans in the San Joaquin Valley of California had reduc ed air emissions by 34 tons daily, or more than 20% of the total emissions attributed to agricultural practices ( Baker et al., 2005 ; USDA

- NRCS, 2006a )

. 2.3.4 Climate Change Agriculture, including land

- use changes 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 tractors) include carbon monoxide, nitrogen oxides, reactive organic gases, particulate matter, and sulfur oxides ( US

EPA, 2010b )

. Agricultural soil management practices, including nitrogen

- based fertilizer application and cropping pra ctices, are the largest source of U.S. nitrous oxide ( N 2 O ) emissions; croplands account for 69% of the total N 2 US

EPA, 2010b O emissions attributable to agricultural land uses ( )

. Agriculture sources of methane emissions are primarily associated with enteric emissions of gas from cattle and manure management. Carbon dioxide is also a significant GHG gas associated with several agricultural practices, including land uses and energy consumption ( US

- EPA, 2010b )

. Tillage contributes to the release of GHG because of the loss of carb on dioxide ( CO 2 Baker et al., 2005 ) to the atmosphere, and the exposure and oxidation of soil organic matter ( ) . The carbon footpr int for cotton production has been estimated to be approximately 300 pounds of carbon equivalent emission per acre ( Nelson et al., 2009 ) . The carbon footprint of cotton is directly affected by the associated cultivation practices. In a no

- tillage production environment, if credit is taken for the sequestration of carbon in soil, cotton actually stores more carbon than is emitted from the production ( Cot ton Incorporated, 2010d ) . Reduced tillage increases sequestration rates of carbon emissions in soils, with 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 dep endent on the production practices employed to grow various commodities, the region in which the commodities are grown, and the individual choices made by growers. For example, emissions of nitrous oxide, produced naturally in soils through microbial nitr ification and denitrification, can be dramatically influenced by fertilization, introduction of grazing animals, cultivation of nitrogen

- fixing crops and forage, retention of crop residues (i.e., no

- till conservation), irrigation, and fallowing of land ( US

- EPA, 2010b ) . These same agricultural practices can influence the decomposition of carbon

-BAYER TWINLINKTM COTTON

20 containi ng organic matter sequestered in soil, resulting 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 sequestration in soils ( US

- EPA, 2010b )

. The EPA has identified regional differences in GHG emissions associated with agricultural practices on different soil types, noting that carbon emission rates differ between mineral soils and organic soils ( US

- EPA, 2010b )

. Mineral soils contain from 1 to 6% organic carbon by weight in their natur al state, whereas organic soils may contain as much as 20% carbon by weight ( US

- EPA, 2010b )

. In mi neral 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 balance between carbon inputs from decaying plant matter and organic
a mendments and carbon losses from microbial decomposition ( US

- EPA, 2010b ) . Organic soils, with thei r 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 US

- EPA, 2010b Eq ( )

. The adoption of conservation tillage, particularly in the Midwest regions with mineral soil shows the highest rates of carbon sequestration ( US

- EPA, 2010b )

. Changes in a griculture

- related GHG production will not be significant unless large amounts of crop plantings produce changes in measureable concentrations ( USDA

- APHIS, 2010a )

. For example, the E PA has identified a net reduction in the sequestration of carbon in soil over an 18

- year time scale, which it attributes to the declining influence of the Conservation Reserve Program which had encouraged growers to take marginal lands out of production ( US

- EPA, 2010b ) . To a certain extent, the EPA also noted that adoption of conservation tillage resul ted in increases in carbon sequestration on those croplands ( US

- EPA, 2010b )

. The highest rates of carbon sequestration in mineral soils 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 noted in the southeastern coastal region, the areas around the Great Lakes, and the central and northern agricultural areas along the West Coast ( US

- EPA, 2010b )

. One outcome of the potential effects of agriculture o n 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 practices will be required to change in respon se to these changes in the ranges of weeds and pests of agriculture ( IPCC, 2007 )

. Climate change may potentially provide a positive 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 observed across all growing regions. The IPCC report notes that certain regions of the U.S. will be negatively im pacted because the available water resources may be substantially reduced. Note that the extent of climate change effects on agriculture is highly speculative. Nevertheless, North American production is expected to adapt to climate change impacts with im proved cultivars and responsive 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), and better control of irrigation 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 e xample, herbicide tolerance allows cultivation with minimal tillage required t o control volunteers and weeds ( Towery and Werblow, 2010 ) . This subsection provides an overview of the relationships of these practices to the biotic community.
2.4.1 Animals Intensive agricultural lands, such as those used in crop production , 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 strategies can increase the value of crop fields to wildlife ( Sharpe, 2010 ) . Some of these strategies are noted below.

**•** Conservation tillage and no

- till practices have a positive impact on wildlife ( Towery and Werblow, 2010 )

. Benefits include improved water quality, retention of cover, availability of waste grain on the soil surface for feed, and increased populations of invertebrates as a food source for turkey, quail, and songbirds ( Sharpe, 2010 )

.

**•** Crop rotations reduce the likelihood of crop disease, insect pests, weed pests, and the need for pesticides ( University of California IPM Online, 2008a )

. 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 for insectivores ( Palmer and Bromley, 2010 ; Sharpe, 2010 )

. Crop rotations with legumes and small grains have been shown to provide excellent wildlife nesting cover, food, and brood

- rearing habitat for quail in North Carolina ( Sharpe, 2010 )

.

**•** Field edges can be managed to promote wildlife. 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 produced ( Sharpe, 2010 ) . Allowing field edges to return to volunteer vegetation does not contribute to major pest problems in the crop field itself ( Sh arpe, 2010 )

. Volunteer border 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 ) . Research conducted at North Carolina State University and the North Carolina Wildl ife 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 strips of row crops with strips of solid stand crops (i.e., grasses, legumes, or small grains) with the str i ps following the contour of the land ( Sharpe, 2010 )

. The primary purpose of contour

- strip cropping is to reduce soil erosion and water runoff, b ut the solid stand 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. Ditch banks, for example, function as narrow wetlands that provide nesting sites and cover, serve as wildlife corridors, and provide areas for the wildlife to occupy when crop fields lack cover ( Sharpe, 2010 ) . Ditches have been shown to support birds, rodents, reptiles, furbearers, amphibians, fish, and aquatic organisms ( Sharpe, 2010 )

. Minimizing pest icide exposure of ditches, aquatic habitats, border areas, strip

- crop areas, and 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 bollworm and tobacco budworm, most invertebrates are considered beneficial ( University of Arkansas 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 body of other insects during at least one stage of their life cycle. Other beneficial insects are the pollinators. Maj or 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 organisms, including earthworms, termites, ants, beetles, millipedes, and others contribute to the decay of organic matter and the 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 surrounde d by woodland, rangelands, and/or pasture/grassland areas. These plant communities may be natural or managed plant habitats for the control of soil and wind erosion and serve as wildlife habitats ( USDA

- APHIS, 2010a )

. Weed control programs are important aspects of cotton cultivation. In this 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 wi th harvesting equ ipment and can stain lint ( USDA

APHIS, 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 i ncludes herbicides, crop rotation, weed surveillance, and weed monitoring ( Univer sity of California IPM Online, 2009b )

. Crop rotation is an important component of a successful weed management program in cotton cultivation ( 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 cultural practices , as well as chemical controls ( University of California IPM Online, 2009b )

. The types of weeds in and around a cotton field depend on the immediate area in which the cotton is planted. Those weed species also vary dep ending 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 per ennial grasses (monocots), broad

- leaf weeds (dicots), and sedges ( Cyperus spp. )

. To assist growers in managing weeds, individual states, typically through their state agricultural 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 herbicidal control can be found at Unive rsity of California IPM Online ( 2009b )

. Herbicide

- tolerant cotton provides several weed management advantages to the growers. Broad spectrum post

- emergent herbicides, such as glufosinate ammonium, provide control of weeds early in the cultivation cycle, minimizing competition in the fields , and providing optimal conditions for cotton growth ( University of California IPM Online, 2009b )

. Applications of
over

- the

- top post

- emergent broad spectrum herbicides to an herbicide

- tolerant crop allow the grower to decrease th e overall use of herbicides before cultivation, reduce use of soil

- applied herbicides, and streamline field cultivation activities for weed control ( Marra and Martin, 2007 ; O'Sullivan and Sikkema, 2004 )

. Production costs decrease because growers apply fewer herbicides and field cultivation requirements are reduced ( University of California IPM Online, 2009b )

. Herbicide

- tolerant cotton has the potential to impact the cultivation of other crops in the same fields in later years. Cotton is periodically seen as a volunteer in soybeans, either when cotton seeds have successfully overwintered or in field s where a failed cotton crop is replanted to soybeans ( Stewart et al., 2003 )

. Cotton volunteers 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 ( Scott, 2008 ) . These cotton volunteers typically do not reduce crop yield , but can act as rese rvoirs for insect pests of cotton ( Stewart et al., 2003 ) . Successful control of cotton volunteers, includin g herbicide

- resistant varieties, is accomplished through using various combinations of herbicides ( Miller et al., 2004 ; Stewart et al., 2003 )

.BAYER TWINLINKTM COTTON

24 2.4.3 Biological Diversity Biodiversity refers to all plants, animals, and microorganisms interacting in an ec osystem ( Wilson, 1988 )

. Among other benefits, bi odiversity provides valuable genetic resources for crop improvement ( Harlan, 1975 ) and also provides other functions beyond food, fiber, fuel, and income. These include pollination, genetic introgression, biological control, nutrient recycling, competition against natural enemies, soil s tructure, 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 provide these functions to the crop ( Altieri, 2000 )

. The degree of biodiversity in an agroecosystem depends on four primary characteristics : 1) diversity of vegetation within and around the agroecos ystem; 2) permanence of various crops within the system; 3) intensity of management; and 4) extent of isolation of the agroecosystem from natural vegetation ( Altieri, 1999 )

. The reintroduction of woodlots, fencerows, he dgerows, wetlands, etc. is one way to reintroduce biodiversity into large scale monocultures

. Some enhancement strategies include intercropping (the planting of two or more crops simultaneously to occupy the same field), agroforestry, crop rotations, cove r crops, no

- tillage, 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.), and h edgerows and windbreaks ( Altieri, 1999 )

. To some degree these practices are being utilized by cotton growers to increase biodiversity ( Cotton Incorporated, 2010e )

. The use of genetically modified cotton affects biological diversity by : 1) providing the opportunity to use ecological management practices that enhance habitat creation and 2) decreasing the use of pesticides which in turn, limits the impact on the floral and faunal communities that can potentially inhabit the fields and immediate surroundings. The use of no

till, cover crops, crop rotation, intercropping, and good ditch/border/hedgerow management contributes to biodiversity in and around cotton fie lds ( Palmer and Bromley, 2010 ; Sharpe, 2010 )

. Habitat preservation and biodiversity, as well as cotton pro duction, have benefited from modern cotton technology. It is now possible to grow 50% more cotton on the same land required 40 years ago ( Cotton Incorporated, 2010e )

. Increased productivity on th e same amount of land allows growers to preserve habitat while maintaining food and fiber security ( Cotton Incorporated, 2010e )

. Various methods for promoting animal and plant biodiversity in and around cotton fields have already been discussed above. H abitat preservation and plant biodiversity have a large impact on insect populations. Research conducted by Altieri ( 1999 ) and Altieri and Letourneau ( Altieri and Letourneau, 1982 , 1984 ) indicates: 1) Maintaining some weeds harbors and supports beneficial arthropods that suppress herbivore insect pests; 2) Polycultures of plants support lower herbivore populations because they provide a more stable and continuous availability of food and habitat for beneficial insects; and 3) Adjacent wild vegetation provides alternate food and habitat for natural enemies to p est herbivores.BAYER TWINLINKTM COTTON

25 The use of broad

- spectrum insecticides is one of the most severe constraints for biological diversity in crops ( Cattaneo 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 the 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

- spectrum insecticide use caused a 48% reduction in 13 of the 22 taxa that were studied ( Naranjo, 2005a )

. A comparison 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 ( Cattaneo et al., 2006 )

. Soil microorganisms play a key role in soil structure formation, decomposition of organic matter, toxin removal, nutrient cycling, and most biochemical soil processes ( 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 include plant type (providers of specific carbon and energy sources into the soil), soil type (texture, structure, organic matter, aggregate stabilit y, pH, and nutrient content), and agricultural management practices (crop rotation, tillage, herbicide and fertilizer application, and irrigation) ( Garbeva et al., 2004 )

. Plant roots, including cotton, release a large variety of compounds into the soil creating a unique environment for micr oorganisms 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 cotton has been raised as an issue of concern ( US

- EPA, 2010a ) . This subsection provides a basis for evaluating the weediness potential of cotton as well as the potential for outcrossing and gene escape in cotton. The potential for outcrossing o r gene escape is defined as the ability of the gene to escape 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 that 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 the field is rarely successful ( Scott, 2008 ) . Pollen dispersal is the most likely potential source of gene movement ( Scott, 2008 )

. The cotton flower is perfect, containing both male and female parts, and can self

- or cross

pollinate ( Meyer et al., 2007 )

. However, the flowers of most cot ton species, including G. hirsutum , are generally considered to be self

- pollinating ( Meyer et al., 2007 ; Scott, 2008 )

. Cotton pollen is heavy and somewhat sticky and does not lend itself to wind dispersal and
s ubsequent pollination ( Scott, 2008 ) . Cotton can, 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 collect mainly the nectar from the cotton plants, and rarely the pollen. In addition, physical isolation from plants attractive to the beesBAYER TWINLINKTM COTTON

26 significantly reduces t he potential for pollen movement, as cotton flowers have a nectar high in glucose and low in sucrose, which probably makes it less attractive to bees. McGregor ( 1976 ) traced the movement of pollen from a cotton field surrounded by a large number of honeybee colo nies using fluorescent particles on source cotton plants. McGregor 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 the bees showed the presence of t he fluorescent particles. By comparison, the isolation distances for Foundation, Registered, and Certified seeds in 7 CFR Part 201 are 1320, 1320 , and 660 feet, respectively ( USDA

- AMS, 2010c )

. Despite these characteristics of cotton pollen which limit wind dispersal and bee pollination, it is common practice to maintain isolation distances to prevent pollen movement from other cotton
sources an d to plant border or barrier rows to intercept pollen to maintain and protect cotton seed characteristics for seed certification programs ( Wozniak, 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 so urce mixing during planting, harvesting, transporting, storage, cleaning, and ginning ( Bradford, 2006 )

. A majority of field

- based research with cotton shows an outcrossing rate of 10% or less within 1 meter of the pollen source ( USDA

- APHIS, 2010a )

. In u pland 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 transferred beyond 12 meters ( McGregor, 1976 ; Scott, 2008 ; Vaissiere and Vinson, 1994 )

. Reports of outcrossing of a bromoxynil

- tolerant cotton are described in the USDA petition for CalGene’s GlyTol cotton 06

- 332

- 01p and in a study of pollen mediated gene flow by Van Deynze et al

. ( 2005 )

. Seed samples were collected in the border rows of Calgene's winter nursery sites in Catamarca, Argentina , and Pongola, Republic of South Africa, as well as in Stoneville, MS, USA. Sampling distance was one to 20 meters away from the bromoxynil

tolerant cotton. The frequency of outcrossing was determined by the crop and the poll inator. It is interesting to note that although the rate is higher for Argentina and South Africa (most likely due to the behavioral differences between European 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 directions over two years from commercial seed fields of bromoxynil

- tolerant (BXN ) and Roundup Ready ®

1994 (RR ) cotton in the California cotton growing region at various distances from non

- transgenic cotton fields . T heir results confirm ed, as well as refine d, those of Kareiva et al

. ( ) , as larger distances were studied. In spite of variations due to the respective cardinal positions of the fields, the same decline with distance was observed. Van Deynze et al. further report ed 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 limitatio ns identified by M cGregor.
There are few wild relatives of cultivated cotton where cotton is cultivated. Two species of cotton are cultivated in the U.S. : u pland cotton ( G

. hirsutum ) and Pima cotton ( G. barbadense ) ( USDA

- ERS, 2009 ) . There are two wild relatives 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 t o 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 numbers 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 characteristics , and is considered a rare plant in Hawaii ( Pleasants and Wendel , 2010 ; University 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 commercially 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 laboratory studies demonstrated that the historic literature is incorrect, and that these three species share common 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 Islands ( US

- EPA, 2008a ) . Where feral populations of cotton species similar to cultivated cotton exis t, EPA has prohibited the sale or distribution of Bt cotton in those areas, on the premise that these containment measures would prevent the movement of the registered Bt endotoxin from Bt cot ton 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 cotton cultivation occurs ( Coile and Garland, 2003 ; USDA

NASS, 2007 ; Wunderlin and Hansen, 2010 )

. Horizontal gene transfer and consequent expression of DNA from a plant species to bacteria is unlikely to occur ( Keese, 2008 )

. Many bacteria (or parts thereof) that are closely associated with plants have be en sequenced, including Agrobacterium and Rhizobium ( Kaneko et al., 2000 ; Kaneko et al., 2 002 ; 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 t ransfer 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 , f ocus primarily on human and animal consumption. Although c otton is grown primarily for textileBAYER TWINLINKTM COTTON

28 production, valuable food o il is produced from c otton seed . T he remaining seed residue (cottonseed meal and hulls) is used as livestock fee d and as a high

- prot ein flour ( OECD, 2009 )

. Cotton seed meal animal feed provides protein, fat, and energy ( National Cottonseed Products Association, 2002 )

. Non

- GE cotton varieties, both those developed 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 orga nisms for food and feed may undergo a voluntary consultation process with the FDA prior to release onto the market. Although a voluntary process, thus far , all applicants who have wish ed 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 commercialize a bioengineered food meets with the agency to identify and discuss relevant saf ety, 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 evaluates the submission and responds to the developer by letter ( US

- FDA, 2010 )

. Bayer has provided the FDA with information on the identity, function, and characterization of the genes for TwinLinkTM Cotton , including expression of the gene products. The FDA is currently reviewing the information submitted by the applicant. In the absence of a completed FDA determination, APHIS takes into consideration prior FDA reviews of comparable products to make a preliminary assessment of the potential food and safety impacts

. Note that with regard to the expression of glufosinate ammonium tolerance, the gene construct in the Bayer TwinLinkTM Cotton is the same as that approved by the FDA in June 2003 for the LibertyLink ® US

- FDA, 2003 Cotton product ( ) . In that approval, the FDA noted tha t the transformational event in LibertyLink ® US

- FDA, 2003 Cotton was not materially different in composition, safety, or any other relevant parameter than other cotton g rown, marketed and consumed at that time ( )

. This previous FDA review will be used by APHIS to analyze the food and safety impacts associated with the incorporation and expression of glufosinate ammonium tolerance in TwinLinkTM Cotton. The FDA’s oversight of the food and safety impacts associated wit h the incorporation and expression of pesticidal substances, in this case, the Cry proteins associated with Bt, are more limited. T he EPA is the primary authority for the review of plant

- incorporated protectants.
EPA Section 3 FIFRA registration applicati ons 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 produ ct. APHIS has been advised that the product will need 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 preliminary assessment of the potential impacts

. 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 assessment, the Bt BRAD found no unreasonable adverse effects associated with exposure to the Bt products. The Bt protein 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 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 PAT protein ( 40 CFR §174.522; US

- EPA, 2010g )

. A temporary exemption from the requirement of a tolerance has been issued for Cry2Ae ( 40 CFR §174.530; US

- EPA, 2010e )

. This previous E PA review will be used by APHIS to analyze the food and safety impacts associated with the incorporation and expression of Cry proteins in TwinLinkTM Cotton. Glufosinate ammonium is currently registered under various trade names for control of weeds in cott on and other crops ; the Ignite ® /Liberty ® trade name is registered for use on LibertyLink ® BCS, 2010a corn, cotton, soy, rice and canola varieties ( ; Scott, 2008 )

. EPA’s core pesticide risk assessment and regulatory processes ensure that protections are in place for all populations of nonta rget species. These assessments provide EPA with information needed to develop label use
r estrictions for the pesticide. G rowers are required to use pesticides, such as glufosinate ammonium, consistent with the application instructions provided on the EP A

- approved pesticide label ( see, e.g., BCS, 2010a )

. These label restrictions carry the weight of law and are enforced by EPA and the states (Federal Insecticide, Fungicide, and Rodenticide Act 7 USC 136j (a)(2)(G) Unlawful A cts) . Therefore, it is expected that glufosinate ammonium use on the TwinLinkTM Cotton product would be consistent with the EPA

- approved label.
Glufosinate ammonium acts in plants by inhibiting the enzyme glutamine synthase cau sing a toxic buildup of ammonia within the treated plant ( Scott, 2008 )

. Glufosinate ammonium is a nonselective herbicide for both non

- crop and crop uses. 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 environment. Soil microorganisms, bees, earthworms, birds , and mamma ls are unaffected by glufosinate ammonium ( Scott, 2008 )

. 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 utilized in the animal feed industry as sources of protein, fiber and energy ( National C ottonseed 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 other plant and animal protein sources ( National Cottonseed Products Association, 2002 ) . Cottonseed hulls are the outer covering of the cotton seed which are removed prior to oil extraction ( National Cottonseed Products Association, 2002 )

. The hulls provide a source of fiber to animal feed rations ( National Cottonseed Products Association, 2002 ) . Whole cottonseed is the seed remaining 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 animal fee d ( Blasi and Drouillard, 2002 ; Calhoun, 2011 ) , gossypol, one of the chemical constituents of cottonseed meal, is potentially toxic ( Gadberry, 2011 )

. Ruminants (i.e. cattle) have the ability to detoxify gossypol dur ing the fermentation process in the gut ( Gadberry, 2011 ) . Gossypol has been identified as potentially causing reduced fert ility in young developing bulls, resulting in recommendations to limit cottonseed meal in the diet for developing bulls ( Blasi and Drouillard, 2002 ; University of Arkansas Cooperative Extension Service, 2011 )

. Non

- GE cotton varieties, both those developed for conventional use and for use in organic prod uction systems, are not routinely required to be evaluated by any regulatory agency in the U.S. for animal feed safety prior to release in the market. Under the FFDCA, it is the responsibility of food and feed manufacturers to ensure that the products the y market are safe and properly labeled. F eed derived from TwinLinkTM Cotton must be in compliance with all applicable legal and regulatory requirements. GE organisms for feed may undergo a voluntary consultation process with the FDA prior to release onto the market. Although a voluntary process, thus far , all applicants who have wish ed 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 commercial ize a bioengineered food meets with the agency to identify and discuss relevant safety, 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 evaluates the submission and responds to the developer by letter ( US

- FDA, 2010 )

. Bayer has provided the FDA with information on the identity, function, and characterizatio n of the genes for TwinLinkTM Cotton , including expression of the gene products. The FDA is currently reviewing the information submitted by the applicant. In the absence of a completed FDA determination, APHIS takes into consideration prior FDA reviews o f comparable products to make a preliminary assessment of the potential food and safety impacts

. Note that with regard to the expression of glufosinate ammonium tolerance, the gene construct in the Bayer TwinLinkTM Cotton is the same as that approved by the FDA in June 2003 for the LibertyLink ® US

- FDA, 2003 Cotton product ( ) . In that approval, the FDA noted that the transformational event in LibertyLink ® US

- FDA, 2003 Cotton was not materially different in composition, safety, or any other relevant parameter than other cotton g rown, marketed and consumed at that time ( )

. This previous FDA review will be used by APHIS to analyze the food and safety impacts associated with the incorporation and expression of glufosinat e ammonium tolerance in TwinLinkTM Cotton.BAYER TWINLINKTM COTTON

31 The FDA’s oversight of the food and safety impacts associated with the incorporation and expression of pesticidal substances, in this case, the Cry proteins associated with Bt, are more limited. T he EPA is the pr imary authority for the review of plant

- incorporated 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 advised that the product will need three separate new active ingredient registrations under Section 3 of FIFRA:

**•** One for Cry1Ab f rom 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 preliminary assessment of the potential impacts

. 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 assessment, t he Bt BRAD found no unreasonable adverse effects associated with exposure to the Bt products. The Bt protein 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 p lant 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 PAT protein ( 40 CFR §174.522; US

- EPA, 2010g )

. A temporary exemption from the requirement of a tolerance has been issued for Cry2Ae ( 40 CFR §174.530; US

- EPA, 2010e )

. This previous EPA review will be used by APHIS to analyze the food and safety impacts associated with the incorporation and expression of Cry proteins in TwinLinkTM Cotton.
Glufosinate ammonium is curr ently registered under various trade names for control of weeds in cotton and other crops ; the Ignite ® /Liberty ® trade name is registered for use on LibertyLink ® BCS, 2010a corn, cotton, soy, rice and canola varieties ( ; Scott, 2008 )

. EPA’s core pesticide risk assessment and regulatory processes ensure that protections are in place for all populations of nontarget species. These assessments provide EPA with information needed to develop label use
r estrictions for the pesticide. G rowers are required to use pesticides, such as glufosinat e ammonium, consistent with the application instructions provided on the EPA

- approved pesticide label ( see, e.g., BCS, 2010a )

. These label restrictions carry the weight of law and are enforced by E PA and the states (Federal Insecticide, Fungicide, and Rodenticide Act 7 USC 136j (a)(2)(G) Unlawful A cts) . Therefore, it is expected that glufosinate ammonium use on the TwinLinkTM Cotton product would be consistent with the EPA

- approved label.
Glufosinat e ammonium acts in plants by inhibiting the enzyme glutamine synthase causing a toxic buildup of ammonia within the treated plant ( Scott, 2008 )

. Glufosinate ammonium is a nonselective herbicide for both non

- crop and crop uses. It is highly b iodegradable, has no residual activity, and has very low toxicity for humans. 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 environment. Soil microorganisms, bees, earthworms, birds , and mammals are unaffected by glufosinate ammonium ( Scott, 2008 )

. 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 into textiles ( OECD, 2009 )

. Cotton seed also produces valuable food oil and the remaining seed residue (cottonseed meal and hulls) is used as livestock feed and as a high

- protein flour ( OECD, 2009 )

. The remainder of the plant is left in the field for decomposition as fertilizer ( OECD, 2009 )

. In the U.S. for the 2010 production year, cotton 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 Car olina (0.545 million acres) and T ennessee (0.387 million acres) ( USDA

- NASS, 2010c )

. Texas, Arkansas, Georgia, North Carolina , and Mississippi collectively produced over 58% of t he cotton in 2010 ( USDA

- NASS, 2010e ) . In 2007, total cotton cultivated land in the U.S. was about 10,500,000 acres (slightly larger than the combined size of the states of Maryland and Connecticut) with a market value of a bout $5 billion ( USDA

- NASS, 2007 )

. Cotton acreage in the U.S. rose slightly during the first half of the 2000s, with a correspondi ng increase in production ( USDA

- ERS, 2009 )

. The increase in production has been attributed to advances in technology (seed varieties, fertilizers, pesticides, and machinery) and production practices ( reduced tillage, irrigation, crop rotations, and pest management systems) ( USDA

- ERS, 2009 )

. The impact of these changes has been particularly 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 alternative crops, such as corn and soybeans ( USDA

- ERS, 2009 ) . All regions of the Cotton Belt have experienced sign ificant declines compared with the first half of the 2000s ( USDA

- ERS, 2009 )

. Although U.S. cotton production decreased considerably following the area reductions of the late 2000s, consistently higher yields helped limit the effect of these acreage 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 number 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 increase , but the share of small cotton farms (under 100 acres) has decline d ( USDA

- ERS, 2009 )

. The U.S. cotton industry generates about 200,000 jobs among the various sectors from farm to textile mill, and accounts for more than $25 billion in products and services annually ( USDA

ERS, 2009 )

. Total 2010 cotton production is forecast at 18.9 million 480

- pound bales, up 55 % from 2009 production of 12.2 million bales ( USDA

- 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 forecast 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 sh are than for wheat, corn, soybeans, or rice ( USDA

- ERS, 2009 )

. Through trade in yarn, fabric, and clothing, 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 peaked in 1997; since then, U.S. mill use of cotton has plummeted, dropping about 50% by 2005 and nearly 70% b y 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 before then . T he MFA was a multilateral agreement signed in 1974, but it began in the 1930s as a mechanism to protect domestic textile markets, establishing strict limits o n imports from Japan, Hong Kong, Pakistan and India, among many others ( Meyer et al., 2007 ) . Under the MFA, Canada, the U.S., and the EU could set limits on the amount of foreign

- made apparel and textiles they would allow into their countries ( Meyer et al., 2007 )

. The MFA ended after the 1995 Uruguay 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 finished textiles ( Meyer et al., 2007 ) . U.S. consumer demand for cotton products remains strong, but imported clothing now accounts for most purchases by U.S.
consumers ( USDA

- ERS, 2006 ) . The changes after the end of the MFA is reflected in the importance of exports for cotton growers . The USDA ERS reports that U

. S

. c otton mills consumed 60% of the domestic cotton through the 1990s, but not long after 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 demand concerns confronting other field crops. However, because cotton is used primarily in manufactured products, such as clothing and home fu rnishings, the industry faces additional challenges associated with the economic well

- being of downstream manufacturing industries , as well as the economic well

being of the final consumer ( USDA

- ERS, 2009 )

. The USDA ERS notes that U.S. cotton productivity has increased in recent years as a result of technological advancement s, including biotechnology

- derived varieties ( Meyer et al., 2007 )

. The introduction of cotton providing insect resistance or herbicide tolerance also has glob al implications. Herbicide

- tolerant cotton is one of the most widely and rapidly adopted crops in the U.S., followed b y insect

- resistant cotton ( USDA

- ERS, 2010a ) . In 2010, GE varieties of cotton were c ultivated 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 cotto n has been widely adopted in India and China, two major cotton producers ( 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 producing countries like Australia, Argentina, Mexico, and South Africa ( Meyer et al., 2007 )

.BAYER TWINLINKTM COTTON

34 

### 3 ALTERNATIVES This document analyzes the potential environmental consequences of a determination of nonregulated status of BCS TwinLinkTM Cotton. To respond favorably to a petition for nonregulated status, APHIS 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 subject to 7 CFR part 340 or the plant pest provision s of the Plant Protection Act.

T wo 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 Alternative, APHIS would deny the petition

. TwinLinkTM Cotton and progeny derived from TwinLinkTM Cotton would continue to be regulated articles under the regulations at 7 CFR Part 340

. Permits issued or notifications acknowledged by APHIS would still be required for introductions of TwinLinkTM Cotton and measures to ensure physical and reproductive confinem ent would continue to be implemented

. APHIS might choose this alternative if there were insufficient evidence to demonstrate the lack of plant pest risk from the unconfined cultivation of TwinLinkTM Cotton

. This alternative is not the Preferred Alternativ e because APHIS has concluded through a Plant Pest Risk Assessment ( USDA

- APHIS, 2010b ) that 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 responding 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 ) . Permits issued or notification s acknowledged by APHIS would no longer be required for introductions 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 for nonregulated status based o n the requirements in 7 CFR part 340 and the agency’s authority under the plant pest provisions of the Plant Protection Act. Because the agency has concluded that TwinLinkTM Cotton is unlikely to pose a plant pest risk, a determination of nonregulated sta tus of TwinLinkTM Cotton is a response that is consistent with the plant pest provisions of the PPA, the regulations codified in 7 CFR part 340, and the biotechnology regulatory policies in the Coordinated Framework. Under this alternative, growers may have future access to TwinLinkTM Cotton and progeny derived from this event if the developer decides to commercialize TwinLinkTM Cotton.BAYER TWINLINKTM COTTON

35 3.3 ALTERNATIVES CONSIDERED BUT REJECTED FROM FURTHER CONSIDERATION APHIS assembled a list of alternatives that might be cons idered for TwinLinkTM Cotton

. The agency evaluated these alternatives , in light of the agency's authority under the plant pest provisions of the Plant Protection Act, and the regulations at 7 CFR part 340, with respect to environmental safety, efficacy, an d practicality to identify which alternatives would be further considered for TwinLinkTM Cotton . Based on this evaluation, APHIS rejected several alternatives. These alternatives are discussed briefly below along with the specific reasons for rejecting eac h.
3.3.1 Prohibit A ny TwinLinkTM Cotton from Being Released In response to public comments that stated a preference that no GE organisms enter the marketplace, APHIS considered prohibiting the release of TwinLinkTM Cotton , including denying any permits associated with the field testing. APHIS determined that this alternative is not appropriate given that APHIS has concluded that TwinLinkTM Cotton is unlikely to pose a plant pest risk ( USDA

- APHIS, 2010b )

. In enacting the Plan t Protection 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 Heads of Ex ecutive Departments and Agencies, the White House Emerging Technologies Interagency Policy Coordination Committee developed broad principles, consistent with Executive Order 13563, to guide the development and implementation of policies for oversight of em erging technologies (such as genetic engineering) at the agency level. In accordance with this memorandum, agencies should adhere to Executive Order 13563 and, consistent with that Executive Order, the following principle, among others, to the extent perm itted by law, when regulating emerging technologies: “[D]ecisions should be based on the best reasonably obtainable scientific, technical, economic, and other information, within the boundaries of the authorities and mandates of each agency”

Based on ou r 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 petition 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 associated with some, but not all lines described in a petition. Because APHIS has concluded that TwinLinkTM Cotton is unlikely to pose a plant pes t risk,BAYER TWINLINKTM COTTON

36 there is no regulatory basis under 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 respons e 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, because APHIS has concluded that TwinLinkTM Cotton i s unlikely to pose a plant pest risk (USDA

- APHIS, 2010b) , an alternative based on requiring isolation distances would be inconsistent with the statutory authority under the plant pest provisions of the Plant Protection Act and regulations in 7 CFR part 340. APHIS also considered geographically restricting the production of TwinLinkTM C otton based on the location of production of non

- GE cotton in organic production systems in response to public concerns regarding possible gene movement between GE and non

- GE plants. However, as presented in APHIS’ plant pest risk assessment for TwinLinkTM Cotton , there are no geographic differences associated with any identifiable plant pest risks for TwinLinkTM Cotton (USDA

APHIS, 2010b). This alternative was rejected and not analyzed in detail because APHIS has concluded that TwinLinkTM Cotton does not p ose a plant pest risk, and will not exhibit a greater plant pest risk in any geographically restricted area. Therefore, such an alternative would not be consistent with APHIS’ statutory authority under the plant pest provisions of the Plant Protection Act and regulations in Part 340 and the biotechnology regulatory policies embodied in the Coordinated Framework. Based on the foregoing, the imposition of isolation distances or geographic restrictions would not meet APHIS’ purpose and need to respond approp riately to a petition for nonregulated status based on the requirements in 7 CFR part 340 and the agency’s authority under the plant pest provisions of the Plant Protection Act. Nevertheless, APHIS is not expecting significant effects. However, individua ls might choose on their own to geographically isolate their non

GE 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 testing to identify GE products in non

- GE production systems. APHIS notes there are no nationally

- established re gulations involving testing, criteria, or limits of GE material 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 the Plant Protection Act, the regulations at 7 CFR part 340 and the biotechnology regulatory policies embodied in the Coordinated Framework. Therefore, imposing such a requirement for 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 impacts 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 Impacts 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 Production Unchanged Unchanged Cropping P ractices Unchanged Unchanged Seed Production Unchanged Unchanged Organic Farming Unchanged Unchanged Specialty Cotton Production 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 Environment 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 deregulated GE organisms FDA consultation pending , EPA tolerance exemptions and conditional pesticide registrations being reviewed
Compliance with Other Laws CWW, CAA, EOs Fully compliant Fully compliantBAYER TWINLINKTM COTTON

38 N otes :

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 ENVIRONMENT AL CONSEQUENCES This analysis of potential environmental consequences addresses the potential impact to the human environment from the alternatives analyzed in this EA, namely taking no action and a determination by the agency that TwinLinkTM Cotton does no t pose a plant pest risk.

4.1 SCOPE OF THE ENVIRONMENTAL ANALYSIS 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 environme ntal issue and includes a discussion of the c umulative impacts that are associated with the Preferred Alternative , when combined with other recent past, present and reasonably foreseeable future actions within the affected environment

. Certain aspects of this product and its cultivation would be no different between the alternatives; those instances are described below

.

For the discussion of environmental consequences, 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 Communities ( 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 Species ( Subsection 4. 9 ); and

**•** Consideration o f Executive Orders, Standards, a nd Treaties Relating t o Environmental Impacts ( Subsection 4. 10 ).

Although the Pref erred Alternative would allow for new plantings of TwinLinkTM C otton to occur anywhere in the U.S., APHIS is limit ing 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, Arkansas, California, Florida, Georgia, Kansas, Louisiana, Mississippi, Missouri, New Mexico, North Carolina, Oklaho ma, South Carolina, Tennessee, Texas, and Virginia ( USDA

- NASS, 2010c )

. The environmental consequences of the No Action and Preferred Alternatives are analyzed under the assumpt ion that farmers who produce conventional cotton, TwinLinkTM C otton , or cotton using organic methods are using reasonable, commonly accepted best management practices specific to their agricultural cotton production

. However, APHIS recognizes that not all farmers follow these best management practices for cotton

. 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 A GRICULTURAL PRODUCTION OF COTTON

One of APHIS’s missions is to improve American agricultural productivity

. Best management practices are commonly accepted, practical ways to grow cotton, regardless of whether the cotton farmer is using conventional practices with non

- GE or GE varieties, or organic practices . These management practices consider crop

- specific planting dates, seeding rates, and harvest times , among others. Over the years, cotton production has resulted in well

- established, widely

practiced management practices that are availa ble through local Cooperative Extension Service offices and their respective websites

. The National Information System for the Regional Integrated Pest Management Centers publishes crop profiles for major crops on a state

- by

- state basis

. These crop profi les provide production guidance for local growers, including recommended practices for specific pest control

. Crop profiles for many of the cotton production states can be reviewed at ( www.ipmcenters.org/cropprofiles/index.cfm )

. BCS’ studies demonstrate that agronomic characteristics and cultivation practices required for TwinLinkTM Cotton are essentially indistinguishable from other cotton varieties , including other herbicide

- tolerant and insect

- resistant varieties ( Scott, 2008 ; USDA

- APHIS, 2010b )

. Although TwinLinkTM Cotton might be expected to replace other varieties of cotton currently cultivated, new acreage is not expected to be developed to accommodate the cultivation of TwinLinkTM Cotton ( Scott, 2008 ) . None of the best management practices currently employed for cotton production is expected to change if TwinLinkTM Cotton is deregulated. Accordingly, the potential impacts on agricultural production of TwinLink TM Cotton resulting from management practic es 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 development

. Cotton acreage 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 considerably lower as market prices have favored the planting of alternative 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 th e GE cotton acreage was GE insect

resistant (Bt) cotton and 78% was herbicide

- tolerant ( USDA

- ERS, 2010a , 2010c )

. Most cotton is planted on farms that have been in cotton production for at least five years ( USDA

- NASS, 2007 )

.

Based on current acreage trends, conventional cotton production practices with GE varieties will likely continue to dominate or perhaps increase in acreage under the No Action Alternative

. Cotton is currently produced commercially in 17 states ( No Action : Acreage and Areas of Cotton Production

USDA

- NASS, 2010c ) and under the No Action Alternative, this range of production is not expected to change.

BCS’ studies demonstrate that agronomic characteristics and cultivation practices required for TwinLinkTM Cotton are essentially indistinguishable from ot her cotton varieties, including other herbicide

- tolerant and insect

- resistant varieties ( Preferred Alternative : Acreage and Areas of Cotton Production

Scott, 2008 ; USDA

- APHIS, 2010b )

. BCS positsBAYER TWINLINKTM COTTON

41 that although TwinLinkTM Cotton might be expected to replace other varieties of cotton currently cultivated, new acreage is not expected to be developed to accommodate the culti vation of TwinLinkTM Cotton ( Scott, 2008 )

. The introduced lepidopteran resistance trait and the herbicide tolerance trait do not confer any competitive advantage in terms of weediness or to extend the range of cultivation outside of existing c ultivation areas ( Scott, 2008 ) . T he Preferred Alternative, a determination of nonregulated status of TwinLinkTM C otton , is thus not expected to increase cotton production, either by its availability alone or accompanied b y other factors, or cause an increase in overall GE cotton acreage

. Impacts would be similar to the No Action Alternative

.

Cumulative effects of a determination of nonregulated status of TwinLinkTM C otton are unlikely

. Neither the No Action Alternative nor a determination of nonregulated status of TwinLinkTM C otton is expected to directly cause an increase in agricultural acreage devoted to cotton production or those cotton acres devoted t o GE cotton cultivation

. The availability of TwinLinkTM Cotton will not change cultivation areas for cotton production in the U.S. and there are no anticipated changes to the availability of GE and non

- GE cotton varieties on the market under either alternative

. Cumulative Effects : Acreage and Areas of Cotton Production

4.2.2 Cropping Practices : Crop Rotation and Pesticide Use Crop rotation in cotton is conducted to optimize soil nutrition and fertility, reduce pathogen loads, and control certain cotton pests ( Gazaway et al., 2007 ) ; ( University of California IPM Online, 2008a )

. Crop rotation practices have been described previously in Secti on 2. Crop rotation practices are not expected to change with the planting and production of TwinLink TM Cotton. In the U.S., the cotton industry has consistently relied heavily on insecticide use strategies to manage arthropod pests ( Gianessi and Carpenter, 1999 ) . Insecticides were applied to 66 % of the cotton acreage in 2007 ( USDA

- NASS, 2008 ) . Of thirty listed insecticides, Acephate was the most utilized insecticide, with 26% of the planted 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 the acreage at an average rate of 0.565 pounds per year ( USDA

- NASS, 2008 ) . Other insecticides appl ied in the 2007 growing year included Acetamiprid (6% of acreage), Cyfluthrin (8% of acreage), Cypermethrin (7% of acreage), Imadacloprid (6% of acreage), Lambda

- cyhalothrin (5% of acreage), Malathion (5% of acreage), and Thiamethoxam (11% of acreage).

R esistance to commonly used insecticides (pyrethroids, organophosphates, and carbamates) has led to a need for new insecticide chemistries or other novel approaches such as the use of sterile insects, pathogens, and transgenic cotton ( Gianessi and Carpenter, 1999 ) . Cotton varieties expressing the Bt endotoxin have emerged as a viable product replacing many of these commonly used pesticides ( Scott, 2008 )

. One of the documented benefits of Bt cotton has been the reduction in the use of broad

- sp ectrum insecticides during cotton production ( Benbrook, 2009 ; Fernandez

- Cornejo and Caswell, 2006 )

.BAYER TWINLINKTM COTTON

42 Each acre planted in Bt cotton is presumed to displace the average pounds of insecticide previously sprayed on an acre of conventional cotton for lepidopteran control ( Benbrook, 2009 )

. In a comprehen sive evaluation of 13 years of USDA NASS data (1996 through 2008) on pesticide usage (including both herbicides and insecticides), Benbrook found that pesticide application rates for managing lepidopteran pests in cotton changed 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 table presents an extracted summary to show use trends.

Source : Benbrook ( 2009 ) Supplemental Table 12. Insecticides 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 insec ticides to control lepidopteran pests in cotton directly associated 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 ( Scott, 2008 ) . In 2007, 35 different herbicides were applied to 97% of the cotton acreage ( USDA

- NASS, 2008 ) . Glyphosate isopropylamine salt was the most commonly applied herbic ide, 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 basis were Triflurali n and Diuron, at 29 and 26 % with average application rates of 0.921 and 0.499 pounds per acre per crop year, respectively ( USDA

- NASS, 2008 )

. Other herbicides applied to cotton incl uded 2,4

- D (6% of the acreage), Carfentrazone

- ethyl (10% of the acreage), Flumioxazin (6% of the acreage), Pendimethalin (17% of the acreage), Prometryn (7% of the acreage) , Pyraflufen

- ethyl ( 8 % of the acreage) , pyrithiobac

- sodium (1 0 % of the acreage) , and S

- metolachlor ( 6 % of the acreage) ( USDA

- NASS, 2008 )

. The consequences of adoption of herbicide tolerance show a contrasting trend. Herbicide

tolerant cotton was first grown commercially in the U.S. in 1997 and by 2008 , herbicide

- tolerant cotton was planted on 68% of total cotton plantings ( Brookes and Barfoot, 2010 ) . Stacked varieties, providing both herbicide tolerance and insect resistance, comprised nearly 93% of the total plantings in 2008 ( Benbrook, 2009 ) . Benbrook has reported that the adoption of herbicide

tolerant crops has resulted in an increase in the volume of herbicides applied to crops ( Benbrook, 2009 ) . Benbrook notes that herbicide use declined between 1996 and 2 001, apparently in direct response to the adoption of herbicide

- tolerant cotton ( Benbrook, 2009 ) . However, since that time, herbicide use has increased. Reported increases in herbicide use reflect an increase in use of conventional herbicid es, as well as an increase in glyphosate applications ( Benbrook, 2009 )

. Benbrook concludes that this reported increase in herbicide use is a direct consequence of theBAYER TWINLINKTM COTTON

43 development of glyphosate

- resistant weeds in cotton. As of 2008, nine wee ds were identified as glyphosate

- resistant, with three of these weeds, horseweed (marestail), Johnsongrass, and pigweed (Palmer amaranth) specifically identified as difficult to control weeds in cotton.

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 deregulated GE lepidopteran

- resistant and her bicide

- tolerant cotton varieties, as well as conventional cotton varieties . It is likely that growers would continue to experience the continued emergence of glyphosate

- tolerant weeds, requiring modifications of crop management practices to address these weeds. These changes may involve all of the techniques identified by Benb rook ( No Action : Cropping Practices : Crop Rotation and Pe sticide Use

2009 ) , including use of other herbicides for weed control.

BCS’ s tudies demonstrate TwinLinkTM Cotton is essentially indistinguishable from other cotton varieties used in terms of agronomic characteristics and cultivation practices ( Preferred Alternative : Cropping Practices : Crop Rotation and Pesticide Use

Scott, 2008 ; USDA

- APHIS, 2010b ) . Therefore, cotton crop rotation practices are expected to be unchanged
u nder the Preferred Alternative

. If TwinLinkTM Cotton is adopted, a continued reduction in the use of budworm/bollworm insecticides applications and the number of acre

- treatments per year as reported in Benbrook’s trend analysis is expected to occur

. The deregulation of TwinLinkTM Cotton will provide growers with another alternative Bt cotton variety to cultivate

. Herbicide use patterns have the potential to change as well. The introduction of TwinLinkTM Cotton provides a stacked variety, expressing Bt

- based lepidopteran resistance combined with tolerance to glufosinate ammonium, an alternative herbicide. TwinLinkTM Cotton provides growers with an alternative to those cotton varieties resistant to glyphosate, thus expanding options in the field for weed control. In those fields where glyphosate

- resistant weed s 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 glufosinate ammonium could reduce applications of those other herbici des needed to manage glyphosate

- resistant weeds.

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 foreseeable actions that would aggregate with effects of the proposed action to affect changes in crop rotation

. Studies demonstrate TwinLinkTM Cotton is essen tially indistinguishable from other cotton varieties used in terms of agronomic characteristics and cultivation practices ( Cumulative Effects : Cropping Practices : Crop Rotation and Pesticide Use

Scott, 2008 )

. It is anticipated that the trend of reduced broad

- spectrum insecticide use by cotton growers will contin ue due to the adoption of Bt cotton and other cultural practices . There is the potential that the introduction of glufosinate ammonium tolerance may result in a reduction in total herbicide use as growers adopt different herbicide treatment strategies inv olving glufosinate ammonium asBAYER TWINLINKTM COTTON

44 a post

- emergent crop treatment, thereby reducing the use of some of the other herbicides which have been required in response to the emergence of glyphosate

- 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 isolation dis tances to prevent pollen movement from other cotton sources ; 2) planting border or barrier rows to intercept pollen ; 3) employing natural barriers to pollen ; and 4) f ield monitoring for off

- types, other crops, weeds, and disease

.

Under the No Action Alternative, current cotton seed production practices are not expected to change. No A ction : Seed Production

The production of cotton seed for TwinLinkTM Cotton will be conducted consistent with these standard practices; consequently, no changes to seed production practices are required. Based on the data provided by BCS for TwinLinkTM Cotton ( Preferred Alternative : Seed Production

Scott, 2008 ) , as well as previous experience with o ther Bt cotton varieties that have been widely adopted by growers since their introduction in 1996 ( USDA

- ERS, 2010a ) , APHIS has concluded that the availability of TwinLinkTM Cotton would not alter the agr onomic practices, locations, and seed production and quality characteristics of conventional and GE seed production ( USDA

- APHIS, 2010b ) . The deregulation of TwinLinkTM Cotton will not require a change to seed production practices.

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 TwinLinkTM 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 agronomic practices and locations for cotton seed production using TwinLinkTM Cotton are not expected, no cumulative effects have been identified for seed production. Cumulative Impacts : Seed Production

4.2.4 Organic Farming

O rganic production plans prepared pursuant to t he National Organic Program (NOP) include practical methods to protect organically

- produced crops from accidental contaminati on with GE materials. Typically, organic growers use more than one method to prevent unwanted material from entering their fields including : isolation of the farm, physical barriers or buffer zones between organic production and non

- organic production, a s well as formal communications between neighboring farms ( Baier, 2008 ; Guerena and Sullivan, 2003 ; NCAT, 2003 ) . These practices follow the same system utilized for th e cultivation of certified seed under the AOSCA procedures.BAYER TWINLINKTM COTTON

45 APHIS recognizes that producers of non

- GE cotton, particularly producers who sell their products to markets sensitive to genetically engineered traits (e.g., organic or some export markets), can b e reasonably assumed to be using 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 pra ctices as APHIS’s baseline for the analys i s of the alternatives

.

Current availability of seed for conventional (both GE and non

- GE) cotton varieties, and those cotton varieties that are developed for organic production, is expe cted to remain the same under the No Action Alternative

. Commercial production 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 organi c cotton will continue to fluctuate with market demands , as it has over the last 10 years, and these markets are likely to continue to fluctuate under the No Action Alternative ( No Action : Organic Farming

USDA

- ERS, 2010c , 2010e )

.

Transgenic cotton lines including those that are resistant to lepidopteran insects and herbicide

tolerant are already in widespread use by farmers

. TwinLinkTM Cotton should not present any new and different issues and impacts for organic and other specialty cotton producers and
consumers

. Preferred Alternative : Organic Farming

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

. The trend in the cultivation of GE cotton, non

GE, and organic cotton varieties, and the corresponding production systems to maintain 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. demonstrated that TwinLinkTM Cotton is not significantly different in plant growth, yield, and reproductive capacity from its nontransgenic counterpart ( USDA

- APHIS, 2010b )

. No differences were observed in pollen diameter, weight, and viability

. Therefore, TwinLinkTM Cotton is expected to pres ent a similar risk of cross

- pollination as existing cotton cultivars. The practices currently employed to preserve and maintain purity of organic production systems would not be required to change to accommodate the production of TwinLinkTM Cotton.
Histo rically, organic cotton production represents a small percentage (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 commercial cotton production

. Accordingly, the deregulation of TwinLinkTM Cotton is not expected to have a significant impact
on organic cotton production.BAYER TWINLINKTM COTTON

46 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 nonregulat ed 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 systems to maintain their marke t share. Although 12 GE cotton events or lines were deregulated pursuant to Part 340 and the Plant Protection Act , between 2000 and 2008, the total acreage associated with the organic production of cotton remained at slightly above 15,000 acres ( Cumulative Effects : Organic Farming

USDA

- ERS, 2010e )

. 4.2.5 Specialty Cotton Production Specialty crop growers employ practices and standards for seed production, cultivation, and product handling and processing to ensure that their products are not pollinated by or commingled with conventional or genetically engineered crops ( Bradford, 2006 ) . These management practices include maintaining isolation distances to prevent pollen movement from other cotton sources, planting border or barrier rows to intercept pollen, and employing natural barriers to pol len ( Bradford, 2006 ; NCAT, 2003 ; Wozniak, 2002 )

. The se management practices allow the grower to meet standards for the production of specialty crop seed , m aintain genetic purity , and protect the ge netic diversity of cotton ( Bradford, 2006 )

.

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

. No Action : Specialty Systems

As noted in the discussion of Seed Production, no changes in the production of seed or cultivation of cotton are required to accommodate TwinLinkTM Cotton. Preferred Alternative : Specialty Systems

It is unlikely that specialty system farmers , or other farmers who choose to plant non transgenic varieties or sell non transgenic seed, will be substantially impacted by the expected commercial use of TwinLinkTM Cotton

. Transgenic cotton lines including those that are resistant to lepidopteran insect s and express herbicide tolerance 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 TwinLinkTM Cotton is not significantly different in plant growth, yield, and reproductive capacity from its nontransgenic counterpart ( USDA

- APHIS, 2010b )

. No differences were observ ed in pollen diameter, weight, and viability

. Therefore, TwinLinkTM Cotton is expected to present a similar risk of cross

- pollination as existing cotton cultivars including other GE cotton varieties. The practices currently employed to preserve and mainta in 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 would not change the availability andBAYER TWINLINKTM COTTON

47 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 orga nic cotton farming, are in place to maintain the genetic diversity of cotton

. Cotton growers 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.

A determination of nonregulated status of TwinLinkTM Cotton is not expected to change the market demands for GE cotton or cotton produced using specialty systems

. A determination of nonregulated status of TwinLinkTM Co tton will add another GE cotton variety to the cotton market

. Between 2000 and 2009, 12 GE cotton events or lines were deregulated pursuant to Part 340 and the Plant Protection Act

. Based on demonstrated agronomic characteristics and cultivation practices, and because the market share of specialty cotton varieties is unlikely to change by the introduction of TwinLinkTM Cotton , APHIS has determined that there are no past, present, or re asonably foreseeable changes that would impact specialty cotton producers and
consumers

. C umulative Effects : Specialty Systems

4.3 PHYSICAL ENVIRONMENT
4.3.1 Water Resources Although c otton has been developed as a drought

- tolerant crop ( Cotton Inco rporated, 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 factor in the reduction of acres of cotton grown in California, Arizona, and New Mexico in the past decade ( Cotton Incorporated, 2010b )

. In these
ar eas, cotton has been displaced by higher value crops and land uses ( Cotton Incorporated, 2010b )

. Water quality is also preserved in modern cotton productions systems

. The increase in conservation t illage practices has resulted in a reduction of runoff from agricultural lands, decreasing non

- point source pollution of suspended sediment, fertilizer, and pesticides

. Intensive monitoring of surface water and groundwater proximate to agricultural fields has demonstrated the benefits of no

- till cotton in protecting both ground and surface water resources University o f Tennessee Agricultural Extension Service, 2010 ( )

. Better nutrient management and precision technologies are ensuring inputs are used by the crop and are not entering ground or surface waters ( Cotton Incorporated, 2010a )

.

Under the No Action Alternative, current l and acreage and agronomic practices , including irrigation, tillage, and nutrient management associated with cotton production would not be expected to change. Therefore, no expected changes to water u se associated with cotton production is expected for this alternative. No Action : Water UseBAYER TWINLINKTM COTTON

48 In 2010, GE cotton occupied 93% of the cotton acreage ( Preferred Alternative : Water Use
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 production for years ( USDA

- APHIS, 20 10a ) . The introduction of GE cotton has resulted in the adoption of conservation practices ( Cotton Incorporated, 2010a ) . These conservation practices, including reduced tillage and precision agriculture play a significant role in water conservation and maintaining water quality by minimizing soil erosion ( USDA

- NRCS, 2006b )

. TwinLinkTM Cotton would not change cultivation practices for cotton product ion, nor would it increase the total acres and range 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.

No cumulative eff ects on water use have been identified for a determination 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

. Cumulative Effect : Water Use

4.3.2 Soil This section discusses the potential consequences of the No Action and the Preferred Alternatives on soil. T he USDA Natural Resources Conservation Service (NRCS) has identified significant reductions in the loss of soil from croplands in the U.S

. , finding that t otal 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 herbicide

tolerant cotton has contributed to this reduction in soil loss through a shift towards conservation tillage and the use of cover crops ( Cotton Incorporated, 2010c )

. I ncreases 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 ; McClelland et al., 2000 ; Towery and Werblow, 2010 ; USDA

- NRCS, 2010 )

. Additional s oil quality benefits from adoption of GE cotton may also be realized by reducing the risks associa ted with environmental spills or misapplications of chemical insecticides to the soil , and reductions in the frequency with which insecticides must be applied.

Land acreage and agronomic practices associated with traditional and existing non

- regulated GE cotton production would not be expected to change in response to the N o A ction A lternative

. No Action : SoilBAYER TWINLINKTM COTTON

49 Continued adoption of existing deregulated GE c otton varieties 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 production for years ( USDA

- APHIS, 2010a )

. C urrent agronomic practices associated 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 Cotton is still a regulated article. To the extent that currently available GE cotton varieties can result in the grower’s adoption of conservation tillage, soils would be positively affe cted by that adoption.

No changes to agronomic practices typically applied in the management of conventional cotton, including other commercially available GE cotton varieties, are required for TwinLinkTM Cotton. Preferred Alternative : Soil

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 ( Scott, 2008 )

. No increases in fertilizers and pesticides were required, nor were any changes in cultivation, planting, harvesting, and volunteer control required ( Scott, 2008 )

. It is expected that similar agronomic practices that are currently used for com mercially 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 reduc tion 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 disturbance because of fewer passes by heavy farm equipment. It is expected tha t only glufosinate ammonium would be applied as a post

- emergent herbicide
on TwinLinkTM Cotton. Glufosinate ammonium has been shown to rapidly dissipate from most agricultural ecosystems across a wide range of soil and climatic conditions, with a median so il half

- life (the time it takes for half of the glufosinate ammonium to dissipate in the soil) of 7.5 days ( PPDB, 2010 )

. When used in accordance with the label, glufosinate ammonium accumulation in soil has not been shown to be significant

. The bar gene which has been introduced into TwinLinkTM Cotton was originally isolated from Streptomyces hygroscopius , a common soil bacteria ( Scott, 2008 )

. The PAT enzyme produced by the TwinLinkTM Cotton is naturally occurring in the soil ( CFIA, 2004 )

. The cultivation of TwinLinkTM Cotton and th e attendant production of the PAT enzyme will therefore not cause an impact to the physicochemical characteristics of the soil. As explained above, if TwinLinkTM Cotton is adopted and cultivated, there may be positive impacts to soil when compared with trad itional cotton cultivation practices.

APHIS has not identified any cumulative effects of the use of TwinLinkTM Cotton to soils. Bayer has compared phenotypic, agronomic, and cultivation characteristics between TwinLinkTM Cumulative Effects : SoilBAYER TWINLINKTM COTTON

50 Cotton and control cottons ( Scott, 2008 )

. TwinLinkTM Cotton required the same soil, fertilizer, water and pest management practices (except for lepidopteran pest control) as non

- GE cotton ( Scott, 2008 )

. Consequently, the phenotypic, agr onomic, and ecological data presented by BCS ( Scott, 2008 ) 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

. Bas ed 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 include smoke from agricultural burning, tillage, heavy equipment emissions, pesticide drift from spraying, and nitrous oxide emissions from the use of nitrogen fertilizer ( Aneja et al., 2009 ; Hoeft et al., 2000 ; USDA

- NRCS, 2006c )

. The adoption of GE cotton has reduced air emissions from several of these sources. Conservation practices, including conservation t illage associated with GE cotton production requires fewer tractor passes across a field , thus decreas ing dust generation and tractor emissions; surface residues and untilled organic matter physically hold the soil in place thus decreasing wind erosion of soils and pesticide drift in wind

- eroded soils ( Baker et al., 2005 ; Cotton Incorporated, 2010d ; USDA

- NRCS, 2006c )

. Reduced tillage also increases sequestration rates of carbon emissions in soils ( Causarano et al., 2006 ) . This section discusses the potential consequences of the No Action and the Preferred Alternatives on air quality.

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

. No Action : Air Quality

A doption of GE c otton varieties would be expected 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 with these practices would be expected to follow. Air quality would continue to be affected by a gronomic practices associated with conventional methods of cotton production such as tillage, cultivation, pesticide and fertilizer applications, and the use of agricultural equipment

.

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 cotton cultivars

. It is expected that similar agronomic practices that are currently used for commercially available Bt cotton or herbicide

tolerant cotton will also be used by growers of TwinLinkTM Cotton. Preferred Alternative : Air QualityBAYER TWINLINKTM COTTON

51 If TwinLi nkTM Cotton is adopted and replaces non

- GE cotton varieties, air quality issues associated with pesticide application and use in non

- GE cotton production may benefit from the use of TwinLinkTM Cotton due to a reduction in the application of lepidopteran targ eted insecticides and similar reductions in herbicide applications required as growers take advantage of the post

- emergent herbicide glufosinate ammonium

. The collective impact is a reduction in the number of acre

- treatments per year requir ing the us e of heavy farm equipment, with a positive impact on air quality associated with the decrease in equipment

related emissions , as well as a decrease in dusts and pesticide drift s. Based on this information, APHIS concludes that the production of TwinLinkTM Cotton is not expected to adversely affect air quality and, in fact, may provide some benefit. However, considering the wide adoption of similar GE cotton lines, o verall impacts are likely to be similar to the No Action Alternative

.

Based on the findings described above, APHIS has determined that there are no past, present, or reasonably foreseeable actions that would aggregate with effects 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. Cumulative Effects : Air Q uality

4.3.4 Climate Change Agriculture, including land

- use changes for farming, is responsib le 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 US

- EPA, 2010b O emissions ( ) . A more comprehensive discussion of the contribution of agricultural practices to GHGs is provided in Subsection 2.3.4 , Affect ed Environment

– Climate Change. Agriculture

- related GHG emissions include carbon dioxide ( CO 2 ) , N 2 O, and methane ( CH 4 )

, produced through the combustion of fossil 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 consequences of the No Action and the Preferred Alternatives on climate change.

Under the No Action Alternative, environmental releases of TwinLinkTM Cotton would be under APHIS regulation

. Due to the limited size of these field trials, there would be no measurable effect on climate change from these confined env ironmental releases

. Land acreage and cultivation practices associated with cotton production would not be affected . Current
a gronomic practices associated with conventional cotton production and current GE cotton varieties which contribute to GHG emissi ons , including tillage, cultivation, irrigation, pesticide application, fertilizer applications and use of agriculture equipment, would not be expected to change if TwinLinkTM Cotton remains a regulated article. To the extent that the adoption and cultivat ion of GE cotton varieties allows the grower to implement conservation practices, GHG No Act ion : Climate ChangeBAYER TWINLINKTM COTTON

52 emissions would be expected to be reduced commensurate with the air quality improvement s anticipated from adoption of conservation tillage practices.

A determination of nonregulated status of TwinLinkTM Cotton w ould not change the cultivation or agronomic practices, or agricultural land acreage associated with growing cotton, and thus is expected to have the same effect on climate change as the No Action Alternative. Preferred Alternativ e : Climate Change

APHIS has not identified any cumulative effects for this issue

. The use of TwinLinkTM Cotton in commercial cotton production is not exp ected to cause any cumulative 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 TwinLinkTM Cotton deregulation

. T he consequences of the Preferred Action Al ternative on commercial cotton production and acreage are the same as for the No Action Alternative

. Cumulative Effects : Climate Change

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 insects inhabiting cotton fields. In addition, m ammal and bird species use cotton fields and adjacent vegetation for food and habitat throughout the year . This section discusses the potential consequences of the No Action and the Prefe rred 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.

Under the No Action Alternative, conventional and GE transgenic cotton production will continue while TwinLinkTM Cotton remains a regulated article

. Potential impacts of GE and
non

- GE cotton production practices on non

- target species would be unchanged

. No Action : Animals

BCS data indicates that t he a gronomic practices used to produce TwinLinkTM Cotton will be the same as those used to produce other conventionally grown GE and non

- GE cotton

. TwinLinkTM Cotton production will not change land acreage or any cultivation practices for cotton production

. Preferred Alternative : Animals

Since the mid

- 1990s, transgenic cotton lines have been commercialized without substantiated reports of significant deleterious impacts on non

- target organisms ( Mendelsohn et al., 2003 ; OECD, 2007 ; US

- EPA, 2008a ; USDA

- APHIS, 2010a ) . Evidence submit ted by BCS fromBAYER TWINLINKTM COTTON

53 laboratory and field studies as well as other peer reviewed studies demonstrate a lack of toxicity of the TwinLinkTM Cotton on non

- target representative species ( Scott, 2008 )

. BCS has presented laboratory test data with indic ator test species to determine potential toxicity of Cry enzyme toxin at doses higher than would be anticipated under field conditions ( Rose et al., 2007 )

. BCS’ selection of representati ve indicator test species was based upon the ir 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 invertebrate (water flea , Daphnia magna ) ( Scott, 2008 )

. The data submitted in the petition indicate that no s ignificant adverse effects were observed at the maximum test 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 studies with trans genic plants expressing Cry proteins ( Marvier et al., 2007 ; Naranjo, 2009b ; Romeis et al., 2006 ; Shelton et al., 2009 )

. The non

- target risks associated with e x posure of aquatic organisms is likely very low because cotton pollen is large, sticky, and is not transported 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 endotoxin are not present in vertebrates, these insecticidal proteins are not expected to adversely affect these organisms ( Hofmann et al., 1988a ; Hofmann et al., 1988b ; Shimada et al., 2006b ; US

- EPA, 2008a ; Van Rie et al., 1989 ; Van Rie et al., 1990 )

. An additional advantage of t he use of transgenic cotton producing the Bt proteins is the corresponding reduc tion in use of broad spectrum insecticides 1 Cattaneo et al., 2006 and the consequent reduction of potential impact on diversity of non

- target insects ( ; 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 conventionally grown crops ( Hardee et al., 2001 ; Moar et al., 2003 ; Sanvido et al., 2006 ; Sisterson et al., 2007 )

. BCS has filed applications with the FDA and the EPA for the TwinLinkTM Cotton variety. The FDA and EPA are c urrently reviewing the information submitted by the appli cant. Both agencies have previously reviewed and approved comparable products, including several crop varieties expressing the Bt

- based Cry proteins as plant

- incorporated protectants, as well as the bar gene, resulting in the expression of the PAT enzyme providing tolerance to glufosinate

1 Broad spectrum insecticides are chemical insecticides which ki ll 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” insects. Because the Cry proteins are specif ic 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 ammonium. In the absence of a completed FDA and EPA determination s , APHIS takes into consideration prior agency reviews of comparable products to conduct its assessment of the potential impacts.
EPA has issued a toleranc e exemption 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, 2010g )

. A temporary exemption from the requirement of a tolerance has been issued for Cry2Ae ( 40 CFR §174.530 ; US

- EPA, 2010e )

. The PAT enzyme bar gene construct was deregulated by APHIS in 2003 in the BCS LibertyLink ® USDA

- AP HIS, 2003 Cotton ( ) . In 2003, the FDA completed its food safety consultation on LibertyLink ® US

- FDA, 2003 Cotton and granted tolerance exemptions for both the plant

incorporated protectant, as well as the genetic material necessary for its production ( )

. The EPA granted exemptions from the requirement of a tolerance for residues of glufosinate ammonium, as well as the plant

- pesticide PAT in or on food and feed commodities 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 environmental impacts associated with exposure to and consumption of plant material incorporating the bar gene and expressing the PAT enzyme. In both cases, these agencies determined that the cottonseed, refined oil, and cotton seed meal derived from the LibertyLink ®

Similar international reviews and approvals have been published for the LibertyLink variety was not materially different in composition, safety and nutritional value from other cotton varieties then on the market.

® 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 ® ANZFA, 2005 Cotton was comparable to non

- genetically engineered (non

- GE) cotton in terms of their safety and nutritional adequacy ( ) . In 2004, the Plant Biosafety Office (PBO ) of the Canadian Food Inspection Agency (CFIA ) also concluded that BCS’ LibertyLink ®

CFIA, 2004 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 s ubstantial negative effect on the environment ( ) . The CFIA specifically noted that t he PAT enzyme responsible for glufosinate ammonium tolerance has very specific enzymatic activity associated with the acetyl ation 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 displaces conve ntional, 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. As previously discussed in the analysi s of water, soil, in those fields where TwinLinkTM Cotton is cultivated, growers would be expected to reduce their use of pesticides for control of lepidopteran pests and take advantage of the weed control offered by glufosinate ammonium, thus allowing for a shift to increased conservation practices.BAYER TWINLINKTM COTTON

55 The use of insecticides other than Bt crops may affect non

- target organisms including honey bees, soil invertebrates, or culturable microbial flora ( Dively and Rose, 2003 ; US

- EPA, 2010a )

. The cultivation of insect

- resistant cr ops allows a reduction and replacement of broad

- spectrum insecticides, with a corresponding positive effect on biodiversity, including non

- target insects ( Cantrell, 2006 )

. A notable advantage of GE insecticidal (Bt) crops over conventional insecticides is the high specificity of the Bt toxins, which mini mize the potential toxic effects on
non

- target insects ( Bigler et al., 2008 ; Sanvido et al., 2006 )

. The incorporation of glufosinate ammonium tolerance provides several potential adva ntages over conventional weed management programs, including increased flexibility to manage problem weeds, reduced use of soil

- applied 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 soil

- applied herbicides have the potential to positively impact animal populations in fields planted with TwinLinkTM Cotton ( see, e.g., Eggert et al., 2004 )

. Based on the above information, APHIS concludes that the deregulation 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 cotton (or any other cro p) fields or may also be surrounded by woodland, rangelands, 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 habitats for a variety of transient and non

- transient wildlife species

. The potential impacts to off

- site plant communities is discussed in Subsection 4.4.3, Biological Diversity. Weed control programs are important aspects of cotton cultivation

. In this context, weeds are those plants whi ch, 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 cotton field will vary depending on the geographic region where the cotton is grown

. Common weeds in cotton include grasses, broad

leaf weeds, and sedges ( Cyper us spp.) . Some of these 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, Plants, and Biodiversity” in Subsection 4.4.3.

Under the No Action Alternative, environmental releases of TwinLinkTM Cotton would be under APHIS regulation

. Plant species (i.e., weed s) that typically inhabit GE and non

- GE cotton production systems will continue to be managed through the use of mechanical, cultural, and chemical control methods. No Action : PlantsBAYER TWINLINKTM COTTON

56 I n the event of a determination of nonregulated status of TwinLinkTM Cotton , the risks to wild plants and agricultural productivity from weedy cotton populations are low; volunteer cotton populations are easily managed and feral populations occur rarely in the U.S. Cotton Belt ( Preferred Alternative : Plants

Wozniak, 2002 )

. Agronomic studies conducted by BCS tested the hypothesis that the weediness potential of TwinLinkTM Cotton is unchanged with respect to conventional cotton ( Scott, 2008 )

. No differences were detecte d between TwinLinkTM Cotton and nontransgenic cotton in growth, reproduction, or interactions with pests and diseases, other than the intended effect of protection from lepidopteran pests

. The main natural controls on feral populations of cotton are poor s eed dispersal, poor seed germination, competition from other plants, and a high requirement for moisture ( US

- EPA, 2001b )

. Therefore, the incorporation of either the insect resistance trait or the herbicide tolerance trait is unlikely t o appreciably improve seedling establishment and increase weediness potenti al. Any TwinLinkTM Cotton volunteers could be readily controlled by other readily available herbicides, including glyphosate ( Miller et al., 2004 ; Stewart et al., 2003 )

. The incorporation of glufosinate ammonium tolerance provides several potential advantages over conventional weed management programs, including increased flexibility to manage problem weeds, reduced use of soil

- applied herbicides, reduced total use of herbicides, and more wide spread 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 areas adjacent to those plant ed 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 impacts would be similar to the No Action Alternative.
4.4.3 Biological Diversity

B iodiversity in an agroecosystem depends on four pr imary characteristics : 1) diversity of vegetation within and around the agroecosystem; 2) permanence of various crops within the system; 3) intensity of management , including selection and use of insecticides and herbicides ; and 4) extent of isolation of the agroecosystem from natural vegetation ( Altieri, 1999 )

. The introduction of woodlots, fencerows, hedgerows, 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

- tillage, composting, green manuring (growing a crop specifically for the purpose of inc orporating 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 windbreaks ( Altieri, 1999 )

. To some degree these practices currently are being utilized by cotton growers to increase biodiversity ( Cotton Incorporated, 2010a ) . The adoption of GE crops, with the concomitant reduction in insecticide use and enhanced soil c onservation practices, has also contributed to the increase in biodiversityBAYER TWINLINKTM COTTON

57 of s oil microorganisms , beneficial organisms, and plants ( Dively and Rose, 2003 ; Naranjo, 2009a ; Sanvido et al., 2006 )

.

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 existing deregulated GE lepidopteran

- resistant (Bt) cotton products, herbicide

- tolerant cotton products, stacked varieties presenting both, and conventional cotton varieties . The implications of a gronomic practices associated wi th cotton production , whether traditional or GE varieties, woul d not change. No Action : Biological Diversity

The use of genetically modified cotton varieties currently available affects biological diversity by : 1) providing the opportunity to use conservation practices that enhance habitat creation; and 2) decreasing the use of pesticides which in turn, limits the impact on the floral and faunal communities that can potentially inhabit the fields and immediate surroundings ( Preferred Alternative : Biological Diversity

Cotton Incorporated, 2010a ; Naranjo, 2009a ; Sanvido et al., 2006 ) . Incorporation of herbicide tolerance in the crop allows the grower to adopt t he use of no

- till, cover crops, crop rotation, 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 constraints for biological diversity in crops ( 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 )

. A comparison of non

- transgenic and transgenic cotton production revealed no substantial impact on ant and beetle diversity in cotton fields ; by contrast, broad

- spectrum insecticide use demonstrated considerably reduced diversity in these populations ( Cattaneo et al., 2006 ) . Cotton expressing the Bt Cry proteins allows for the reduction in use of these broad spectrum insecticides, and is not expected to present a risk to b eneficial 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 ; o n the other hand, broad

- spectrum insecticide use caused a 48% reductio n in 13 of the 22 taxa that were studied ( Naranjo, 2005a , 2005b )

. Bt toxin has been found to be present in every major 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 ( Va dakattu and Watson, 2004 )

. The effects of Bt on non

- target soil microorganisms in Bt maize and Bt cotton cultivation found that microbial biodiversity and activity was not different from that of their non

- Bt counterparts ( EFSA, 2008 ; Hönemann et al., 2008 ; Icoz et al., 2008 ; Shen et al., 2006 )

. Plant biodiversity associated with cotton cultivation is also an issue of concern. Although cotton fields are cultivated as monocultures to optimize cotton yield, areas surrounding a cotton field may harbor a wide variety of plants. Broad spectrum herbicides have the potential to impact off

site plant communities. Glufosinate ammonium provides opportunities to the grower to controlBAYER TWINLINKTM COTTON

58 weeds in the field with fewer applications of herbicides than in traditional cultivation syste ms or those systems where glyphosate resistance has emerged ( University of California IPM Online, 2008b ) . Accordingly, adoption of a glufosinate ammonium protocol could minimize off

- site impacts. The application processes for glufosinate ammonium allow the grower to apply fewer herbicides and reduce field cultivation

. Reducing the number of applications o f herbicides during the cotton production cycle minimizes instances of unintentional drift of these chemicals to adjacent fields, buffer z ones, ditches, and fallow areas. Although TwinLinkTM Cotton does provide the grower with a new choice for a herbicide

- t olerant 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 varietie s providing either insect resistance, herbicide tolerance , or stacked with both ( Scott, 2008 )

. The refore, the effect of cotton production on plant communities is likely to be unchanged by the introduction of TwinLinkTM Cotton

. A determinatio n of nonregulated status of TwinLinkTM Cotton will not change the cultivation or agronomic practices, or agricultural land acreage associated with growing cotton . TwinLinkTM Cotton would be an additional Bt cotton variety providing a different option for h erbicide use ( glufosinate ammonium) for growers to use and therefore is expected to have the same effect on biological diversity as the No Action Alternative. Therefore, the deregulation of TwinLinkTM Cotton for the control of lepidopteran pests and tolerance of glufosinate ammonium is highly 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 cotton managed with conventional broad

- s pectrum insecticides

.

APHIS has determined that there are no impacts from past, present, or reasonably foreseeable actions that would aggregate with effects of the proposed action to create cumulativ e 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 evidence, t he diversity and abundance of non

- target organisms in lepidopteran

- resistant cotton is at least as high as in conventional cotton ; and in many studies , higher biodiversity is associated with transgenic cotton ( Cumulative Effects : Animals, Plants, and Biodiversity

Cattaneo et al., 2006 ; Head et al., 2005 ; Naranjo, 2005a , 2005b ; Torres and Ruberson , 2007 ; Torres et al., 2006 ; Whitehouse et al., 2005 )

. Cultivation o f 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 biodive rsity compared with conventionally managed cotton

. Therefore, the likelihood of adverse cumulative effects on non

target organisms and biodiversity following the introduction of TwinLinkTM Cotton is minimal.
4.4.4 Gene Movement Gene flow is the movement of genes from one organism to another. Vertical gene flow, or introgression, is the movement of genes to sexually compatible relatives ( Ellstrand, 2003 ; Quist,BAYER TWINLINKTM COTTON

59 2010 )

. In asses sing the risk of gene introgression from TwinLinkTM Cotton to its sexually compatible relatives, APHIS considered two primary issues : 1) the potential for gene flow and introgression to feral u pland cotton and cotton relatives and 2) the potential impact of introgression . This is discussed below under the “Preferred Alternative” heading. Horizontal gene flow is the stable movement of genes from one organism to another without reproduction or human intervention ( Keese, 2008 ; Quist, 2010 )

. There is no evidence of naturally occurring t ransgene movement from transgenic crops to sexually incompatible species ( Scott, 2008 ; Stewart, 2008 )

. Horizontal gene transfer and consequ ent expression of DNA from
one plant to another plant or other phyla (e.g., species of bacteria ) is unlikely to occur ( Keese, 2008 ) . This event would require physical relocation of the complete genetic material from the transgenic plant to the new location, including not only the genes which code for the production of specific proteins, bu t also those portions of the genome which regulate the activity of those genes ( Keese, 2008 ; Scott, 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 evidence that eukaryotic cells are 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 t herefore constrained as to the type and size of novel genetic material which can be acquired by horizontal gene transfer ( Stewart, 2008 )

. Two bacteria species commonly associated with plants, Agrobac terium 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 (ab normal outgrowth) ( University of Illinois, 2010 )

. Rhizobium aids in nitrogen fixation in legume nodules ( Wilkinson and Elevitch, 2011 ) . The genomes of both bacteria have been sequenced , and the sequenced genes evaluated for exogenous genes ( Kaneko et al., 2000 ; Kaneko et al., 2002 ; Wood et al., 2001 ) . Despite what would appear to be millennia of symbiotic relationships between these bacteria and their host plants, t here is no evidence that these organisms contain genes derived from plants ( USDA

APHIS, 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 bacterial expression ( USDA

- APHIS, 2010a ) . Horizontal gene flow, resulting in the relocation of entire transgenes including the regulatory portions of the DNA (those parts of the DNA which code for the production of the specific proteins in that relocated transgene) has never be en shown to occur in nature ( Clarke, 2007 ; Stewart, 2008 )

. Thus, even if horizontal gene transfer occurred, proteins associated with the se transgenes are not likely to be produced in the new host organism ( Stewart, 2008 ; USDA

- APHIS, 2010a ) . Based on this information, APHIS considers the horizontal gene flow from TwinLinkTM Cotton to unrelated species to be highly unlikely, and the same as potential horizontal gene flow from existing GE cotton varie ties.

Under the No Action Alternative, conventional and GE transgenic cotton production will No Action : Gene MovementBAYER TWINLINKTM COTTON

60 continue while TwinLinkTM Cotton will remain a regulated article

. Cotton land acreage and cultivation practices for both conventional and GE transgenic cotton varieties will remain the
same

. In 2010 , Bt cotton varieties occupied 93% of the cotton acreage ( USDA

- ERS, 2010c )

. Gene flow from current commercially available GE cultivars t o non

- GE cotton cultivars, feral cotton populations, and wild sexually compatible relatives will remain unchanged from the current condition.

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

. Preferred Alternative : Gene Movement

The passage of the transgene to a cross

- compatible wild relative could allow the wild relative to become more abundant in the habitat or to invade new habitats because of its lepidopteran resistance ( Wilkinson and Ford, 2007 )

. This increased resistance of the wild relative to one group of insects could alter the balance of other insect herbivores, predators, and parasites ( Wilkinson and Ford, 2007 )

. The introgressed wild crop relative could also impact other plants through interspecific (between species) competition and indirectly affect the insect pollinators, herbivores , predators, and parasites of those species ( Wilkinson and Ford, 2007 ) . Vertical gene flow, or introgression, is influenced 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 barriers to pollen movement ( Daniell, 2002 ; Eastham and Sweet, 2002 ; Messeguer, 2003 ; Quist, 2010 )

. Two species of cotton are cultivated in the U.S. : u pland cotton ( G. hirsutum ) and Pima cotton ( G. barbadense ) ( USDA

- ERS, 2009 )

. There are two wild relatives of interest to U.S. growers , G. thurberi and G. tomentosum, found in the mo untains of southern Arizona and in Hawaii, respectively . APHIS has considered the potential for gene flow from TwinLinkTM Cotton, a GE variety of G. hirsutum , to other u pland cotton varieties, as well as these other three species. With regard to gene flow to other u pland 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 standards for identity protection of seeds. These measures have been developed based on research on the po llination of the u pland cotton flower. Although u pland cotton is considered self

- pollinating ( Hartman and Kester, 1 975 ; Scott, 2008 ) , 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 pollen particles . V ery little pollen is transferred beyond 12 meters ( McGregor, 1976 ; Scott, 2008 ; Vaissiere and Vinson, 1994 )

. By comparison, the isolatio n distances for Foundation, Registered, and Certified seeds specified in 7 CFR Part 201 are 1320, 1320 , and 660 feet, respectively . If these identity practices are adhered to by growers, it is unlikely that TwinLinkTM Cotton will outcross to other upland c otton varieties.BAYER TWINLINKTM COTTON

61 Upland cotton and Pima cotton are frequently cultivated in the same regions ( Hutmacher et al., 2006 ) . Although cross pollinat ion has been demonstrated between these species, Van Deynze and others have found that the isolation distances currently recommended to ensure seed quality for Foundation and Registered seed production are 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 numbers precludes sexual compatibility of G. hirsutum with G. thurberi ( OECD, 2008 )

. G. tomentosum is native to the Hawaiian Islands ( Pleasants and Wendel, 2010 ) . Historic literature suggests that differences in flowering patterns and pollinators presented barriers to cross pollination between G. tomentosum and either G. hirsutum or G. barbadense ( Pleasants and Wendel, 2010 )

. Recent f ield and laboratory studies contradict this historic literature , demonstrating that these three species share common pollinators, and further that differences in flower structure and flowering habit s 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 analyses 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 ) . T he terms and con ditions of EPA’s conditional registration for Cry1Ab in cotton prohibits commercial cultivation south of Route 60 near Tampa ( US

- EPA, 2008a )

. TwinLinkTM Cotton is neither expected to be planted 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 sale and distribution of Bt cotton for commercial planting where it could outcross with the wild cotton species G. tomentosum

. These restrictions in Hawaii also apply to Bt cotton test plots and breeding nurseries ( US

- EPA, 2001b )

. Because of these restrictions, 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 introgression to occur from TwinLinkTM Cotton to sexually compatible wild relatives and considered whether such introgression would result in increased weediness

. Based on the plant pest risk assessment, APHIS has conclude d that TwinLinkTM Cotton is not a plant pest and that gene flow between this product and wild or feral relatives will not occur in the 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 characteristics of weedy or wild relatives if gene flow or introgression were to occur ( USDA

- APHIS, 2010b )

. TwinLinkTM Cotton would provide growers with an additional cotton variety combining a Bt

- based lepidopteran protection , as well as herbicide tolerance, and is expected to have the same effect on gene movement as the No Action Alternative.BAYER TWINLINKTM COTTON

62 Based on available scientific evidence, APHIS has not identified any cumulative effects on gene movement that would occur from a determination of nonregulated status of TwinLinkTM Cotton. Cumulative Ef fect : Gene Movement

Gene movement between sexually compatible cotton species is no greater for TwinLinkTM Cotton than it is for other non

- GE or GE cultivars

. Many factors limit the likelihood of gene movement between cotton species

. These include :

**•** C otton is considered to b e a self

- pollinating crop with a very low frequency of outcrossing from insect pollinators or wind;

**•** T he Cry1Ab , Cry2Ae , and bar genes in TwinLinkTM Cotton and other GE cottons impart no agronomic advantages whereby a greater potential for weediness or invas iveness would result should introgression occur;

**•** N either GE or non

- GE cotton cultivars form self

- sustaining populations outside of cultivation because of limitations in seed dispersal, germination, and high moisture requirements ( US

- EPA, 2001b , 2008a ) ;

**•** T he cotton industry has measures in place to restrict pollen movement and gene flow between cotton fields through the 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

**•** W here 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 pr oduction ( US

- EPA, 2001b , 2008a )

. In addition, there is no evidence that horizontal gene transfer 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, proteins corresponding to the transgenes are not likely to be produced

. 4.5 PUBLIC HEATH
4.5.1 Human Health
Under FFDCA , it is the responsibility of food and feed manufacturers to ensure that the products they market are safe and properly labeled

. Food and feed derived from TwinLinkTM Cotton must be in compliance with all applicable legal and regulatory requirements

. GE o rganisms for food and feed may undergo a voluntary consultation 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 properties such as Bt toxin. 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, a s well asBAYER TWINLINKTM COTTON

63 a registration review to the EPA for this product. The FDA and EPA are c urrently reviewing the information submitted by the applicant.

Under the N o A ction A lternative, TwinLinkTM Cotton 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 alternative. No Action : Human Health

APHIS considers the FDA and the EPA regulatory assessment in making its determination of the potential impacts of deregulation of the new agricultural product. BCS has submitted a food and feed safety and nutritional assessment t o the FDA, as well as a registration review to the EPA for this product. The FDA and EPA are c urrently 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 human health impact determination. Preferred Al ternative : Human Health

Similar products have been deregulated and approved for general cultivation beginning in 1996 with the introduction of Bt products and followed shortly after by the introdu ction 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 p roducts 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 expression of glufosinate ammonium tole rance , the gene construct in the Bayer TwinLinkTM Cotton is the same as that approved by the FDA in June, 2003 for the LibertyLink ® US

- FDA, 2003 Cotton product ( )

. In that approval, the FDA noted that the transformational event in LibertyLink ® US

- FDA, 2003 Cotton was not materially different in composition, safety, or any other relevant parameter in cotton then g rown, marketed, and consumed ( )

. 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 assessment, the Bt BRAD found no unreasonable adverse effects associated from exposure to the Bt products

. The Bt protein is a registered microbial pesticide which is approved for use on food crops

. The EPA c oncluded in the BRAD that the Bt products incorporated as plant protectants would be expected to behave as ex pected 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, 2010g )

. A temporary exemption from the requirement of a tolerance has been issued for Cry2Ae ( 40 CFR §174.530; US

- EPA, 2010e )

. F or the majority of Bt proteins currently registered, the source bacterium has been a registered microbial pesticide, which has been approved for use on food crops without specific restrictions

.BAYER TWINLINKTM COTTON

64 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 epidemiological studies to gether, over the years, indicate that biopesticides pose minimal risks to humans and animals ( Betz et al., 2000 ; US

- EPA, 2001b , 2008a )

. T he Cry proteins present 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 ha s 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 this 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 ) . APHIS deregulated the PAT enzy me bar gene construct in cotton in 2003 with the BCS LibertyLink ® USDA

- APHIS, 2003 Cotton ( ) . In 2003, the FDA completed its f ood safety consultation on LibertyLink ® US

- FDA, 2003 Cotton and granted tolerance exemptions for both the plant

incorporated protectant, as well as the genetic material necessary for its production ( ) . The EPA granted exemptions from the requirement of a tolerance for residues of g lufosinate ammonium, as well as the plant

- 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 evaluation s based on Codex Alimentarius Commission procedures to assess any potential adver se effects to humans or animals resulting from environmental release s of TwinLinkTM Cotton containing the Cry1Ab, Cry2Ae , and PAT proteins see ( FAO, 2001 ; Scott, 2008 ) . These safety studies included evaluating prot ein 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 ( Scott, 2008 )

. The results of studies conducted by BCS confirm that the crops containing these proteins can be safely used as food or feed ( Scott, 2008 )

. The Australia and New Zealand Food Authority (ANZFA) also completed a Toxicological Review and Risk Assessment, and a Technical 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 adequacy. In 2004, the Plant Biosafety Office (PBO) of the Canadian Food Inspection Agenc y (CFIA) concluded that BCS’s LibertyLink ®

CFIA, 2004 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 environment ( )

. CFIA further noted that the PAT enzyme responsible for glufosinate ammonium tolerance has very specific enzymatic activity associated with the acetylation of glufosinate ammonium, and is rapi dly inactivated in mammalian stomach and intestinal fluids by enzymatic degradation and pH mediated proteolysis ( CFIA, 2004 )

. As with other Bt cotton and herbicide

- tolerant cotton products previously deregulated and
c ommercialized , including the LibertyLink ® variety in 2003 , TwinLinkTM Cotton is expected toBAYER TWINLINKTM COTTON

65 be used throughout cotton producing areas of the country

. Based on the analysis of field and laboratory data and scientific literature provided by BCS ( Sc ott, 2008 ) , and safety data available on other GE cotton, APHIS has concluded that a determination of nonregulated status of TwinLinkTM Cotton would have no adverse impacts on human health

. Overall impacts are similar to the No Action Alternative

.

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. Cumulative Effects : Human Health

4.5.2 Worker Safety

E PA’s Worker Protection Standard (WPS ) (40 CFR Part 170) was published in 1992 to require actions to reduce the risk of pesticide poisonings and injuries among agricultural workers and pesticide handlers

. The WPS offers protections to more than two and a half million agricultural workers who work with pes ticides 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 protective equipment, restricted entry intervals fo llowing pesticide application, decontamination supplies, and emergency medical assistance

.

During agricultural production of cotton, agricultural workers and pesticide applicators may be exposed to a variety of EPA registered p esticides ( No Action : Worker Safety

US

- EPA, 2001b )

. Under the No Action Alternative, agricultural workers and pesticide applicators may be exposed to these agricultural chemicals during cotton production . Such chemicals would be expected to include those prod ucts currently used for insect pest and plant pest management in both GE and non

- GE cotton cultivation.

Before the introduction of Bt cotton varieties in 1996, chemical pesticides were widely used to control lepidopteran pests of cotton

. Organophosphate, carbamate, and pyrethroid products accounted for a substantial percentage of the insecticides used ( Preferred Alternative : Worker Safety

USDA

- NASS, 2008 )

. However, these three classes of pesticides require numerous safety warnings and extensive use restrictions, which raise concerns for worker safety ( US

- EPA, 2001b )

. Bt cotton varieti es have offered a very effective and environmentally benign alternative to chemical insecticides for lepidopteran insect pest control

. As a result, Bt cotton varieties have been extensively adopted by cotton farmers and now represent 73 % of the commercial cotton planted ( USDA

- ERS, 2010a )

. Herbicide

- tolerant cotton varieties have seen similar adoption within the industry, with a 78% adoption rate in 2010 ( USDA

- ERS, 2010a )

. The adoption of herbicide tolerance has allowed growers to shift their herbicide use to products like glufosinate ammonium which has a low toxicity to humans

. Avoidance of other herbicides provides corresponding benefits

.BAYER TWINLINKTM COTTON

66 Similar to the No Action Alternative, it is expected that EPA registered pesticides that are currently used for cotton production will continue to be used by growers

. If TwinLinkTM Cotton is adopted, agricultural workers and pesticide applicators may benefit from the use of TwinLinkTM Cotton due to a reduction in the use of budworm/bollworm insecticide applications and the use of the low

- toxicity herbicide glufosinate ammonium. The adoption of glufosinate ammonium

- tolerant cotton allows growers to change cultivation practices in those fields where glyphosate resistance has emerged, with the resulting potential reduction in herbicide treatments required in those current glyphosate

- based systems. A reduction in insecticide and herbicide use would be nefit worker safety.

A determination of nonregulated status of TwinLinkTM Cotton is not expected to increase the total acreage of cotton production or the use of Bt cotton, herbicide

- tolerant cotton, or stacked varieties presenting both traits

. BCS anticipates that TwinLinkTM Cotton will primarily replace some of the insect

- resistant and herbicide

- tolerant cotton cultivars already 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 u se of lepidopteran pest insecticides . Similarly, replacement of herbicide

- tolerant cotton currently based on expressing glyphosate tolerance with TwinLinkTM Cotton has the potential to reduce herbicide use to manage weedy species which are glyphosate

- resis tant. This has been the case with the adoption of other GM cotton cultivars ( Cumulative Effects : Worker Safety

Benbrook, 2009 )

. A reduction in pesticide use and exposure should positively benefit worker safety

. 4.6 ANIMAL FEED Cotton is grown primarily for its lint, but the s eed residue, called cottonseed meal, provides a valuable animal feed ( Meyer et al., 2007 )

. Cotton seed meal animal feed provides protein, fat, and energy ( National Cottonseed Products Association, 2002 )

. Under FFDCA, it is the responsibility of feed manufacturers to ensure that the products they market are safe and properly labele d

. Feed derived from TwinLinkTM Cotton must be in compliance with all applicable legal and regulatory requirements

. GE organisms for feed may undergo a voluntary consultation process with the FDA prior to release onto the market

. 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. In the absence of these completed agency reviews, AP HIS takes into consideration prior reviews of comparable products as a starting point for its animal health impact determination.

Under the No Action A lternative, TwinLinkTM Cotton will remain a regulated product and will not be available as an animal feed. No Action : Animal FeedBAYER TWINLINKTM COTTON

67 Cotton

- based animal feed will still be available from currently cultivated cotton crops, both conventional varieties as well as deregulated GE cotton 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.

BCS has submitted compositional and nutritional characteristics of TwinLinkTM Cotton seed to APHIS ( Preferred Alternative : Animal Feed

Scott, 2008 )

. The nutritional components of TwinLinkTM Cotton were compared with the non

- transgenic cotton cultivar, Coker 315. The compositional comparison was based on analysis of products collected from seven field trials ( Scott, 2008 )

. Chemical analysis of the cotton samples included the nutritional factors proximate (moisture , crude protein, lipids, ash, carbohydrates, and energy); minerals, vitamins, amino acids , and fatty acids (cottonseed oil) ; and the a ntinutri ent factors of cotton includ ing the gossypols, phytic acid, and the cyclic fatty acids ( Scott, 2008 )

. The field trials were conducted to simulate modern cotton cultivation techniques, so that the replicates evaluated included fields treated with glufosinate ammoni um in accordance with the label provisions and untreated fields ( Scott, 2008 )

. The results provided by BCS show no differences between TwinLinkTM Cotton and the non

transgenic Coker 315 cultivars ; reported results for each parameter were also within the reference ranges for cotton composition ( Scott, 2008 )

. All the analytes measured fell within the normal range of natural variation in cotton ( ILSI, 2008 ; O ECD, 2004 )

. T he EPA has established a permanent exemption from the requirement of a tolerance for the PAT enzyme and the genetic material necessary for its production in plants ( 40 CFR 174.522; US

- EPA, 2010g )

. A similar permanent exemption has been published for the Cry1Ab 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 cotton has been published at 40 CFR 174. 5 30, in accordance with Expe rimental Use Permit 264

- EU

- 143 ( US

- EPA, 2010e , 2010f )

. APHIS’s assessment of the safety of this product for animals focuses on p lant pest risk ( USDA

APHIS, 2010b ) and effects on wildlife and threatened and endangered species and those analyses are based on the comparison of the GE TwinLinkTM Cotton to its non

- GE counterpart

. N o new issu es appear to be associated with the Cry and the PAT proteins in TwinLinkTM Cotton

. Based on th is information APHIS has concluded that a determination of nonregulated status of TwinLinkTM Cotton would have no significant impacts on animal feed o r animal health

. Overall impacts are similar to the No Action Alternative.

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

. Cumulative Effects : Animal FeedBAYER TWINLINKTM COTTON

68 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 billion pounds of cotton ( US

- EPA, 2009 )

. The USDA ERS notes that U.S. cotton productivity has increased in recent years as a result of technological advancements, including biotechnology

- derived varieties ( Meyer et al., 2007 )

. Herbicide

tolerant cotton is one of the most widely and rapidly adopted crops in the U.S., followed by insect

- resistant cotton ( Fernandez

- Cornejo and Caswell, 2006 ; USDA

- ERS, 2010a )

. By th e adoption of new technologies, including agronomic 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 protection from the damage incurred by insect pests and an economical alternative to broad

- spectrum insecticides . In addition, the adoption of post

emergent herbicide

- tolerant varieties provide opportunities to apply fewer herbicides and to reduce field cultivation ( University of California IPM Online, 2009c ; USDA

- APHIS, 2010a )

.

Under the No Action Alternative, TwinLinkTM Cotton would continue to be regulated art icles under the regulations at 7 CFR Part 340

. Growers and other parties who are involved in production, handling, processing, or consumption of cotton would not have access to TwinLinkTM Cotton , but would continue to have access to existing deregulated GE lepidopteran

resistant or herbicide

- tolerant ( or stacked) cotton varieties as well as other deregulated GE and conventional cotton varieties

. Domestic growers will continue to utilize currently available traditional and GE cotton varieties based upon availability and market demand . Consequently, the economic profile of cotton production and marketing is not expected to change. No Action : Domestic Economic Environment

BCS anticipates that TwinLinkTM Cotton would replac e currently available Bt cotton varieties and may be selected by growers of conventional 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 ( Preferred Alternative : Domestic Economic Environment

Scott , 2008 )

. 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 pub lic interest 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, based on an analysis of biotechnology

- derived crops planted in 2004, the National Center f or Food and Agricultural Policy estimated that products providing protection 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 provide d a summary of other studies that characterize effects on yield, pesticide use, and grower returns , suggesting that in addition to the economic benefits to the GE developers andBAYER TWINLINKTM COTTON

69 seed firms associated with technology fees and seed premiums, both U.S. and foreign consumers indirectly benefit through lower commodity prices resulting from increased supplies ( Fernandez

Cornejo and Caswell, 2006 )

. Market ben efits were estimated at $230 million for herbicide

tolerant cotton and $210 million for insect

- resistant cotton (Bt cotton) after only two years of availability ( Fernandez

- Cornejo and Caswell, 2006 )

. BCS’ field trials demonstrated that TwinLinkTM Cotton was effective in contr olling 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 ( Scott, 2008 )

. These pests cause severe economic damage to the cotton crop if not controlled

. If controlled by chemical pesticides, there is the need for large insecticide input annually to control these pests ( Scott, 2008 )

. The incorporation of the two Bt

- based Cry protein s obviates the need for the input of these broad spectrum insecticides, with a commensurate economic benefit (reduced 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 200 9 , 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 20 09 cost and loss estimates for arthropod cotton pests were $ 502 million ( Williams, 2010 )

. BCS anticipates that TwinLinkTM Cotton will replace or displace existing GE cotton varieties currently in the market

. Specific economic projections 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 utilization of glufosinate ammonium allows growers to reduce or eliminate multiple herbicides to control glyphosate

resistant weeds, farms adopting TwinLinkTM Cotton might experience an increase in net income

. However, net income differentials cannot be projected. T he net income of cotton producers just growing Bt co tton varieties or herbicide

- tolerant varieties is not available ( USDA

- NASS, 2007 )

. However, as BCS anticipates that TwinLinkTM Cotton would only replace existing GE cotton varieties and not result in increased cotton acreage , resulting socioeconomic impacts are expected to be similar to the N o A ction A lternative.
Cumulative Effects : Domestic Economic Environment
Based on the information described above, APHIS concludes that a determination of non

regulated status of TwinLinkTM Cotton in itself will have no for eseeable adverse cumulative domestic economic effects . TwinLinkTM Cotton would simply replace or displace existing cotton varieties for those growers seeking a new combination of insect resistance and herbicide tolerance. TwinLinkTM Cotton is not expected to lead to an expansion of U.S. cotton acreage

. No net cumulative effects have been identified associated with a determination of non

- regulated status and cultivation of TwinLinkTM Cotton

. 4.7.2 Trade Economic Environment at Risk
Cotton is th e single most important 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 theBAYER TWINLINKTM COTTON

70 leading supplier in the international market ( Meyer et al., 2007 )

. However, the U.S. cotton sector has faced a number of challenges as it shifts from a domestic

- oriented market to one focused largely on the global marketplace

. Domesti c 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 has increased rapidly with the recent expansion of global textile prod uction ( Meyer et al., 2007 )

. The USDA ERS reports that U.S. c otton mills consumed 60% of the domestic cotton through the 1990s, but by the mid

- 2000s 70% of th e 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 general equilibrium model assessed the impacts of interna tional Bt cotton adoption on cotton and related 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 ben efits were 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 pri ce

. Employment and trade balances in the textile and apparel sectors increased for China and India, but generally declined elsewhere

. Individual countries obtained greater economic welfare gains upon adoption of Bt cotton ; n on

- adopting regions lost cotto n market share to adopting regions ( Frisvold and Reeves, 2007 )

. U.S. cotton production reached consecutive records in 2004 and 2005 sea sons, with rising global cotton demand providing a market for much of the increased output ( Meyer et al., 2007 )

. However, the growing use of better crop production technologies 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 crops 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 ( USDA

- FAS, 2010a )

. Meanwhile, debate over trade policy and the sustainability of current farm programs are a source of uncertainty for U.S. agricultural commodities in general and the cotton sector in particular ( 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 f armers’ 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 producing countries like Australia, Argentina, Mexico, and South Africa ( Meyer et al., 2007 ; USDA

- ERS, 2009 )

.

The cropping and marketing decisions made by cotton growers are unlikely to be influenced by the selection of this alternative and it is expected that approximately 93% of the cotton produced will continue to be planted with the currently available G E cotton ( No Action : Trade Economic Environment

USDA

- ERS, 2010c ) . U.S. cotton will continue to play a role in global cotton production, and will continue to be a supplierBAYER TWINLINKTM COTTON

71 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, and 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 millions of hectares of cotton land back into production in China and has revolutionized India’s cotton industry ( Meyer et al., 2007 )

. Economic evidence shows that international trade and productivity is enhanced by the adoption of Bt cotton and that these products can be channeled into suitable export markets. Increased global production of Bt cotton ado ption 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 than if they did not adopt it, and further, found non

- adopting regions to lose cotton market share to the adopting regions ( Frisvold and Reeves, 2007 ) . These conditions are not expected to change if TwinLinkTM Cotton remains a regulated article.

BCS intends to submit do ssiers to the proper regulatory authorities of foreign governments to allow the importation of U

. S . c ottonseed and to have regulatory processes in place so as to allow for the international movement of this product ( Preferred Alternative : Trade Economic Environment

Scott, 2008 )

. These may include submissions to the relevant r egulatory a uthorities in Canada, Mexico, the European Union ( EU ) , and Japan, among others

. TwinLinkTM Cotton has been, or is currently, in field trials in cotton growing region s around the world ( Scott, 2008 )

. 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 TwinLinkTM Cotton line which could provide another lepidopteran insect resistance and herbicide

- tolerant 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 the deregulation of TwinLinkTM Cotton are anticipated to be very similar to the No Action Alternative.

Current and historic economic evidence indicate s 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 negatively impact the trade economic environment

. Cumulative Effects : Trade Economic Environment

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

. According to data from the 2003 Agricultural Resource Management Survey (ARMS ), farms gr owing cotton tend to be larger than those growing other crops, with above average gross farm incomes, government payments, farm expenses, netBAYER TWINLINKTM COTTON

72 incomes, farm asset values, and debt

- to

- asset ratios ( Meyer et al., 2007 )

. Large farm operations are more likely to be organized 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 farm 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 cash 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 cotton farms is mainly due to their larger farm operations

. Cotton farms averaged 1,199 acres per farm, compared with 376 acres for non

cotton farms ( USDA

- ERS, 2009 )

. Cotton farms’ average 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 expen se compared to $91 of cash expense for a non

- cotton farm)

. Larger farms can achieve 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 averaged $142,463 in 2003

. In comparison, the household income for non

- cotton farms in cotton

- producing states averaged $71,447 ( USDA

ERS, 2009 )

. For most farm households, income from off

- farm sources exceeds income from the farm operation

. However, cotton producers derive the majority of their family income from the farm . Eleven percent of cotton p roducers listed nonfarm jobs or businesses as their main occupation, compared with 48% of non

- cotton farm operators ( USDA

- ERS, 2009 )

.

Under the No Action Alternative, no direct impact on the social environment surrounding cotton farming is expected. The cropping and marketing decisions made by cotton growers are unlikely to be influenced by the selection of this alternative. No Action : Social Environment

Most of the cotton acreage in the U.S. is planted 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 insect

resistant (Bt) cotton and 78% was herbicide

- tolerant ( Preferred Alternative : Social Environment

USDA

- ERS, 2010a , 2010c )

. Most cotton is planted on farms that have been in cotton production for at least five years ( U SDA

- NASS, 2007 )

. 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 cultivati on areas

. A determination of nonregulated status of TwinLinkTM Cotton by APHIS is not expected to significantly expand the number of cotton acres, and cotton acreage is expected to remain relatively stable

. Overall impacts are similar to the No Action Alternative

.BAYER TWINLINKTM COTTON

73 Based on the information described above, APHIS has determined that there are no past, present, or reasonably foreseeable actions that in aggregate with effects of the proposed action would impact the social environment surrounding cotton farming. Cumulative Effects : Social Environment

4.8 OTHER CUMULATIVE EFFECTS The p otential cumulative effects regarding specific issues have been analyzed and addressed above

. No further potential cumulative effects have been identified

. To date, none of the GE cotton varieties that have been deregulated pursuant to Part 340 and the Plant Protection Act and used for commercial cotton production or cotton breeding programs have been subsequently 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 herbicides, or may present a different combination of insect pest

- resistant , but may also have progeny w ith no transgenes at all

. APHIS ’ current regulations at 7 CFR Part 340 do not provide for Agency oversight of GE cotton varieties previously deregulated pursuant to Part 340 and the Plant Protection Act , nor over stacked varieties combining deregulated GE varieties 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 deregulated GE variety, as company plans and market demands pl ay a significant role in those business decisions

. P redicting all potential combinations of stacked varieties that could be created using both deregulated GE cotton varieties and also non

- GE cotton varieties is hypothetical and purely speculative . No further analysis 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 extinctions 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 implement 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 add ed 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 destruction, modification, or curtailment of its habitat or range;BAYER TWINLINKTM COTTON

74 **•** Overutilization for commercial, recreational, scientific, or educational purposes;

**•** Disease or predation;

**•** The inadequacy of existing regulatory mechanisms; and

**•** The natural or manmade factors affecting it s survival. Once an animal or plant is added to the list, in accordance with the ESA, protective measures apply to the species and its habitat. These measures include protection from adverse effects of Federal activities. Section 7 (a)(2) of the ESA r equires 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 species or result in the destruction or adverse modificatio n of designated critical habitat. It is the responsibility of the Federal agency taking the action to assess the effects of their action and to consult with the USFWS and NMFS if it is determined that the action “may affect” listed species or critical hab itat. To facilitate APHIS’ ESA consultation process, APHIS met with the USFWS to discuss factors relevant to their effects analysis for petitions for nonregulated status and developed the following process for conducting an effects determination for biote chnology regulatory actions. Appendix A provides a copy of the 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 necessary, the biological assessment. For each transgene(s)/transgenic plant the following information, data, and questions are considered by APHIS:

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

**•** Cha racterization 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 transgene and its products (if any) are produced in the plant and their quantity;

**•** A revie w of the agronomic performance of the plant including disease and pest susceptibilities, weediness potential, and agronomic and environmental impact;

**•** Determination of the concentrations of known plant toxicants (if any are known in the plant); and

**•** Analysis to determine if the transgenic plant is sexually compatible with any threatened or endangered plant species (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 F ederally 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 secti on 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 Conservation Online System (ECOS ; see Appendix B ).BAYER TWINLINKTM COTTON

75 4.9.1 Potential Effects of TwinLinkTM Cotton and Agricultural Practices on TES BCS anticipates that TwinLinkTM Cotton would replace currently available Bt cotton varieties and may be selected by growers of conventional cotton varieties mak ing a change to a GE variety; however, availability of TwinLinkTM Cotton is not expected to result in an increase in cotton acreage ( Scott, 2008 ) . Accordingly, t he issues discussed herein focus on the potential environmental consequences of th e deregulation of TwinLinkTM Cotton on TES species in the areas where cotton is grown

. APHIS therefore focuses its TES review on the 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

- target organisms and the natural environment.

BC S has presented data evaluating the agronomic and morphological characteristics of TwinLinkTM Cotton, including compositional and nutritional characteristics, safety evaluations and toxicity tests, comparing the product to a conventional cotton variety (i.e. , Coker 315) ( Scott, 2008 ) . 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 previous discussions on Human Health and Animal Feed for an expanded discussion of BCS’ comparison

. )
The agronomic and morphologic characteristics data provided by BCS were used in APHIS analysis of the weediness potential for TwinLinkTM Cotton , and evaluated for the potential to impact TES. Agronomic studies conducted by BCS tested the hypothesis that the
we ediness potential of TwinLinkTM Cotton is unchanged with respect to conventional cotton ( Scott, 2008 ) . No differences were detected between TwinLinkTM Cotton and nontransgenic cotton in growth, reproduction, or interactions with pests and disea ses, other than the intended effect of protection from lepidopteran pests. The main natural controls 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 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 c ould be readily controlled by other readily available herbicides, including glyphosate ( Miller et al., 2004 ; Stewart et al., 2003 ) . Based on this analysis, APHIS has concluded that deregulation of TwinLinkTM Cotton has no potential to affect TES , as a result of weediness

. In a ddition 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 assessment in making its determination of the potent ial impacts of deregulation 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 tolerance and registration reviews to the EPA for TwinLinkTM Co tton. 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 th e potential impacts

. As discussed above in Animal and Plant Communities ( Subsection 4.4)BAYER TWINLINKTM COTTON

76 and Public Health ( Subsection 4.5), both agencies have previously reviewed and approved comparable products, including several crop varieties expressing the Bt

- based Cry proteins as plant

- incorporated protectants, as well as the bar gene, resulting in the expression of the PAT enzyme providing tolerance to glufosinate ammonium. Data regarding the environmental safety of PAT proteins have been extensively reviewed in international scientifi c, 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 consumed 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 mechanism to transfer this gene to unrelated plant species, APHIS limits its analysis of the implications of exposure to the PAT protein to the pot ential 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 listings ( Arizona, 2009 ; Hawaii, 2001 ) with the exception of G. hirsutum in the State of Florida , where w ild populations of u pland cotton, G. hirsutum have been listed as endangered by the s tate ( 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 southern Florida where it i s found ( Coile and Garland, 2003 ; US

- EPA, 2008a ; USDA

- NASS, 2007 ; Wunderlin and Hansen, 2010 )

. Acco rdingly, the deregulation of TwinLinkTM Cotton is not expected to impact state endangered feral cotton populations. Evaluation of the potential exposure of threatened and endangered species to the Bt

- based Cry proteins focuses on the route of exposure. Cry 1Ab and Cry2Ae proteins are highly specific to Lepidoptera. The specificity of Bt crystalline proteins to lepidopteran insect larvae, but not for other insects, birds, and mammals results from the highly specific receptors for these proteins in the larvae midgut ( Lemaux, 2009 ) . Once activated by insect

specific proteases in the insect midgut, Cry proteins bind to receptors in the midgut. Such binding leads to the formation of pores in the midgut membrane s and ultimately to cell lysis and death. The specific binding of Bt

- based Cry proteins to midgut membrane receptors is a key determinant of pest specificity ( Showalter et al., 2009 ) . Accordingly, the APHIS review of the potential impacts is focused on Federally listed threatened and endangered lepidopterans in the cotton growing regions of the U.S.

APH IS obtained and reviewed the USFWS Federal List of listed (or proposed) TES and considered the potential impacts of a determination of nonregulated status of TwinLinkTM Cotton on lepidopterans (Appendix B ). There are currently 21 species of butterflies, mo ths, 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 specie s to the Bt

- based Cry proteins in TwinLinkTM Cotton is only likely if the species occur in the areas where cotton is grown, as cotton plant parts (seeds, pollen, and crop debris) are not readily transported long distances without human involvement.BAYER TWINLINKTM COTTON

77 APHIS has thoroughly examined all listed and proposed threatened and endangered lepidopterans and compared their habitats to those habitats present in counties where cotton is grown. APHIS has determined that the breeding habitats of listed Lepidoptera do not
o verlap 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 limited 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 Californi a, Florida, and North Carolina ( US

- FWS, 2011b ) (see Appendix B : Threatened 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
m itchellii francisci ) are known to occur in counties were cotton is grown. The Quino Checkerspot butterfly is found in coastal 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 southe rn 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 b e a facultative fire

- follower in non

- desert areas and in desert plant communities 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 slo pes, and flats of foothill woodlands, chaparral margins, and coniferous forests ( US

- FWS, 2003 ) . Areas where 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 Checkers pot 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 larvae 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 ) . Since listing, the primary threats to the Kern 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 primrose, 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. Because cotto n 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 larv al host plants.BAYER TWINLINKTM COTTON

78 The Saint Francis’ satyr is known only in the sand hills of North Carolina, i n Cumberland and Hoke Counties ( US

- FWS, 2010a ) . The species is known only to be located on government land (Fort Bragg U.S. Army installation). The habitat occupied by this satyr consists primarily of wide, wet meadows dominated by a high diversity of sedges ( Carex ) and other wetland graminoids. Saint Francis’ satyr has also been observed in pitcher plant ( Sarracenia flava ) swales, with cane ( Arundinaria tecta ), rough

- leaved loosestrife ( Lysimachia asperulaefolia ), 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 Panicum grass. Areas in Hoke and Cumberland County where cotton is grown are likely to have been used in agricultural production for many years and would be unsuitable habitat fo r the Saint Francis’ satyr. As noted above, the EPA conducts a comprehensive assessment of the implications of pesticide registrations on TES. EPA’s evaluations include exposure assessments and ecological effects data for each registered pesticide. The EP A ( 2001b , 2008a ) has published the following specific statements regarding potential effects of Bt

- based Cry p roteins incorporated in cotton

— ...The larvae of endangered Lepidoptera species in cotton growing counties (Quino Checkerspot butterfly, Saint Francis' s atyr butterfly and Kern p rimrose
s phinx moth) are not going to be exposed 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 p rimrose s phinx moth (threatened, no t endangered) is found only on a privately owned ranch in the Walker Basin, Kern County California. Finally, the only known population of Saint Francis’ s atyr butterfly is found in wetlands dominated by sedges and grasses on Department of Defense 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, would 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 containing the Cry1Ac protein. In addition, because EPA is imposing conditions for geo graphic areas (Hawaii and Florida) that have sexually compatible wild or weedy 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 plants, and because the most probable exposure scenario does not appear to affect listed species, E PA believes that this action will have no effect on listed species ( US

- EPA, 2001b ; see also US

- EPA 2008a )

.BAYER TWINLINKTM COTTON

79 Based on the information above, APHIS has determined there would be no effects to the three listed lepidopteran species as a result of a determination of nonregulated status of TwinLin kTM Cotton

. 4.9.2 Potential Effects of the Use of Glufosinate Ammonium As the action agency for pesticide registrations, EPA has the responsibility to conduct an assessment of effects of a registration action on endangered species. The EPA Endangered Species Pro tection Program web site, http://www.epa.gov/espp/

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 through and break down in the environment. Risks to birds, fish, invertebrates, m ammals , and plants are routinely assessed and used in EPA’s determinations of whether a pesticide may be licensed for use in the U.S. , describes the EPA assessment process for endangered species. Some of the elements of that process, generally taken from the web site, are summarized below.

EPA’s core pesticide risk assessment and regulatory 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 are enforced by EPA and the states (Federal Insecticide, Fun gicide, and Rodenticide Act 7 USC 136j (a)(2)(G) Unlawful acts). Because TES may need specific protection, EPA has developed risk assessment procedures described in the Overview of the Ecological Risk Assessment Process ( US

- EPA, 2004 ) to determine whether individuals of a listed species 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 ammonium 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 registration review of this product, and has published a preliminary work plan for this process ( US

- EPA, 2008b )

. The EPS’ preliminary work plan provides for an ecological risk ass essment (ERA ) to allow the EPA to determine whether glufosinate ammonium use has “no effect” or “may affect” F ederally listed threatened or endangered species (listed species) or their designated crit ical habitat ( US

- EPA, 2008b )

. If the ERA indicates that glufosinate ammonium “may affect” a listed species or its designated critical habitat, the assessment will be refined 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 affect” a listed species or its designated critic al habitat, the Agency will consult with the U.S. Fish and Wildlife Service and National Marine Fisheries Service, as appropriate.BAYER TWINLINKTM COTTON

80 To mitigate potential adverse effects to TES, EPA has imposed specific label use restrictions for glufosinate ammonium use w hen 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, non

target crops) is min imal (e.g., when wind is blowing away from the sensitive areas)” ( BCS, 2010a )

. To facilitate pesticide applicators adherence to EPA label use restrictions for glufosinate ammonium , BCS’ label for Ignite, the commercial form of glufosin ate ammonium, provides specific instructions for managing spray drift and cautions pertaining to applications around water so as to minimize potential effects to non

- target organisms ( BCS, 2010a ) . L abel requirements for BCS’ Ignite formu lations prohibit application in conditions or locations where adverse impact on F ederally listed endangered/threatened plants or aquatic species is likely ( BCS, 2010a )

. BCS has also provided a technical 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 appl ication to TwinLinkTM Cotton ; EPA has considered potential impacts to TES as part of their registration and labeling process for glufosinate ammonium ; and adherence to EPA label use restrictions by the pesticide applicator will ensure that the use of glufos inate ammonium will not adversely affect TES or critical habitat. Based on these factors and the legal requirements for pesticide applicators to follow EPA label use restrictions, APHIS has determined that the use of EPA registered glufosinate ammonium fo r TwinLinkTM Cotton production will not adversely impact listed species or species proposed for listing and would not adversely impact designated critical habitat or habitat proposed for designation.
Based on the above information, APHIS has determined that the Preferred Alternative, a determination of nonregulated status of TwinLinkTM Cotton, would have no effect on F ederally listed TES and species proposed for listing, or on designated critical habitat or habitat proposed for designation. Consequently, a written concurrence or formal consultation with the USFWS is not required for this action.
4.10 CONSIDERATION OF EXECUTIVE ORDERS, STANDARDS, AND TREATIES R ELATING TO ENVIRONMENTAL IMPACTS
4.10.1 Executive Orders with Domestic Implications The following two executi ve orders require consideration of the potential impacts to minority and low income populations and children:

**•** Executive Order (EO ) 12898 ( US

- NARA, 2010 ) , "Federal Actions to Address Environmental Justice in Minority Populations and Low

- Income Populations," requires F ederal agencies to conduct their programs, policies, and activities that substantially affect human health or the environment in a manner so as not to exclude persons and populations from participation in or benefiting from such programs

. ItBAYER TWINLINKTM COTTON

81 also enforces existing statutes to prevent minority and low

- income communities from being subjected to disp roportionately high and adverse human health or environmental effects

.

**•** EO 13045, “Protection of Children from Environmental Health Risks and Safety Risks,” acknowledges that children may suffer disproportionately from environmental health and safety risk s because of their developmental stage, greater metabolic activity levels, and behavior patterns, as compared to adults

. The EO (to the extent permitted by law and consistent with the agency’s mission) requires each F ederal agency to identify, assess, and address environmental health risks and safety risks that may disproportionately affect children. Each alternative was analyzed with respect to EO 12898 and EO 13045

. Neither alternative is expected to have a disproportionate adverse effect on minorities, low

- income populations, or children

. Available mammalian toxicity, along with the history of safe use of microbial Bt products and other cotton varieties expressing Bt and PAT proteins, establishes the safety of TwinLinkTM Cotton and its products to huma ns, including minorities, low income populations, and children who might be exposed to them through agricultural production and/or processin g. No additional safety precautions would need to be taken

. None of the impacts on agricultural practices expecte d to be associated with deregulation of TwinLinkTM Cotton are expected to have a disproportionate adverse effect on minorities, low income populations, or children

. As noted above, the cultivation of previously deregulated cotton varieties with similar ins ect

- resistant traits has been associated with a decrease and/or shift in pesticide applications for those who adopt these varieties that is either favorable or neutral with respect to environmental and human toxicity

. If pesticide applications are reduced , there may be a beneficial effect on children and low income populations that might be exposed to the chemicals

. These populations might include migrant farm workers and their families, and other rural dwelling individuals who are exposed to pesticides t hrough groundwater contamination or other means of exposure

. It is expected that EPA and USDA Economic Research Service would monitor the use of this product to determine impacts on agricultural practices , such as chemical use , as they have done previousl y for other products. The following executive order addresses Federal responsibilities regarding the introduction and effects of invasive species:
EO 1311 ( US

- NARA, 2010 ) , “Invasive Species,” states that F ederal agencies take action to prevent the introduction of invasive species, to provide for their control, and to minimize the economic, ecological, and human health impacts that invasive specie s cause

. Non

- engineered cotton as well as other Bt and herbicide

- 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 reviewed by APHIS, TwinLinkTM Cotton pl ants are sufficiently similar inBAYER TWINLINKTM COTTON

82 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 protection of migratory bird populations:
EO 13186 ( US

- NARA, 2010 ) , “Responsibilities of Federal Agencies to Protect Migratory Birds,” states that F ederal agencies taking actions that have, or are likely to have, a measurable negative effect on migratory bird populations are directed to develop and implement, within two years, a Memorandum of Understanding (MOU ) with the Fish and Wildlife Service that shall promote the conservation of migratory bird populations

. Data submitted by the applicant has shown no difference in compositional and nutritional quality of TwinLinkTM Cotton compared with oth er GE cotton or non

- GE cotton, apart from the presence of the two Cry proteins and the PAT protein

. The migratory birds 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 counts in North American cotton fields

. Comparing bird populations from 1991 to 1995 and 1996 to 2000 , EPA ( 2001b ) showed that bird counts increased an d were positively correlated with Bt cotton adoption, the reduction in insecticide use , and the relative presence of the species in cotton fields prior to the introduction of Bt cotton ( US

- EPA, 2001b )

. B road

- spectrum i ns ecticides, which are considered to be one of the largest impediments to agroecosystem biodiversity, have been reduced by the commercial use of Bt cotton ( Benbrook, 2009 ; USDA

- 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 effects 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 demonstr ated 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 ( Naranjo, 2009a )

. The history of safe use of crops expressing the PAT enzyme strongly suggests that TwinLinkTM Cotton will not present a risk to birds. APHIS deregulated the PAT enzyme bar gene construct in cotton in 2003 with the BCS LibertyLink ® USDA

- APHIS, 2003 Cotton ( ) . In 2003, the FDA completed its food safety consultation on LibertyLink ®

US

- FDA, 2003 Cotton, and granted tolerance exemptions for both the plant

- incorporated protectant as well as the genetic material necessary for its production ( ) . The EPA granted exempti ons from the requirement of a tolerance for residues of glufosinate ammonium as well as the plant

- pesticide 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 does not present a risk to migratory birds.BAYER TWINLINKTM COTTON

83 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 In ternational Implications
EO 12114 ( US

- NARA, 2010 ), “Environmental Effects Abroad of Major Federal Actions” requires F ederal officials to take into consi deration any potential environmental effects outside the U.S., its territories, and possessions that result from actions being taken

. APHIS has given this EO careful consideration and does not expect a significant environmental impact outside the U.S. in the event of a determination of nonregulated 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 i nternationally apply equally to those covered by an APHIS determination 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 developed under the International Plant Protection Convention ( IPPC, 2010 )

. Th e purpose of the IPPC “is to secure a common and effective action to prevent the spread and introduction of pests of plants and plant products and to promote appropriate measures for their control” ( IPPC, 20 10 )

. 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 reciprocal acceptance of phytosanitary certification among the nations that have signed 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 Measure 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 assessment procedures for genetically engineered organisms are consi stent with the guidance developed under the IPPC

. In addition, issues that may relate to commercialization and transboundary movement of particular agricultural commodities produced through biotechnology are being addressed in other international forums a nd through national regulations.
The Cartagena Protocol on Biosafety is a treaty under the United Nations Convention on Biological Diversity (CBD ) that established a framework for the safe transboundary movement, with respect to the environment and biodiversity, of LMOs, which include those modified through biotechnology. The Protocol came into force on September 11, 2003, and 160 countries are Pa rties 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.BAYER TWINLINKTM COTTON

84 exporters will still need to comply with those r egulations 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 req uire consent from the importing country under an advanced informed agreement (AIA ) provision, which includes 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 database on domestic use of LMOs for FFP that may be subject to transboundary movement. To facilitate 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.g ov/ ) . These data will be available to the Biosafety Clearinghouse. APHIS continues to work toward harmonization of biosafety and biotechnology consensus documents, guidelines, and regulations, including within the North American Plant Protection Organ ization (NAPPO ), which includes Mexico, Canada, and the U.S., and within the Organization for Economic Cooperation and Development (OECD ). NAPPO has completed three modules of the Regional Standards for Phytosanitary Measures (RSPM ) No. 14, Importation and Release into the Environment of Transgenic 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 agricultural biotechnology issues for the U.S., Mexico, and Canada

. In addition, bilateral discussions on biotechnology regulatory issues are held regularly with other countries including Argentina, Brazil, Japan, China, and Korea.
4.10.3 Compliance with Clean Water Act and Clean Air Act This EA evaluated the changes in cotton 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

. There is no expected change in water use and quality due to the cultivation of TwinLinkTM Cotton compared with current cotton seed and cotton production . Also, t here is no ex pected change in air quality associated with the cultivation of TwinLinkTM Cotton

. Based on this review, APHIS concludes that the cultivation of TwinLinkTM Cotton would comply with the Clean Water Act and the Clean Air Act.
4.10.4 Impacts on Unique Characteristics of Geographic Areas

A determination of non

- regulated status of TwinLinkTM Cotton is not expected to impact unique characteristics of geographic areas such as park lands, prime farm lands, wetlands, wild and scenic areas, or ecologically critical areas

.BAYER TWINLINKTM COTTON

85 T he common agricultural practices that would be carried out in the cultivation of TwinLinkTM Cotton are not expected to deviate from current practices

. The product is expected to be deployed on agricultural land currently suitable for production of cotton a nd replace existing varieties, and is not expected to increase the acreage of cotton production

. There are no proposed major ground disturbances; no new physical destruction or damage to property; no alterations of property, wildlife habitat, or landscap es; and no prescribed sale, lease, or transfer of ownership of any property

. This action is limited to a determination of non

regulated status of TwinLinkTM Cotton

. This 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 preparation, planting, irrigation, and harvesting of plants would be used on agricultural lands planted to TwinLinkTM 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 Act (NHPA ) of 1966 as A mended The NHPA of 1966 and its implementing regulations (36 CFR 800) require F ederal agencies to : 1) determine whether activities they propose constitute "undertakings" that ha ve the potential to cause effects on historic properties and 2) if so, t o evaluate the effects of such undertakings on such historic resources and consult with the Advisory Council on Historic Preservation (i.e ., State Historic Preservation Office, Tribal Historic Preservation Officers), as appropriate

. APHIS’ proposed actio n, a determination of nonregulated status of TwinLinkTM Cotton, is not expected to adversely impact cultural resources on tribal properties

. Any farming activity that may be taken by farmers on tribal lands would only be conducted at the tribe’s request; t hus, the tribes would have control over any potential conflict with cultural resources 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 National 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 protected under the NHPA

. In general, common agricultural activities conducted under this action do not have the potential to introduce visual, 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 example, 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 . Neverthel ess, it is expected that this noise would only be temporary and short

- term. The cultivation of TwinLinkTM Cotton is not expected to change any of these agronomic practices that would result in an adverse impact under the NHPA.BAYER TWINLINKTM COTTON

86 

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- 377. Ruiz, N., Lavelle, P., & Jimenez, J. (2008). Soil Macrofauna Field Manual: Technical Level Retrieved January 17, 2011, from ftp://ftp.fao.org/docrep/fao/011/i0211e/i0211e.pdf Sankula, S., Marmon, G., & Blumenthal, E. (2005). Biotechnology

- Derived Cro ps Planted in 2004

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- 29). Raleigh: North Carolina Wildlife Resources Commission. Shelton, A. M., Naranjo, S. E., Romeis, J., Hellmich, R. L., Wolt, J. D., Federici, B. A., Albajes, R., Bigler, F., Burgess, E. P., Dively, G. P., Gatehouse, A. M., Malone, L. A., Roush, R., Sears, M., & Sehnal, F. (2009). Setting the record straight: a rebuttal to an erroneous analysis on transgenic insecticidal crops and natural enemies. [Letter]. Transgenic research, 18 (3), 317

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- 49. Showalter, A. M., Heuberger, S., Tabashnik, B. E., & Carrière, Y. (2009). A primer for using transgenic insecticidal cotton in developing countries. Journal of Insect Science, 9 (22), 39. Sisterson, M. S., Biggs, R. W., Manhardt, N. M., Carrière, Y., Dennehy, T. J., & Tabashnik, B. E. (2 007). Effects of transgenic Bt cotton on insecticide use and abundance of two generalist predators. Entomologia Experimentalis et Applicata, 124 (3), 305

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- 7458.2007.00584.x Stewart, C. N. (2008). Gene Flow and the Risk of Transgene S pread Retrieved November 30, 2010, from http://agribiotech.info/details/Stewart

- GeneFlow%20Mar%208%20

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94 Stewart, S. D. (2007). Bt Cotton The University of Tennessee Extension Publication W129 , 3. Sundstrom, F. J., Williams, J., Van Deynze, A., & Bradford, K. J. (2002). Identity Preservation of Agricultural Commodities. [Accessed November 29, 2010]. Agricultural Biotechnology in California Series, Publication 8077 . Retrieved from http://anrcatalog.ucdavis .edu/pdf/8077.pdf Tabashnik, B. E., Gassmann, A. J., Crowder, D. W., & Carriere, Y. (2008). Insect resistance to Bt crops: Evidence versus theory. Nature Biotechnology, 26 (2), 199

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- 202. Towery, D., & Werblow, S. (2010). Facilitating Conservation Farming Practice s and Enhancing Environmental Sustainability with Agricultural Biotechnology (pp. 28). West Lafayette, IN: Conservation Technology Information Center,. University of Arkansas Cooperative Extension Service. (2006). Cotton Insect Management: Beneficial Insects and Diseases of Cotton Retrieved November 9, 2010, from http:/ /www.aragriculture.org/insects/cotton/beneficial.htm#Spiders University of Arkansas Cooperative Extension Service. (2011). Beef Cattle Nutrition and Feeding: Frequently Asked Questions, Feeding Cottonseed Meal to Developing Bulls Retrieved March 28, 2011, from www.aragriculture.org/livestock/beef/nutrition/frequent_questions/cottonseed.htm University of California IPM Online. (2008a). Cotton: Crop Rotation Retrieved November 9, 2010, from http://www.ipm.ucdavis.edu/PMG/r114900611.html University of California IPM Online. (2008b). Cotton: Transgenic Cotton 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: University of California Agriculture and Natural Resources. Univer sity of California IPM Online. (2009c). Cotton: Integrated Weed Management 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/hortanswers/det ailproblem.cfm?PathogenID=23 University of Tennessee Agricultural Extension Service. (2010). Beaver Creek Study Final Report: Making a Splash AE03

- 63 Retrieved January 20, 2011, from http://economics. ag.utk.edu/bcstudy.htmlBAYER TWINLINKTM COTTON

95 US

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- 5: Notice to Manufacturers, Formulators, Producers and Registrants of Pesticide Products. Washington, DC: U.S. Environmental Protection Agency, Office of Pesticide Programs U S

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- Incorporated Protectants October 15, 2001 Biopesticides Registration Action Document Retrieved January 22, 2010, from http://www.epa.gov/opp00001 /biopesticides/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 Endangered and Threatened Species Effects Determinations (pp. 92). Wash ington, D.C.: U.S. Environmental Protection Agency, Office of Prevention, Pesticides and Toxic Substances, Office of Pesticide Programs. US

- EPA. (2008a). Biopesticides Registration Action Document: Bacillus thuringiensis modified Cry1Ab (SYN

- IR67B

- 1) and V ip3Aa19 (SYN

- IR102

- 7) insecticidal proteins and the genetic material necessary for their production in COT102 X COT67B cotton. In U.S. Environmental Protection Agency, Office of Pesticide Programs (Ed.). US

- EPA. (2008b). Glufosinate Final Work Plan Registr ation Review August 2008. In S. Bradbury (Ed.), (Vol. Docket Number: EPA

- HQ

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- 0190): U.S. Environmental Protection Agency. US

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- reg

- prod.htm US

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- 2008 Agriculture (Chapter 6) Washington, DC: United States Environmental Protection Agenc y. US

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- EPA. (2010d). Bacillus thuringiensis Cry1Ab Protein in all Plants; Exemption from the Requirement of a Tolerance, 40 CFR §174.511. US

- EPA. (2010e). Bacillus thuringiens is 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): E xemption from the Requirement of Tolerance. 40 CFR §174.522, first published at 40 CFR §180.1151. US

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- Foods Derived from New Plant Varieties. Retrieved January 18, 2011, from http://www.fda.gov/Food/GuidanceComplianceRegulatoryInformation/GuidanceDocume nts/Biotechnology/ucm096095.htm US

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- FWS. (2003). Recovery Plan for the Quino Checkerspot Butterfly ( Euphydryas editha quino ) (pp. 179). Portland, OR: U.S. Fish and Wildlife Service. US

- FWS. (2007a). Kern Primrose Sphinx Moth ( Euproserpinus euterpe ) 5

- Year Review: Summary and Evaluation (pp. 21). Sacramento, CA: U.S. Fish and Wildlife Service.BAYER TWINLINKTM COTTON

96 US

- FWS. (2010a). Saint Francis' Satyr Butterfly ( Neonympha mitchellii francisci ) Retrieved January 25, 2011, from http://ecos.fws.gov/speciesProfile/profile/speciesProfile.a ction?spcode=I0MZ#crithab US

- FWS. (2011a). Endangered Species Program 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/SpeciesReport.do?groups=I&listingType=L&mapstatus= 1 US

- NARA. (2010). Executive Orders Disposition Tables Index Ret rieved January 31, 2011, from http://www.archives.gov/federal

- register/executive

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collections/conservation/Gossypium

- tomentosum USDA

- AMS. (2010a). Cotton Varieties Planted 2010 Crop. Memphis, TN: U. S. Department of Agriculture, Agricultural Marketing Service. USDA

- AMS. (2010b). National Organic Program Retrieved November 23, 2010, from http://www.am s.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 CropScience USA LP Petition (02

- 042

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- regulated Status for Glufosin ate

- ammonium Herbicide

tolerant Cotton Transformation Event LLCotton25 (Environmental Assessment): U.S. Department of Agriculture, Animal and Plant Health Inspection Service, Biotechnology Regulatory Services. USDA

- APHIS. (2010a). Draft Environmental Asses sment, Syngenta Biotechnology, Inc. Petition (07

- 108

- 01p) for Determination of Nonregulated Status for Lepidopteran

Resistant Event COT67B Cotton. Riverdale, MD: U.S. Department of Agriculture, Animal and Plant Health Inspection Service. USDA

- APHIS. (2010b ). Determination of Plant Pest Risk for Insect

- Resistant and Glufosinate Ammonium

- Tolerant (TwinLinkTM) Cotton, Gossypium hirsutum , events T304

- 40 x GHB119 Petition 08

- 340

- 01p : U.S. Department of Agriculture, Animal and Plant Health Inspection Service. USDA

- ARS. (2010a). Taxon: Gossypium thurberi Tod. Retrieved November 3, 2010, from National Germplasm Resources Laboratory [Online Database] http://www.ars

grin.gov/cgi

- bin/npgs/html/taxon .pl?17947 USDA

- ERS. (2005a). Agricultural Chemicals and Production Technology: Pest Management Retrieved November 3, 2010, from http://www.ers.usda.gov/Briefing/AgChemicals/sust ainability.htm USDA

- ERS. (2005b). Agricultural Chemicals and Production Technology: Sustainability and Production Systems Retrieved November 3, 2010, from http://www.ers.usda.go v/Briefing/AgChemicals/sustainability.htm USDA

- ERS. (2006). U.S. Textile and Apparel Industries and Rural America Retrieved December 5, 2010, from http://www.ers.usda.gov/briefing/cotton/ustextileapparel.htm USDA

- ERS. (2009). Cotton: Background Retrieved November 2, 2010, from http://www.ers.usda.gov/Briefing/Cotton/background.htm USDA

- ERS. (2010a). Adoption of Genetically Engineered Crops in the U.S. Retrieved November 2, 2010, from http://www.ers.usda.gov/data/BiotechCrops/BAYER TWINLINKTM COTTON

97 USDA

- ERS. (2010b). Cotton production costs and ret urns per planted acre, excluding Government payments, 2008

- 2009 [Data table]. Retrieved December 3, 2010, from http://www.ers.usda.gov/data/costsandreturns/testpick.htm#re cent USDA

- ERS. (2010c). Adoption of Genetically Engineered Crops in the U.S.: Upland Cotton Varieties Retrieved December 3, 2010, from http://www.ers.usda.gov/Data/BiotechC rops/ExtentofAdoptionTable2.htm USDA

- ERS. (2010d). Conservation Policy: Compliance Provisions for Soil and Wetland Conservation Retrieved December 3, 2010, from http://www.er s.usda.gov/Briefing/ConservationPolicy/compliance.htm USDA

- ERS. (2010e). Table 3

-- Certified Organic and Total U.S. Acreage, Selected Crops and Livestock, 1995

- 2008 Retrieved November 5, 2010, from http:/ /www.ers.usda.gov/Data/Organic/ USDA

- FAS. (2010a). Cotton: World Markets and Trade Retrieved November 2, 2010, from http://www.fas.usda.gov/cotton/circular/2010/octobe r/Cotton_full10

- 08.pdf USDA

- FS. (2010). Gossypium hirsutum L. Wild cotton . San Juan, PR: Forest Service, International Institute of Tropical Forestry. 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 R etrieved November 6, 2010, from http://usda.mannlib.cornell.edu/usda/current/AgriChemUsFC/AgriChemUsFC

- 05

- 21

- 2008.pdf USDA

- NASS. (2010a). U.S. All Cotton Pr oduction Retrieved October 29, 2010, from http://www.nass.usda.gov/Charts_and_Maps/graphics/cotnprod.pdf USDA

- NASS. (2010b). U.S. All Cotton Acres Retrieved October 29, 2010, fr om http://www.nass.usda.gov/Charts_and_Maps/graphics/cotnac.pdf USDA

- NASS. (2010c). 2010 Upland Cotton Harvested Areas Retrieved November 2, 2010, from http://www.nass.usda.gov/Charts_and_Maps/graphics/cotnacm.pdf USDA

- NASS. (2010e). Crop Production Retrieved November 5, 2010, from http://usda.mannlib.cornell.edu/usda/nass/CropProd//2010s/2010/CropProd

- 10

- 08

- 2010.pdf USDA

- NRCS. (2006a). Conservation Resource Brief: Soil Quality, Number 0601 Retrieved November 9, 2010, from http://www.nrcs.usda.gov/feature/outlook/Soil%20Quality.pdf USDA

- NRCS. (2006b). Conservation Resource Brief: Soil Erosion, Number 0602 Retriev ed November 9, 2010, from http://www.nrcs.usda.gov/feature/outlook/Soil%20Erosion.pdf USDA

- NRCS. (2006c). Conservation Resource Brief: Air Quality, Number 0605 Retrieved November 9, 2010, from http://www.nrcs.usda.gov/feature/outlook/Air%20Quality.pdf USDA

- NRCS. (2010). 2007 National Resources Inventory: Soil Erosion on Cropland: National Resources Conservation Service. Vadakattu, G., & Watson, S. (2004). Ecological Impacts of GM Cotton on Soil Biodiversity: Below Ground Production of Bt by GM Cotton and Bt Cotton Impacts on Soil Biological Processes (pp. 72): CSIRO Land and Water.BAYER TWINLINKTM COTTON

98 Vaissiere, B. E., & Vinson, B. (1994). Pollen morphology 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, F. J., & Brad ford, K. J. (2005). Pollen

- mediated gene flow in California cotton depends on pollinator activity. Crop Science, 45 (4), 1565

- 1570. doi: 10.2135/cropsci2004.0463 Van Rie, J., Jansens, S., Höfte, H., Degheele, D., & Van Mellaert, H. (1989). Specificity of Ba cillus 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., & 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, J. R., Majewski, M. S., & Capel, P. D. (2008). Pe sticides in rain in four agricultural watersheds in the United States. Journal of Environmental Quality, 37 (3), 1101

- 1115. Whitehouse, M. E. A., Wilson, L. J., & Fitt, G. P. (2005). A Comparison of Arthropod Communities in Transgenic Bt and Conventional C otton in Australia. Environmental Entomology, 34 (5), 1224

- 1241. Wilkinson, K. M., & Elevitch, C. R. (2011). The Overstory #65: Biological Nitrogen Fixation. Agroforestry Ejournal Retrieved February 24, 2011, from http://www.agroforestry.net/overstory/overstory65.html Wilkinson, M. J., & Ford, C. S. (2007). Estimating the potential for ecological harm from gene flow to crop wild relatives. Collection of Biosafety Reviews, 3 , 42

- 63. Willi ams, M. R. (2010). Cotton Insect Loss Estimates

- 2009. Paper presented at the 2010 Beltwide Cotton Conferences, New Orleans, LA. Wilson, E. O. (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., Karp, 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., Tingey, S., Tomb, J., Gordon, M., Olson, M., & Nester, E. (2001). The genome of the natural genetic engineer Agrobacterium tumefaciens C58. Science, 294 (5550), 2317

- 2323. Wozniak, C. A. (2002). Gene Flow Assessment for Plant

- Incorporated Protectants by the Biopesticide and Pollution Prevention Division, U.S. EPA. Paper presented at the Scientific Methods Workshop: Ecological and Agronomic Consequences of Gene Flow from Transgenic Crops to Wild Relatives, Columbus, Ohio. Wunde rlin, R., & Hansen, B. (2010). Atlas of Florida Vascular Plants Retrieved December 5, 2010, from http://www.florida.plantatlas.usf.edu/BAYER TWINLINKTM COTTON

99 

### 6 L IST 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 ENDANGERED SPECIES DECISION TREE FOR FWS CONSULTATIONSBAYER TWINLINKTM COTTON

4/11/2011 A

- 1 

### APPENDIX A
APHIS THREATENED AND ENDANGERED SPECIES DECISION TREE FOR FWS CONSULTATIONS

DECISION TREE ON WHETHER SECTION 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 Information Systems for Biotechnology, at http://isb.vt.edu .) APHIS will re

- evaluate an d 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 following information, data, and questions will be
a ddressed by APHIS, and the EAs on each petition will be publicly available. APHIS review will encompass:

**•** 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 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, and agronomic and environmental impact;

**•** Determination of the concentrations of known plant toxicants (if any are known in the plant),

**•** Analysis to determine if the transgenic pla nt 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 standards as is required of all other foods. Many of the food crops currently being developed using biotechnology do not contain substances that are significantly different from those already consumed by human and thus do not require pre

market ap proval. Consistent with its 1992 policy, 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 status and conclusion of the FDA consultations in its EAs.BAYER TWINLINKTM COTTON

2 Below is a description of our review process to whether a consultation 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

1. Are naturally occurring plant toxins (toxicants) or allelochemicals increased over th e normal concentration range in parental plant species? without human intervention?

2. Does the transgene product or its metabolites have any significant similarities to known toxins 2 3. Will the new phenotype(s) imparted to the transgenic plant allow the plant to be grown or employed in new habitats (e.g., outside agro

- ecosystem) ?

3 4. Does the pest resistance . 4

A. The transgene acts only in one or more of the following ways: gene act by one of the mechanisms listed below? If the answer is YES then a consultation with U.S. Fish and Wildlife Service is NOT necessary.

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 plan t 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 sequence 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 t he 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 pr ocess mutually agreed upon with the U.S. Fish and Wildlife Service to determine 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 concurrence or formal consultation with the Fish and Wildlife Service is not required for this EA.
________ BRS has reviewed the data in accordanc e with a process mutually agreed upon with the U.S. Fish and Wildlife Service to determine when a consultation, as required under Section 7 of the Endangered Species Act, is needed. APHIS 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 consequently obtained written concurrence from the Fish and Wildlife Service. ________ BRS has reviewed the data in accordance with a process mutually agreed upon with the U.S. Fish and Wildlife Service to determine when a consultation, as required under Section 7 of the Endangered Species Act, is needed

. APHIS reached a determination that the release following a determination of non

- regulated status i s likely to affect adversely one or more listed threatened or endangered species and has initiated a formal consultation 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, H abitat, and F ood S ources of T hreatened and E ndangered Lepidoptera ( US

FWS, 2011b ) Given the specificity of activity of the Cry proteins, species outside the insect order Lepidoptera should not be affected ( Naranjo, 2009a ; Sanvido et al., 2006 )

. APHIS has thoroughly examined all FWS threatened and endangered lepidopteran species to determine if there could be possible effects from the proteins found in TwinLinkTM C otton . Threatened and endangered lepidopterans in the U.S. have restrictive habitats; and their larvae typically feed on specific host plants, none of which includes cotto n or its sexually compatible relatives, or plants expected to be found in cultivated agricultural land. Furthermore, an examination of county distribution of endangered lepidopterans shows that most are not known to occur in counties where cotton is grown , with the exception of the Kern primrose sphinx moth, the Quino C heckerspot butterfly and the Saint Francis ’ satyr butterfly

. The EPA ( 2001b ) has already concluded that the habitats of these species do not overlap with cotton f ields 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 poll en 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 plants fed upon by en dangered/threatened species, would be very small compared to the levels fed to test species ( Scott, 2008 ; 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 con stituent 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 plant s; 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 )

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- 2 Species Description Butterfly, El Segundo blue Euphilotes battoides allyni Now limited to one 302

- acre parcel (owned by the Los Angeles Airpor t) and one 2

- acre parcel (owned by the Standard 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 remnant inhabited by a m etapopulation, any patch of foodplant in open to semi

shaded 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 ( NatureServe, 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, 2005 ; Szymanski et al., 2004 ; US

- FWS, 1998a , 2010c )

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- 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 scrub 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 ( US

FWS, 1998b )

.
Butterfly, Oregon silverspot Speyeria zerene hippolyta Known to occur in Del Norte County, CA; in Clatsop, Lane, Lincoln, Tillamook and Counties, OR and Pacific County, WA in salt

- spray meadows with its host Viola adunca ( US

- FWS, 2001b )

.
Butterfly, Palos Verdes blue Glaucopsyche lygdamus pa losverdesensis 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 scopa rius ) ( 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 Chaparral, coastal sage scrub, with primary host plants : dwarf plantain ( Plantago erecta ), wooly plantain ( Plantago patagonica ), the white snapdragon ( Antirrhinum coulterianum ), thread

leaved 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 that receive little direct sunlight

. It lives near prolific growths of the larval food plant, stonecrop ( Sedum spathulifolium ), which is a low

- growin g succulent

. Stonecrop is associated with rocky outcrops that occur at 900

- 1075 feet elevation ( Butterfly Conservation Initiative, 2006b ; NatureServe, 201 0i ; 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 whi ch is torchwood Amyris elemifera

. Adults do stray into other 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 sand dune strand vegetation dominates

. One population found in chaparral

woodland 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 )

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- 4 Species Description Butterfly, Uncompahgre fritillary Boloria acrocnema Restrict ed 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 , 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 and a few introduced Solanaceae (tobacco family)

. However, adults do wander and larvae have turned up on cultivat ed Solanaceae

. Larvae of Blackburn's sphinx 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 cons ume leaves, stems, flowers, and buds

. However, many of the host plants recorded for this species are not native to the Hawaiian Islands, and include Nicotiana tabacum (commercial tobacco), Nicotiana glauca (tree tobacco), Solanum melongena (eggplant), Lyc opersicon 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, B. J., & Bach, C. E. (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. Butterfly Conse rvation Initiative. (2006a). Palos Verdes Blue Butterfly Retrieved January 31, 2011, from http://www.butterflyrecovery.org/species_profiles/profiles.php?id=13 Butterfly Con servation Initiative. (2006b). San Bruno Elfin Butterfly Retrieved January 31, 2011, from http://www.butterflyrecovery.org/species_profiles/profiles.php?id=16 Butterfly Con servation Initiative. (2006c). Callippe Silverspot Butterfly Retrieved March 8, 2011, from http://www.butterflyrecovery.org/species_profiles/profiles.php?id=2 Khan, A. H., & Afzal, M. (1950). Natural crossing in cotton in Western Punjab IV. Agents of natural crossing. Agronomy Journal, 42 (5), 236

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- 5 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 Bl ue Retrieved January 31, 2011, from http://www.natureserve.org/explorer/servlet/NatureServe?searchName=Euphilotes+battoi des+all NatureServe. (2010d). Plebejus icarioides fenderi : Fender's Blue Retrieved January 31, 2011, from http://www.natureserve.org/explorer/servlet/NatureS erve?searchName=Icaricia+icarioide s+fend NatureServe. (2010e). Plebejus melissa samuelis : Karner Blue Retrieved January 31, 2011, from http://www.nature serve.org/explorer/servlet/NatureServe?searchName=Lycaeides+meliss a+sam NatureServe. (2010f). Plebejus anna lotis : Lotis Blue Retrieved January 31, 2011, from ht tp://www.fws.gov/sacramento/es/animal_spp_acct/langes_metalmark_butterfly.pdf NatureServe. (2010g). Plebejus icarioides missionensis : Mission Blue Retrieved January 31, 2011, from http://www.natureserve.org/explorer/servlet/NatureServe?searchName=Icaricia+icarioide s+missi NatureServe. (2010h). Neonympha mitchellii francisci : Mitchell's Satyr Retrieved January 31, 2011, from http://www.natureserve.org/explorer/servlet/NatureServe?searchName=Neonympha+mit chellii+francisci NatureServe. (2010i). Callophrys mossii bayensis : San Bruno Elfin Retri eved January 31, 2011, from http://www.natureserve.org/explorer/servlet/NatureServe?searchName=Callophrys+moss ii+bayensis NatureServe. (2010j). Papi lio aristodemus ponceanus : Schaus' Swallowtail Retrieved January 31, 2011, from http://www.natureserve.org/explorer/servlet/NatureServe?searc hName=Heraclides+aristo demus+ponceanus NatureServe. (2010k). Euphilotes enoptes smithi : Smith's Blue Retrieved January 31, 2011, from http://www.nat ureserve.org/explorer/servlet/NatureServe?searchName=Euphilotes+enopt es+smithi NatureServe. (2010l). Boloria improba acrocnema : Uncompahgre Fritillary Retrieved January 31, 2011, from http://www.natureserve.org/explorer/servlet/NatureServe?searchName=Boloria+acrocne
ma NatureServe. (2010m). Manduca blackburni : Blackburn's Sphinx Moth Retrieved January 31, 2011, from http://www.natureserve.org/explorer/servlet/NatureServe?searchName=Manduca+blackb urniBAYER TWINLINKTM COTTON

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- 6 Sanvido, O., Stark, M., Romeis, J., & Bigler, F. (2006). Ecological Impa cts of Genetically Modified Crops: Experiences from Ten Years of Experimental Field Research and Commercial Cultivation. Zürich, Switzerland: Agroscope Reckenholz

- Tänikon Research Station ART. Scott, A. L. (2008). Petition for Determination of Nonregulated Status for Insect

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- Tolerant Cotton: TwinLinkTM Cotton (Events T304

- 40 x GHB119). Research Triangle Park, NC: Bayer CropScience. Szymanski, J., Shuey, J. A., & Oberhauser, K. (2004). Population structure of the endangered M itchell's Satyr, Neonympha mitchellii mitchellii (French): Implications for conservation. The American Midland Naturalist, 152 (2), 304

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- Incorporated Protectants October 15, 2001 Biopesticides Registration Action Document Retrieved January 22, 2010, from http://www.epa.gov/opp0000 1/biopesticides/pips/bt_brad2/5

- benefits.pdf US

- EPA. (2008). Biopesticides Registration Action Document ( Bacillus thuringiensis Cry2Ab2 Protein and its Genetic Material Necessary for its Production in Cotton) (Chemical PC Code 006487) Amended: U.S. Environ mental Protection Agency, Office of Pesticide Programs. US

- FWS. (1980). 50 CFR Part 17 Listing the Palos Verdes Blue Butterfly as an Endangered Species with Critical Habitat. Federal Register, 45 (129), 44939

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- FWS. (1994). Uncompahgre Fritillary B utterfly Recovery Plan (pp. 20). Denver, CO: U.S. Fish and Wildlife Service. US

- FWS. (1998a). Recovery Plan for Mitchell's Satyr Butterfly Neonympha mitchellii mitchellii French (pp. 71). Fort Snelling, MN: U.S. Department of the Interior, Fish and Wildlif e Service. US

- FWS. (1998b). Seven Coastal Plants and the Myrtle's Silverspot Butterfly Recovery Plan (pp. 141). Portland, OR: U.S. Fish and Wildlife Service. US

- FWS. (1999). Schaus Swallowtail Butterfly Heraclides aristodemus ponceanus Mult

- Species Recover y Plan for South Florida: Invertebrates (pp. 743

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- 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

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- FWS. (2000b). 50 CFR Part 17 Endangered and Threatened Wildlife and Plants; Deter mination of Endangered Status for Blackburn's Sphinx Moth from the Hawaiian Islands. Federal Register, 65 (21), 4770

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- FWS. (2001a). 50 CFR Part 17 Endangered and Threatened Wildlife and Plants; Final Determination of Critical Habitat for the Bay Ch eckerspot Butterfly ( Euphydryas editha bayensis ); Final Rule. Federal Register, 66 (83), 21450

- 21489. US

- FWS. (2001b). Oregon Silverspot Butterfly ( Speyeria zerene hippolyta ) Revised Recovery Plan (pp. 113). Portland, OR: U.S. Fish and Wildlife Service. US

- FWS. (2003). Recovery Plan for the Quino Checkerspot Butterfly ( Euphydryas editha quino ) (pp. 179). Portland, OR: U.S. Fish and Wildlife Service.BAYER TWINLINKTM COTTON

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- 7 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 (pp. 21). Sacramento, CA: U.S. Fish and Wildlife Service. US

- FWS. (2007b). Endangered and Threatened Wildlife and Plants; Initiation of 5

- Year Reviews of 58 Species in California and Nevada; Availability of Completed 5

- Year Reviews in California and Nevada. Federal Register, 72 (30), 7064

- 7068. US

- FWS. (2008). La nge's Metalmark Butterfly: Apodemia mormo langei Retrieved January 31, 2011, from http://www.fws.gov/sacramento/es/animal_spp_acct/langes_metalmark_butterfly.pdf US

- FWS. (2010b). Callippe Silverspot Butterfly ( Speyeria callippe callippe ) Retrieved January 31, 2011, from http://ecos.fws.gov/speciesProfile/profile/speciesProf ile.action?spcode=I019 US

- FWS. (2010c). Mitchell's Satyr Butterfly ( Neonympha mitchellii mitchellii ) Retrieved January 31, 2011, from http://ecos.fws.gov/speciesPro file/profile/speciesProfile.action?spcode=I00K US

- FWS. (2010d). Quino Checkerspot Butterfly ( Euphydryas editha quino ) Retrieved January 31, 2011, from http://ecos.f ws.gov/speciesProfile/profile/speciesProfile.action?spcode=I00P US

- FWS. (2010e). San Bruno Elfin Butterfly: Callophrys mossii bayensis Retrieved March 8, 2011, from http://www.fws.gov/sacramento/es/animal_spp_acct/san_bruno_elfin_butterfly.pdf US

- FWS. (2010f). Endangered Species in the Pacific Islands 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/SpeciesReport.do?group s=I&listingType=L&mapstatus= 1 USDA

- APHIS. (2010b). Determination of Plant Pest Risk for Insect

- Resistant and Glufosinate Ammonium

- Tolerant (TwinLinkTM) Cotton, Gossypium hirsutum , events T304

- 40 x GHB119 Petition 08

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- 01p : U.S. Department of Agriculture, Animal and Plant Health Inspection Service.