# Syngenta Biotechnology, Inc. Insect Resistant MIR162 Corn

OEeD 

## Unique Identifier: SYN-IR162-4



## Final Environmental Assessment

March 2010 Agency Contact CynthiaEck Document Control Officer Biotechnology Regulatory Services USDA,APHIS Riverdale, MD 20737

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1Table ofContents

I. SUMMARY 3

II. PURPOSE & NEED 3

III. INTRODUCTION 6

IV. AFFECTED ENVIRONMENT 7

A. Corn 7 B. Agricultural Production ofCorn 8 C. CornLepidopteran Pests 9

V. ALTERNA TIVES 10

A. No Action: Continuation as a Regulated Article 10 B. Grantnonregulated status to MIR162 corn, "inwbole"­ Preferred Alternative: Determination tbatMIR162 Corn is No Longer a Regulated Article 11

VI. ENVIRONMENTAL CONSEQUENCES 11

A. No Action 12 1. Corn 12 2. Agricultural Production ofCorn 12 3. CornLepidopteran Pests 12

B. Preferred Alternatives 13 1. Corn 13 2. Agricultural Production ofCorn 14 3. Potential Impacts ofLine MIR162 Corn on Insect Control Practices 14 4. Organic and OtherNon-transgenic Corn Production 15 5. Potential Impact on Non-target Organisms, Including

Beneficial Organisms and Threatened orEndangered Species 17 6. Cumulative Effects 20 7. Socioeconomic Analysis 22

C. Consideration ofExecutive Orders, Standards and Treaties Relating to Environmental Impacts 27

VII. REFERENCES 30

VIII. PREPARERS, REVIEWERS, CONSULTAnONS, AGENCY CONTACT 36

IX. APPENDIX Appendix I. Environmental and Human Health 37 Appendix II. Syngenta MIR162 Corn Forage and GrainCompositional Analysis SafetyofVip3Aa20 Protein
39

2

## I. Summary

The Animal and Plant Health Inspection Service (APHIS), United States Department of Agriculture (USDA), has prepared an Environmental Assessment (EA) in response to a petition (APHIS Number 07-253-01p) from Syngenta Biotechnology, Inc. (Syngenta) regarding theregulatory status ofgenetically engineered (transgenic) corn resistant to lepidopteran insect feeding from transformation event MIR162. This corn is currently a regulated article under USDAregulations at 7 CFRpart 340, and as such, interstate movements, importations, and field tests ofMIR162 corn have been conducted under permits issued ornotifications acknowledged byAPHIS. Syngentapetitioned APHIS requesting a determinationthat MIR162 corn does not present a plant pest risk, and therefore MIR162 corn and itsprogeny derived from crosses with othernonregulated
corn should no longerberegulated articles under these APHIS regulations. 

## II. Purpose and Need
"Protecting American agriculture" is the basic charge oftheUSDA-APHIS. APHIS provides leadership in ensuring the health and care ofplants and animals. The agency improves agricultural productivity and competitiveness, and contributes to the national economy and the public health. USDA asserts that all methods ofagricultural production (conventional, organic, orthe use ofgenetically engineered varieties) can provide benefits to the environment, consumers, and farm income.


### FederalRegulatoryAuthority

In 1986, the Federal Government's Office ofScience and Technology Policy (OSTP) published a policy document known as the Coordinated Framework for the Regulation of Biotechnology. This document specifies three Federal agencies that areresponsible for regulatingbiotechnology in the U.S.: USDA's APHIS, the Environmental Protection Agency (EPA), and the U.S. Department ofHealth and Human Services' Food and Drug Administration (FDA). APHIS regulates genetically engineered (GE) organisms under the Plant Protection Act of2000. 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). FDA regulates GE organisms underthe authority ofthe Federal Food, Drug, and Cosmetic Act. The FDApolicy statement concerning regulation ofproducts derived from newplant varieties, including those genetically engineered, was published in the Federal Register
on May 29, 1992 (57 FR 22984-23005). Underthis policy, FDA uses what is termed a consultationprocess to ensure that human food and animal feed safety issues or other regulatory issues (e.g., labeling) are resolved priorto commercial distribution of bioengineered food. Together, these agencies ensure that the products ofmodern biotechnology are safe to grow, safe to eat, and safe for the environment. USDA, EPA, and FDA enforce agency-specific regulations to products ofbiotechnology that are based
on the specific nature ofeach GE organism. Products are regulated according to their

3intended use and some products are regulated bymore than one agency.


### USDA RegulatoryAuthority

The APHIS Biotechnology Regulatory Service's (BRS) mission is to protect the United States' agriculture and environment using a dynamic and science-based regulatory framework that allows for the safe development and use ofgenetically engineered organisms. APHIS regulations at 7 Code ofFederal Regulations (CFR) part 340, which
were promulgatedpursuant to authority granted bythe Plant Protection Act, as amended (7 United States Code (U.S.c.) 7701-7772), regulate the introduction (importation, interstate movement, orrelease into the environment) ofcertain GE organisms and products. A GE organism is considered a regulated article ifthe donor organism, recipient organism, vector, orvector agent used in engineering the organismbelongs to
one ofthe taxa listed in the regulation (7 CFRpart 340.2) and is also considered a plant pest. A GE organism is also regulated underpart 340 when APHIS has reason to believe that the GE organism may be a plantpest orAPHIS does not have sufficient information to determine ifthe GE organism is unlikely to pose a plantpest risk.

A person maypetition the agency to evaluate submitted data and determine that a particularregulated article is unlikelyto pose a plantpest risk, and, therefore, should no longerberegulated under 7 CFRpart 340.6 entitled "Petition for Determination of Nonregulated Status." The petitioner is required to provide information under 

### § 340.6(c)(4) related to plant pestrisk that the agency uses to determine whether the regulated article is unlikely to present a greaterplantpest risk than the unmodified organism. A GE organism is no longer subject to the regulatoryrequirements of7 CFR part 340 when APHIS determines that it is unlikely to pose a plant pest risk. Syngenta Biotechnology, Inc. (hereafter "Syngenta") ofResearch Triangle Park, NC submitted a petition to APHIS seeking a determination ofnonregulated status for their transgenic event MIR162 com (hereafter "MIR162 com"). The MIR162 comhas been engineered to express a bacterial protein Vip3Aa20 from Bacillus thuringiensis that is toxic to a certain lepidopteran insect pests. This com is also engineered to express anotherprotein, phosphomannose isomerase (PMI) from Escherichia coli, which was used as a selectable markerto identify com seedlings containing Vip3Aa20 gene during the development ofMIR162 com. The MIR162 com is currently regulated under 7 CFR part 340. This comhas been considered a regulated article because it was genetically engineered with regulatory sequences derived from plant pests and because a plant pest
was used as a vector agent to deliver those sequences to the plant. Interstate movements and field trials ofthe MIR162 com have been conducted underpermits issued or notifications acknowledged byAPHIS.

Underthe authority of7 CFRpart 340, APHIS has the responsibility for the safe development and use ofgenetically engineered organisms under the provisions ofthe Plant Protection Act. APHIS must respond to petitioners that request a determination of the regulated status ofgenetically engineered organisms, including genetically engineered crop plants such as MIR162 com. Ifa petition for nonregulated status is

4submitted, APHIS must detennine whetherthe genetically engineered organism is unlikely to pose a plant pest risk.

As a Federal agency subject to compliance with the National Environmental Policy Act (NEPA)! (42 U.S.C. 4321 et seq.), APHIS has prepared this EA to consider the potential environmental effects ofthis proposed action (granting nonregulated status) and the reasonable alternatives to that action consistent with NEPA implementing regulations (40 CFR §§ 1500-1508, 7 CFRpart 1(b), and 7 CFRpart 372) and the USDA and APHIS NEPA implementing regulations and procedures. This EAhas been prepared in orderto specifically evaluate the effects on the quality ofthe human environment! that mayresult from the deregulation ofthe MIR162 com.


# u.s. EnvironmentalProtectionAgencyandFoodandDrug AdministrationRegulatoryAuthority

The MIR162 comis also subject to regulation by other agencies. The U.S. Environmental Protection Agency (EPA) is responsible for regulation ofpesticides under the Federal Insecticide, Fungicide, and Rodenticide Act (FIFRA) (7 U.S.C. §136 et seq.). FIFRA requires that all pesticides beregistered before distribution, sale, and use, unless exempted byEPA regulation. Before a product is registered as a pesticide under FIFRA, it must be shown that when used in accordance with the label, it will not result in unreasonable adverse effects on the environment. Accordingly, EPA grants pennits to allow a pesticide producer to test a new pesticide product outside the laboratoryunder Experimental Use Pennits (EUPs), which are used for large-scale (more than 10 acres of land or 1 acre ofwater) testing ofefficacy and gathering ofenvironmental fate, ecological effects, and crop residue chemistry (40 CFRpart 172). Syngenta obtained an experimental use pennit from EPA that allowed for broad-scale field testing ofthe MIR162 com; this pennitwas granted on March 26, 2007 and was in effect through March 31, 2008 (72.FR 34009-34010). On July 23,2008 EPA announced receipt ofa petition from Syngenta to conditionally register three pesticide products containing the new active ingredient Vip3Aa20 and the genetic material necessary for its production in com (73 FR42799-42801). These pesticide products included MIR162 and the two comhybrids Btl1 x MIR162 and Btl 1 x MIR162 x MIR604 (Btl 1 and MIR604 contain the additional insecticidal protein active ingredients Cry1Ab and Cry3A, respectively, and bothhave previously been deregulated by APHIS and registered as plant-incorporated protectants by EPA). On April 30, 2009, EPA announced the approval ofthese conditional registrations involving MIR 162 and the hybrids (74 FR 19956-19957).

Under NEPA regulations, the "human environment" includes "the natural and physical environment and the relationship ofpeople with that environment" (40 CFR §150S.14)
2 Cryproteins are crystal proteins that are produced withinthe spores ofBtbacteria. A majority of deregulated Bt crops currently available inthe U.S. market express Cryproteins. I

5Under the Federal Food, Drug, and Cosmetic Act (FDCA) (21 U.S.C. §301 et seq.), pesticides added to (or contained in) raw agricultural commodities are prohibited unless a tolerance or exemption from tolerance has been established. EPA establishes residue tolerances for pesticides under the authority ofthe FDCA. The FDA enforces the tolerances set by the EPA. On April 4, 2007 EPA established a temporary exemption from the requirement ofa tolerance for Vip3Aa20 residues in maize commodities, pursuant to §408(d) ofthe Federal Food, Drug, and Cosmetic Act, 21 U.S.C. §346a(d). On August 6, 2008, EPA granted exemption from the requirement ofa tolerance for residues ofB. thuringiensis Vip3Aaproteins (including the Vip3Aa20 variant) in or on food and feed commodities ofcom (73 FR45620-45624). OnMay 14, 2004, EPA granted an exemption from the requirement ofa tolerance for residues in or onplant commodities ofphosphomannose isomerase and the genetic material necessary for its production in all plants when applied/used as plant-incorporated protectant inert ingredients (69 FR26770-26775). With the publication ofEPA'sregistration document, APHIS will use this finalized information to provide additional scientific support to its consideration ofpotential environmental impacts.
FDA, which has primaryregulatory authority over food and feed safety, published a policy statement in the Federal Register on May 29, 1992 (57 FR22984-23005) concerning regulation ofproducts derived from new plant varieties, including those genetically engineered. Under this policy, FDA uses what is termed a consultation process to ensure that human food and animal feed safety issues or otherregulatory issues (e.g. labeling) are resolved priorto commercial distribution ofa bioengineered food. Syngenta submitted a summary oftheir safety assessment to FDA on August 3, 2007, and additional information on December 17, 2007 and March 31, 2008. Syngenta's submissions to FDA indicated that food and feed derived from com event MIR162 are as safe (Appendix I in this EA) and nutritious as food and feed derived from conventional
com (Appendix II in this EA). FDAcompleted their consultation onMIR162 on December 9,2008 and concluded that it had "no further questions concerning grain and forage derived from com event MIR162" (FDA BNF No. 000113).

Public Involvement

APHIS-BRS routinely seeks public comment on draft environmental assessments prepared in response to petitions to deregulate GE organisms. APHIS-BRS does this through a notice published in the Federal Register. This EA, the petition submitted by Syngenta, and APHIS's plant pest risk assessment, were made available for public comment for a period of60 days. Comments that were received within the 60-day commentperiod were fully analyzed and used by APHIS to determine ifthe petition to deregulate the MIR162 com shouldbe granted.

Decision to Be Made

APHIS will use the information from this EA, and the comments received, to assist APHIS' decisionmaker to determine whether to grant nonregulated status, or to continue to regulate MIR162 comunder the regulations at 7 CFRpart 340, or that an

6Environmental Impact Statement is necessaryprior to the decision to grant nonregulated status to this com variety. 

## III. Introduction

Com is susceptible to attack by a variety ofinsects (Table 1, pg. 12 in petition) from the time it is planted until it is consumed as food or feed. Syngentahas developed a GE com hybrid, named MIR162, that is resistant to the feeding damage caused by com earworm (Helicoverpa zea), fall armyworm (SpodopteraJrugiperda), black cutworm (Agrotis ipsilon), and westernbean cutworm (Striacosta albicosta) larvae that are not controlled well with existing technology. This insect resistance in MIR162 comes from a bacterial gene called Vip3Aa20 (Vip = Vegetative insecticidal protein). The MIR162 com also contains manA gene from E. coli encoding the enzyme phosphomannose isomerase (PMI), whichwas used as a selectable marker during transformant selection. The manA gene expression confers no otherbenefit to the regenerated transformed complant.

The family ofVip3Aaproteins, inwhichVip3Aa20 belongs, are produced bythe bacteriumB.thuringiensis (hereafter "Bt") (Estruch et al. 1996) that act as toxins to kill insect prey (Estruch et al. 1996; Schnepfet al. 1998). Vip3Aaproteins are similarto certain Cryproteins2 (Hafte and Whiteley 1989) and are demonstrated to have toxic effects only on certain insects (Table 2.1 on p. 23 in Carozzi and Koziel 1997). The mechanism bywhich Vip proteins exert their insecticidal activityhas been studied and found to be similar, but not identical, to that which has been described for the Bt Cry proteins that are contained in several commercial insecticide formulations and APHIS deregulated GE plants engineered for insect resistance (USDA-APHIS, 2010, see Transgenic phenotype column in Table: examples include coleopteran-, or lepidopteranresistant, comborer (ECB) resistant, comrootworm resistant, Colorado potato beetle resistant). The Vip and Cryproteins bind to different receptors in the insect (Lee et al. 2003), and the insecticidal activity ofVip3Aaproteins is limited to species within selected families ofthe order Lepidoptera (Table 27, pg. 74-75 in Syngenta, 2007). For example, MIR162 alone has no activity against European comborer (Ostrinia nubilalis) but is efficacious in limiting feeding damage caused bythe other four insect pests (com earworm, fall armyworm, black cutworm, western bean cutworm) (Figure 21, pg. 76 in petition); whereas the Btl 1 GE comvariety (containing a Cryprotein) is highly efficacious against European comborer, but it has limited orno activity against the other four insects. USDA, APHIS has previously granted nonregulated status to 11 insect resistant GE com varieties containing Cryproteins from Bt (USDA-APHIS 2009).

The MIR162 com has been field tested in the United States since 1999 as authorized by APHIS. Associated notifications acknowledged and permits issued byAPHIS are listed in Appendix A ofthe petition (pg. 127-128). The list compiles more than 20 test sites in diverse regions ofthe U.S. including themajor com growing area ofthe Midwest and winternurseries inHawaii. Field tests conducted under APHIS oversight allow for evaluation in agricultural settings under confinement measures designed to minimize the likelihood ofpersistence in the environment after completion ofthe field trial. Under confined field trial conditions, applicants gather data for agronomic characteristics and

7product perfonnance in response to insects, disease orother stresses. These data are also valuable to APHIS as the agency assesses the potential for a new com varietyto pose a plant pest risk. APHIS' evaluation ofthis data maybe found in the APHIS plant pest risk assessment (USDA-APHIS 2009). 

## IV. Affected Environment
A. Corn

Com is primarily grown in wann temperate climates (Nonnan et al. 1995). Field com is the leading agricultural production crop globally, with the 2009 growing season expected to yield 789 million metric tons ofgrain (lCG 2009). Com is grown for animal feed, human food, vegetable oil, high fructose com syrups, starch, fennentation into ethanol, and a multitude ofindustrial uses (Hoeft et aI., 2000).

Zea mays L. subsp. mays, known as maize throughout the world, and as com in the U.S., is a member oftheMaydeae tribe ofthe grass family, Poaceae. Itis an annual plant with separate male and female flowers on eachplant (monoecious) that requires human intervention for its seed dispersal and propagation. Additional infonnation onthe biology ofcom canbe found within the Organisation for Economic Co-Operation and Development consensus document (OECD 2003).

Com is predominantly a wind-pollinated outcrossing species (OECD 2003). Transgenes in crops have the potential to move between sexually compatiblepopulations, and more so in com being a wind-pollinated plant with separate male and female flower bearing structures (inflorescences). Gene flow rate between compopulations is extremely variable depending onthe spatial, temporal, genetic and environmental factors (Brookes and Barfoot et ai. 2004; Messegue et ai. 2006). Yet, available experimental evidence indicates that gene flow rates drop substantially (1 %) beyond 20 meters (Henry et ai. 2003; Ma et ai. 2004; Messeguer et ai. 2006). To maintain varietal purity, the AOSCA (Association ofOfficial Seed Certifying Agencies) recommends 200 meters isolation for nearby compopUlations as the foundation class ofcertified seed production (AOSCA 2003).

The insect resistance trait ofMIR162 has the potential to enhance the fitness ofwild and weedyrelatives ifgene flow occurs betweenthe MIR 162 com crop and wild or weedy
compopulations. However, there are no large populations orwidely distributed wild
complants (teosinte) inthe U.S., and eventhe few non-weedy feral popUlations inthe U.S. have limited opportunity for outcrossing with transgenic com cultivars (see USDAAPHIS 2009).

Com is not weedy, and does not persist outside cultivated areas (USDA-APHIS 2009). APHIS knows ofno reports in which compropagated vegetatively under field conditions, since the onlyknown propagation method for com is through seed gennination. Com seed is sensitive to cold and typically does not survive freezing winter conditions. Consequently, comhas no innate donnancy (Simpson 1990; Table 18, pg. 61 Syngenta

82007). Even ifcom seeds from a previous year's crop overwinter and germinate the following year, manual or chemical measures are available and are often applied to
remove these volunteers (see Table 1in Wright et al. 2009).


### B. AgriculturalProduction ofCorn The U.S. accounts for about 41% ofglobal comproduction (Bange 2007). Com is the largest crop grown in the U.S. in terms ofboth volume and value. Approximately 86 million acres were planted in 2008 growing season, yielding 12 billionbushels (305 millionmetric tons) with a gross crop value of$47 billion ($3.9Ibushel) (USDA-NASS 2008a; USDA-NASS 2008b). The upper Midwest region ofthe U.S. provides an ideal combination oftemperature, rainfall, and soil type for the cultivation ofcom. Iowa, Illinois, Nebraska, Minnesota, Indiana, Ohio, Wisconsin, Missouri, Kansas, and South Dakota are major com growing states. Production in these ten states accounts for 77% of total annual production (USDA-NASS 2008b).

The use ofcom as a source offuel ethanol has increased dramatically over the past two years and is expected to continue to increase as the U.S. focuses on employing renewable
sources ofenergy. The Federal Energy Act of2005 includes a nationwide renewable fuels standard (RFS) that will result in the use ofmore than 7.5 billion gallons ofethanol and biodiesel by 2012 (42 USC 15801, page 1069). Over 20% ofcommodity com in 2007 was used for ethanol production (Trostle 2008; USDA-ERS 2008). By 2010, U.S. ethanol production could displace the equivalent of311 ,000 barrels ofimported crude oil per day (GAO, 1996).

The U.S. is by far the world's largest exporter ofcom, and in 200312004 through 2007/2008 accounted for on average 60% ofworld com exports (USDA-ERS 2009a). Total U.S. agricultural exports in 2006 were valued at $71 billion, 10% ofwhich was attributable to com (Brooks 2007). Agricultural exports generate employment, income, and purchasing powerin both farm and nonfarm sectors ofthe economy. Production from almost one-third ofU.S. cropland moved into export channels in 2005 and generated $166.1 billion in business activity (Food Institute 2007). Technology advances, such as those attributed to GE crops, increase agricultural productivity and keep domestic growers competitive in the global market (NCRA 2010).

Based on USDA survey data, adoption ofgenetically engineered insect-resistant com increased from zero percent ofthe U.S. com acreage in 1996 to 63 percent in 2009 (USDA-ERS 2009). The rapid commercialization ofGE insect-resistant com (IR com) varieties by com growers is attributed to benefits offered by those com varieties in terms ofreduced conventional insecticide use, increased profits, and improved grain quality (Fernandez-Conejo and Caswell 2006).

In addition to insect resistant (IR) com cultivation, U.S. farmers have also planted GE herbicide tolerant (HT) com varieties since 1996. A few GE com cultivars contain both IR (European comborerresistance, comrootworm, etc.) and HT traits (glyphosate tolerance, imidizolinone tolerance). Among GE varieties ofcom (IR and HT com cultivars), 68% ofall GE com varieties planted contained a herbicide tolerant (HT) trait
9(USDA-ERS 2009b). Herbicides were applied to 97 percent ofthe GE and non-GE com planted acreage in 2005, with atrazine, glyphosate, S-metolachlor and acetochlorbeing applied to 66%, 31%,23% and 23% ofplanted com acres, respectively (USDA-NASS 2006). MIR162 com is not expected to alter current or future comweed control practices. The main introduced trait in MIR162 is expected to provide resistance to certain groups ofinsect pests. Therefore, except for change in insect resistance management, all other agricultural practices ofthe MIR162 com, including com weed control practices, are not expected to be different from those ofconventional com cultivation. According to USDA-ERS (2009) report, 15%**(~13** million acres) ofthe U.S. com acreage was planted with the non-GE com varieties in 2009. Likewise, according to USDA-ERS' latest data on organic comproduction, less than 1percent (0.16%) ofcom crop area in 2005 was devoted to organic com (USDA-ERS 2009c). Under USDA National Organic Program regulations (USDA-AMS 2010), the use ofsynthetic pesticides, fertilizers, and genetically engineered crops is strictly limited (7 CFRpart 205.105 and definition, "excluded methods"). As a result, MIR162 com is not approved for use in organic production systems because it is genetically engineered. Maintaining the integrity ofthe organic production process is important to producers oforganic com.
There are manypractices organic producers use to prevent movement ofGE com or the pollen from GE com into their organic production fields (Bradford 2006, Schienmann 2003, Ziegler 2000). Growers may chose to plant only organic seed; plant earlier or later than neighboring farmers who maybeusing GE crops, ensuring that the flowering times between GE and organicallyproduced crops will differ, thus minimizing the change of pollen movement between fields; and also employ adequate isolation distances between the organic field and the fields ofneighbors to minimize the chance that pollen will be carried between the fields. Additionally, organic growers must maintain records to show that production and handling procedures comply with USDA organic standards (7 CFR part 205).


### C. 

### Corn LepidopteranPests

Com crop is susceptible to attackby a variety ofinsects throughout its life cycle (see pg. 12 in Syngenta 2007). Two ofthe five most widespread and damaging insects ofcom in the U.S. ComBelt are the European com borer and comrootworms (Hoeft, et al. 2000). Although a few conventional insect control practices (chemical and microbial insecticides, crop rotation etc.) are available for cominsect pests, the stalk boring insects such as the European comborerhave been difficult to control and in some areas, it is not profitable to use chemical control against such insect pests (Martin and Hyde 2001). Conventional insecticide and crop rotation practices have been proven effective in controlling the damage caused by comrootworms (Ma et aI., 2009). Prior to the introduction ofGE rootworm-protected Bt varieties in 2003, an estimated 14 million
acres ofcom were treated annually with conventional insecticides to control com rootworms (Ward et al. 2005); insecticides for rootworm control accounted for the largest single use ofinsecticides in the U.S. Treatment ofcomrootworm with chemical

10pesticides mayhave decreased by34%, the difference in acreage between use of European com borerprotected com and comrootworm protected Bt combetween 2003 (when CRW comwas first available) and 2009 (USDA-ERS 2009c). The use of conventional insecticide treatment is less effective for some cominsect pests, such as com earworm (Hoeft et a1. 2000), as some ofthese compests may enter areas shielded from aerial chemical applications; com earworm follows the silk channel and enters the protected recesses ofthe com ear (Burkness et al. 2009)

.
In addition to direct damage caused by feeding on plant tissue, cominsect pests are also known to play an important role inthe transmission and dissemination ofpathogenic organisms during com development (Dowd 1998). For example, it has been shownthat insect feeding damage enhances mycotoxincontamination ofcom crop (Williams et a1. 2002) that have toxic and carcinogenic effects inhumans and animals (see Wu 2006 for details). The introduction ofGE Bt comvarieties has provided growers solutions to
some ofthe above-mentioned pest problems by limiting damage caused by certain lepidopteran insectpests (Hurley et a1. 2006) and fungal diseases (Wu 2006) without posing any significant risk to the environment orto human health (Mendelsohn et a1. 2003). 

# v. Alternatives

This EA analyzes the potential environmental consequences ofa proposal to grant nonregulated status to the MIR162 com. In order forMIR162 comto be granted nonregulated status, APHIS must determine that this GE comvarietyis unlikelyto pose a plantpest risk. The analysis byAPHIS in its plant pest risk assessment (USDAAPHIS, 2009) demonstrates that there were sufficient data to determine that the MIR162
comis unlikely to pose a plantpest risk and therefore is eligible for nonregulated status.
The regulations at 7 CFRpart 340.6(d)(3)(i) state thatAPHIS may "approve the petition in whole orin part." Because APHIS has found that the MIR162 comis unlikely to pose a plantpest risk, the only action alternative considered in this EA is to grant nonregulated status "in whole" to the com line under consideration. An "inpart" deregulation canbe given ifthere is a plantpest risk associated with some, but not all lines requested in a petition. Thepetition for the MIR162 com onlyrequested APHIS to grant nonregulated status to a single com event, therefore, an "inpart" determination is not an appropriate consideration. Thus, onlytwo alternatives will be considered inthis EA: (1) no action, or (2) to grant nonregulated status to MIR162 com "inwhole." APHIS has assessed the potential for environmental impacts for each alternative in the "Environmental Consequences" sections below.


### A. No Action: Continuation asaRegulatedArticle

Underthe "no action" alternative, APHIS would deny the petition. MIR162 com and its progeny would continue to be regulated under 7 CFRpart 340. Permits issued or notifications acknowledgedby APHIS would still berequired for introductions of MIR162 com and measures to ensure physical and reproductive confinementwould
11continue to be implemented. APHIS might choose this alternative ifthere were insufficient evidence to demonstrate the lack ofplant pest risk from the unconfined cultivation ofthe MIR162 com and its progeny.
Under this no action alternative, growers and otherparties who are involved in production, handling, processing or consumption ofcom would continue to have access to existing deregulated GE insect resistant com as well as conventional com varieties. However, growers would not have widespread access to the MIR162 com since it would continue to be regulated underPart 340. This alternative is not the preferred alternative because APHIS' evaluation ofMIR162 data inthe plant pestrisk assessment demonstrates that the MIR162 com is unlikely to pose a plant pest risk (USDA-APHIS, 2009). Choosing this alternative would hinder the purpose and need ofAPHIS to allow for the safe development and use ofGE organisms given that the MIR162 com is unlikely to pose aplantpest risk.


### B. Grant nonregulatedstatus to MIR162 corn, "in whole"- Preferred Alternative:Determination thatSyngenta MIR162 Corn isNo Longera RegulatedArticle

Under this alternative, MIR162 com and its progeny would no longerbe considered regulated articles under 7 CFRpart 340. Permits ornotifications acknowledged by APHIS would no longer berequired for introductions in the United States and its territories ofthe MIR162 com orits progeny. MIR162 com is eligible for nomegulated status because APHIS has determined that this GE organism is unlikelyto pose a plant pest risk (USDA-APHIS 2009). APHIS might choose this alternative ifthere was sufficient evidence to demonstrate the lack ofplant pest risk associated from the unconfined release ofthis insect resistant com event.
Under this alternative, growers mayhave future access to MIR162 com and progeny derived from this variety ifthe developer decides to commercialize this insect resistant
comvariety. Inaddition, growers and otherparties who are involved inproduction, handling, processing or consumption ofcomwould continue to have access to existing deregulated GE insect resistant com as well as conventional comvarieties. If commercialized, MIR162 comwill likelybe introduced in areas where comis currently grown and is not expected to alter the current range ofcom cultivation in the US.

APHIS has chosen Alternative B as the preferred alternative because APHIS has determined that MIR162 comis unlikely to pose a plantpest risk (USDA-APHIS 2009). By granting nomegulated status to MIR162 com, the purpose and need to allow the safe development and use ofGE organisms is met. 

### Alternatives Considered but Rejected from Further Consideration
Geographic restrictions APHIS considered geographic restrictions based upon geographic variation in plant pest risk. As presented in APHIS plant pest risk assessment for MIR162 com, there is no 12geographic differences in the plantpest risks for MIR162 com (USDA-APHIS 2009). This alternative was rejected and not analyzed in detail because MIR162 com is unlikely to pose aplant pest risk and therefore, APHIS will have no regulatory authority over MIR162 com and will be unable to impose regulatory restrictions on this GE com variety.



## VI. Environmental Consequences

According to APHIS regulations at 7 CFRpart 340, an organism is no longer subject to regulatoryrequirements when it is demonstrated not to present a plant pest risk. Under the regulations, APHIS is required to render a determination on a petition for nonregulated status. The analysis ofpotential environmental consequences in the following sections address the potential impact to the human environment from the alternatives analyzed in this EA, namely taking no action and granting nonregulated status to MIR162 com, "in whole."

SCOPE OFTHE ENVIRONMENTAL ANALYSIS Although the preferred alternative would allow for new plantings ofMIR162 com to
occur anywhere in the U.S., APHIS limited the environmental analysis to those areas that currently support com production. To determine areas ofcomproduction, APHIS used data from the National Agricultural Statistics Service (NASS) 2009 Census of Agriculture to determine where comis produced in the United States (USDA-NASS 2009), accessed 3/5/2010). According to the 2007 Censuses ofAgriculture, 49 states produce com grain in the US.



### A. NoAction

Underthe "no action" alternative, MIR162 comhybrids would continue to be a regulated article. APHIS' assessment ofenvironmental consequences underthe no action alternative is describedbelow.

A-I. Corn

Underthe 'no action' alternative, conventional and GE transgenic comhybrids crop husbandry will remain unchanged and MIR162 com hybrids will remain a regulated article.

The food/feed nutritional and safety assessment for the MIR162 com has been reviewed bythe FDA. Under Federal Food, Drug, and Cosmetic Act (FFDCA), it is the responsibility offood and feed manufacturers to ensure that the products they market are safe and properly labeled. Food and feed derived from the MIR162 commust be in
13compliance with all applicable legal and regulatory requirements. FDA completed their consultation on MIR162 on December 9,2008 and concluded that it had "no further questions concerning grain and forage derived from com event MIR162" (FDA BNF No. 000113).

APHIS' assessment ofthe safety ofthis product focuses on its potential to pose a plant pest risk, and that analysis isbased on the comparison ofthe GE com to its non-GE counterpart (USDA-APHIS 2009). Based on the assessment offield and laboratory evidence provided in Syngenta's petition, accompanying scientific literature and safety data available on earlier insect-resistant GE comhybrids, APHIS has concluded that MIR162 comwould have no significant impacts on human or animal health.

A-2. AgriculturalProduction of Corn

Conventional and GE comproduction occurs on land that is dedicated to crop production. Most com is planted in agricultural fields that have been in crop production for years. Most ofthe com acreage in the U.S. is planted to GE comhybrids. Ofthe total
comacres planted in 2008, 85% were GE comhybrids that were eitherherbicide tolerant, insect resistant, orboth (USDA-ERS 2009). Likewise, according to USDA-ERS latest data on organic comproduction, in 2005 less than 1percent (0.16%) ofcomcrop area was devoted to organic com (USDA-ERS. 2009c).

Conventional production practices that use GE varieties will likely still dominate in terms ofacreage, orperhaps increase in acreage (Fernandez-Cornejo Caswell 2006; APHIS received four new petitions for non regulated status in 2008/2009 for com varieties), without granting nonregulated status to MIR162 comunder the "no action" alternative. The availability ofconventional, GE and organic com seed varieties will likelyremain the same under the "no action" alternative, including MIR162 comhybrids remaining unavailable for commercial use. Com is currently produced in 49 US states (USDA­ NASS 2008), and under the "no action" alternative, based upon current com production practices and available information and trend dataprovide by USDA-ERA (2009 a-c) and USDA- NASS (2008), it isreasonable to expect this range ofproduction wi11likely remain unchanged.

Yield losses due to weeds and diseases were substantial until the introduction ofcrop protection chemicals in the 1960s (Perrin, 1997; Giannessi, 2008). Weeds compete with crops for light, nutrients, water, and other growth factors. The large-scale commercial cultivation ofGEherbicide tolerant (glyphosate tolerant) com crop acreage has steadily increased from 1996 accounting for nearly 68 percent ofall com acreage in 2009 (USDA-ERS 2009). Glyphosate is a highly effective, nonselective, broad-spectrum herbicide and in general, considered "environmentally friendly" when compared to other herbicides (Cerdeira and Duke 2006). Herbicides were applied to 97 percent ofall com acreage, GE and non-GE in 2005, with atrazine, glyphosate, S-metolachlor and acetochlorbeing applied to 66%, 31%, 23% and 23% ofplanted com acres, respectively (USDA-NASS 2006). In addition, com crops are also susceptible to attack by a variety ofinsects from the time ofplantinguntil consumed as food or feed. Based on USDA

14survey data, adoption ofgenetically engineered insect-resistant comincreased from zero percent oftheU.S. com acreage in 1996 to 63 percent in 2009 (USDA-ERS 2009). Conventional insect control practices (chemical and microbial insecticides, crop rotation etc.) are also available to control com insect pest damage to GE and non-GE com varieties.

Underthe "no action" alternative, herbicides and insecticides will still be used alone orin combination and selected based on their effectiveness on the different weed and insect species in the cornfield. Human and environmental exposure to insecticides and herbicides will continue to occur. Different herbicides have different modes ofaction; the correct herbicide rate mustbe used for each in order to obtain good weed control results and to minimize complant injury. APHIS has no authority under the Plant Protection Act to regulate pesticide (herbicide orinsecticide) use. The use ofpesticides is regulatedbythe Federal Insecticide, Fungicide, and Rodenticide Act (FIFRA) restrictions administered bythe EPA, which mandate registration for use ofall pesticides. EPA includes instructions and restrictions onhow pesticides can be applied, and has determined that there is no unreasonable environmental risk ifthe user adheres to the directions. Directions include application restrictions that minimize impacts on nearby environments. Violators ofthe regulations are liable for all negative consequences of their actions; therefore, farmers who use pesticides are verylikelyto follow its label restrictions, and thereby limiting any potential adverse impacts.

IfAPHIS chooses the no action alternative there wouldbe no direct impact on organic or othernon-transgenic com farmers. The current cultivationpractices are not likelyto change and 85% ofthe comproduced would likely continue to be plantedwith current GE comvarieties (USDA-ERS 2009b).

A-3. Corn Lepidopteran Pests

Comis susceptible to damage by a variety ofinsect pests throughout its developmental cycle. Cominsectpests are categorized as major and consistent pests, major and sporadic pests, and moderate to minorpests based on annual destructiveness and their geographic distribution (pg. 12 in Syngenta 2007). Yield losses due to insect pests are unpredictable and challenging for conventional (non-GE) com farmers and insectpest problems have the potential to substantially reduce crop yield and quality. Crop losses attributable to the European comborer (Martin and Hyde 2001) and comrootworm infestations (Ma et aL 2009) have been well characterized and are significant.

The introduction ofGE Bt com cultivars which encode proteins (Cryproteins from B. thuringiensis) that are toxic to these species have providedU.S. com growers with a powerful tool for effectivelyprotecting crop yields and environmental benefits (Marvier et al. 2007). The large-scale commercial cultivation ofboth insect resistant (Bt resistant) andherbicide tolerant (glyphosate tolerant) com crop acreage has steadily increased from 1996, accounting for 85% ofcom acreage in 2009 (USAD-ERS 2009). Growers have substantially switchedto GE comhybrids because they protect the inherent yield potential ofcom crops byreducing the grower's input costs. Underthe "no action"



### 15alternative, GE Bt com varieties and EPA approved insecticides win remain available for
use based on the need and effectiveness against different insect species infesting commercial cornfields.
Furthermore, the planting ofinsect-protected com hybrids benefits the environmentby decreasing the use ofconventional pesticide applications bymore than 20millionpounds annually (Figure 8 in Fernandez-Cornejo and Caswell 2006; Benbrook 2004). APHISderegulated GE Bt comvarieties are expected to remain available for commercial use. Therefore, this environmental benefit is expected to continue under the no action alternative.


### B. PreferredAlternative

Underthis alternative, MIR162 com would no longerbe a regulated article under 7 CFR part 340. Permits issued and/ornotifications acknowledgedbyAPHIS would no longer berequired for introductions ofMIR162 com. APHIS has chosenthe preferred alternative for the proposed actionbecause MIR162 com lacks plantpest characteristics,
as determined in APHIS' Plant PestRisk Assessment (USDA-APHIS 2009). APHIS' assessment ofenvironmental consequences under thepreferred alternative is described below.

B-1. Corn

Underthis alternative, conventional and GE transgenic com hybrid cropproduction will remain unchanged and MIR162 com wouldbe available to growers for commercial production. Similarto anycommercially-available com hybrid, a potential impact of planting this GE insect resistant com hybrid maybe gene introgression ofMIR162 com into other sexually compatible orrelated species. APHIS evaluated thepotential for gene introgression ofMIR162 com to sexually compatible wild relatives and considered whether such introgression would result inincreased weediness in wildrelatives. APHIS assessed various morphological and agronomic traits, such as seed dormancy, vegetative and reproductive traits, volunteerpotential, disease and pest susceptibility, and the fitness advantage ofVip3Aa20 gene ofMIR162 com (USDA-APHIS 2009). Based onthe scientific analysis ofdatapresented in APHIS' plantpest risk assessment, APHIS has determined that the MIR162 com is no more likelyto become a weed than other cultivated com varieties; it is not a plantpest; and gene flow between the MIR162 com and weedy and wild relatives will not occur inthe United States (USDA-APHIS 2009). Based on the above considerations, APHIS decision to grant nonregulated status to the MIR162 comwill not adversely impact sexuallycompatible wildrelatives ortheir weediness potential. Overall impacts wouldbe similarto the no-action alternative.

A food and feed nutritional and safety assessment ofthe MIR162 com has been completed bythe FDA. Under Federal Food, Drug, and Cosmetic Act (FFDCA), it is the responsibility offood and feed manufacturers to ensure thattheproducts theymarket are safe and properly labeled. Food and feed derived from the MIR162 com mustbe in compliance with all applicable legal and regulatory requirements. FDA completedtheir

16consultation on MIR162 on December 9,2008 and concluded that it had "no further questions concerning grain and forage derived from com event MIR162" (FDA BNF No. 000113).

APHIS focused on the potential ofMIR162 comto pose a plant pest risk, and compared the GE com to its non-GE counterpart (USDA-APHIS 2009). From the assessment of laboratory evidence provided in Syngenta'spetition and the accompanying scientific literature, APHIS has concluded that MIR162 comwould have no significant impacts on human oranimal health. Overall impacts would be similarto the no action alternative.

B-2. AgriculturalProduction of Corn

In 2009, GE insect-resistant com varieties represented 63 percent ofthe com acreage in the US (USDA-ERS 2009). Ifderegulated byAPHIS, MIR162 com would be an additional GE insect resistant com variety available to growers for commercial production. This comvariety will likelybe introduced to areas where com is currently grown as a replacement product for other varieties (conventional and GE) already available in the market. For example, fall armyworm damage can be suppressed using the Bt-expressing variety Herculex, but other Bt varieties such as YieldGard and KnockoutlNatureGard have less activity. However, when armyworm infestations are high all three varieties mayrequire supplemental insecticidal treatment (Bessin 2004). The proposed MIR162 variety (including varieties stacked to include the trait) would provide growers an alternative to these GE insect resistant com lines iffall armyworm damage is expected to be high.

Similarto the no action alternative, com will continue to beproduced in 49 states (USDA-NASS 2008) and the range ofcomproduction wi11likely be unchanged as a result ofAPHIS' deregulating MIR162 com. MIR162 com does not enhance any other agronomic traits, besides insect resistance. Insects targeted byMIR 162 are commonly found in areas currently under com cultivation (University ofIllinois Extension, 2004; Penn State University, 2010) and the pest insects that MIR162 controls are not a limiting factor that prevents com from being produced in any specific US location. Therefore, control ofthe susceptible pest insects will not open any new areas to com cultivation. Syngentahas field tested MIR162 com since 1999 underpermits issued ornotifications acknowledged byAPHIS across 20 representative com growing areas (pg. 127 in Syngenta 2007). The majority ofagronomic data were collected during the 2005 and 2006 growing seasons across 6-10 locations representative ofthe major com-growing
areas ofthe upper mid-west U.S. Except for test weight, grain moisture at maturity and plant emergence, the traits ofMIR162-derived hybrids were not statistically significant compared with their control (non-GE) counterparts (Tables 22 & 23, pg. 65-66 in Syngenta 2007). APHIS also assessed whetherthe MIR162 com is any more likelyto become a weed thanthe isogenic nontransgenic com line, or other comvarieties currently under cultivation (USDA-APHIS 2009). APHIS thoroughly considered the basic biology ofcom and evaluated the unique characteristics ofthe MIR162 comunder field conditions (USDA-APHIS 2009). Based on the agronomic field data and a literature

17survey ofcom's weediness potential, APHIS concluded that MIR162 com lacks abilityto persist as a troublesome weed (USDA-APHIS 2009).

The main introduced trait in MIR162 is expected to providepest resistance to certain insects. Therefore, except for change in insect pest management practices, all other agricultural practices for MIR162 com, including conventional com weed control practices and herbicide use, are not expectedto be different from those for conventional
com cultivation. These impacts, including the use ofEPA registered herbicides would be similarto the no action alternative. The use ofMIR162 com should reduce human and environmental exposure to insecticides used for insect pest control in other, existing com varieties.

B-3. PotentialImpacts of LineMIRJ62 Corn on Insect ControlPractices

Underthis alternative, in addition to MIR162 com, insect control options including the
use ofconventional insecticide applications, microbial insecticide applications, crop rotation, and planting ofGE insect resistant cultivars will remain available to com growers. Before the introduction ofGE comvarieties, com growers had difficulty controlling European comborer, which caused up to $1 billion ofannual economic loss in the U.S. including costs ofpesticide treatment and lost yield (Martin and Hyde, 2001). The introduction ofthe first GE Bt comhybrids in 1996 provided growers with an effective means oflimiting damage caused byEuropean comborer. GE Bt com use (both Bt only and stacked) grew from zero percent ofcom acreage in 1996 to 63 percent in 2009 (USDA-ERS 2009). These GE comhybrids express either a cryJAh orcryJF gene from B. thuringiensis, which encode proteins that are highlytoxic to European com borer and cry3BhJ orcry34AhJ, and cry35AhJ that are toxic to comrootworm. Based on the effectiveness ofcurrently available GE Bt comvarieties (Marvier et al. 2007), it is reasonable to assume that farmers usingMIR162 will observe similarpositivebenefits. Controlling above-ground insects presents a challenge for com growers, as manypests
are shielded from aerial chemical applications ortreatment may notbe economically feasible. As a result, the majority ofcom fields are not treated with pesticides for leaf-, stalk-, and ear-feeding insects. MIR162 comhas thepotential to control above-ground insectpests including com earworm, black cutworm, western bean cutworm, and fall armyworm that are not controlled by some Bt com varieties expressing Cryproteins. Doane Marketing Research AgroTrak studies (Doane Marketing Research 2006) indicate that growers in 2005 and 2006 were treating approximatelythree million acres a year with conventional insecticides for control ofthese insects with an estimated grower cost of20to 23 million dollars (Table 32, pg. 94 inpetition). Compared to the total number ofcom acres planted annually inthe U.S. (86 million acres in 2008), this represents a relatively small use ofconventional pesticides «3.5% ofthe total com acreage).

In additionto direct damage caused byfeeding on plant tissue, insects play an important role inthe transmission and dissemination ofpathogenic organisms during com development. FeedingbyDiabrotica rootworms has been associated with increased frequencies ofFusarium fungal infection (Dicke and Guthrie 1988), and rootworm

18feeding may also lead to increased incidences ofstalk rots. Likewise, com earworm feeding is associated with Fusarium infestation (Smeltzer, 1949). Ear, kernel, and cob rots occur wherever com is grown and result in reduced test weight, poor grain quality, and mycotoxin contamination offood and feed. Fusarium kernel or ear rot is the most widespread disease ofcom ears and is frequently associated with insect feeding damage. These pathogenic infections can lead to reduced crop quality, ability to harvest, and yield. Mycotoxin contamination ofcom grain presents a potential threat to livestock health and it has been found worldwide in animal feed (Placinta et ai. 1999; Monbaliu et ai. 2010). Since some current Bt varieties with partial resistance to com earworm are partially successful in reducing fumonisin content (Clements et aI., 2003), the availability of MIR162 will provide farmers with an additional management option that will likelyhelp farmers increase their capacity to alleviate adverse affects ofmycotoxins on crops and animals.

Under this alternative, EPA approved insecticides will remain available for use based on the need and effectiveness against different insect species infesting commercial cornfields. However, growers mayhave only a narrow time window during which insecticides can be applied to com crops. For example, optimal insecticidal application for com earworm infestation is the period of 1-3 days during 90-100% silking stage (Burkness et ai. 2009). After earworms enter into enclosed parts ofthe com ear, earworms are shielded from contact with the insecticides rendering them ineffective. MIR162 comprovides excellent protection against feeding damage caused by com earworm, black cutworm, western bean cutworm, and fall armyworm without the application limitations associated with the use ofinsecticides. Forthis reason, the introduction ofMIR162 com will have a positive impact on current com insect control practices. This product has the potential to displace many conventional insecticide applications on com (see pg. 92-96 in Sygenta 2007) resulting in a reduction in the number ofpounds ofinsecticides that maybe used to protect com from insect damage.

B-4. Organic and OtherNon-transgenic Corn Production

The National Organic Program (NOP) is administered byUSDA's Agricultural Marketing Service (AMS). Organic farming operations as described bythe National Organic Program requires 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 bytheir 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 ofexcluded methods. Excluded methods include a variety ofmethods used to geneticallymodify organisms orinfluence their growth and development bymeans that
are notpossible under natural conditions orprocesses. Organic certification involves oversight by an accredited certifying agent ofthe materials and practices used to produce orhandle an organic agricultural product. This oversight includes an annual review ofthe certified operation's organic system plan and on-site

19inspections ofthe certified operation and its records. Although the National Organic Standards prohibit the use ofexcluded methods, they do not require testing ofinputs or products for the presence ofexcluded methods. The presence ofa detectable residue ofa product ofexcluded methods alone does not necessarily constitute a violation ofthe National Organic Standards (USDA-AMS 2007). The unintentional presence ofthe products ofexcluded methods will not affect the status ofan organic product or operation when the operation has not used excluded methods and has taken reasonable steps (such
as isolation zones, use ofbufferrows surrounding the organic crops or adjustingplanting dates and appropriate cleaning ofplanting and harvesting equipment) to avoid contact with the products ofexcluded methods as detailed in their approved organic system plan. Organic certification ofa production orhandling operation is aprocess claim, not a product claim.

Itisnot likely that organic farmers, orother farmers who choose not to plant transgenic varieties or sell transgenic grain will be significantly impacted bythe commercial use of MIR162 com. Nontransgenic comwill likely stillbe sold and will bereadily available to those who wish to plant it. Despite the introduction and adoption oftransgenic com cultivars overthepast decade, includingmultiple varieties ofBtcom, non-GE specialty and organic comremain readily available. In 2006, there were at least 18 seed companies in the U.S. specializing in organic com seed (see pg. 110-111 in Syngenta 2007). Organic and otherfarmers have expressed concern that the widespread planting ofBt
complants will hasten the development ofpestresistance to pesticidal Bt endotoxins. Farmers purchasing seed will know this product is transgenic because it will be marketed
as Vip3aA20 lepidopteran resistant; and based on the EPA insect resistance management (IRM)2 policy (BPPD-EPA 2001), farmers will be educatedbythe Syngenta's stewardship plan about recommended management practices on MIR162 com cultivation. Transgenic com lines resistant to lepidopteran insects, and/or tolerant to specific herbicides are already in widespread use byfarmers. This particularproduct should not present new and different issues than existing insect resistant Bt com cultivars with respect to impacts onorganic farmers.

APHIS recognizes that comis open-pollinating and 

### it is possible that the engineered genes could move viawind-blown pollen to an adjacent field. All com, whether genetically engineered ornot, can transmit pollen to nearby cornfields. However, an influx ofpollen originating from a given com variety may not appreciably change the characteristics ofcomin adjacent fields because gene flow declines as a power of**1I~** and other factors such as wind speed, host variety and temperature also affect the results (Aylor et aI., 2003; Jones and Brooks, 1950). For example, assessing observations ofa large number ofcommercial canola fields, incidence oftransgene flow was on the order

2 Insect resistance management (IRM) is the term used to describe practices aimed at reducing the potential for insect pests to become resistant to a pesticide. Specific IRM strategies, such as the high dose/structured refuge strategy, developed by EPA are expected to mitigate insect resistance to specific Bt proteins produced in corn 20of0.015 % at 500m (see Fig 2 in Rieger, 2002) and in a smallercom study, 0.05% at 100m (Goggia et aI., 2006).

Methods ofspatial and temporal isolation are widely used and accepted when seed producers are seeking to minimize the influx ofpollen from sources outside the seed production field. To maintain varietal purity, the AOSCA (Association ofOfficial Seed Certifying Agencies) recommends 200 meters isolation for nearby compopulations to produce foundation class ofcertified seed (AOSCA 2003). These methods are readily applicable to the production ofcertified organic com seed. Gene flow rate between com populations is extremely variable depending onthe spatial, temporal, genetic and environmental factors (Brookes and Barfoot et al. 2004; Messegue et ai. 2006). Yet, available experimental evidence indicates that gene flow rates drop substantially (1%) beyond 20 meters (Henry et al. 2003; Maet al. 2004; Messeguer et al. 2006).

Dataprovided in the petition from agronomic trials conducted in 2005 and 2006 in a variety oflocations in the U.S. demonstrated that the MIR162 comis not significantly different inyield from its nontransgenic counterpart (Tables 22 and 23, pg. 65-66 in Syngenta 2007), and the MIR162 comhybrids were not significantly different from control lines (non-GE) in terms ofpollen viability, morphology, and diameter (Table 24, pg. 67 in Syngenta 2007). Therefore, MIR162 comhybrids are not expected to have an increased abilityto cross-pollinate other comvarieties when compared to conventional varieties that are currently available for commercialplanting.

IfSyngentareceives regulatory approval from all appropriate agencies, it will likely make MIR162 com available to growers and breeders. Itis not likelythat otherfarmers who choose not to plant or sell MIR162 com,such as organic producers, orthat other transgenic com varieties will be significantly impactedbythe expected commercial use ofthisproduct as (a) non-transgenic comvarieties will likely still be sold and will be readily available to those who wish to plant them; (b) Syngenta's stewardship planwill provide farmers thatpurchase MIR162 comrecommended management practices for MIR162 com cultivation; (c) methods ofspatial and temporal isolation are widely used and accepted and com seed producers employing them can minimize the influx ofpollen from sources outside the seed production field; (d) 85% ofthe 2008 com acreage in the United States is alreadyplanted to transgenic herbicide tolerant and/or insect resistant varieties; and (e) APHIS expects that MIR162 mayreplace some ofthepresently available GE comvarieties without significantly affecting the overall total com acreage. APHIS concludes that organic farmers will be able to coexistwith biotech comproducers
as they do now.
B-5. PotentialImpact on Non-target Organisms, IncludingBeneficial Organisms and Threatened orEndangered Species

APHIS evaluated the potential for MIR162 complants and theirproducts to have damaging ortoxic effects directly or indirectly on non-target organisms (USDA-APHIS 2009). Non-target organisms considered were those representative ofthe agricultural environment, including those that are recognized as beneficial to agriculture (Table 31,

21pg. 89 in Syngenta 2007) oras threatened orendangered in the U.S. APHIS also considered potential impacts on other "non-target" pests, since such impacts could potentially change agricultural practices. The technical details ofthe experiment onnontarget organisms havebeendescribed inthe Plant Pest Risk Assessment ofMIR162 com (USDA-APHIS 2009; see also pg. 85-88 in petition for details).

Different types (variants) ofVip proteins occur in nature, and three variants ofVip protein (Vip3Aal, Vip3Aal9, Vip3Aa20) are used for the nontarget impact investigations. The three protein variants (Vip3Aal, Vip3Aal9, Vip3Aa20) differ from each other by 1-2 amino acids (Table 12, pg. 48 in Syngenta 2007), and all three are found bebiochemically and functionally equivalent (see Table 13 and explanation thereof
onpg. 50 in Syngenta2007). The Vip3Aal9 variant is present in Syngenta's deregulated cotton event COT102. Likewise, the Vip3Aa19 variant was also present in com cultivar Pachamaize. Syngenta discontinued Pachamaize due to agronomic performance reasons and replaced its commercial development byMIR162 maize. Therefore each one ofthe nontarget exposure investigation, detailed in the following paragraphs, was carried out using one ofthe three Vip variant proteins (Table 30, pg. 84 in Syngenta 2007).

Potential impacts ofVip3Aa on higheranimals. In thebird (bobwhite quail, Colinus virginianus), mammal (mouse, Mus musculus), and honeybee (Apis mellifera) study conducted by Syngenta, there were no observable adverse effects ordifferences in survival noted at doses ofVip3Aproteins that were well above those expected from exposure to the Vip3Aa20protein from the MIR162 complanted in the field (Table 31, pg. 89 in Syngenta2007 ).
Potential impacts ofVip3Aa on above-groundarthropods. Adult pink spotted ladybird beetle (Coleomegilla maculata), seven-spot ladybird beetle (Coccinella septempunctata), second-instarminutepirate bug (Orius insidiosus), adult green lacewings (Chrysoperla carnea), and two- to three-day old C. carnea larvae were exposed to Vip3Aal9protein. The difference in survival ofthe beetles inthe treatment and control groups was not statistically significant (seepg. 86 in Syngenta 2007). Although not an endangered orthreatened species, Danausplexippus (monarch butterfly) is a species ofhigh conservation interest, and there has been concern that itmaybe harmedbyconsumingpollen from transgenic insect-protected com. Studies ofa similar Bt protein, Cry lAb, have shown that exposure ofMonarch butterfly larvae to this protein in the vicinity ofcom fields is nothigh enough to cause mortality (Anderson et aL 2004). However, Vip3A has no toxicity to monarch butterflies (Lee et al. 2003). APHIS concludes thatbecause ofapparent tolerance to the toxin (D. plexippus) and limited actual exposure, the larval monarch butterflies are not likely to be impacted bythe cultivation ofthis Vip3Aa20-expressing crop. Effects ofaVip3A X CrylAb Bt com variety on non target arthropods were assessed byDively (2005) and compared to an isogenic control line. Populations ofmost insect taxa in both varieties were not significantly different, although some increases ordecreases were caused byprey density responses, the absence ofplant injury or some plantbased factor, ratherthan toxicity of the host plant.

22Potential impacts on threatenedandendangeredarthropods. With respect to the Endangered Species Act and oversight bythe U.S. Fish and Wildlife Service (FWS), APHIS has established an agreement with FWS specifyinghow APHIS should review dataprovided byapplicants. The process includes a framework to detennine ifa FWS consultation maybe required under section 7 ofthe Endangered Species Act, before APHIS may grant nonregulated status to products such as MIR162 corn. APHIS has obtained and reviewed the list offederally listed threatened and endangered species, species proposed for listing, and designated critical habitat orhabitat proposed for designation as part ofthe review process. Based upon this review and adherence to the established process identified in the FWS agreement, APHIS has concluded that a consultation with FWS was not necessary for MIR162 corn.
Given the narrow specificity ofthe Vip3Aa20 activity, species outside the insect order Lepidoptera are not expected to be affected byVip3Aa20 protein toxicity. Its receptormediated mechanism ofaction and absence ofactivity in bioassays with multiple species outside ofthe orderLepidoptera, as discussed in preceeding paragraphs, supportthis conclusion. Furthennore, Syngenta observed no hannful effects ofVip3Aaproteins in nontarget organism hazard identification studies that used a wide range oftaxa at expected environmental concentrations. The test results indicated a lack ofrisk associated with exposure to Vip3Aa20 in the MIR162 corn (Table 31, pg. 89-90 in Syngenta2007).

The Vip3Aa20 protein is selectivelytoxic to a few species ofinsect pests belonging to the order Lepidoptera. The onlythreatened or endangered lepidopteran species with potential for exposure to insecticidal proteins in corn is the Karnerblue butterfly (Lycaeides melissa samuelis) (EPA 2001; USFWS 2007). The Karner bluebutterfly requires wild lupine (Lupinusperennis) as an oviposition substrate and larval food source, while the adults feed on wild flowers. The potential route ofexposure is consumption ofmaize pollen that has settled on the leaves ofits food plant, the wild lupine (Lupinusperennis). Karnerblue butterfly is known to exist along the northern extent ofthe range ofwild lupine, where there are prolonged periods ofwinter snowpack, in parts ofWisconsin, Michigan, Minnesota, Indiana, New Hampshire, New York, and Illinois (Haack 1993). Although there are two counties in Wisconsin that have been identified as having apotential overlap between cornpollen shed and the presence of Karnerblue larvae (peterson et al. 2006 ), there is no evidence ofKarnerbluebutterfly exposure to corn pollen in these locations. According to Peterson et al. (2006) the exposure ofthe Karner blue butterflyto maize pollen was minimal in all other locations because most lupine popUlations are separated from maize fields byat least 500metres, and because maize anthesis (flowering) usually occurs after the Karnerblue larvae have finished feeding.

APHIS coordinates review ofpetitions with other agencies that have regulatory oversight for these same products. With respect to threatened and endangered species, EPA also plays a specific role inthe evaluation ofGE insecticideresistant crops, including MIR162
corn. The Endangered Species Protection Program (ESPP) ofEPA places geographically specific use limitations on pesticides in orderto protect threatened and endangered

23species from pesticides (EPA 2009). Anyuse restrictions are identified on the EPA approved label associated with a specific EPA-registered product.

The EPA on August 6, 2008 approved a tolerance for Vip3Aa20 in comor cotton, and on February 13,2009 conditionally approved the use ofVip3Aa20 for use with two otherBt proteins (EPA 2009a, 2009b). In these assessments, EPA stated that the agency was not
aware ofany adverse effects ofVip3Aaproteins on the abundance ofnon-target beneficial systems in anypopulation in a field environment, inclusive ofpest parasites, pest predators orpollinators. EPA also concluded that the hybrid will produce no unreasonableharm to the environment orany federally listed threatened and endangered species. Although refuges were required for commercial use ofthe Vip3Aaprotein, EPA did not specify any other geographic limits for use in production agriculture. EPA also concluded that the use ofthe pesticidal protein contained inMIR162 corn will not cause any unreasonable adverse effects onthe environment during the time ofconditional registration (EPA 2009a,b). APHIS concurs withthe EPA assessment (EPA 2009) that the proteinproduced bythis variety will have no adverse effects in the environment.

As previously discussed, APHIS concludes that MIRI62 will not adversely impact arthropods which maybe exposed to the hostplant. APHIS concludes that for other endangered orthreatened animals orplant species inthe U.S., MIRI62 com is not expected to have any harmful effects because toxicity ofVip3Aa20isrestricted to Lepidoptera. Based upon the information available in the literature, on the EPA assessment and on the data supplied by Syngenta, APHIS determined that the unconfined release following deregulation ofMIRI62 comwould have no effect on federally listed threatened and endangered species, species proposed for listing, orondesignated critical habitat orhabitat proposed for designation (USDA-APHIS 2009).
Environmental fate in soilandeffects on soildwelling organisms. The purpose ofa Syngenta (2007) soil fate studywas to test the inherent degradability ofVip3Aa20 in a soil typical ofcom-growing areas with healthy microbial activity. Most proteins do not persist oraccumulate in soil because they are inherently degradable in soils (Bums 1982; Marx et al. 2005). Multiple investigations have demonstrated that Bt Cryproteins are rapidly degraded in a variety ofsoil types and that the proteins do not accumulate (EPA 2001; Head et al. 2002; Dubelman et al. 2005). Vip proteins are similarto Cryproteins in that they are also found in naturally occurring soil bacteria and commercial microbial insecticides (de Maagd et at. 2001). Under the activity ofsoil proteases, the Vip3Aa protein would be expected to be susceptible to degradation, unless specificallyprotected from such enzymatic hydrolysis (Marx et at 2005) Syngenta (2007) conducted a laboratory study to determine the degradability of Vip3Aal9protein (Vip3Aal9 and Vip3Aa20 are functionally equivalent, see earliertext) in five soils (clay, sandy clay, loam, sandy loam, silt loam). A rapid decline inthe levels ofVip3Aa19 was observed in all soil types, wherein degradation was measured as loss of insecticidal activity. The time to 50% dissipation (DT50) was estimated to bebetween 6.0 and 12.6 days across soil types and test concentrations. The results ofthis study showed that Vip3Aaprotein is rapidly degradable innormal soils. Thus, the Syngenta soil study data (Syngenta 2007) indicate that that there will not be a significant environmental
24impact on the soil environmentby granting nonregulated status to MIR162 due to limited persistence ofVip3Aa20 in the soil.

Residual complant debris is oftentimes incorporated into the soil after harvest, and Syngenta (2007) assessed impacts ofthe MIR162 comorits Vip3Aaprotein on representative soil dwelling organisms. Data onthe effects ofVip3Aaprotein on earthworms, Eisenia foetida; a collembolan, Folsomia candida; and rove beetles, Aleochara bilineatwere analyzed (see p. 87 in Syngenta 2007). These were variously incorporated as isolated protein orlyophilized leafmaterial as appropriate. Toxicity exposure ratios for these organisms was >9, >114 and>1316, respectively, and indicate a lack ofrisk for soil organisms (Syngenta 2007).

B-6. Cumulative Effects

APHIS considered whethertheproposed action could lead to significant cumulative impacts, when considered in light ofotherpast, present, and reasonably foreseeable future actions, regardless ofwhat agency orperson initiated such actions. APHIS has evaluated the potential cumulative impacts ofgranting nonregulated status to MIR162
com. As discussed in the following paragraphs, bothpetitioner's data and USDA-APHIS review do not indicate any cumulative impact on the environment as aresult ofAPHIS deregulation ofMIR 162 com (seepg. 104-108 in Syngenta 2007).

Corn Production GE insect resistant com acreage has been steadilyincreasing from 0% to 63% overthe last 13 years in the US (USDA-ERS 2009). MIR162 is not the first Bt comproduct to be granted nonregulated status. APHIS has previously made determinations ofnonregulated status for several other Btcom cultivars including petition numbers: 94-319-01p at http://www.aphis.usda.govIbrs/aphisdocs2/94 _31901P _corn.pdf; 95-093-01p at http://www.aphis.usda.govlbrs/aphisdocs2/95_09301p_com.pdf; 95-195-01p at http://www.aphis.usda.govlbrs/aphisdocs2/95_19501p_com.pdf; 96-291-0Ip at http://www.aphis.usda.govlbrs/aphisdocs2/96_2910Ip_com.pdf; 97-013-01p at http://www.aphis.usda.govlbrs/aphisdocs2/97 _0130Ip_com.pdf; 97-265-01p at http://www.aphis.usda.govlbrs/aphisdocs2/97_26501 p_com.pdf; 00-136-Olp at http://www.aphis.usda.govlbrs/aphisdocs2/00_13601p_com.pd; 01-137-0Ip at http://www.aphis.usda.govlbrs/aphisdocs2/0l_13701p_com.pdf; 03-l81-0Ip at http://www.aphis.usda.govlbrs/aphisdocs2/03_181Olp_com.pdf; 04-125-Olp at http://www.aphis.usda.govlbrs/aphisdocs2/04_12501p_com.pdf; 04-362-01p at http://www.aphis.usda.govlbrs/aphisdocs2/04_36201p_com.pdf; 06-298-01p at http://www.aphis.usda.govlbrs/aphisdocs2/06_29801p_com.pdf; According to Syngenta, the combined-traitBt11xMIR162 hybrids are efficacious against five major insectpests (European comborer, com earworm, fall armyworm, black cutworm, western bean cutworm) and such hybrids have the potential to provide growers the means ofprotecting their com crops from damage caused byabroaderrange of lepidopteran pests (USDA-APHIS 2009). Based upon the increasing trend in GE insect resistant com acreage and the number ofGE insect resistant com varieties that are

25commercially available to growers, it is reasonable to assume that the commercial availability, acceptance and use ofGE insect resistant com crops will continue into the foreseeable future with orwithout APHIS deregulation ofMIRl62 com. Comwill continue to be produced in 49 US states (USDA-NASS 2009) and the range ofcom production will likelybe unchanged as a result ofAPHIS deregulating MIR162 com. Non-transgenic com, including organic varieties will likely still be sold and will be readily available to those who wish to plant it. Despite the introduction and adoption of transgenic com cultivars overthe past decade, including multiple varieties ofBtcom, non-GE specialty and organic com remain readily available.

Geneticpurity of corn germplasm. APHIS does not foresee a cumulative impact on the genetic purity and diversity ofnon-GE com cultivars and germplasm from granting nonregulated status to MIR162 com (see pg. lOS in Syngenta 2007). Genetic purity and diversityhas been a feature ofcom improvement cultivation for decades as part ofhybrid seed and specialty comproduction. Many agricultural practices are effectively used by certified seed producers to grow and produce qualityhybrid seed, including maintaining isolation distances to preventpollen movement from othercom; planting borderor barrierrows to intercept pollen; employing natural barriers to pollen movement such as treelines, manual or mechanical detasseling; genetic male sterility; and staggered planting dates (see for example, Bradford 2006; MRSC 2009). These widespread management practices remain in use today and have served to ensure that the broad adoption of transgenic com in the U.S. (including the sale and cultivation ofmultiple Bt com varieties over more than a decade) has had no significant impact, even in the aggregate,
on the production ofcom seed and specialty comproducts. APHIS expects that certified growers ofMIR162 and future hybrid com seed producers will continue to follow similar types ofmanagementpractices to maintain genetic purity and diversity ofcom germplasm (see discussion in Syngenta, 2007 at IX C.7, Potential cumulative impacts.)

In general, all management practices used in conventional hybrid seed production to
ensure quality standards are also used for the production ofspecialty and organic com seed and typically are sufficient to meet assigned standards (see for example, USDAAMS 201Oa). In addition, certifiers oforganic standards assess farm plans and production methods, and make inspections to determine whether grower-proposed methods for organicallyproduced crops are accomplished and standards are maintained.

Genetic diversity of corn. APHIS does not foresee significant cumulative impacts on the genetic diversity oron the availability ofdiverse com germplasm resources because of the adoption ofmultiple varieties oftransgenic com.
Itis common practice in com seed production systems (GE and non-GE) to develop genetically distinct comhybrids for various geographies and purposes, and to continually improve these varieties through establishedplant breeding practices and techniques. A variety ofGE and non-GE com cultivars have been and wi11likely continue to remain available to those who wish to plant them. Despite the introduction and adoption of transgenic com cultivars overthe past decade, including multiple varieties ofBtcom, a variety ofnon-GE comincluding specialty and organic varieties remain readily available.

26In 2006, there were at least 18 seed companies in the U.S. specializing in organic com seed (see pg. 110-111 in Syngenta 2007).

In addition, the adoption ofgenetically engineered comwas preceded by a worldwide effort to identify and preserve sources ofcom genetic diversity, and to make these
resources available for utilization bypublic and private combreeders. Among these efforts are the Germplasm Enhancement ofMaize program ("GEM"), a cooperative effort undertakenby USDA, public and privateplant sectorbreeders, NGOs (Non Governmental Organizations) and international public cooperators. GEM was established to further identify com genetic diversity and to provide it in useful form in order to broaden the genetic base ofthis crop. Multiple Btcorn events and insect resistance developing in the field. APHIS does not foresee significant cumulative impacts resulting from multiple Bt com events and insect resistance developing in the field.

MIR162 comprovides no protection against feeding damage caused byEuropean com borer. Syngenta has indicated that they intend to commercialize MIR162 com as a combined-trait hybrid with Syngenta's Btl 1com event (an APHIS deregulated GE insect resistant comvariety that contains CrylAbBtprotein targeting European comborer) to control a variety oflepidopteran insect pests, such as European comborer, com earworm, black cutworm, western bean cutworm, and fall armyworm. According to Syngenta, the combined-trait hybrid (Bt1lxMIRI62) has a unique benefit. In this case, the combinedtrait containing CrylAb and Vip3Aa20 proteins have been demonstrated to provide highdose control ofEuropean comborer, com earworm, and fall armyworm. Becausethe type oftoxicity ofVip proteins to insects seems to differ from those ofCry proteins (Lee et aI, 2003), a strategy ofcombining such toxins would beeffective in reducingrisk of
cross- resistance developing in lepidopterous pests that are affected byboth (Tabashnik et 

### aI. 2009). Sygenta also intends to commercialize MIR162 as a combined traits hybrid with Btl 1 and MIR604, and increase its targeted insects to include comrootworm control (EPA 2009b) Syngenta has submitted an Insect Resistant Management (IRM) plan to the EPA for Bt1lxMIRI62 comthatrequires growers to plant a 20% structured refuge (see BBPDEPA 2001 for details) that can beplanted as strips within or surrounding the Btcom field
or as ablockwithin, adjacent to, or up to 0.5 mile away. The proposed refuge requirements are the same in the ComBelt and cotton growing areas (see BBPD-EPA 2001 for details). Use ofthese refuge strategies by growers have been shown to either delay orprevent the development ofcertain lepidopteran insect pest developing resistance to Bt (Bates et al. 2005). Although confirmed evidence ofinsect resistanceto Bt com has developed in the field in Puerto Rico (Spodopterajrugiperda to Cry 

### IF), and in South Africa (Buseolajusca to CrylAB) and to Bt cultivars ofcotton in the Southeast US, most pest populations remain susceptible to Bts (Tabashnik et al. 2009). Crossresistance is possible, as between CrylAc and Cry2Ab in one cotton pest, and should be considered in assessing effectiveness offuture strategies to avert development of resistance to Bt crops (Tabashnik et al. 2009a).

27Potential impacts on threatenedandendangeredspecies. APHIS does not foresee significant cumulative impacts on threatened and endangered species resulting from APHIS deregulating MIR162 com. Therestriction oftoxicity ofVip3Aa20 to Lepidoptera, and theminimal exposure ofengendered Lepidopterato com, indicates that planting ofthe MIR162 com is expected to have no harmful effects on any endangered or threatened species in the U.S (see lack ofeffects onwide range oftest organisms in Syngenta 2007).

Potential impacts on biodiversity. APHIS has determined that there are no past, present,
orreasonably foreseeable actions that would aggregate with effects ofthe proposed action to create cumulative impacts orreduce the long-term productivity or sustainability ofanyofthe resources associated with the ecosysteminwhich the MIR162 comis planted.

The importance ofcom as a food crop, andits dependence onhuman management, has produced a long history ofdiligence to protect germplasm lines ofcom (see Genetic purity ofcom germplasm section above). Decades priorto the introduction oftransgenic
comproducts, the com industrydeveloped effective methods and means to maintain product segmentation and genetic purity standards. Specialty corns, for example, were successfully isolated for years and continue tobe grown today, even with transgenic com widely adopted in the U.S. Moreover, with respect to bothconventional and transgenic com, the ability to protect and maintain the genetic purity ofbreeding lines is critical to seed companies and developers ofnew varieties such as MIR162. Consequently, seed companies routinely apply standard breeding techniques includingphysical and temporal isolation thathave proven effective at maintaining the genetic purity ofbreeding lines. Genetically engineered cornlines with Bttraits (both Cry and Vip proteins) have been available on the market since 1994 and the bodyofevidence in peer-reviewed literature does not suggest anynegative effect onbiodiversity. APHIS review and analysis of Syngenta's data (USDA-APHIS 2009) indicate that the line MIR162 com exhibits no traits that would cause increased weediness, that its unconfined cultivation should not lead to increased weediness ofothercultivated corn orother sexually compatible relatives. Consequently, MIR162 is not likely to harm non-target organisms commonto the agricultural ecosystem orthreatened or endangered species recognizedbythe U.S. Fish and Wildlife Service.

B-7. SocioeconomicAnalysis

The mainbody ofinformation on socioeconomic analysis described in the following paragraphs comes from Syngenta's socioeconomic analysis (see pg. 108-112 and Appendix 1 in Syngenta 2007); the Environmental Protection Agency's (EPA) Biopesticide Registration Action Document, Vip3Aa20 com (EPA 2009c); USDAEconomic Research Service' (USDA-ERS) report, "The First Decade ofGenetically Engineered Crops inthe United States" (Fernandez-Cornejo and Caswell 2006); and BioTech InfoNet Technical Paper, "Genetically Engineered Crops and Pesticide Use in

28the United States" (Benbrook 2004). APHIS assessment on the petitioner's socioeconomic analysis is restricted to the MIR162 com and its stacked hybrids with otherBt traits (BtllxMIRI62,BtllxMIR162xMIR604). AgriculturalBenefits. The increased adoption ofBtcom cultivars since its introduction in the mid 1990s (USDA-ERS 2009) could implythat insect resistance varietiesprovide benefits to com farmers. This is more evident since the early 2000's whenthe Btcom varieties that are effective against European corn borer were complemented with a second generation Bt com cultivar that provide protection against com root worm. According to USDA's Agricultural and Resource Management Surveys (ARMS) conducted in 2001-03 (see Figure 7 in Fernandez-Cornejo and Caswell 2006) most ofthe farmers adopting GE com, cotton, and soybeans indicated that they did so mainlyto increase yields through improvedpest controL Because the MIR162 com is effective against two majorcom insect pests, com earworm and western bean cutworm, which are not effectively controlled by earlier Bt com cultivars, it is reasonable to assume that likewise, potential benefits do exist for farmers adopting the MlR162 com cultivars. Although itis difficult at this time to accuratelypredict the magnitude ofeconomic benefits ofthe MIR162 com hybrids in the marketplace, because such hybrids are not currently in commercial production, the improved pestprotection profile ofBt11xMIR162 commay translate into correspondinglyhigheroverall economic benefits to growers, consumers, and other downstream users ofcom products. Syngenta has suggested that their stacked hybrids Btl 1 x MIR162 x MIR604 will provide unsurpassed control oftarget pests and that the product's "broad-lepidopteran control, particularly for com earworm and western bean cutworm, potentiallyresults inbetter performance than those ofcompetitors. However, EPA's Biopesticides and Pollution Prevention Division (BPPD) notes that these statements are unverified assumptions. Although the data supportthat the stack containing the Btl 1 , MIR162, and MIR604 traits produces reasonably good efficacy against western bean cutworm, the MIR604 trait when combined with the MIR162 trait showed some evidence ofa possible synergistic effect in the control ofcom rootworm. BPPDreasons that the sample size used by Syngenta for the investigation is too small to delineate impacts ofindividual Bt traits in stacked hybrids. According to BPPD, Syngenta's specific economic benefits are based onbestcase assumptions (i.e., quick and broad adoption ofthe product in the marketplace). Competition from previouslyregistered Bt comproducts (already established in the market) and grower familiarity with these products mayreduce the overall adoption, and the potential benefits for MIR162 com and its associated products. Despite this shortcominghowever, BPPD notes that both the stack and pyramid products, and the single-trait product appear to provide good protection against European com borer and
com earworm. Formany growers, the potential broad lepidopteran control expected by Bt11xMIR162 hybrids mayprovide added benefits than currently available Btcom cultivars, as com growers with multiple pestproblems are expected to be protected from major compests.
So far, Bthybrids have mixed responses when it comes to monetary benefits to com growers. In 2006, USDA-ERS published arep011 on "The First Decade ofGenetically

29Engineered Crops in the United States" focusing on GE crops and their adoption in the United States over the past 10 years (Fernandez-Cornejo and Caswell 2006). The economic analysis in that report mainly focused onthe field data from 1997 and 1998 (see Table 1). On average, BTtechnologybenefitted com farmers with 5% higher yields in the United States, and yield effects were larger inyears with highpestpressures as noted earlierbyother investigators (Carpenteret al. 2002). Many field tests and farm surveys have also examined the yield and cost effects ofusing Bt com crops (Table 1). The majority ofthe results show Bt com crops produce higher yields than conventional crops. A more recent ERS studyusing 2001 survey data found that, on average, actual
com yield was 12.5 bushels per acre higher for Bt comthan for conventional com, an increase of9% (Fernandez-Cornejo and Li 2005).

The economic benefits ofgrowing Btcom do not appear to be consistent across growing
seasons. There was a negative association between adoption ofBtcom and producer net returns in 1998. According to Fernandez-Cornejo and Caswell (2006) this negative trend suggests that Bt com mayhave been used on some acreage where the value ofprotection against the European comborerwas lowerthan the premium paid for the Bt seed. Becausepest infestations vary from one region to another and from one yearto another, the economic benefits ofBtcom are likelyto be greatest where pest pressures are most
severe (Carpenter et al. 2002; Shelton et al. 2002). Farmers must decide to use Btcom before theyknow what the European comborerpestpressure will be that year. Forthat
reason managementpractices aretailored accordingly (Mason et al. 1996). Because of this unpredictable variation, many farmers generally ignored European cornborer infestation and accepted the losses it caused (Shelton et al. 2002). According to Gurian-Sherman (2009) there was a 3-4 percent yield advantage for Bt com varieties in the U.S. for combining the benefits ofEuropean comborer and comroot
worm resistance. However, Gurian-Shel111an's analysis also showed that Btcorn yield benefits were not much different from what was achieved through traditional breeding. Forexample, com yield has been increasing on average 1percent peryear over the past several decades, and Bt com crops had the same yearly improvement in the last 14 years since the introduction offirst Bt crop in 1996. Several approaches, such as organic cultivation, wheat-com rotation etc, otherthan current pesticide regimes and GE have the potential to reduce yield loss from comborerand rootworm incom. These approaches also have other associated benefits such as lower levels ofpesticide use, improved soil qualities, increased carbon sequestration, and improved waterquality (but also see the comments on the report by Sheridan 2009). Adoption ofGE crops is associated with reduced pesticide use. Insecticide use on fields planted to 8tcom substantially decreased since its introduction in mid 1990s (Figure 1 and Figure 8 in Fernandez-Cornejo and Caswe112006) and use rates oncom (in tel111s of active ingredient) have declined since the introduction ofGE comin 1996. More recently, using 2001 data, USDA-ERS found that insecticide use was 8 percent lowerper planted acre for adopters ofBt com than for nonadopters (Fernandez-Cornejo and Li 2005). The USDA-ERS results generally agree with field-test and other farm surveys that have examined the effects ofusing GE crops (Table 1).

30Table I. Summary ofprimary studies onthe effects ofgenetically engineered Bt com on yields, pesticide use, and returns (Modified from Fernandez-Conejo and Caswe1l2006).

Reference Data Source Effects on (NA = Not analyzed in the study) Yield Pesticide Use Returns

Rice and Pilcher, 1998 Survey Increase Decrease Depends on infestation Marra et aI., 1998 Survey Increase Decrease Increase Benbrook, 2001 Survey Increase 

### NA Decrease MeBride & El-Osta, 2002 Survey 

### 'NA NA Decrease Duffy, 2001 Survey Increase 

### NA Same Pilcheret aI., 2002 Survey Increase Decrease 

### NA Baute, Sears, and Schaafsma, 2002 Experiments Increase 

### NA Depends on infestation Dillehay et aI., 2004 Experiments Increase 

### NA NA Fernandez-Cornejo & Li, 2005 Survey Increase Decrease 

### NA

The MIRI62 com and stacked hybrids may further reduce the insecticide use ifthe currenttrend in insecticide usage continues (Figure 8 in Fernandez-Cornejo and Caswell 2006). But according to Benbrook (2004) the insecticide reduction rate appears to have plateaued (Figure 1), and any further reduction in insecticide use from new Bt com cultivars maybe marginal. Forexample, the amount ofinsecticide saved per acre ofBt
com in 1996 was 0.16 pounds ofactive ingredient. As more acres ofBt com were planted, insecticide use was reduced on a smaller share ofthese Bt acres, leading to a lower average reduction in insecticide use across all acres planted to Bt com. Inrecent years, the reduction has been only 0.02 pounds per acre (see Figure 1in Benbrook 2004). Besides the monetary benefits, Fernandez-Cornejo and Caswell (2006) noted that there
are several otherbeneficial factors, such as ease ofoperation and time savings, which mayhave made GE crops attractive to farmers. Despite the mixed results on Btbenefits, from the com growers' perspective, as reflected in the increased rate ofadoption ofBt
corn cultivars ever since 1996, the farm profitability has graduallybeen increasing through higher yields and/or lower costs (e.g., operator labor, energy savings, pesticide purchases) by growing Bt com crops.

31Olart2. TrendsinInsedicideApplicationsto Managethe ElropeanOlm BorerandSoutI1v.esb!mComBorerandtheReductioninInsed:iddel.l!ie

### onl BtOlmAaes

1996 1997 1998 1999 2I.XlO 2001 2002 2003 2OCI4 Years**~** 0.25 a:: Q/ 0.2 a::n til "0**~ ~**  0.15 

### «Q.**"O~** 0.1 

### .a '§, 0.05

### ~ 0

Figure 1. Reproduced from Benbrook 2004 "Chart 2. Trends in Insecticide Applications to Manage the European Com Borer and Southwestern ComBorer and the Reduction in Insecticide Use on Bt Com Acres".

Anotherpotential economic benefit for growers and downstream consumers is increased competition in the marketplace for pest-control products, including hybrid seed from multiple marketers oflepidopteran-tolerant Bt comvarieties. The commercial availability ofMIR162 hybrid com seed mayrepresent a important newpest control option and tool for growers. Increased grower choice mayexert downward pressure on the cost ofproducts that offer control oflepidopteranpests (see pg. 110 in Syngenta 2007).

Human Health andEnvironmentalBenefits. There is no human health concerns with respect to toxicityor allergenicity and no unreasonable environmental concerns with respectto toxicity ofthe insecticidalproteins expressed in the MIR162 com (Appendix I in this EA; 2001). Also, use ofBtcom can decrease farm worker exposure to Bt sprays and chemical insecticides and reduce the mold infestation on com seeds (Carpenter et al. 2002). Consequently, anyreduction inmoldtoxins resulting from use of Bt com can provide direct benefits to people and com-fed livestock. In a variety offield studies, otherinsect protected com expressing Btproteins have been shown to have extensively lower levels ofcornmon mycotoxins that are produced by fungal pathogens (Wu 2006); however, the mycotoxin levels in MIR 162 comincommercial cultivation is not available at this time.

InsectResistanceManagementBenefits. According to EPAthe MIR162 com and its stacked Bt hybrids have the potential to delay development ofresistance in other com varieties expressing Crytoxins. The introduction ofMIR162 com and its stacks or

32pyramids may have an additional benefit ofprolonging the lifetime ofother com Plant Incorporated Protectants (PIP) technologies byproviding another mode ofaction for European com borer, com earworm, fall armyworm, and comrootworm. Effects on the ExportMarket. Syngenta does not expect any effects on the United States
com export market bythe cultivation ofthe MIR162 cultivars since Syngenta is actively pursuing regulatory approvals for the MIR162 comincountries that import comfrom the United States or Canada. Regulatory filings for the MIR162 com are in process for Colombia, Japan, South Korea, Taiwan, China, the Philippines, Australia and New Zealand, South Africa, the European Union, Russia, and Switzerland. Syngenta's stewardship agreements with growers will include a term requiring growers to divert this product away from export markets (i.e. channeling) where the grain has not yet received regulatory approval for import. Syngenta will communicate these requirements to growers using a wide-ranging grower education campaign (e.g., grower Stewardship Guide (see pg. 111 in Syngenta 2007).


### C. 

### Consideration of Executive Orders, Standards and Treaties Relatingto EnvironmentalImpacts

Executive Order (EO) 12898, "Federal Actions toAddress EnvironmentalJustice in Minority Populations and Low-Income Populations," requires Federal agencies to conduct theirprograms, policies, and activities that substantially affect humanhealth or the environment in a manner so as not to exclude persons and populations from participation in orbenefiting from such programs. It also enforces existing statutes to prevent minority and low-income communities from being subjected to disproportionately high and adverse human health or environmental effects.



### EO 13045, "Protection ofChildren from Environmental Health Risks and Safety Risks," acknowledges that children may suffer disproportionately from environmental health and safetyrisks because oftheir developmental stage, greater metabolic activity levels, and behaviorpatterns, as compared to adults. The EO (to the extent permitted by law and consistentwith the agency's mission) requires each Federal 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 13045. Based on the information submitted bythe applicant and assessed byAPHIS, MIR162 com is not expected to have a disproportionate adverse effect onminorities, low-income populations, or children. MIR162 is not significantlydifferent than conventional com and has successfully completed the FDAvoluntary consultation for food and feed use. Collectively, the available mammalian toxicity, along with the history ofsafe use of microbial Btproducts and other com varieties expressing Btproteins, establishes the safety ofthe com line MIR162 and its products to humans, including minorities, low income populations, and children who might be exposed to them through agricultural production and/orprocessing.



### 33None ofthe impacts on agricultural practices expected to be associated with deregulation ofthe com line MIR162 are expected to have a disproportionate adverse effect on minorities, low income populations, or children. As noted throughout the EA, the cultivation ofpreviously deregulated com varieties with similar insect resistant traits has been associated with a decrease or shift in pesticide applications for those who adopt these varieties that is either favorable orneutral with respect to environmental and human toxicity. Ifpesticide applications are reduced, there maybe 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 otherrural dwelling individuals who are exposed to pesticides through ground-water contamination
orothermeans ofexposure. Itis expected that EPA and USDA Economic Research Service would monitor the use ofthis product to determine impacts on agricultural practices such as chemical use as theyhave done previously for Btproducts.

EO 13112, "Invasive Species", states that Federal agencies take action to prevent the introduction ofinvasive species and provide for their control and to minimize the economic, ecological, and human health impacts that invasive species cause. Non-GE
com as well as GE Bt and herbicide tolerant com varieties that have been granted nonregulated status are widely grown inthe United States. Based onhistorical experience with these varieties and the data submitted bythe applicant and assessed by APHIS, MIR162 comis sufficiently similar in fitness characteristics to other com varieties currently grown, and it is not expected to have an increased invasive potential (see USDA-APHIS 2009).

INTERNATIONAL IMPLICATIONS

EO 12114, "EnvironmentalEffects Abroad ofMajorFederalActions" requires Federal officials to take into consideration any potential environmental effects outside the U.S., its territories and possessions that result from actions being taken. APHIS has given this due consideration and does not expect a significant environmental impact outside the U.S. should nonregulated status be determined for the com line MIR162 or if the other alternatives are chosen. All the considerable, existing national and international regulatory authorities and phytosanitary regimes that currently applyto introductions of
new com cultivars internationally, apply equallyto those coveredby an APHIS determination ofnonregulated status under 7 CFRpart 340. AnyMIR162 comin international commerce subsequent to a determination ofnon-regulated status for the line MIR162 wouldbe fully subject to national phytosanitaryrequirements and be in accordance with phytosanitary standards developed underthe International Plant Protection Convention (IPPC).

The purpose ofthe IPPC "is to secure a common and effective action to prevent the spread and introduction ofpests ofplants and plant products and to promote appropriate
measures for their control" (IPPC 2010)). The protection it affords extends to natural flora and plantproducts and includes both direct and indirect damage bypests, including weeds. The IPPC has set a standard for the reciprocal acceptance ofphytosanitary certification among the nations that have signed or acceded to the Convention (173

34countries as ofAugust 2009). In April, 2004, a standard for pestrisk analysis ofliving modified organisms (LMOs) was adopted at a meeting ofthe governingbodyofthe IPPC
as a supplement to an existing standard, International Standard for PhytosanitaryMeasure No. 11 (ISPM-ll; PestRisk Analysis for Quarantine Pests). The standard acknowledges that all LMOs will not present a pestrisk, and that a determination needs to be made early in the PRA for importation as to whetherthe LMOposes a potentialpestriskresulting from the genetic modification. APHIS pestrisk assessment procedures for bioengineered organisms are consistent with the guidance developed under the IPpc. In addition, issues that mayrelate to commercialization and transboundarymovement ofparticular agricultural commodities produced through biotechnology are being addressed in other international forums and through national regulations.

The Cartagena Protocol on Biosafety is a treaty underthe United Nations Convention
on Biological Diversity (CBD) that established a framework for the safe transboundary movement, with respect to the environment and biodiversity, ofLMOs, which includes those modified through biotechnology. The Protocol came into force on September 11, 2003 and 156 countries are parties to it as ofJune 24,2009 (see CBD 2010). Although the U.S. is not a partyto the CBD, and thus not a partyto the CartagenaProtocol on Biosafety, U.S. exporters will still need to complywith domestic regulations that importing countries that are parties to the Protocol haveput in placeto complywith their obligations. The first intentional transboundarymovement ofLMOs intended for environmental release (field trials orcommercial planting) willrequire consent from the importing country under an advanced informed agreement (AlA)provision, which includes a requirement for a risk assessment consistent with Annex III ofthe Protocol, and the required documentation. LMOs imported for food, feed orprocessing (FFP) are exempt from the AlAprocedure, and are covered underArticle 11 and Annex II ofthe Protocol. UnderArticle 11 Parties must post decisions to the Biosafety Clearinghouse database on domestic use ofLMOs for FFP that maybe subject to transboundary movement. To facilitate compliance with obligations to this protocol, the US Government has developed a website that provides the status ofall regulatory reviews completed for different uses ofbioengineeredproducts (http://usbiotechreg.nbii.gov). These datawill be available to the BiosafetyClearinghouse.

APHIS continuesto worktoward harmonization ofbiosafety and biotechnology
consensus documents, guidelines andregulations, including within the North American Plant Protection Organization (NAPPO), which includes Mexico, Canada, and the U.S. and in the Organization for Economic Cooperation and Development (OECD). NAPPO has completed three modules ofa standard for theImportation andRelease into the Environment of Transgenic Plants in NAPPOMember Countries (see http://www.nappo.orglStandards/Std-e.html). APHIS also participates inthe North American Biotechnology Initiative (NABI), a forum for information exchange and cooperation on agricultural biotechnologyissues for the U.S., Mexico and Canada. In addition, bilateral discussions onbiotechnologyregulatory issues are held regularly with othercountries including: Argentina, Brazil, Japan, China, and Korea. Manycountries, e.g. Argentina, Australia, Canada, China, Japan, Korea, Philippines, SouthAfrica, Switzerland, the United Kingdom, and the European Union have already approved Bt

35com varieties to be grown or imported for food or feed (AgBios 2010). There should be
no effects on the U.S. com export market since Syngenta is activelypursuing regulatory approvals for the MIR162 com in countries with functioning regulatory systems for geneticallymodified organisms and that import com from the U.S. or Canada. Regulatory filings for the MIR162 com are inprocess for Colombia, Japan, South Korea, Taiwan, China, the Philippines, Australia and New Zealand, South Africa, the European Union, Russia, and Switzerland.

COMPLIANCE WITH CLEAN WATERACT AND CLEAN AIRACT This EA evaluated the changes in comproduction due to the unrestricted use ofMIR162
com. MIR162 comwill not lead to the increased production ofcom in U.S. agriculture. The inclusion ofthe MIR162 com into comhybrids and their use in agriCUlture is not expected to have an adverse effect onwater quality, water use, or air quality. There is no expected change inwater use due to the production ofMIR162 com compared to current
comproduction regimes, nor is it expected that air quality will change due to do the production ofMIR162 com. The commercial availability ofMIR162 is expected to continue to provide improvements in water quality similar to other commercially available GE insecticide resistant crops due the potential for continued reduction in use of
more hazardous chemical pesticides, many ofwhich are toxic to aquatic organisms.

36

## VI. REFERENCES

Agbios 2010. GM Database. AgBios. Merrickville, Ontario, Canada http://www.agbios.comldbase.php
Anderson, P.L., Hellmich, RL., Sears, M.K., Sumerford, D.V., and Lewis, L.C. 2004. Effects ofCry1Ab-ExpressingCom Anthers on Monarch Butterfly Larvae. Environ. EntomoL 33(4):1109-1115

AOSCA (Association ofOfficial Seed Certifying Agencies). 2003. Operational Procedures, Crop Standards and Service Programs Publications. Meridian, Idaho. Aylor, D.E., Schultes, N.P., and Shields, 2003. Review. An aerobiological framework for assessing cross-pollination inmaize. Agric. Forest Meteorology 119,111-129 Bange, G.A. (2007). The situation and outlook for world com, soybean and cotton markets. Chairperson, World Agricultural Outlook Board Office ofthe Chief Economist, USDA. Presentation to National Grain and Oils Infonnation Center Beijing, China, July 2,2007. http://www

.usda.gov/oce/speeches/2007 /GeraldBange--ChinaNGOIC2007.doc
Bates, S., Zhao, J-Z, Roush, RT., and Shelton, A.M. 2005. Insect resistance management in GM crops: past, present and future. Nature BiotechnoI. 23: 57-62.
Bradford, K.J. 2006. Methods to maintain genetic purity ofseed stocks. Genetic Engineering ProducerFact Sheet 2. Publication 8189. University ofCalifornia, Division ofAgriculture and Natural Science. http://www.plantsciences.ucdavis.edulbradford/8189.pdf
Brookes, G. and Barfoot, P. 2004. Coexistence ofGM and Non GM Crops: Case Study ofMaize Grown in Spain. Dorchester, lJK: PGEconomics Ltd. http://www.pgeconomics.co.uk

. Burkness, E.C., Galvan, T. L. and Hutchison, W. D. 2009. OptimizingHelicoverpa zea (Lepidoptera: Noctuidae): Insecticidal efficacy in Minnesota sweet com: a logistic regression to assess timingparameters. J. Econ. EntomoL 102,677-684.
Bums, RG. 1982. Enzyme activity in soil: location and possible role in microbial ecology. Soil BioI. andBiochem. 14: 423-427.
Benbrook, C. M. 2004. Genetically Engineered Crops and Pesticide Use inthe United States: BioTech InfoNet Technical Paper Number 7, October 2004. The First Nine Years (http://www.biotcch-info.net/full version first nine.pdf).

37Bessin, R. 2004. Fall Armywonn in com. ENTFACT-IIO. University ofKentucky, College ofAgriculture http://www.ca.uky.edulENTOMOLOGY/entfacts/efllO.asp

BPPD-EPA. 2001. Insect Resistance Management. Pp. IIDI-153 In, BtPlantIncorporated Protectants, October 15,2001. Biopesticides Registration Action Document, Biopesticides and Pollution Prevention Division, Office ofPesticide Programs, U.S. Environmental Protection Agency. ( http://www

.epa.govIoppbppd1 Ibiopesticides/pipslbt_ brad214-inn.pdf) (Last updated: October 15th, 2008).

BPPD-EPA. 2009. Benefits and public interest finding. Pp. 115-134In, Bacillus thuringiensis Vip3Aa20 Insecticidal Protein and the Genetic Material Necessary for Its Production (via Elements ofVectorpNOV1300) in Event MIR162 Maize (OECD Unique Identifier: SYN-IR162-4), Biopesticides Registration Action Document, Biopesticides and Pollution Prevention Division, Office ofPesticide Programs, U.S. Environmental ProtectionAgency. ( http://www.epa.gov/oppbppdIlbiopesticides/ingredients/tech docs/ brad_006599.pdf) (Last updated: March 2009).
Brittan, K. 2006. Methods to Enable the Coexistence ofDiverse Com Production Systems. University ofCalifornia, Division ofAgriculture and Natural Resources. Publication 8192. http://ucbiotech.orglresources/factsheets/8192.pdf
Burkness, E.C., Galvan, T. L. and Hutchison, W. D. 2009. OptimizingHelicoverpa zea (Lepidoptera: Noctuidae): Insecticidal efficacy inMinnesota sweet com: a logistic regression to assess timing parameters. 

### J. Econ. Entomol. 102,677-684.
Carozzi, N., and M. Koziel. 1997. Advances in Insect Control. Taylor & Francis Ltd., Bristol, PA. Carpenter, J., Felsot, A., Goode, T., Hammig, M., Onstad, D., and Sankula, S. 2002. ComparativeEnvironmental Impacts ofBiotechnology-derived and Traditional Soybean, Com, and Cotton Crops. Council for Agricultural Science and Technology, Ames, Iowa. www.cast-science.org. Sponsored bythe United Soybean Board (www.unitedsoybean.org). http://www.soyconnection.com/soybean

_oil/pdf/EnvironmentalImpactStudyExecutiveSummary-English.pdf

CBD 2010. The CartegenaProtocol onBiosafety. The Convetion on Bioversity. http://www

.cbd.intlbiosafetyl
Cerdeira, A.L. and Duke, S.O. (2006). The current status and environmental impacts of glyphosate-resistant crops: A review. J. Environ. Qual. 35, 1633-1658. doi: 10.2134/jeq2005.03

38Clements, M. J., Campbell, K.W., Maragos, C. M., Pilcher, 

## c., Headrick, J. M., Pataky, 1. K., and White, D. G. 2003. Influence ofCrylAb protein and hybrid genotype on fumonisin contamination and fusarium earrot ofcom. Crop Sci. 43:1283-1293

de Maagd, R. A, Bravo, A and Crickmore, N. 2001. HowBacillus thuringiensis has evolved specific toxinsto colonizethe insect world. Trends in Gen. 17: 193-199.
Dicke, F. F. and Guthrie,W. D. 1988. Themost important com insects. Pp. 767-867In Sprague, G. F. and J. W. Dudley (eds.) Com and Com Improvement (3fd edition). American Society ofAgronomy-Crop Science Society ofAmerica-Soil Science Society ofAmerica, Madison, Wisconsin.
Dively, G.P. 2005. Impact ofTransgenic VIP3A X CrylAb Lepidopteran-resistant Field Com onthe Nontarget Arthropod Community. Environ. Entomol. 34, 1267-1291

DoaneMarketing Research. 2006. U.S. Seed com TraitTrak Study. A DoaneMarketing Research Syndicated Tracking Study. St. Louis, Missouri.
Dowd, P. F. ]998. Involvement ofarthropods in the establishment ofmycotoxigenic fungi under field conditions. Pp. 307- 350In K. K. Sinha and D. Bhatnagar (eds.) Mycotoxins inAgriculture and Food Safety. Marcel Dekkar, Inc. New York.
Dubelman, S., Ayden, B. R., Bader, B. M., Brown, C. R., Jiang, C. and Vlachos, D. 2005. CrylAb protein does notpersist in soil after 3 years ofsustainedBtcom use. Environ. Entomol. 34: 915-921.
Estruch, J.1., Warren, G.W., Mullins, M.A, Nye, G.J., Craig, J.A. and Koziel, M. G. 1996. Vip3A, a novelBacillus thuringiensis vegetative insecticidalprotein with a wide spectrum ofactivities against lepidopteraninsects. Proc. Nat. Acad. Sci. (USA) 93: 5389-5394.

EPA. 2001. Biopesticides RegistrationAction Document Bacillus thuringiensis Plant IncorporatedProtectants. United States Environmental Protection Agency. (http://www.epa.gov/pesticides/biopesticides/pipslht_brad.htm) (Last updated: 1011512008)

EPA 2008. Bacillus thuringiensis CrylAb Delta-EndotoxinProtein and the Genetic Material Necessaryfor Its Production (via Elements ofVectorpZ01502) in Event Btll Com (OECDUnique Identifier: SYN-BT011-1)(006444) & Bacillus thuringiensis Vip3Aa20 Insecticidal Protein and the Genetic Material Necessary for Its Production (via Elements ofVectorpNOV1300) in Event MIRl62 Maize (OECD Unique Identifier: SYN-IR162-4)(006599) Fact Sheet. http://www.epa.gov/pesticides/biopesticides/ingredients/factsheets/factsheet_0065 99-006444.html
EPA 2009. Pesticides: Endangered SpeciesProtection Program. United States Environmental ProtectionAgency.

39http://www.epa.gov/espp/coloring!especies.htm (Last updated: 2/912009)

EPA 2009a. Bacillus thuringiensis CrylAb Delta-Endotoxin Protein and the Genetic Material Necessary for Its Production (via Elements ofVectorpZ01502) in Event Btl 1 Corn (OECD Unique Identifier: SYN-BT011-1)(006444) & Bacillus thuringiensis Vip3Aa20 Insecticidal Protein and the Genetic Material Necessary for Its Production (via Elements ofVectorpNOV1300) inEvent MIR162 Maize (OECD Unique Identifier: SYN-IRI62-4)(006599) Fact Sheet. http://www.epa.gov /pesticides/biopesticides/ingredients/factsheets/factsheet

_0065 99-006444.html
EPA 2009b. Bacillus thuringiensis CrylAb Delta-Endotoxin Protein and the Genetic Material Necessary for Its Production (via Elements ofVectorpZ01502)in Event Btl 1 Corn (OECD Unique Identifier: SYN-BT011-1)(006444) & Bacillus thuringiensis Vip3Aa20 Insecticidal Protein and the Genetic Material Necessary for Its Production (via Elements ofVectorpNOV1300) in Event MIR162 Maize (OECD Unique Identifier: SYN-IRI62-4)(006599) & Modified Cry3A Protein and the Genetic Material Necessary for Its Production (via Elements ofVector pZM26) in Event MIR604 Corn (OECD Unique Identifier: SYN-IR604­ 5)(006509) Fact Sheet. US Environmental Protection Agency. http://www

.epa.gov/oppbppd1 /biopesticides/ingredients/factsheets/factsheet

_0065 99-006444-006509.htm

EPA2009c. Biopesticides Registration Action Document. Bacillus thuringiensis Vip3Aa20 Insecticidal Protein and the Genetic Material Necessary for Its Production (viaElements ofVectorpNOV1300) inEvent MIR162 Maize (OECD Unique Identifier: SYN-IRI62-4) PC Code: 006599. http://www.epa.gov/oppbppdl/biopesticides/ingredients/tech_docs/brad_006599

. pdf

Fernandez-Cornejo, J., and Li, J. 2005. The Impacts ofAdopting GeneticallyEngineered Crops in the USA: The Case ofBt Com. Paperpresented at the American Agricultural Economics Association meetings. Providence. (http://www.ers.usda.gov/publications/eibll1eibIt.pdf) Fernandez-Cornejo, J., andCaswell, M

.. 2006. First Decade ofEngineered Crops in the United States. USDA-ERS Economic Information Bulletin 11, April 2006. (http://www.ers.usda.gov/publications/eib111eibIt.pdf)

Food Institute. 2007. U.S. agriculture ends year with record trade. (WASHINGTON) (Statistical data) The Food Institute Report. February 26th

, 2007.

GAO (1996). Motor Fuels: Issues Related to Reformulated Gasoline, Oxygenated Fuels, and Biofuels. RCED-96-121 June 27, 1996. http://www.gao.gov/productsIRCED96-121

40Gianessi, L., and Reigner, N. 2006. Pesticide use in

## u.s. crop production:2002. A report preparedbyCropLife Foundation (www.croplifefoundation.org), Washington, D.C. (http://www.croplifefoundation.org/Documents/PUDINPUD%202002/ Inst%20and%200P%20Data%20Report.pdf)
Gianessi, L.P. 2008. The value ofherbicides in U.S. crop protection. Proceedings ofthe 60th Conference ofthe California Weed Science Society. 2008. http://www.cwss.orglproceedingsfilesI2008/24

_2008.pdf.**Goggi~,** A.S., Caragea, P., Lopez-Sanchez, H., Westgate, M.D., Arritte, R and Clark, C. 2006. Statistical analysis ofoutcrossingbetween adjacent maize grainproduction fields. Field Crops Res. 99, 147-157.
Gurian-Sherman, D. 2009. Failure to yield: evaluatingtheperformance ofgenetically engineered crops. UCS (Union ofConcerned Scientists) Publications, Cambridge, MA (http://www.ucsusa.orglassets/documents/food _and_agriculture/failure-toyield.pdf)
Haack, RA, 1993. The Endangered Karner Blue Butterfly (Lepidoptera: Lycaenidae): Biology, management considerations, and data gaps. Pp 83-110In Gillespie, AR., G.R Parker, P.E. Pope and G. Rink (eds). Proceedings 9th Central Hardwood Forest Conference. General Technical Report NC-161. USDA-Forest Service, North Central Forest Experiment Station, 

## st. PauL MN.
Head, G., Surber, J. B., Watson, J. A., Martin, J. W., and Duan, J. J. 2002. No detection ofCrylAcprotein in soil after multiple years ofBt cotton (Bollgard) use. Environ. EntomoL 31 :30-36.
Henry, C., Morgan, D., Weekes, R, Daniels Rand Boffey C. 2003. Farm Scale Evaluations ofGM Crops: Monitoring Gene Flow from GM Crops to Non-GM Equivalent Crops inthe Vicinity. Contract Reference EPG 115/138. Final Report 2000/2003. (http://www.defra.gov.uk/environment/gm/research/pdfi'epg_1-5­ 138.pdf).
Hoeft, R.G., Nafziger, E.D., Johnson, RR and Aldrich, S.R 2000. Modern Corn and Soybean Production. 1st ed., Chapter 1, Corn as a Crop. MCSP Publications, Champaign, IL.
Hafte, H., and White, H. Roo 1989. Insecticidal crystal proteins ofBacillus thuringiensis. Microbiol. Molec.BioL Rev. 53: 242-255. Hurley, T. M., Langrock, 1., and Ostlie, K. 2006. Estimating the benefits ofBt corn and cost ofinsectresistance management ex ante. J. Agric. Resource Econ. 31: 355­ 375.

41ICG (International Grains Council). 2009. Grain Market Report (GMR No. 394,29 Oct 2009). (http://www.igc.org.uk/enldownloads/gmrsummary/gmrsumme.pdf).

IPPC 2010. Home page. International Plant Protection Convention. Protectingthe World's Plant Resources from Pests. http://www.ippc.intlIPPlEnidefault.jsp

. Jones, M.D., and Brooks, J.S., 1950. Effectiveness ofdistance and border rows in preventing outcrossing in corn. Oklahoma Agricultural Experiment Station Technical Bulletin No. T-38, Stillwater, OK, 18 pp. Lee, M.K., Walters, F.S., Hart, H., Palekar, N., and Chen, J-S. 2003. The mode ofaction ofthe Bacillus thuringiensis vegetative insecticidal protein Vip3A differs from that ofCry1 Ab-endotoxin. AppI. and Environ. MicrobioL 69, 4648-4657
Ma, B.L., Meloche, F. and Wei, L. 2009. Agronomic assessment ofBttrait and seed or soil-applied insecticides on the control ofcorn rootworm and yield. Field Crops Res. 111, 189-196
Ma, B.L., Subedi, K. D. and Reid L. M. 2004. Extent ofcrossfertilization in maize by pollen from neighbouringtransgenic hybrids. Crop Sci. 44: 1273-1282.
Marra, M., Carlson, G. and Hubbell, B. 1998. Economic impacts ofthe first crop biotechnologies. (http://www.ag.econ.ncsu.edu/faculty/marra/firstcrop/impOOl.gif)
Martin, M.A. and Hyde, 

### J. 2001. Economic considerations for the adoption oftransgenic crops: the case ofBtcorn. J. Nematology 33, 173-177.
Marx, M-C., Kandeler, E., Wood, M., Wermbter, N., and Jarvis, S.c. 2005. Exploring the enzymatic landscape: distribution and kinetics ofhydrolytic enzymes in soil particle-size fractions. Soil BioI. and Biochem. 37,35-48.
Marvier, M., McCreedy, c., Regetz, J., and Kareiva P. 2007. A meta-analysis ofeffects of Bt cotton and maize on nontarget invertebrates. Science 316:1475-1477.
Marx, M-C., E. Kandeler, M. Wood, N. Wermbter, and S. C. Jarvis. 2005. Exploring the enzymatic landscape: distribution and kinetics ofhydrolytic enzymes in soil particle-size fractions. Soil BioI. and Biochem. 37:35-48.
Mason, C. E., Rice, M. E., Calvin, D. D., Van Duyn, J. W., Showers, W. B., Hutchison, W. D., Witkowski, J. F., Higgins, R. A., Onstad, D. W. and Dively, G. P. 1996. European cornborer ecology and management. North Central Regional Extension Publication No 327. Iowa State University, Ames, Iowa. 57 pp.

42Messeguer, J., Pefias, G., Ballester, J., Bas, M., Serra, J. and Salvia, J

.. 2006. Pollenmediated gene flow in maize in real situations ofcoexistence. Plant Biotechnol. J. 4: 633-645.
Mendelsohn, M., Kough, 

### .T., Vaituzis, Z. and Matthews, K

.. 2003. AreBt crops safe? Nature Biotechnol. 21: 1003-1009.
Monbaliu, S., Van Poucke, C., Detavemier, e, Dumoulin, F., Van De Velde, M., Schoeters, Van Dyck, S., Averkieva, 0., Van Peteghem, C., and De Saeger, S. 2010. Occurrence ofmycotoxins in feed as analyzed by a multi-mycotoxin LCMSIMS method. J. Agric. Food Chern. 58, 66-71.

MRSC 2009. Field standards for hybrid com seed. WAC 16-302-295. Municipal Research and Service Center for Washington. StatutoryAuthority: RCW 15.49.005, 15.49.081, 15.49.310, 15.49.370(3) and chapter 17.24 RCW. 00-24­ 077, § 16-302-295, filed 12/4/00, effective 114/01. http://www.mrsc.org!

NCRA. 2010. Impact Analysis and Decision Strategies for Agricultural Research. Statement ofIssues and Justification. NC1 003 North Central Region of Administrators ofAgricultural Research Experiment Stations

.. http://nimss.umd.edulhomepageslhome.cfm?trackID=515
Nonnan, M. 1. T., Pearson, C. J., and Searle, P. O. E

.. 1995. The Ecology ofTropical Food Crops (2nd edition). Cambridge University Press, Cambridge, 

### u.K., pp 126-144.

OECD (Organization for Economic Co-operation and Development). 2003. Consensus document on the biology ofZea mays subsp. mays (maize). Series on Harmonisation ofRegulatory Oversight in Biotechnology, No. 27, OECD, Paris. (http://www.0Iis.oecd.org!0Iis/2003doc.nsf/LinkToINT0000426E/$FILE/JT00147 699.PDF. (Last Modified: 23 July, 2003)

Penn State University (2010). Agronomy Guide 2009-2010. Part 2, Section 2: ComPest Management. INSECTS. Crop Management Extension Group (CMEG). Penn State University, Penn State Cooperative Extension. http://agguide.agronomy.psu.edulpm/sec2/sec23d.cfm
Perrin, M. 1997. Crop protection: taking stock for the new millennium. Crop Prot. 16, 449-456.
Peterson, R. K., Meyer, S. J., Wolf, AT.,Wolt, J. D. and Davies, P. M

.. 2006. Genetically engineeredplants, endangered species, and risk: A temporal and spatial exposure assessment for Kamer blue butterfly larvae andBtmaize pollen. Risk Anal. 26: 845-858.
Placinta, eM., D'Mello, J.P.F. and Macdonald, AM.e 1999. A review ofworldwide contamination ofcereal grains and animal feed with Fusarium mycotoxins. Anim. Feed Sci. and TechnoL 78,21-37.
43Rieger, M. A., Lamond, M., Preston C,Powles, S.B., and Roush, R. T. 2002. Pollenmediated movement ofherbicide resistance between commercial canola fields Science 296, 2386-2388
Schienmann, J. 2003. Co-existance ofgenetically modified crops with conventional and organic farming. Environ. BiosafetyRes. 2: 213-217. Schnepf, E., Crickmore, N., Van Rie, J., Lereclus, D., Baum, J., Feitelson, J. Zeigler, D. R. and Dean, D. H

.. 1998. Bacillus thuringiensis and its pesticidal crystal proteins. MicrobioL and MoL BioL Rev. 62: 775-806.
Shelton, A. M., Zhao, J.-Z., and Roush, R. T

.. 2002. Economic, ecological, food safety, and social consequences ofthe development ofBt transgenic plants. Annu. Rev. Entomol. 47: 845-881.
Sheridan, C 2009. Report claims no yield advantage for Bt crops. Nature Biotechnol. 27: 588-589. Simpson, G. M. 1990. Seed Dormancyin Grasses. Cambridge UniversityPress, New York.
Smeltzer, D.G. 1959. Relationship between Fusarium earrot and com earworm infestation. Agron. J. 51:53-54.-14
Tabashnik, B.E., Van Rensburg, J.BJ., and Carriere, Y. 2009. Field-evolved insect resistance to Bt crops: Definition, theory, and data. J. Econ. EntomoL 102,2011­ 2025.
Tabashnik, B.E., Unnithan, G.C, Masson, L., Crowder, D.W., Li, X., and Carrierre, Y. (2009a). Asymetrical cross-resistancebetweenBacillus thuringiensis toxins Cry1Ac and Cry2Ab inpinkbollworm. P. Nat. Acad. Sci. 106, 11889-11894.
Trostle, R. 2008. Global agricultural supply and demand: factors contributing to the recent increase in food commodityprices. United States Department of Agriculture, Economic Research Service. University ofIllinois Extension 2004. Integrated Pest Management. Com: Insects http://ipm.illinois.edulfieldcrops/corn/insects.html

USDA-AMS. 2007. National Organic Program Q&A http://www.ams.usda.gov/nop/Q&A.html

USDA-AMS 2010a. National Organic Program. US Department ofAgriculture, Agricultural Marketing Service. http://www.ams.usda.gov/AMSv1.0fNOP

44USDA-AMS. 2010. National Organic Program. Resources: Regulations. http://www.ams.usda.gov/AMSvI.O/ams.fetchTemplateData.do?template=Templ ateF&navID=RegulationsNOPNationaIOrganicProgramHome&rightNavi=Regul ationsNOPNationaIOrganicProgramHome&topNav=&leftNav=&page=NOPReso urceCenterRegulations&resultType=&acct=noprulemaking

USDA-APHIS. 2009. Plant Pest Risk Analysis for SyngentaMIRI62 com. USDA, APHIS, Biotechnology Regulatory Service. Riverdale, MD (URL: http://www.aphis.usda.gOYIbrs/notJeg.html)

USDA-APHIS. 2010. Petitions ofNonregulated Status Granted orPendingbyAPHIS as ofFebruary 2,2010. http://www.aphis.usda.govlbrs/notJeg.html

USDA-ERS. 2008. Feeds grains database. United States Department ofAgriculture, Economic Research Service. http://www.ers.usda.gov/datalfeedgrains/. Access date: August 25, 2008.

USDA-ERS. 2009a. BriefingRoom: Com: Trade. http://www.ers.usda.goviBriefingiCorn/trade.htm

. Access date: March 8, 2010.

USDA-ERS. 2009b. Adoption ofGenetically Engineered Crops inthe U.S.: Com Varieties. Data Sets. USDA Economic Research Service. http://www.ers.usda.govlDataiBiotechCrops/ExtentofAdoptionTablel.htm

USDA-ERS. 2009c. Adoption ofGenetically Engineered Crops inthe U.S.: Extent of Adoption. USDA Economic Research Service. http://www.ers.usda.govlDataibiotechcrops/adoption.htm

USDA-ERS. 2009. Adoption ofGenetically Engineered Crops inthe U.S.: Com Varieties. United States Department ofAgriculture, Economic Research Service, (UpdatedJuly 1, 2009) (Access date: August 10, 2009) (http://www.ers.usda.gov/dataiBiotechCrops/ExtentofAdoptionTableI.htm)

USDA-ERS. 2009c. Organic Production. National Tables. http://www.ers.usda.gov/datalorganiclDataiCertified%20and%20total%20US%20 acreage%20selected%2Ocrops%20Iivestock%2095-05.xls

USDA-NASS. 2006. Agricultural Chemical Usage 2005 Field Crops Summary. United States Department ofAgriculture, National Agricultural Statistics Service, May 2006. (http://usda.mannlib.comell.eduiusdainass/AgriChemUsFC/12000sl2006/AgriChe mUsFC-05-17-2006.pdf)

USDA-NASS.2007. Crop Production. United States Department ofAgriculture, National Agricultural Statistics, February2007.

45(http://usda.mannlib.comell.eduiusdainass/CropProdl 12000s/2007ICropProd-02­ 09-2007.pdf)

USDA-NASS. 2008a. Statistics ofGrain and Feed. United States Department ofAgriculture, National Agricultural Statistics Service, Washington, D.C. (http://www.nass.usda.govlPublications/Ag_StatisticsI200S/ChapOl.pdf )

USDA-NASS. 2008b. Acreage. United States Department ofAgriculture, National Agricultural Statistics Service, Washington, D.C. (http://usda.mannlib.comell

.edu/usdalnassl Acrell2000s/20081Acre-06-30­ 2008.pdf)

USDA-NASS. (2009) Census ofAgriculture. Table 26. Field Crops 2007 and 2002. http://www.agcensus.usda.govlPublicationsI2007IFuILReportNolume_l,_Chapte
r 2_ US_State

_Levellst99_ 2 _026_026.pdf

USDA-NASS.20l0. Crop Production 2009 Summary. National Agricultural Statistics Service. CrPr 2-1 (10).

USFWS.2007. Threatened and endangered species system. A database developed by the U.S. Fish and Wildlife Service. (http://ecos.fws.gov/tessyublic/SpeciesReport.do?groups=I&listingType=L&ma pstatus=l)
Ward, D., T. De Gooyer, T. Vaughn, G. Head, M. McKee, J. Astwood, and J. Pershing. 2005. Genetically enhanced maize as a potential management option for com rootworm: YieldGard® rootworm maize case study. Pp 239-262 In S. Vidal, U. Kuhlmann, and C. Edwards (eds.)Western Com Rootworm: Ecology and Management.CAB International, Wallingford, U.K.
Williams, P. W., P. M. Buckley, and G. L. Windham. 2002. Southwestern comborer (Lepidoptera: Crambidae) damage and aflatoxin accumulation in maize. J. Econ. Entomol. 95: 1049-1053.
Wright, B., M. Bernards, and L. Sandell. 2009. Volunteer com presents new challenges. Crop. Crop Watch News Service, University ofNebraska-Lincoln Extension, Institute ofAgriculture and Natural Resources, Lincoln, Nebraska. (http://cropwatch.unLedu/archives/2009/crop22/weeds

_volunteer

_corn.htm)
Wu, F. 2006. Mycotoxin reduction in Bt com: potential economic, health, and regulatory impacts. Transgenic Res. 15:277-289. Ziegler, K. E. 2000. Popcorn. Pp. 199-234. In A. R. Hallaur (ed.) Specialty Corns. CRC Press, BocaRaton, Florida.

46

## VII. PREPARERS, REVIEWERS, CONSULTANATIONS, AGENCY CONTACT
Biotechnology Regulatory Services (BRS) Michael Gregoire, Deputy Administrator Beverly Simmons, Associate Deputy Administrator for Emerging and International Programs SidneyAbel, Assistant Deputy Administrator
BRS, Environmental Risk Analysis Division Michael Watson, Ph.D., Division Director Susan M. Koehler, Ph.D., Branch Chief, Plants Branch (Reviewer) John Cordts, M.S., M.B.A., Biotechnologist (Reviewer) Subray Hegde, Ph. D., Biotechnologist (Preparer ofEnvironmental Assessment)
BRS, Policy and Coordination Division David Reinhold, Branch Chief, Communication & International Affairs Branch (Reviewer) Andrea Huberty, Ph. D., Branch Chief, Regulatory & Environmental Analysis Branch (Reviewer) Richard Coker, Regulatory Analyst

Consultations Michael Mendelsohn, Senior Regulatory Specialist Office ofPesticide Programs/ Biopesticides & Pollution PreventionDivision U.S. Environmental Protection Agency Agency Contact Ms. Cynthia Eck, Document Control Officer USDA, APHIS, BRS 4700 RiverRoad, Unit 147 Riverdale, MD 20737-1237 Phone: (301) 734-0667 Fax: (301) 734-8669 Cynthia.A.Eck@aphis.usda.gov

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## VIII. APPENDIX Appendix

### 1. EnvironmentalandHuman Health Safety ofVip3Aa20 Protein
Previously deregulated Bt comcultivars have resulted inreduced conventional pesticide use, as farmers find Btproducts more effective in mitigating lepidopteran insect feeding damage. It is reasonable to expectthat deregulation and commercialization ofthe MIR162 comwill result in further reductions in the use ofconventional pesticides. This reduction inconventional pesticide use would diminish the environmental risks of chemical pesticide insect control, as the chemical alternatives to MIR162 present wellcharacterizedrisks to humans and otherwildlife, whereas Vip3Aa20 presents no such risk. Substantial data support a conclusion that Vip3Aa20 toxicitywill be limited to sensitive lepidopteran species that are sufficiently exposed to the protein (USDA-APHIS 2009).

The toxicity ofinsecticidal Btproteins, such as Vip3a20, depends ontheirbinding to specific receptors present in the insect midgut. Research demonstrates that this specificity limits the proteins' toxic effect to certain lepidopteran species. A discussion
on the mechanism ofaction for Vip3Aa20, its spectrum ofactivity, and its lack of toxicityto non-lepidopteran species is presented in the petition (pg. 47-50).

Health and safety studies have been conducted with the novel proteins contained in MIR162 com. A comprehensive assessment ofthe safety ofthe introduced proteins, Vip3Aa20 and PMI, demonstrate that bothproteins are nontoxic to mammalian species and are unlikelyto be food allergens (69 FR26770-26775; 73 FR45620-45624; FDA BNFNo. 000113). The Vip3Aa20protein is considered nontoxic because it does not share significant amino acid homology with known protein toxins, is nontoxic to mice at
a veryhigh dose of 1250 mg Vip3Aa20lkgbw, is rapidly degraded in simulated mammalian gastric fluid, and its insecticidal mode ofaction for Vip3Aa20 is not relevant to mammals. Vip3Aa20 is also not likelyto be a food allergen because it is not derived from a known
source ofallergenic proteins, it does not have any significant amino acid sequence identityto known allergenic proteins, it is rapidly degraded in simulated mammalian gastric fluid, and it is labile uponheating at temperatures of65°C and above. The PMI protein is considered nontoxic because it does not share significant amino acid homology with known protein toxins, it is nontoxic to mice at a very high dose of3030mg PMIlkg bw, and it is rapidly degraded in simulated mammalian gastric fluid. PMI is not likelyto be a food allergen because it is not derived from a known source ofallergenic proteins, it does nothave any significant amino acid sequence identity to known allergenic proteins with implications for its allergenic potential, itis rapidly degraded in simulated mammalian gastric fluid, and it is labile upon heating at temperatures of37°C and above.

48A pennanent exemption from the requirement ofa food tolerance currently exists under 73 FR 45620-4562440 for Vip3Aa20 in maize and under 40 CFR §180.1252 for PMI in all plants.

USDA-APHIS. 2009. Plant Pest Risk Analysis for Syngenta MIR162 corn. USDA, APHIS, BiotechnologyRegulatory Service. Riverdale, MD (URL: http://www.aphis.usda.govIbrs/notJeg.html)

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### AppendixIL Syngenta MIR162 Corn ForageandGrain Compositional Analysis

The data for the discussion and interpretation on the composition analysis ofthe MIR162
corn come from Syngenta's petition (Petition Appendix E, pages 158-180 for experimental details and statistical analyses, and pages 69-73 for the interpretation and discussion ofresults) and was reviewed byAPHIS. The rationale for the composition analysis offorage and grain is to identify any changes in nutrient or anti-nutrient content ofthe new crop in the context ofits use as food or feed and to assess its biochemical equivalence and similarity to conventional maize. Assessment ofthe plant's composition also allows the developer and APHIS to evaluate possible unintended effects that might arise from insertion ofthe Vip3Aa20 and PMI genes into the plant's genome. This assessment was undertakenbyperforming quantitative analyses of65 components (Table 25, pg. 70 inpetition) bothfrom MIR162 hybrid corn and a nontransgenic control variety. The analytes measured in this studywere selected based on recommendations of the Organisation for Economic Co-operation and Development (OECD, 2002) for comparative assessment ofcomposition ofnew varieties ofmaize. The plant materials for the analysis come from six diverse corn growing regions ofthe U.S. (Appendix Table E1, pg. 158 in petition) during 2005. Plants were self-pollinated byhand and the developing ears were bagged to avoid cross-pollination. All analyses were conducted using methods published and approved bythe Association ofAnalytical Communities (AOAC) International or other industry-standard analytical methods.

Nine components ofcorn forage were measured; the difference between MIR162 and control mean values was found to be statistically significant for one ofthese analytes. Fifty-six components ofgrain were measured; the difference between MIR162 and control mean values was found to be statistically significant for 13 ofthese analytes. The results for these 14 analytes that had a statistically significant outcome for genotype effect (Table 26, pg. 72 inpetition) havebeendiscussed in the following paragraphs.

The forage compositional analyses for proximates and minerals revealed a single statistically significant difference between MIR162 and control mean values. The mean value for MIR162 Neutral Detergent Fiber (NDF) was 11.34%higherthan the corresponding control value. This difference is considered relatively small and the MIR162 mean falls well within the range ofnormal values reported byInternational Life Sciences Institute (ILSI) (Ridley et al. 2004; ILSI 2006) and Organisation for Economic Co-operation and Development (OECD). No statistically significant genotype by location interactions were noted for the forage compositional analyses. Compositional analyses ofgrain revealed no statistically significant differences between MIR162 and control means for 43 ofthe 56 analytes examined in across-location comparisons. Statistically significant differences were noted for levels ofthe proximates ash, NDF, starch, three grain minerals (calcium, iron, and phosphorus), levels ofvitamin A**(~-carotene),** vitamin B6 (pyridoxine), and vitamin E (a-tocopherol), linoleic and linolenic fatty acids. These differences were small 

### « 8%) and the MIR162 mean values

50were well within the ranges ofnonnal values for the control maize. Additionally, the average values for all proximates were within the ranges reported byILSI and OECD. Statistically significant differences were notedbetween MIR162 and control mean levels ofvitamin A**(~-carotene),** vitamin 

### B6 (pyridoxine), and vitamin E (a-tocopherol). These differences were small 

### « 7%) and the meanvalues observed for these vitamins in the MIR162 grain were well within the range ofvalues observed for the control grain. Additionally, the MIR162 means for all vitamins fell within the nonnal range ofvalues reported for conventional maizebyILSI and OECD. For vitamin A and vitamin 

### B9 a statistically significant genotype-by-Iocation interaction was noted, which suggests that the effect ofgenotype was not consistent across locations, hence, the comparison of genotypes averaged across locations may not be valid. Individual location means for the two analytes are provided inTable E-8 (Appendix E). The vitamin A and vitamin 

### B9 levels at all locations were within the ranges reported in the literature.

There were no significant differences noted for any ofthe 18 amino acids or anti­ nutrients measured and all average values were within the ranges reportedbyILSI and OECD (Appedix E, pg. 158-180 inpetition). Statistically significant differences were noted for linoleic and linolenic fatty acids. These differences were very small 

### « 4%) and the MIR162 mean values observed for these fatty acids were within the ranges ofvalues observed for the control grain. Furthennore, the average values for all fatty acids were within the range ofnonnal values reported for conventional maize byILSI and OECD. Statistically significant differences were noted in the secondary metabolites ferulic acid and p-cournaric acid. These differences were relatively small 

### « 15%) and the MIR162
mean values for these secondary metabolites were within the ranges ofvalues observed for the control grain. Additionally, the mean values for all MIR162 secondary metabolites and anti-nutrients were within the nonnal range ofvalues reported for conventional maize by ILSI and OECD. Collectively, the observed differences between MIR162 and control means are considered ofno biological significance and represent typical random variance. The magnitude ofthe differences was small, all MIR162 values fell within normal ranges for conventional maize, and the MIR162 and control dataranges significantly overlapped. MIR162 is therefore, not compositionally different from conventional maize.

References

ILSI. 2006. International Life Sciences Institue Crop Composition Database, 3.0. International Life Sciences Institute. http://wvv,v.cropcomposition.org/. Access date: 7/2612009.

OECD (Organization for Economic Co-operation and Development). 2002. Consensus document on the compositional considerations for new varieties ofmaize (Zea

51mays): Key food and feed nutrients, anti-nutirents and secondary metabolites. Series on the Safety ofNovel Foods and Feeds, No.6. OECD Environment Directorate. Joint meeting ofthe chemicals committee and the working party on chemicals, pesticides and biotechnology. Paris, 42 pp. Ridley, W. P., R. D. Shillito, I. Coats, H.-Y. Steiner, M. Shawgo, A. Phillips, P. Dussold, and L. Kurtyak. 2004. Development of the International Life Sciences Institue Crop Composition Database. Journal ofFood Composition and Analysis. 17:423­ 438.

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