BIOPESTICIDES REGISTRATION ACTION DOCUMENT

DRAFT



## Cydia pomonella Granulovirus Isolate V22 (CpGV-V22)

Pesticide Chemical (PC) Code: 128090

U.S. Environmental Protection Agency Office of Pesticide Programs Biopesticides and Pollution Prevention Division

December 19, 2012Cydia pomonella Granulovirus isolate V22 DRAFT Biopesticides Registration Action Document Page 2 of 29

TABLE OF CONTENTS

I. EXECUTIVE SUMMARY ...................................................................................................... 4

II. ACTIVE INGREDIENT OVERVIEW ................................................................................ 5

III. REGULATORY BACKGROUND ...................................................................................... 5

A. Application for Pe sticide Registration .................................................................................... ............................. 5

B. Food Tolerance Exemption .................................................................................................. ................................. 5

IV. RISK ASSESSMENT .......................................................................................................... .. 5

A. Product Analysis Asse ssment (40 CFR § 158.2120) ........................................................................... ................ 5

B. Human Health Risk As sessment (40 CF R 158.2140) ............................................................................ .............. 6

C. Environmental Assessm ent (40 CFR § 158.215 0) .............................................................................. ............... 11

V. ENVIRONMENTAL JUSTICE .......................................................................................... 18

VI. RISK MANAGEMENT DECISION ................................................................................. 18

VII. ACTIONS REQUIRED OF THE REGISTRANT ......................................................... 19

A. Final Prin ted Labeling .................................................................................................... .................................... 19

B. Terms of Registra tion ..................................................................................................... .................................... 19

C. Reporting of Adverse Effects and Hypersensitivi ty Incidents ............................................................... .......... 19

VIII. BIBLIOGRAPHY ........................................................................................................... . 20

A. Studies Submitted to Support the Regist ration of Pesticide Products Containing Cydia pomonella granulovirus isolate

V22.................................................................................................................................. ......................................... 20

B. Environmental Protection Agen cy Risk Assess ment Memoranda ................................................................. . 22

C. Other Referen ces .......................................................................................................... ....................................... 22

APPENDIX A. MICROBIAL PESTICIDES DATA REQUIREMENTS ............................ 23

APPENDIX B. PESTICIDE PRODUCT ................................................................................. 29Cydia pomonella Granulovirus isolate V22 DRAFT Biopesticides Registration Action Document Page 3 of 29

BIOPESTICIDES REGISTRATION ACTION DOCUMENT TEAM

Office of Pesticide Programs Biopesticides and Pollution Prevention Division Microbial Pesticides Branch (MPB)

Science Review

John Kough, Ph.D. Product Analysis and Human Health Shannon Borges, M.S. Nontarget Effects

Regulatory Review

Ann Sibold, M.A. Regulatory Action Leader, MPB Shannon Borges, M.S. Team Leader, MPB Sheryl Reilly, Ph.D. Acting Chief, MPB Kimberly Nesci, M.S. Acting Chief, MPBCydia pomonella Granulovirus isolate V22 DRAFT Biopesticides Registration Action Document Page 4 of 29 I. EXECUTIVE SUMMARY

Description of the Active Ingredient: Cydia pomonella Granulovirus isolate V22 is a naturally occurring insect virus that is widespread in the environment. It is a granul ovirus (also known as granulosis virus or GV), which is one of two genera taxonomically classified in the family Baculoviridae. The other member of this family is the Nucleopolyhedrovirus (also known as nuclear polyhedrosis virus or NPV). Granuloviruses are known to only infect insects of the order Lepidoptera, which includes moths and but terflies. Granuloviruses were first registered for use as pesticides in 1995. For more information on granuloviruses registered as pesticides, see NPV and GV Insect Viruses

.

Cydia pomonella granulovirus isolate V22 (also referred to in this document as CpGV-V22) is specifically active against larvae of the apple codling moth, Cydia pomonella, and the Oriental fruit moth, Grapholita

molesta. After ingestion by a moth larva, the virus multiplie s rapidly within its host, causing an infection resulting in cell destruction and death, typically within 3-7 days.

One microbial pesticide product cont aining CpGV-V22, Madex Twin Insecticidal Virus (EPA file symbol 69553-R) is proposed for regist ration. Madex Twin Insecticidal Virus is a dilutable end-use product containing 0.06% CpGV-V22. It is labeled for applic ation to apple, quince, pear, peach, nectarine, apricot, plum, prunes, cherry, walnut, almond, pluot, and aprium in agricultural and residential sett ings to control apple codling moth and Oriental fruit moth. It is appl ied before the larvae hatch and reapplied as necessary to maintain control.

Assessing Risks to Human Health: Data derived from appropriate tests give no indications that CpGV-V22 is toxic or pathogenic to humans. No harm ful health effects to humans are expect ed from its use as an insecticide.

Assessing Risks to the Environment: Pesticide products containing CpGV-V22 are not likely to pose a risk to the environment, including humans a nd other nontarget organisms.

Public Participation: On October 1, 2009, the U.S. Environmental Protection Agency (EPA or Agency) announced a policy to provide a more meaningful opportunity for the public to participate in major registration decisions before they occur. According to this policy, EPA provides a public comment period prior to making a registration decision for the following types of applications : new active ingredients; first food uses; first outdoor uses; first residential uses; or any other registration acti ons for which EPA believes there may be significant public

interest.

Consistent with the policy of making registration decisions more transparent, the pesticide product containing a new active ingredient, CpGV-V22 , is being subjected to a comment period. In addition to containing a new active ingredient, the registration of this pesticide produc t will result in the first outdoor and food uses for CpGV-V22

. The following documents are being made available for co mment in the docket, identification number EPA-HQOPP-2011-0984, and accessed through either http://www.regulations.gov or http://www.epa.gov/pesticides/regu lating/registrati on-status.html : (1) draft Cydia pomonella granulovirus isolate V22 (CpGV-V22) Biopesticides Registration Ac tion Document (BRAD); (2) dr aft label for the end-use pesticide product, Madex Twin Insecticidal Virus.

The public participation comment period is being init iated on December 19, 2012 and will close on January 3,

2013.

For definitions of scientific terms, please refer to http://www.epa.gov/pesticides/glossary/

.Cydia pomonella Granulovirus isolate V22 DRAFT Biopesticides Registration Action Document Page 5 of 29 II. ACTIVE INGREDIENT OVERVIEW

Biological Name: Cydia pomonella granulovirus isolate V22

Culture Collection: Deposited at the DSMZ Plant Virus Co llection in Braunschweig, Germany under

the accession number GV-0014.

OPP Chemical Code: 128090, formerly 129090

Type of Pesticide: Microbial Pesticide – Insecticide

See Appendix B for specific information (i.e., use sites, application rates, methods of application, formulation types, and target pests) regarding the pesticide products containing this active ingredient.

III. REGULATORY BACKGROUND

A. Application for Pesticide Registration

On October 18, 2011, SciReg, Inc., 12733 Director’s Loop, Woodbridge, VA 22192, agent for Andermatt Biocontrol AG (also referred to as “Andermatt” in th is document), Stahlermatten 6, CH-6146 Grossdietwil, Switzerland, submitted an application for registration of Madex Twin Insecticidal Virus (EPA File Symbol 69553-R), containing the new microbia l pesticide active ingredient, Cydia pomonella granulovirus Isolate V22 (also referred to in this document as CpGV-V22) . The proposed product is a dilutable end-use product (containing 0.06% CpGV-V22) intended to be a preharvest insecticide to control the appl e codling moth, Cydia pomonella and the Oriental fruit moth, Grapholita molesta applied to apple, quince, pear, peach, nectarine, apricot, plum, prunes, cherry, walnut, almond, pluot a nd aprium in agricultural a nd residential settings to control apple codling moth and oriental fruit moth. It is to be applied before larvae hatch and reapplied as necessary to maintain control.

In the Federal Register (FR) of March 9, 2012 (77 FR 14362), EPA a nnounced receipt of an application for registration of this pestic ide product for agricultural and residentia l uses, and no comments were received during the 30-day comment period. (For more information, see the following docket at http://www.regulations.gov/ : EPA-HQ-OPP-2011-0984.)

B. Food Tolerance Exemption

An exemption from the requirement of a tolerance is esta blished for residues of the microbial pest control agent

Occlusion Bodies of th e Granulosis Virus of Cydia pomonella (codling moth) in or on all raw agricultural

commodities 40 CFR 180.1148 . Residues of CpGV-V22 have been determined to be within the scope of 40

CFR 180.1148.

IV. RISK ASSESSMENT

A. Product Analysis Assessment ( 40 CFR § 158.2120 )Cydia pomonella Granulovirus isolate V22 DRAFT Biopesticides Registration Action Document Page 6 of 29 Microbial pesticide product analysis data requirements 1 include product chemistry and composition, analysis and certified limits, and physical and chemical characte ristics data. Product chemis try and composition data include information about the identity of the active ingredient, the manufacturing process, deposition of a sample of the microbial active ingred ient in a nationally reco gnized culture collection, and discussion of the potential for formation of unintentional ingredients. An alysis and certified limits data include information on analysis of samples and certification of limits. Physical and chemical ch aracteristics data describe basic characteristics of the registered pesticide products, in cluding color, physical state, odor, stability, miscibility, pH, corrosion characteristics, viscosity and density.

EPA assigned a unique PC code (128090) to CpGV-V22 to distinguish the ne w active ingredient from the PC code (129090) associated with other Cydia pomonella Granulovirus (CpGV) active ingredients.

All product analysis data required for a time-limited unconditional registration of CpGV-V22 have been fulfilled. Confirmatory data (including storage stability and corrosion characteristics data and data showing how the laboratory colonies of Cydia pomonella and Grapholita molesta are maintained and assured to be diseasefree) will be required as a term of registration.

Refer to Table 1 in Appendix A for a summary of product chemistry and composition, analysis and certified

limits data. Refer to Table 2 in Appendix A for a summary of physical and chemical characteristics data.

B. Human Health Risk Assessment ( 40 CFR 158.2140 )

1. Toxicity

All Tier I mammalian toxicology data requirements for CpGV-V22 have been fulfilled and support the registration of the propos ed pesticide products. The majority of the required data were cited to studies conducted with CpGV-M, the Mexican isolate of CpGV, an d other baculoviruses. Product analysis data were submitted that demonstrated CpGV-V22 is an isolate of CpGV-M, distinguishable only by molecular genetic methods and bioassays on G. molesta members of the baculovirus gr oup that are clos ely related to CpGV-V22. The basis for citing these studies to fulfill the data requi rements for CpGV-V22 is that all baculoviruses behave similarly (insect-specific) and pres ent no risk to mammalian species. Tier II and Tier III studies were not required. The following summarizes the mammalian toxicology data base for Cydia pomonella granulovirus Isolate V22. (For more information, refer to Table 3 in Appendix A .)

a. Acute Toxicity/Pathogenicity – Tier I

Acute Oral Toxicity/Pathogeni city (OPPTS Harmonized Guide lines 885.3050 & 870.1100) (MRID No. 42517202): In lieu of testing CpGV-V22, Andermatt cited da ta conducted with closel y related CpGV (Mexican Isolate 1964). A distinct clearance pa ttern was evident in the feces and heart/lungs through day 7 when dosed

with 2 x 10 12 granulosis inclusion bodies/ml. CLASSIFICATION: Acceptable, Toxicity Category IV.

Acute Pulmonary Toxicity/Pathogenicity (OPPTS Harmonized Guideline 885.3150) (MRID No. 42517203): In lieu of testing CpGV- V22, Andermatt cited da ta conducted with closel y related CpGV (Mexican Isolate 1964) were cited for Acute Pulm onary Toxicity/Pathogenicity in th e Rat. A distinct clearance pattern was evident in the feces and heart/lungs through day 7 when dosed with 1 x 10 8 granulosis inclusion bodies/ml.

1 For information on microbial data requirements, refer to http://www.epa.gov/fedrgstr/EPAPEST/2007/October/Day-26/p20828.htm

.Cydia pomonella Granulovirus isolate V22 DRAFT Biopesticides Registration Action Document Page 7 of 29 CLASSIFICATION: Acceptable, Toxicity Category IV.

Acute Injection Toxicity/Pathogenicity (Intra venous) – Rat (OPPTS Harmonized Guideline 885.3200) : These data are not required for a virus.

Cell Culture (OPPTS Harmonized Guideline 885.3500) (MRID Nos. 43161202, 48994601 and 48986801): Andermatt cited a study conducted with the CpGV-Mexican isolate that demonstrated that CpGV-Mexican does not infect, replicate in, transfor m or cause toxicity in mammalian cell lines. EPA requested additional information/data to confirm that CpGV-V22 is similar to the CpGV-M1 (“Mexican”) strain. Andermatt supplied a statement of origin that Cydia pomonella granulovirus Isolate V22 is a selection derived from the Cydia pomonella granulovirus Mexico strain. Th ey also provided sequence alignment information for two genes (the granulin gene ( polh/gran ) and late expression factor gene ( lef8 )) and a Restriction Endo nuclease (REN) profile DNA gel that support their statement of origin. This in formation permits bridging data from the previously accepted cell culture study (MRID 431612-02) that had been conducted with the CpGV-Mexican Isolate.

Acute Oral Toxicity – Rat (O PPTS Harmonized Guideline 870.1100 : Andermatt has provided information on the added inerts that indicate they are approved for food incorporation in other uses and do not present a toxicity hazard.

Acute Dermal Toxicity – Rat (OPPTS Harmonized Guideline 870.120) : Andermatt has provided

information on the added inerts that indicat e they do not present a toxicity hazard.

Acute Inhalation Toxicity – Rat (OPPTS Harmonized Guideline 870.1300) : Andermatt has provided

information on the added inerts that indicat e they do not present a toxicity hazard.

Primary eye irritation (OPPTS Harmoniz ed Guideline 870.2400) (MRID No. 48624716): This cited study was conducted in rabb its with Hoe083311 (the closely relate d Mexican isolate). There were no signs of ocular lesions or iritis duri ng the test period. All animals showed slight to moderate discharge and 1of 3 displayed diffuse crimson color at 1-2 hours after instillation. Diffuse cr imson color was observed in 2 of 3 animals at 24 and 48 hours. Positive signs of irrita tion were not significant by day 3 and cleared by day 7.

Chemosis was visible on all animals at 1 hour , but was not significant and cleared by day 3.

CLASSIFICATION: Acceptable, Toxicity category III.

Primary dermal irritation (OPPTS Harm onized Guideline 870.2500) (MRID No. 48624717): This cited study was conducted in ra bbits with Granupom SC, which contains CpGV isolate Hoe083311 (the closely related Mexican isolat e). No animals died during this study, and none showed any signs of erythema or edema. The study was terminated on day 10 after bandage re moval. The lack of any si gns of irritation indicates Granupom SC is Toxicity categor y IV for dermal irritation.

CLASSIFICATION: Acceptable, Toxicity Category IV.

Hypersensitivity Incidents (OPPTS Harmonized Guideline 885.3400) : EPA regulations re quire reporting of hypersensitivity incidents, including immediate and delayed-type reactions in humans and domestic animals, that occur while testing the technical grade of the active ingredient or manufacturing-use product, or are otherwise known to Andermatt (refer to test note #3 of 40 CFR § 158.2140(d)).

b. Subchronic Toxicity/Pathogenicity, Ti er II; Reproductive Fe rtility Effects, Carcinogenicity, Immuno toxicity, and Infectivity/Pa thogenicity Analysis, Tier

IIICydia pomonella Granulovirus isolate V22 DRAFT Biopesticides Registration Action Document Page 8 of 29

Tier II and Tier III studies were not required for Cydia pomonella Granul ovirus Isolate V22, based on the lack of acute toxicity in the Tier I studies. For more inform ation regarding these data re quirements, refer to Table 3

in Appendix A

.

c. Endocrine Disruptors

As required by the Administrator under the Federal Food, Drug, and Cosmetic Act (FFDCA) section 408(p), EPA has developed the Endocrine Di sruptor Screening Program (EDSP) and has begun to implement the screening program that is to be used to test all pesticides to determin e whether certain substances (including pesticide active and other ingr edients) may have an effect in humans or wildlife simi lar to an effect produced by a “naturally occurring estrogen, or other such endocrin e effects as the Administra tor may designate.” FFDCA section 408(p)(4), authorizes the Admi nistrator, by order, to exempt from the requirements of the Endocrine Disruptor Screening Program a biologi c substance or other su bstance if a determination is made that the substance is anticipated not to pr oduce any effect in humans similar to an effect produced by a naturally occurring estrogenic substance. Between October 2009 and February 2010, EPA issued test orders/data call-ins for the first group of 67 chemicals, which contain 58 pest icide active ingredients and nine inert ingredients. CpGV-V22 is not among the group of 58 pesticide active ingredie nts on the initial list to be screened under the

EDSP.

The Agency believes that CpGV-V22 would not likely pr oduce any effect in humans similar to an effect produced by a naturally occurring estrogenic substance. As such, and pursuant to Section 408(p)(4), EPA will determine in the future whether it can exempt CpGV-V22 from the requirements of the Section 408(p) EDSP. In the event the Agency does determine to exempt this substance from the EDSP, an order will be issued. For further information on the status of the EDSP, the policies and procedures, the list of 67 chemicals, future lists, the test guidelines and the Tier 1 screen ing battery, please visit our website: http://www.epa.gov/endo/

.

2. Federal Food, Drug, and Cosmetic Act (FFDCA) Considerations

Section 408(c)(2)(A)(i) of FFDCA allo ws EPA to establish an exemption from the requirement for a tolerance (the legal limit for a pesticide chemi cal residue in or on a food) only if EP A determines that the exemption is “safe.” Section 408(c)(2)(A)(ii) of FFDCA defines “safe” to mean that “t here is a reasonable certainty that no harm will result from aggregate exposure to the pesticid e chemical residue, including all anticipated dietary exposures and all other exposures for which there is reliable information.” This includes exposure through drinking water and in residential se ttings but does not incl ude occupational exposure. Pursuant to section 408(c)(2)(B) of FFDCA, in establishing or maintaining in effect an exemption from the requirement of a tolerance, EPA must take into account the factors se t forth in section 408(b)(2)(C ) of FFDCA, which require EPA to give special consideration to exposure of infants and children to the pesticide chemical residue in establishing a tolerance exemption and to “ensure that ther e is a reasonable certainty that no harm will result to infants and children from aggregate exposure to the pesticide chemical residue....” Additionally, section 408(b)(2)(D) of FFDCA requires that EP A consider “available information concerning the cumulative effects of [a particular pesticide’s] . . . re sidues and other substances that have a common mechanism of toxicity.”

EPA performs a number of analyses to determine the ri sks from aggregate exposure to pesticide residues. First, EPA determines the toxicity of pe sticides. Second, EPA examines expos ure to the pestic ide through food, drinking water, and other routes as a result of pesticide use in residential settings.

Consistent with section 408(b)(2)(D) of FFDCA, EPA reviewed the available sc ientific data and other relevant information, and considered its validity, completeness, and reliability, and the relationshi p of this information toCydia pomonella Granulovirus isolate V22 DRAFT Biopesticides Registration Action Document Page 9 of 29 human risk. EPA also considered ava ilable information concerning the variab ility of the sensitivities of major identifiable subgroups of consumers, including infants and children. Base d on the acute toxi city/pathogenicity data and information discussed previ ously and presented in Table 3 in Appendix A of this document, the data requirements for a human health dietary risk assessment for CpGV-V22 have been fulfilled.

a. Aggregate Exposure

In examining aggregate exposure, section 408 of FFDCA directs EPA to consider available information concerning exposures from the pestic ide residue in food and all other non-occupational expos ures, including drinking water from ground or surface water, pesticid e use in gardens, residential and other lawns and landscapes, and buildings (residentia l and other indoor uses). This aggr egate exposure poten tial for CpGV-V22

is discussed below.

Food Exposure and Risk Characterization : Madex Twin Insecticidal Virus is a liquid insecticide containing 0.06% CpGV-V22. This product will be applied to stone frui t crops grown in agricultur al settings, and to fruit and nut trees in residential settings. Madex Twin Insect icidal Virus will be diluted with water prior to application and applied with spray equipment.

Human exposure to residues of CpGV-V22 in food may occur through the c onsumption of raw agricultural commodities and fruit and nuts grown in residential ga rdens that have been treated with Madex Twin Insecticidal Virus. This baculovirus is commonly f ound in soil and on plant undergro wth, so EPA expects that the dietary exposure to food residues of CpGV-V22 resulting from its pesticidal uses would be negligible when compared to levels of baculovirus humans are already e xposed to in the natural envi ronment. Food exposure to pesticidal residues of the active ingredie nt is expected to be very low due to the dilution of the pesticide prior to application, and rainfall and irrigation will further reduce residues of the pest icide from the treated crops. In the unlikely event that any residues of the pesticide remain in or on consumed food, no adverse effects would be expected, based on the lack of toxicity and path ogenicity demonstrated in the submitted studies.

Drinking Water Exposure and Risk Characterization : Although residues of CpGV-V22 may migrate to groundwater (through runoff from treated food crops) or appear in surfac e water (through runoff and spray drift), residues are expected to be ve ry low since the product is diluted prio r to application. In the unlikely event that any residues of the pesticide are transferred to surface or ground waters intended for eventual human consumption, the sanitation procedures (e.g., chlorinati on, pH adjustments, filtration, high temperatures) in drinking water treatment plants would be expected to remove any remaining residues of CpGV-V22 from the finished drinking water. In the unlikel y event that residues of the pesticide occur in drinking water, no adverse effects would be expected, based upon the lack of toxi city and pathogenicity dem onstrated in the submitted studies (refer to Section IV.B.1.a. of this BRAD).

Non-occupational, Residential Ex posure and Risk Characterization : Madex Twin Insecticidal Virus is labeled for use around the home, or in schools, day care facilities or other such settings, a nd its use may result in nonoccupational residential exposure. N on-occupational residential exposure ma y occur though contact with treated plants. EPA expects the aggregate expos ure to residues of CpGV-V22 to be negligible when compared to the ordinary, everyday levels to which individuals are already exposed in the natural environment, since CpGVV22 is commonly found in soil and on plant undergrowth. In the unlikely event that any residues of the pesticide occur in drinking water even after being pro cessed at a water treatment facility, no adverse effects would be expected, based on the lack of toxicity and pa thogenicity demonstrated in the submitted studies (refer

to Section IV.B.1.a. of this BRAD).

b

. Cumulative Effects from Substances with a Common Mechanism of Toxicity.Cydia pomonella Granulovirus isolate V22 DRAFT Biopesticides Registration Action Document Page 10 of 29

Section 408(b)(2)(D)(v) of FFDCA requi res that, when considering whether to establish, modify, or revoke a tolerance exemption, EPA consider “available informati on concerning the cumulative effects of [a particular pesticide’s] . . . residues and other substances that have a common mechanism of toxicity.”

CpGV-V22 is not known to have a common mechanism of toxici ty with other substances ; therefore, cumulative

effects with other substa nces are not anticipated. For information regarding EPA’s efforts to identify chemicals that may have a common mechanism of toxicity and to evaluate the cumulative effects of such chemicals, see

EPA’s website at http://www.epa.gov/pesticides/cumulative

.

c. Determination of Safety for th e U.S. Population, Infants and Children

FFDCA section 408(b)(2)(C) pr ovides that, in considering the estab lishment of a tolera nce or tolerance exemption for a pesticide chemical residue, EPA sha ll assess the available information about consumption patterns among infants and children, speci al susceptibility of infants and child ren to pesticide chemical residues,

and the cumulative effects on infants and children of the residues and other substances with a common mechanism of toxicity. In addition, FFDCA section 408(b)(2)(C) provides th at EPA shall apply an additional tenfold (10X) margin of exposure (safety) for infants and children in the case of threshold effects to account for prenatal and postnatal toxicity a nd the completeness of the database on toxicity and exposure unless EPA determines that a different margin of exposure (safety) will be safe for infants and children. This additional margin of exposure (safety) is commonly referred to as the Food Quality Protection Act Safety Factor. In applying this provision, EPA either retain s the default value of 10X or uses a different additiona l safety factor

when reliable data available to EPA support the choice of a different factor.

Based on its analyses of the submitted toxicity data (refer to Section IV.B.1.a. and Table 3 in Appendix A

, EPA concludes that there are no th reshold effects of concern to infants, children, or adults when Madex Twin

Insecticidal Virus is used as labeled in accordan ce with good agricultural practices . No toxicological effects or endpoints were identified in mammalian studies. The Agen cy does not expect that infants and children will have special sensitivity to residues of CpGV-V22. As a result, EPA also concludes that no additional margin of exposure (safety) is necessary to pr otect infants and children and that not adding any additional margin of exposure (safety) will be safe for infants and children.

Based on the same data and EPA analysis presented di rectly above, the Agency c oncludes that there is a reasonable certainty that no harm will result to th e U.S. population, including infants and children, from aggregate exposure to th e residues of CpGV-V22 when it is used as labeled and in accordance with good agricultural practices as an insecticide. Such exposure in cludes all anticipated diet ary exposures and all other exposures for which there is reliab le information. EPA arrived at this conclusion because, considered collectively, the data available on CpGV-V22 do not demonstrate toxic potentia l to humans, including infants

and children.

3. Occupational Exposure and Risk Characterization

The available data (e.g., lack of toxi city noted for oral, dermal, and inha lation routes of exposure to CpGV-V22 do not demonstrate toxic poten tial to persons occupationally exposed to this microbial pest control agent. Standard personal protective equipment (PPE) on the labe l, including an N95 equivale nt respirator, long-sleeve shirt, long pants, shoes, socks and waterproof gloves, fu rther mitigates unintentional exposures to applicators.

4. Human Health Risk CharacterizationCydia pomonella Granulovirus isolate V22 DRAFT Biopesticides Registration Action Document Page 11 of 29 EPA considered human exposure to CpGV-V22 in light of the standard for registration in FIFRA and the relevant safety factors in FFDCA. Since there were no toxicological effects or endpoints identified in mammalian studies, a determination has been made that no unreasonable adverse eff ects to the U.S. population in general, and to infants and children in particular, will result when CpGV-V22 pesticide products are used in accordance with EPA-approved labeling.

C. Environmental Assessment ( 40 CFR § 158.2150 )

1. Ecological Hazards

Data and other information (e.g., scientific literature ) submitted by Andermatt to support requests to waive nontarget organism testing for CpGV-V22 are sufficient to fulfill the relevant microbial pesticide data requirements and for risk assessment pur poses. Further testing of nontarget or ganisms at higher tier levels (i.e., Tiers II, III, and IV) is not requir ed. EPA performed an environmental ri sk assessment based on data and other information provided by Andermatt, and has determined that adverse effects to non target organisms are not anticipated from the pesticidal uses of Cydia pomonella granulovirus Isolate V-22. Moreover, EPA has made a

“No Effect” determination for direct a nd indirect effects to listed species and their designated critical habitats for the proposed uses of Cydia pomonella granulovirus Isolate V-22.

For a summary of the generic data requirements de scribed in this section, refer to Table 4 in Appendix A

.

a. Terrestrial Animals and Plants

Avian Oral Toxicity (Harmonized Guid eline 885.4050) (MRID Nos. 48624719 and 48624718): To meet this data requirement, Andermatt cited a study from the public literature (Lautenschlage r et al. 1979; MRID 48624719) and an Organization for Economic Deve lopment (OECD) consensus document (OECD 2002; MRID 48624718). In Lautenschlager et al . (1979), the response of birds to Lymantria dispar nuclear polyhedral virus (NPV) was tested with bird cens uses, an experiment with caged birds, and collection of birds for necropsy and histopathological examination. The study area (location), NPV formula tions, and treatment methods were described in a related paper that wa s not included with the submission. The bird census was carried out using “spot mapping” techniques on two NPV- treated plots (one plot treated w ith NPV mixed with an insecticide base, and one treated with NPV mixed with Sandoz Virus Adjuvant) and one was a cont rol plot. Censuses were conducted at least four times each day during six days in May and June, and four days in July. The daily mean number of male birds of each species was determined. In the caged quail experiment, Northern bobwhite ( Colinus virginianus ) quail were placed in cages on the treated and control plots, and were maintained for three weeks following a second aerial application of NPV. Quails th at died before the end of the trial were sent to the laboratory for necropsy. In addition to the quail, free-ranging birds were co llected weekly by shooting beginning one week after the second NPV treatment a nd continuing until August. These birds were also examined by necropsy, samples were taken for histopat hology, and exposure to NPV was also confirmed via bioassay of alimentary canals and co ntents (methods were not described). The results showed no change in the population for most species, and the author s concluded that changes that did oc cur were the result of the change in habitat due to defoliation by the le pidopteran pest. For the caged quail , no mortality results were reported, even though it was apparent that mortality did oc cur based on the description of methods. For the histopathology and necropsy results, fe w statistical comparisons could be made due to small numbers, but no treatment related differences were apparent.

This study was not particularly useful for examining the effects of the treatment on birds, since sample sizes were limited to one or two plots, a nd results of the treatment were conf ounded with effects of defoliation by the

pest. CLASSIFICATION: Unacceptable.Cydia pomonella Granulovirus isolate V22 DRAFT Biopesticides Registration Action Document Page 12 of 29

The OECD document (OECD 2002), provi des support for a conclusion that a dverse effects are not expected on birds, since baculoviruses are not known to affect vertebrates. CLASSIFICATION: Supplemental. This document as a whole is considered supplemental, but pr ovides acceptable rationale to conclude that effects on birds are not expected from exposure to Cydia pomonella GV Isolate V22.

Avian Inhalation Toxicity Pathogenicity (Harmonized Guideline 885.4100): This study was not required as the nature of the microbial pestic ide does not indicate potential toxici ty or pathogenicity to birds.

Wild Mammals Toxicity/Pathogenici ty (Harmonized Guideline 885.4150): Information has been submitted to fulfill data requirements set forth in 40 CFR § 158.2140 for determining toxic ity/pathogenicity of CpGV

- V22

to mammals. Based on the review of this information discussed in Section IV. B. 1. a. , there are no apparent concerns for toxicity/pathogenicity to wild ma mmals that could potentially be exposed to CpGV

- V22.

Nontarget Insects (Harmonized Guide line 885.4340) (MRID Nos. 48624727, 48624728 and 48624726): A study was submitted in which ground beetles ( Poecilus cupreus L.) were exposed to Cydia pomonella granulovirus SC at 900 mL/hectare (MRID 48624727). The expos ure rate is 1.5X the proposed maximum application rate for Madex Twin Insecticidal Virus. A pplications were made to moistened sand in the test chamber, and beetles were exposed to the granuloviru s through contact with the spray and treated sand. No

mortalities were observed over the 15-day study period; however, a 19. 73% reduction in feeding rate was observed in beetles exposed to Cydia pomonella GV SC. It was not clear whether this reduction was statistically significant, and no other measurements related to surviv al or reproduction were taken. Based on mortality, the

LC 50 was > 900 mL/ha, and it can be concluded that Cydia pomonella granulovirus SC does not cause mortality in this species of ground beetle. CLASSIFICATION: Acceptable.

Andermatt also submitted a study in which predatory mites ( Typhlodromus pyri SCHEUTEN) were exposed to Cydia pomonella granulovirus SC at 900 mL/hectare (MRID 48624728). Adu lt mites were placed in glass chambers sprayed with Cydia pomonella granulovirus SC and were fed pollen (untreated). A positive control (dimethoate) was included. Significant mortality or reduc tions in fecundity were not observed. The study also

included a measurement of reduction in beneficial capacity; however, fu rther details of a threshold of significance in this measure were not provided. While th e study report stated that mortality was observed over the 14-day experimental period, result s were provided only for the first seven days. Therefore, the 7-day LC 50 (concentration at which 50% mortalit y is observed) was determined to be > 900 mL/hectare. The NOEC based on reproduction is 900 mL/hectare. The study was determined to be supplemental because adequate exposure cannot be confirmed. No evidence was provided to dete rmine the certainty of oral exposure. The positive control substance likely affected th e mites via exposure to their integum ent (an organism’s outer protective covering, such as its cuticle or shell) so it may not be a sufficient measure of the adequacy of exposure.

CLASSIFICATION: Supplemental.

A third study was submitted in which aphid parasitoids ( Aphidius rhopalosiphi ) were exposed to Cydia pomonella granulovirus SC at 900 mL/hect are (MRID 486247-26). Adult parasitoids were placed in glass chambers sprayed with Cydia pomonella granulovirus SC and were fed an untreated honey solution. Adults remained in the chambers for 48 hours, after which females were removed and placed into separate chambers consisting of a pot containing 8- 10 oat seedlings infested with approximately 50 - 100 aphids ( Rhopaolsiphum padi ), and covered with gauze. Females remained in these chambers for 24 hours, after which they were removed and aphids in the chambers were observed for parasitism 11 days later. No mortality occurred in the initial 48 hour peri od, so the 48-hour LC 50 was determined to be > 900 mL/hectare. The NOEC (concentration at which no effects were observed) based on reproduction is 900 mL/hectare. The study was determined to be supplemental, because adequate exposure cannot be conf irmed and the study is of short duration. As with theCydia pomonella Granulovirus isolate V22 DRAFT Biopesticides Registration Action Document Page 13 of 29 other study described above, no evidence is provided to de termine the certainty of oral exposure, and 48 hours is not sufficient time to observe pathogenicity of the test substance. CLASSIFICATION: Supplemental.

Additionally, a review paper by Grön er (1990; MRID 48624723) was submitted, which discusses the impact of baculoviruses on nontarget insects. With regard to species selectivity, this paper states that cross-infectivity studies show that few baculoviruses are species speci fic. However, most have narrow host ranges, never exceeding the order and usually not the family of the host insect. Host range is usually limited to the genus of the host insect, and is usually wider fo r nuclear polyhedral viruses than gra nuloviruses. This information is also reflected in the OECD consensus document (OECD 2002). Gröner (1990) cautions that cross-infectivity studies must be interpreted with care, however, because many do not confirm the identity of the virus progeny with the original inoculum. This problem is also described in the OECD consensus document. Furthermore, specific

interactions between host a nd virus are required for successful infect ion, some of which are controlled by a single gene. This study also discusses th e results of tests with certain groups of nontarge t insects. Specifically,

results of several studies with honey bees, including a study with Cydia pomonella granulovirus, show that the baculoviruses described in the paper do not have effects on honey bees. The author also reviewed papers in

which various baculoviruses have been tested on predators and parasitoids of lepidopteran larvae. Studies with predators showed no effects, though so me studies with parasitoids did sh ow a reduction in progeny emerging from infected lepidopteran larvae. The consensus between the results described in th is paper and in the OECD

document is that these effects are most likely indirect due to redu ced food quality of the hos t insect that resulted from infection, rather than a di rect cause of viral infection.

Honey Bees (Harmonized Guideline 885.4380) (MRID Nos. 48624729 and 48624730): Andermatt submitted a honey bee ( Apis mellifera ) acute oral and contact te st with Granupom, containing Cydia pomonella granulovirus, (MRID 48624729). Honey bees were orally ex posed via 50% sucrose food solution or dermally via tap water, each containing Granupom. Mortality was observed in the feeding test for up to 72 hours, at which time 20% mortality occurred in the group receivi ng Granupom and 2% mortality occurred in the control group. No mortality was observed in the group receiving Granupom via cont act. This study is supplemental

because it is of too short duration. Observations should have continued beyond 72 hours for the oral test, since mortality had occurred in the group receiving Granupom. CLASSIFICATION: Supplemental.

Andermatt also submitted a paper by Gröner et al. (1978; MRID 48624730) in which honey bees were exposed

to an isolate of Cydia pomonella granulovirus, or to an isolate of Mamestra brassicae nuclear polyhedral virus. Several exposure routes were tested, including through the diet, contact (via direct wetting or on treated filter paper), and in the “gaseous phase.” A 10-day pathogenici ty test was also performed, in which bees were fed

either once at a rate of 5 x 10 10 Cydia pomonella granulovirus, or daily at that ra te. No significant mortality was observed in any of these tests. This information is useful for the risk assessment for Cydia pomonella granulovirus Isolate V22.

Nontarget Plants (Harmonized Guideline 885.4300) : This study was not required as the microbial pesticide is not taxonomically related to known plant pathogens.

b. Aquatic Animals and Plants

Freshwater Fish (Harmonized Guideline 885.4200) (MRID Nos. 48624720, 48624721 and 48624725): A study to test the toxicity of Cydia pomonella granulovirus SC on freshwater fi sh was performed with rainbow trout ( Oncorhynchus mykiss ; MRID 48624720). This study was a 96-hour sta tic study with five concentrations of 0.01, 0.1, 1.0, 10, and 100 mg/L, using Cydia pomonella granulovirus SC as the test material, which

contained 2.2 x 10 13 occlusion bodies/L. No mortality related to the test material was demonstrated; however, the sublethal effect of a change in pigment coloration wa s observed in all fish at th e highest test concentration.Cydia pomonella Granulovirus isolate V22 DRAFT Biopesticides Registration Action Document Page 14 of 29 Therefore, the LC 50 was determined to be >100 mg/L; the LOEC (l owest concentration at which an effect was observed) was 100 mg/L and the NOEC (concentration at which no effect was observed) was 10 mg/L. This study is supplemental because the duration was only 96 hour s and this is not sufficient to test pathogenicity.

CLASSIFICATION: Supplemental.

Banowetz et al. (1976; MRID 48624721) was also cited to provide informati on on the hazard of baculoviruses to fish. This study was conducted with Orgyia pseudotsugata nuclear polyhedral virus (OpNPV) that infects the Douglas fir tussock moth. In this study, salmoni d fish cell lines (taken from Chinook salmon [ Oncorhynchus tshawytscha ], and steelhead trout [ Onchorhynchus mykiss ] embryos) were exposed to a suspension of nonoccluded virions of OpNPV. Coho salmon ( Oncorhynchus kisutch ), Chinook salmon and steelhead trout fingerlings were exposed to OpNPV via injection, in wate r, and in food. Exposure in food and water occurred at 100 acre-dosages, and lasted for 18 and 24 hours for wa ter and dietary exposure, respectively. Fish were observed for 30 days. Histopathological observations were made on these fish at 30 days. An additional experiment was performed to determine whether the baculovirus persisted in fi sh and, if so, in which organs it

would be found if infection occurred. Coho salmon smolts were exposed via the same routes and durations as the salmonid fingerlings. Initially, inf ected larvae were to be provided as food, but they were not accepted by the fish, even when coated with bovine serum albumin to cause them to sink, so the baculovirus was provided in artificial food. Three fish were samp led at time 0, 12, 24, 48, and 96 hours. Pooled kidneys, livers, and spleens from each sample group were assayed for baculovirus polyh edra. No pathological changes were observed in the cell lines exposed to the OpNPV, and no change occurred in the growth rate s of the cells. A dditionally, there was no evidence that the virus entered the cells. Histopa thological examination showed no evidence of infection by the virus in salmonid fingerlings, and no virus was r ecovered from the fish after 30 days. The experiment with salmonid smolts exposed to virus showed that the virus was inactivated in the fish within 24 hours after exposure, and no effects were observed. It was noted also that even though the fish were deprived of food for five days prior to the experiment, they would not eat the treated larvae, but did not re ject earthworms cut to the

same size. This indicates that there may be avoidance behavior that woul d reduce exposure to baculoviruses.

The Banowetz et al. (1976) study is he lpful to understand that OpNPV (of which there are two types, both of which were tested) does not infect fi sh or cause adverse effects; howev er, no information is available to specifically bridge the results from this study to CpGV-V22 or to Cydia pomonella granulovirus in general. Therefore, this study is considered supplemental without that information. CLASSIFICATION: Supplemental.

Additional information is available in a paper by Gröner et al. (1981; MRID 48624725). The paper provides few details of testing with baculoviruses, but it make s the point that high amounts of baculoviruses have been observed after natural epizoot ics, meaning that vertebrates, including fish, have been exposed under natural conditions without reported effects. The OECD consensus document (O ECD 2002) also states that effects of

baculoviruses on vertebrates have not been observed. Take n together, the weight of the evidence provided by all

of the data submitted shows that lethal effect s are not expected in fish from exposure to Cydia pomonella granulovirus Isolate V22.

Freshwater Invertebrates (Harmonize d Guideline 885.4240) (MRID No. 48624722): A study with Daphnia

magna was submitted to demonstrate the hazard of CpGV-V22 for freshwater invertebrates (MRID 48624722). In this study, 20 Daphnia were included in a treatment group, and treatment groups were exposed to four

concentrations of Cydia pomonella granulovirus SC at 0.1, 1.0, 10, and 100 mg/L. Negative and positive (potassium dichromate at 0.9 mg/L and 1.9 mg/L) controls were also included. Daphnia were exposed via test medium for 48 hours, and observations of mortality/imm obility were made at 24 and 48 hours. Temperature, dissolved oxygen, and pH were also observed at 0 and 48 hours. No effect was observed in the Daphnia exposed to Cydia pomonella granulovirus SC at any concentration, so the LC 50 was determined to be >100Cydia pomonella Granulovirus isolate V22 DRAFT Biopesticides Registration Action Document Page 15 of 29 mg/L. Mortality occurred in positive c ontrols as expected. This study shows that adverse effects to freshwater invertebrates are not expected as a result of exposure to Cydia pomonella granulovirus SC; however, the study is rated supplemental because it is of too short duration. CLASSIFICATION: Supplemental.

Additional information on the potential hazard of baculoviruses to freshw ater invertebrates is provided in Gröner et al. (1981). A ce rtain type of baculovirus is known to occu r in certain species of shrimp; however, no

evidence is available to show that baculoviruses th at are specific to Lepid opterans are pathogenic to crustaceans. Moreover, it is known th at the host range of granuloviruses have narrower host ranges that are often restricted to a single speci es (OECD 2002). Therefore, it is un likely that host range of CpGV-V22 would include aquatic invertebrates, and the information provided indicates th at adverse effects to freshwater invertebrates due to exposure of CpGV-V22 are unlikely.

Estuarine/Marine Fish and Invertebrates (Harmonized Guideline 885.4280), (MRID No. 48624724): A study from the literature (Lightner et al. 1973; MRID 48624724) was submitted in which Autographa californica nuclear polyhedral virus was tested in white shrimp ( Penaeus setiferus L.) juveniles and postlarval brown shrimp ( Penaeus aztecus Ives) to determine whether this nuclear polyhedral viru s is pathogenic to these marine invertebrates. White shrimp were wild caught from West Bay n ear Galveston, Texas; brown shrimp were obtained from artificially propa gated laboratory stock. Exposure occu rred via two methods: injection and orally through artificial feed. White shrimp juveniles were injected once between the first and second segments

with 4.7 x 10 11 virus rods/shrimp. Controls received a saline injection. In a second experiment, postlarval brown shrimp and white shrimp juveniles were fed a diet containing 800 to 8000 nuclear polyhedral virus/μL or approximately 8.8 x 10 2 to 2.5 x 10 8 nuclear polyhedral virus per shrim p. Dietary exposure occurred over 10 days. In both experiments, animals were sampled fo r histopathological examina tion via light and electron microscopy on days 5, 10, 20, and 30 of the experiment, and both experiments terminated at 30 days. No mortality attributable to the nuclear polyhedral virus occurred; however, cont rol mortality for the injected white shrimp was high (46%). No explanation was provide d, and the paper was unclear as to how the authors determined how much mortality wa s significant. Virus was not detect ed in any tissues examined via microscopy, and the authors determined that the nuclear polyhedral virus was not toxic or pathogenic to these two species of shrimp. This study is scientifically sound, and is classified as supplemental because it was

conducted with a different baculovirus. CLASSIFICATION: Supplemental.

Gröner et al. (1981) indicate th at baculoviruses do affect certain species of shrimp within U.S. waters (this is also stated in Gröner [1990] describe d below); however, according to this review there is no knowledge that baculoviruses specific for Lepidoptera a ffect marine or estuarine fish and invertebrates. The OECD consensus document (OECD 2002) also states th at granuloviruses generally have smaller host ranges than nuclear polyhedral viruses, so CpGV-V22 is not likely to include marine/est uarine organisms in its host range. Therefore, the weight of the ev idence indicates that CpGV-V22 is not likely to cause adverse effects to

marine/estuarine fish or invertebrates.

2. Environmental Fate Assessment Data

Environmental fate and groundwater data requirements (Tier II testing) were not triggered because there were no significant adverse effects to nontarget organisms in th e Tier I toxicity studies, and because of acceptable

rationale to address certai n nontarget organism data requirements not fulfilled by testing, as discussed in C.1

. above and summarized in Table 4, Appendix A.

3. Ecological Exposure and Risk Characterization

Madex Twin Insecticidal Virus is proposed for applicati on to tree fruit and nut crops via spray at rates of 0.5 –Cydia pomonella Granulovirus isolate V22 DRAFT Biopesticides Registration Action Document Page 16 of 29 3.0 fluid ounces per acre (agricultural us e) or 1⁄2 teaspoon per 5 gallons, applie d to the point of wetting leaves without runoff (homeowner use). The proposed applicatio n instructions for agricult ural use recommend at least two applications of Madex Twin Insecticidal Virus per codling moth or Oriental fruit moth larval generation, beginning shortly before larvae hatch and feed into fruit; reapplication can be made as necessary. Since Madex

Twin Insecticidal Virus breaks down in direct sunlight, the labe l directs reapplication af ter 6 – 8 sunny days if moths are still flying and laying eggs. Madex Twin Insectic idal Virus can be applied up to and including the day of harvest. The label for homeowner use is slightly different, and recommends Made x Twin Insecticidal Virus to be applied first in late spring or early summer, when eggs of the first generation begin to hatch, and at least twice during each larval ge neration. Additional applic ations (a maximum number is not given) are recommended in mid- to late summer if necessary, and reapplications are suggested after 6 – 8 sunny days if moths are still flying and laying eggs. Applicatio n equipment is not described on either label.

The proposed applications are expected to result in contact of the active i ngredient with foliar and other above ground plant surfaces, and some degree of sp ray drift is expected to occur. The a.i. is expected to be deposited on soil at application and with rainfa ll, and some may move offsite due to runoff. Since reapplication is recommended after 6 – 8 sunny days, and because it is know n that sunlight adversely affects the viability of CpGV (OECD 2002), CpGV-V22 is not expected to persis t at high levels on most su rfaces to which it is applied. Baculoviruses may persist in the soil under some circumstances (OECD 2002, Lightner et al. 1973); however, it is not possible to pred ict the exact environmental fate of CpGV-V22, since environmental conditions can vary. Conditions that aff ect the fate of baculoviruses are di scussed extensively in OECD (2002). Based on the type of application an d expected environmental fate of CpGV-V22, exposure to terrestrial and aquatic nontarget organisms is possible as a re sult of the proposed la beled applications.

A. Terrestrial Animals and Plants

Birds and Mammals

Birds and mammals would be exposed to CpGV-V22 primarily through ingestion of treated prey or plants. Contact with these surfaces is not expected to resu lt in significant exposure, though oral exposure may occur incidentally through grooming. The study cited from the litera ture is not useful for risk assessment for birds; however, other information that is availabl e is sufficient to conclude that CpGV-V22 is unlikely to infect birds

or cause adverse effects. Data with laboratory mammals were also submitted ; bridging information is needed for these studies, but concerns are not raised that would i ndicate potential risk to w ild mammals. Additionally, the OECD consensus document (OECD 2002) indicates that risk to wild ma mmals is unlikely. While birds and mammals may be exposed to CpGV-V22, there is sufficien t evidence to show that th is exposure will not result

in adverse direct effects to birds and mammals. Therefore, risk to birds and mammals is expected to be minimal. The proposed uses of CpGV-V22 are also not expected to significantly affect the food supply for these nontarget animals, because the range of species affected is expected to be narrow and widespread reductions in populations of nontarget insect and othe r arthropods are not anticipated.

Nontarget Insects and Honey Bees

Three studies were submitted with whic h to determine the hazard of CpGV-V22 to nontarget insects (ground beetles [ Poecilus cupreus ], predatory mite [ Typhlodromus pyri ], and parasitoid wasp [ Aphidius rhopalosiphi ]).

Each of these studies used Cydia pomonella granulovirus SC at an applicati on rate that is 1.5X the proposed

maximum rate of Madex Twin Insecticidal Virus. This exposure level is sufficient; however, the method in

which the test material was delivered to the test an imals in two of the studies (with predatory mites and parasitoid wasp) exposure to treated su rfaces) leaves uncertainty as to the ad equacy of these studies for testing nontarget effects with a substa nce that must be ingested. Granuloviruses must be eaten to infect an insect, and itCydia pomonella Granulovirus isolate V22 DRAFT Biopesticides Registration Action Document Page 17 of 29 is not clear from the study reports whether the test animal s actually consumed the test material, or if they did, whether they consumed it at the dose tested. However, th ese insects, and many nontarget insect species that will be in the area where CpGV-V22 is to be used, are not expected to consume the treated plant material. Therefore, the exposures in these studies may be more representative of the type of exposure that would occur in the field, and signify the potential effects unde r the proposed uses. The exception would be cases in which the nontarget insect is a predator or parasite of the target pest, which may be infected with Cydia pomonella granulovirus Isolate V22, thus delivering a higher exposure. Based on these assumptions, these studies are indicative of a lack of expected effect in the field. The study with gr ound beetles is an exception, in that it appears from the study report that they and their initial diet ary items were directly sprayed.

A study was also submitted (MRID 48624729), and another wa s cited from the literature (Gröner et al. 1978) to demonstrate the hazard of CpGV-V22 to honey bees. The experimental study that was submitted for review is

of too short duration to determine wh ether the material tested is pathog enic to honey bees; however, the Gröner et al. (1978) paper te sted an isolate of Cydia pomonella for a longer duration. These studies provide evidence

that Cydia pomonella does not produce adverse effects in honey bees.

Additional information described in Section B above also provides evidence that CpGV-V22 is unlikely to

cause adverse effects in nont arget insects in honey b ees. Granuloviruses generally have a very narrow host range, and no effects of baculoviruses have been obs erved in honey bees (OECD 2002, Gröner 1990). Based on the weight of the evidence attributable to all of the information submitted, adverse effects to nontarget insects and honey bees is not expected as a result of the proposed uses of Cydia pomonella granulovirus Isolate V22.

Nontarget Plants

Baculoviruses are not taxonomi cally related to any known plant pathogens, and nontarget plant data were not required for Cydia pomonella granulovirus Isolate V22. Risk to nontarget plants is not anticip ated as a result of the proposed uses of Cydia pomonella granulovirus Isolate V22.

B. Aquatic Animals and Plants

Freshwater Fish and Invertebrates

Two studies were submitted to assess the risk of CpGV-V22 with freshwater fish an d invertebrates. Based on

these studies the LC 50 for Cydia pomonella granulovirus SC is > 100 mg/L fo r both freshwater fish (rainbow trout) and invertebrates ( Daphnia magna ), although sublethal effects possibly attributable to the test material

were observed in the freshwater fish tested. Based on the maximum agricultu ral use applicati on rate for Madex Twin Insecticidal Virus (3 fl. oz per acre), the exp ected concentration in a 15 cm -deep wetland, assuming direct application, would be 4400 CpGV-V22 occlusion bodies (OB)/mL. The potency of the test material is given in OB/L, and the density (g/mL) is not provided. Howeve r, assuming a reasonable range of densities from 0.5 g/mL to 2.0 g/mL, the LC 50 s determined in the freshwater studies amounts to 1.1 x 10 6 – 4.4 x 10 6 OB/mL. Even if the test material density is outside the estimated range, the LC 50 s and NOECs are well above the estimated environmental concentration, and risk to fr eshwater fish and invert ebrates is not expected. Considering these data, plus the addi tional information from the supplementa l studies that were cited, adverse

effects to freshwater fish and invertebrates are not anticipated.

Marine/Estuarine Fish and Invertebrates

Based on the information cited from the literature, which is discussed above, adverse e ffects to marine/estuarine fish and invertebrates are not expected.Cydia pomonella Granulovirus isolate V22 DRAFT Biopesticides Registration Action Document Page 18 of 29

Aquatic Plants

As discussed above, CpGV-V22 is not related to known plant pathogens. Therefore, adverse effects to aquatic plants are not anticipated.

4. Threatened and Endangered Species Assessment

Since EPA has determined that no effects are anti cipated for nontarget species exposed to CpGV-V22 as a result of the proposed labeled applic ations, effects to federally-listed th reatened and endangered species and their designated critical habitats are also not expected. Therefore, a “No Eff ect” determination is made for direct

and indirect effects to fede rally listed endangered and threatened spec ies and their designated critical habitats for the proposed uses of CpGV- V22.

V. ENVIRONMENTAL JUSTICE

EPA seeks to achieve environmental justice—the fair treatment and mean ingful involvement of all people regardless of race, color, national origin, or income —with respect to the development, implementation, and enforcement of environmental laws, regulations, and po licies. Fair treatment mean s that no group of people, including racial, ethnic, or socioe conomic groups, should bear a dispr oportionate share of the negative environmental consequences resulting from industrial, municipal, and comme rcial operations or the execution of federal, state, local, an d tribal environmental programs and policies. Meaningful involvement means that (1) potentially affected community reside nts have an appropriate opportunity to participat e in decisions about a proposed activity that will affect th eir environment and/or health; (2) the public’s contribution can influence the regulatory agency’s decision; (3) the concerns of all participants involve d will be considered in the decisionmaking process; and (4) the decision -makers seek out and facilitate th e involvement of those potentially affected. EPA has this goal for all communities and persons across the U.S.

To help address potential environmenta l justice issues, EPA seeks information on any groups or segments of the population who, as a result thei r location, cultural practices, or other fa ctors, may have atypical, unusually high exposure to CpGV-V22, compared to the general population. Please comment if you are aware of any subpopulations that may have atyp ical, unusually high exposure compar ed to the general population.

For additional information regarding environmental justice issues, please vi sit EPA’s web site at http://www.epa.gov/compliance/en vironmentaljustice/index.html

.

VI. RISK MANAGEMENT DECISION

Section 3(c)(5) of FIFRA provides for the registrati on of a pesticide provided that all the following determinations are made: (1) its composition is such as to warrant the claims for it; (2) its labeling and other materials required to be submitted comply with the re quirements of FIFRA; (3) it will perform its intended

function without unreasonable adverse effects on the e nvironment; and (4) when used in accordance with widespread and commonly recognized practice, it will not generally cause unreasonable adverse effects on the

environment.

To satisfy criterion 1, the composition of Madex Twin Insecticidal Virus containing CpGV-V22 pesticide product has well-known properties. EPA has no knowledge that would contradi ct the claims made on the label for the end-use pesticide product, Madex Twin Insecticidal Virus, and this pesticide product is not expected to

cause unreasonable adverse effects on the environment when used according to its label instructions. Criterion 2Cydia pomonella Granulovirus isolate V22 DRAFT Biopesticides Registration Action Document Page 19 of 29 is satisfied by the current product labels , as well as the data and information presented in this document. It is

believed that this end us e product containing CpGV-V22 will not cause any unreasonable adverse effects on the environment and is likely to control the caterpillar pe sts described on the product labe ls, satisfying criterion 3.

Criterion 4 is satisfied in that this pesticide product containing CpGV-V22 is not expected to cause unreasonable adverse effects when used according to labe l instructions. Therefore, Madex Twin Insecticidal Virus containing CpGV-V22 as a new active ingredient, is eligible fo r registration under FIFRA section 3(c)(5) to control the apple codling moth, Cydia pomonella and the Oriental fruit moth, Grapholita molesta in apple, quince, pear, peach, nectarine, apricot, plum, prunes, cherry, walnut, almond, pluot and aprium in agricultural and residential settings

VII. ACTIONS REQUIRED OF THE REGISTRANT

A. Final Printed Labeling

Before releasing pesticide products containing CpGV-V22 for shipment, the registrant will be required to provide appropriate final printed labeling to EPA.

B. Terms of Registration

As terms of the registration, Ande rmatt must submit the following:

Results of storage stability (Harmonized Guidelin e 830.6317) and corrosion characteristics (Harmonized Guideline 830.6320) to confirm that the product is active against the Oriental fruit moth, Grapholita molesta must be submitted within one year of registration.

The registrant will be required to provide confirmatory data showing how the laboratory colonies of Cydia pomonella and Grapholita molesta are maintained and assured to be disease free.

C. Reporting of Adverse Effects and Hypersensitivity Incidents

Notwithstanding the information stated in the previous sections, it should be clea rly understood that certain specific data are required to be reported to EPA as a requirement for maintaining the federal registration for a pesticide product. A brief su mmary of these types of da ta are listed below.

Reports of all incidents of adverse effects to the environment must be submitted to EPA under the provisions stated in FIFRA section 6(a)(2). Additionally, a ll incidents of hypersensi tivity (including both suspected and confirmed incidents) must be repo rted to EPA under the provisions of 40 CFR § 158.2140(d).Cydia pomonella Granulovirus isolate V22 DRAFT Biopesticides Registration Action Document Page 20 of 29 VIII. BIBLIOGRAPHY

For definitions of scientific terms, please refer to http://www.epa.gov/pesticides/glossary/

.

A. Studies Submitted to Support the Registrati on of Pesticide Products Containing Cydia pomonella granulovirus isolate V22.

MRID Citation Reference

43161202 Falcon, L.; Berlowitz, A.; Weiss, T. (1994) Baculovirus Cydia pomonella: Cell Culture Tests with Viral Pest Control Agents (Tie r 1) Concurrent Dose Bioassays: Lab Project Number: Z3K001G: UCB-CELL: 1B. Unpublished study prepared by Northview Pacific Laboratories; Insect Pathology Group. 35 p. 43449602 Falcon, L.; Berlowitz, A.; Weiss, T. (1994) Baculovirus Cydia pomonella: Cell Culture Test with Viral Pest Control Agents (Tier 1): Amendment No. 1 to MRID No. 43161202: Lab Project Number: Z3K001G: UCB-CELL: 1B. Unpublished study prepared by University of California, Berkeley and Northview Pacific Labs, Inc. 6 p. 48624701 Smith, F. (2011) Cydia pomonella Granulovirus Isolate V22 Technical: Product Identity, Manufacturing Process, Discussion of the Formati on of Impurities, and Analysis of Samples. Unpublished study prepared by SciReg, Inc. 32p. 48624702 Smith, F. (2011) MADEX Twin: Manufacturing Process, Discussion of the Formation of Impurities, and Certified Limits. Unpublishe d study prepared by SciReg, Inc. 28p. 48624703 Smith, F. (2011) Cydia pomonella Gravulovirus Isolate V22 Technical and MADEX Twin: Summary of Physical-Chemical Properties. Unpub lished study prepared by SciReg, Inc. 5p. 48624704 Andermatt, M. (2000) MADEX: Physical a nd Chemical Properties. Unpublished study prepared by Andermatt Biocontrol AG. 5p. 48624705 Walter, D. (2006) MADEX 3 Year Storage Stability at 5 Degrees Celsius. Project Number: 20041161/01/PCTY. Unpublished study prepared by Eurofins - GAB GmbH. 39p. 48624706 Walter, D. (2004) pH of the Formulation MADEX. Project Number: 20041161/01/PCPH.Unpublished study prepared by GAB Biotechnologie Gmbh. 14p. 48624707 Walter, D. (2004) Relative Density of MADEX. Project Number: 20041161/01/PCRD. Unpublished study prepared by GAB Biotechnologie Gmbh. 17p. 48624708 Smith, F. (2011) Chemical Compatibility of MADEX. Unpublished study prepared by Andermatt Biocontrol AG. 8p.

48624709 Groner; Knauf; Reub (1990) Cydia pomone lla Granulosus Virus (CpGV) Hoe 083311 Summary on Chemical and Physical Data. Project Number: A55428. Unpublished study prepared

b y Andermatt Biocontrol AG. 9p.

48624710 Smith, F. (2011) Summary of Acute Toxicology Studies (Madex Twin). Unpublished study prepared by SciReg, Inc. 21p.

48624711 Hollander (1980) Tolerance Testing of AcNPV Nuclear Polyhedrosis Virus following SingleDose Administration to SPF Wistar Rats. Proj ect Number: 234/80. Unpublished study prepared

b y Hoechst Aktiengesellschaft. 121p.

48624712 Ignoffo, C.; Huang, H.; Shapiro, M.; et al. ( 1975) Insusceptibility of the Rhesus Monkey, Macaca mulatta, to an Insect Virus, Baculovirus heliothis. Environmental Entomology 4:

569-573.

48624713 Britschgi, D. (2011) Request for Waiver of the Cell Culture Study for Cydia pomonella Granulovirus Isolate V22. Unpublished study prepared by Andermatt Biocontrol AG. 425p.

48624714 Hofmann, T. (1992) HOE 083311; Water Miscible Suspension Concentrate; 2.2*10 Exp. 13 Viruses/L: Testing for Respiratory Sensitization in the Male and Female Pirbright WhiteGuinea Pig. Project Number: 90/0689. Unpublished study prepared by Hoechst Aktiengesellschaft. 90p.

48624715 Deihl, K.; Leist, K. (1986) HOE 0833II 0I LC 08 A101 Testing for Sensitizing Properties of Pirbright-White Guinea Pigs by the Method of Landsteiner. Project Number: 86/1169. Unpublished study prepared by Hoechst Aktiengesellschaft. 23p.

48624716 Hack, R. (1993) HOE 08331; Water Miscible Suspension Concentrate; 2.2*10 Exp. 13Cydia pomonella Granulovirus isolate V22 DRAFT Biopesticides Registration Action Document Page 21 of 29 Viruses/L: Testing for Primary Eye Irritation in the Rabbit. Project Number: 93/0485. Unpublished study prepared by Hoechst Aktiengesellschaft. 27p.

48624717 Kramer, H. (1998) Acute Dermal Irritation/Co rrosion: Granupom SC (CpGV) (in Rabbits). Project Number: 98/10/42/829. Unpublished study prepared by Biochem Gmbh. 26p.

48624718 Organisation for Economic Co-operation and Development (2002) Consensus Document on

Information Used in the Assessment of Environmental Applications Involving

Baculoviruses. Series on Harmonization of Re gulatory Oversight in Biotechnology 87p.

48624719 Lautenschlager, R.; Rothenbacher, H.; Podgwai te, J. (1979) Response of Birds to Aerial Application of the Nucleopolyhedrosis Virus of the Gypsy Moth, Lymantria dispar. Environmental Entomology 8: 760-765.

48624720 Heintze, A. (1998) Acute Toxicity Testing of Granulosevirus CpGV SC in Rainbow Trout (Oncorhynchus mykiss) (Teleostei, Salm onidae). Project Number: 96272/01/AAOM. Unpublished study prepared by GAB Biotechnologie Gmbh. 23p.

48624721 Banowetz, G.; Fryer, J.; Iwai, P.; et al. (1 976) Effects of the Douglas-Fir Tussock Moth Nucleopolyhedrosis Virus (Baculovirus) on Thr ee Species of Salmonid Fish. USDA Forrest Service Research Paper PNW-214 8p.

48624722 Heintze, A. (1998) Acute Toxicity Testing of Granulosevirus CpGV SC on Daphnia magna Using the 48 h Acute Immobilization Test. Project Number: 96272/01/AADM. Unpublished study prepared by GAB Biotechnologie Gmbh. 22p.

48624723 Groner, A. (1990) Safety of Nontarget Inverteb rates of Baculoviruses. P. 134-147 in Safety of Microbial Insecticides by A. Groner. Boca Raton, FL: CRC Press, Inc.

48624724 Lightner, D.; Proctor, R.; Sparks, A.; et al. (1973) Testing Penaeid Shrimp for Susceptibility to an Insect Nuclear Polyhedrosis Virus. Environmental Entomology 2: 611-

613.

48624725 Groner, A.; Huber, J.; Krieg, A. (1981) Use of Baculoviruses in Crop Protection: Safety to Aquatic Organisms. Fischwirt 31: 1-4.

48624726 Kuhner, C. (2001) Granulosevirus CpGV SC: Acute Toxicity to the Aphid Parasitoid, Aphidius rhopalosiphi (Hymenoptera, Braconidae) in the Laboratory. Project Number: 96272/01/NLAP. Unpublished study prepared by GAB Biotechnologie Gmbh. 29p.

48624727 Kuhner, C. (1997) Granulosevirus CpGV SC: Acute Toxicity to the Ground Beetle, Poecilus cupreus L. (Coleoptera, Carabidae) in the Laboratory. Project Number: 96272/01/NLPC. Unpublished study prepared by GAB Biotechnologie Gmbh. 31p.

48624728 Kuhner, C. (1998) Granulosevirus CpGV SC: Acute Toxicity to the Predatory Mite, Typhlodromus pyri SCHEUTEN (Acari, Phytoseiidae) in the Laboratory. Project Number: 96272/01/NLTP. Unpublished study prepared by GAB Biotechnologie Gmbh. 29p.

48624729 Kling, A. (2002) Assessment of Side Effects of Granupom to the Honey Bee, Apis mellifera L. in the Laboratory. Project Number: 20011323/01/BLEU. Unpublished study prepared by GAB Biotechnologie Gmbh. 25p.

48624730 Groner, A.; Huber, J.; Krieg, A.; et al. (1978) Testing of Two Baculovirus Preparations on Honey Bees. Nachrichtenbl. Deut. Pflanzenschutzd 30(3): 39-41.

48624731 Arriaga, E. (2011) Evaluate the Efficacy of ABC-V22 against 1st Codling Moth Cydia pomonella (CM) Generation on Apple. Pr oject Number: BB10312, ABC/V22/CM/2010. Unpublished study prepared by Besana Consulting S.A.L. 16p.

48624732 Arriaga, E. (2011) Evaluate the Efficacy of ABC-V22 against Oriental Fruit Moth Grapholita molesta (OFM) on Peach. Project Number: BAI0311, ABCIV22/0FM/2010. Unpublished study prepared b y Besana Consulting S.A.L. 15p. 48624733 Smith, F. (2011) Nontarget Organi sm Summary (Madex Twin). Unpublished study prepared by SciReg, Inc. Sp. 48738101 Smith, F. (2011) Cydia pomonella Granulovirus Isolate V22 Technical: Product Identity, Manufacturing Process, Discussion of the Form ation of Impurities, and Analysis of Samples. Unpublished study prepared by SciReg, Inc. 17p. 48738102 Smith, F. (2011) Madex Twin: Manufacturing Process, Discussion of the Formation of Impurities, and Certified Limits. Unpublishe d study prepared by SciReg, Inc. 13p. 48738103 Smith, F. (2012) Nontarget Organism Discussion (Madex Twin). Unpublished studyCydia pomonella Granulovirus isolate V22 DRAFT Biopesticides Registration Action Document Page 22 of 29

48986801

48994601 prepared by SciReg, Inc. 10p. Damico, J. (2012) Madex Twin: Supplemental Information in Support of MRID Numbers 48624701, 48624709, 48624713, 48624714, 48624715 and 48624716. Unpublished study prepared by SciReg, Inc. 14p. Smith, F. (2012) Madex Twin: Supplemental Information in Support of MRID Numbers 48624701, 48624720, 48624722, 48624726, 48624727, 48624728 and 48624729. Unpublished study prepared by SciReg, Inc. 8p.

B. Environmental Protection Agency Risk Assessment Memoranda

U.S. EPA. 2012a. BPPD Review of Andermatt’s MADEX Twin an inse ct control product based on the

Granulosis Virus of Cydia pomonella Isolate V22. Memorandum from J. Kough, Ph.D. to A. Sibold dated October 31, 2012.

U.S. EPA. 2012b. BPPD Review of Andermatt’s MADE X Twin an insect control product based on the

Granulosis Virus of Cydia pomonella Isolate V22. Memorandum from J. Kough, Ph.D. to A. Sibold dated November 27, 2012.

U.S. EPA. 2012c. Environmental risk assessm ent for the FIFRA Section 3 registration of Cydia pomonella granulovirus Isolate V22. Memorandum from S. Borges to A. Sibold dated November 30, 2012.

U.S. EPA, 2011c. Nucleopolyhedrovirus es and Granuloviruses (Insect Viruses) Preliminary Work Plan and Summary Document Registration Revi ew: Initial Docket. September 26, 2011.

C. Other References

Banowetz, G.M., J.L. Fryer, P.J. Iwai, and M.E. Ma rtignoni. 1976. Effects of the Douglas-fir tussock moth nucleopolyhedrosis virus (baculovirus) on three species of salmonid fish. USDA Forest Service Research Paper PNW-214.

Gröner, A. 1990. Safety of nontarget invert ebrates of baculoviruses. Pp. 136-147 In M. Laird, L.A. Larey, and E.W. Davidson (eds.), Safety of Microbial Insecticides . CRC Press, Boca Raton, Florida, USA.

Gröner, A., J. Huber, and A. Krieg. 1981. Anwendung von Baculoviren im Pflanzenschutz: Unbedenklichkeit für aquatische Organismen. Fischwaid 31: 25-2 7. (Translated document provided by Andermatt).

Gröner, A., J. Huber, A. Krieg, and W. Pinsdorf. 1978. Bienenprüfung von zwei Baculovirus-Präparaten.

Nachrichtenbl. Deut. Pflanzenschutzd. 30: 39-41. (Translated document provided by Andermatt).

Lautenschlager, R.A., H. Rothenbacher, and J.D. Podgwa ite. 1979. Response of birds to aerial applica tion of the nucleopolyhedrosis virus of the gypsy moth, Lymantria dispar . Environmental Entomology 8: 760-764.

Lightner, D.V., R.R. Proctor, A.K. Sparks, J.R.Adam s, and A.M. Heimpel. 1973. Testing Penaeid shrimp for susceptibility to an insect nuclear polyhedros is virus. Environmental Entomology 2: 611-613.

Organization for Economic Cooperation and Devel opment (OECD). 2002. Series on Harmonization of Regulatory Oversight in Biotec hnology, No. 20: Consensus document on information used in the assessment of environmental applic ations involving baculoviruses. Publication Number: ENV/JM/MONO (2002)1. Paris, France.APPENDIX A. MICROBIAL PESTICIDES DATA REQUIREMENTS(40 CFR PART 158– SUBPART V)TABLE 1. Product Analysis Data Requirements for Granulosis Virus ofCydia pomonellaGranulovirusIsolate V-22, includingthe End-Use Pesticide Product (EP) Madex Twin (40 CFR § 158.2120)HarmonizedGuidelineNumberData Requirement ResultsMRID No.885.1100Product IdentitySupplemental data submitted by Andermatt were upgraded to acceptable based on additional informationprovided by the Ccmpany. This sequence alignment information suggests that there has been no alteration inthepolh/granorlef 8genes and that Isolate V22, along with the registered CpGV-Mexican isolate 1964, isgenome type A.48624701486247044862470648624707486247134898680148994601885.1200Manufacturing ProcessAnin vivoproduction system was described. No details of how the laboratory colonies ofCydia pomonellaandGrapholita molestaare maintained and assured to be disease-free. Confirmatory data will be required as aterm of registration.885.1250Deposition of a Samplein a NationallyRecognized CultureCollectionThe Andermatt has depositedCydia pomonellaIsolate V-22 in the Deutsche Sammlung vonMikroorganismen und Zellculturen GmbHunder accession number DSMZ GV-0014.885.1300Discussion of Formationof UnintentionalIngredientsThe absence of microbial contaminants at levels appropriate for baculovirus based products is in accordancewith the OECD document on that topic except there is no mention of spore-forming bacterial contaminants,especiallyBacillus cereus.Isolates ofBacillus cereuscan be an important cause of bacterial food poisoningand some measure should be added to the quality control procedures to characterize its presence in thisproduct. Confirmatory data will berequired as a term of registration.885.1400 Analysis of Samples Submitted data fulfill the requirementfor analysis of samples. As a term of registration, EPA isrequiringsubmission of a storage stability and corrosion characteristics study within one year of registration.885.1500 Certification of LimitsCertified Limits are acceptable.TABLE 2. Physical and Chemical Characteristics for the Product Madex Twin, ContainingCydia pomonellaGranulovirusIsolate V22(40 CFR § 158.2120)HarmonizedGuidelineNumberData Requirement ResultsMRID No.Cydia pomonellaGranulovirusisolate V22DRAFT Biopesticides Registration Action DocumentPage 24 of 29TABLE 2. Physical and Chemical Characteristics for the Product Madex Twin, ContainingCydia pomonellaGranulovirusIsolate V22(40 CFR § 158.2120)HarmonizedGuidelineNumberData Requirement ResultsMRID No.830.63021Color GreyBrown4862470348624704486247054862470648624707486247094898680148994601830.63031Physical State Liquid830.63041Odor Odorless830.63131Stability to Normal andElevated Temperatures,Metals, and Metal IonsThe information that was submitted was supplemental, but does not need to be upgraded. As a term of theregistration, EPA is requiring submission of storage stability and corrosion characteristics data within oneyear of registration to confirm thatthe active ingredient maintains its activity against the Oriental fruit flyGrapholita molesta.830.6317 Storage StabilityAResults from a 3-year storage stability study forMADEX were submitted. Minimal container weightchanges (<0.5%) over 24 months probably represent a loss due to evaporation of the contents. The 36 monthsresults were not provided. If there are data that demonstrate no decline in activity for 36 months, then alonger storage stability statement could be considered. As a term of the registration, storage stability andcorrosion characteristics data within one year of registration to confirm that the active ingredient maintains itsactivity against the Oriental fruit flyGrapholita molesta.830.63192Miscibility Only required for emulsifiable liquid formulations of microbial pesticides.830.6320CorrosionCharacteristicsOnly required when microbial pesticides are packaged inmetal, plastic, or paper containers. As a term of theregistration, EPA is requiring submission of a storagestability and corrosion characteristics study within oneyear of the date the product is registered.830.70001pHThe mean pH value of a 1%w/v aqueous dilution is 7.0+0.2.830.71003ViscosityThe viscosity is 1,200 mPascal-second.830.7300Density/RelativeDensity/Bulk Density(Specific Gravity)The relative density was determined to beρ19.94.0= 1.160831According to 40 CFR § 158.2120, these data are only required for the technical grade of the active ingredient.Cydia pomonellaGranulovirusisolate V22DRAFT Biopesticides Registration Action DocumentPage 25 of 292According to 40 CFR § 158.2120, these data are only required for emulsifiable liquid forms of microbial pesticides.3According to 40 CFR § 158.2120, these data are only required for liquid forms of microbial pesticides.TABLE 3. Toxicology Data Requirements for Granulosis Virus ofCydia pomonellaIsolate V-22, includingthe End-Use Pesticide Product (EP) Madex Twin (40 CFR § 158.2140)HarmonizedGuidelineNumberDataRequirementResults MRID No.Tier I885.30501Acute OralToxicity/PathogenicityData conducted with closely related CpGV (Mexican Isolate 1964) were cited for Acute Oral Toxicity/Pathogenicityin the Rat. Adistinct clearance pattern was evident in the feces and heart/lungs through day 7 when dosed with 2 x 1012granulosis inclusionbodies/ml by oral gavage.Classification: Acceptable, Toxicity Category IV.42517202885.31501Acute PulmonaryToxicity/PathogenicityData conducted with closely related CpGV (Mexican Isolate 1964) were cited for Acute Pulmonary Toxicity/Pathogenicity in theRat. A distinct clearancepattern was evident in the feces andheart/lungs through day 7 when 1 x 108granulosis inclusion bodieswere administered intratracheally.Classification: Acceptable, Toxicity Category IV.42517203885.3200Acute InjectionToxicity/Pathogenicity(Intravenous)Not required for a virus (See 40 CFR§ 158.2140, test note 2)N/A885.3400HypersensitivityIncidentsHypersensitivity incidents, including immediate-type and delayed-type reactionsof humans or domestic animals, that occur duringthe testing or production of the TGAI or EP, or are otherwise known to Andermatt, must be reported if they occur (See 40 CFR §158.2140(d), test note 3).N/A885.3500 Cell CultureAndermatt cited a study conducted with theCpGV-Mexican isolate that demonstrated that CpGV-Mexican does not infect, replicatein, transform or cause toxicity in mammalian cell lines. EPA requested additional information/data to confirm that CpGV-V22 issimilar to the CpGV-M1 (“Mexican”) strain. Andermatt supplied a statement of origin thatCydia pomonellagranulovirus IsolateV22 is a selection derived from theCydia pomonellagranulovirus Mexico strain. They also provided sequence alignmentinformation for two genes (the granulin gene (polh/gran) and late expression factor gene (lef8)) and a Restriction Endonuclease(REN) profile DNA gel that support their statement of origin. This information permits bridging data from the previously acceptedcell culture study (MRID 431612-02) that had beenconducted with the CpGV-Mexican Isolate.Classification: Acceptable.48624713431612024898680148994601870.1100Acute OralToxicityAndermatt has provided information on the added inerts that indicate they are approvedfor food incorporation in other uses anddonot present a toxicity hazard. (See 40 CFR 158.2040, test note 1).N/A870.12003Acute dermaltoxicityAndermatt has provided information on the added inerts that indicate they are approvedfor food incorporation in other uses anddonot present a toxicity hazard. (See 40 CFR 158.2040, test note 1).N/A870.13003Acute inhalationtoxicityAndermatt has provided information on the added inerts that indicate they are approvedfor food incorporation in other uses anddonot present a toxicity hazard. (See 40 CFR 158.2040, test note 1).N/ACydia pomonellaGranulovirusisolate V22DRAFT Biopesticides Registration Action DocumentPage 26 of 29TABLE 3. Toxicology Data Requirements for Granulosis Virus ofCydia pomonellaIsolate V-22, includingthe End-Use Pesticide Product (EP) Madex Twin (40 CFR § 158.2140)HarmonizedGuidelineNumberDataRequirementResults MRID No.870.2400 Eye IrritationThis cited study was conducted in rabbits with Hoe083311 (the closely related Mexican isolate). There were no signs of ocularlesions or iritis during the test period. All animals showed slight to moderate discharge and1of 3 displayed diffuse crimsoncolor at1-2 hours after instillation. Diffuse crimson color was observed in2 of 3 animals at 24 and 48 hours. Positive signs of irritationwere not significant by day 3 and clearedby day 7. Chemosis was visible on all animals at 1 hour, but was not significant andcleared by day 3.Classification:Acceptable.Toxicity category III.48624716870.2500 Dermal IrritationThis cited study was conducted in rabbitswith Granupom SC, which contains CpGVisolate Hoe083311 (the closely relatedMexican isolate). No animals died during thisstudy, and none showed any signs of erythema or edema. The study was terminatedon day 10 after bandage removal.The lack of any signs of irritation indicates GranupomSC is Toxicity category IV for dermalirritation.Classification:Acceptable.Toxicity Category IV.48624717Tiers II and IIINot required forGranulosis Virus ofCydia pomonellaIsolate V-22, based on the lack of acute toxicity in the Tier I studies.Table 4. Non-target Organism Toxicity Requirements forCydia pomonellaGranulovirus Isolate V22 (40 CFR 158.2150)Requirement (OPPTSGuideline No.)MRID Toxicity and ClassificationAvian oral toxicity(885.4050)486247194873810348624718A study cited from literature (Lautenschlager et al. 1979) showed no effects ofLymantria disparNPV on wild caught birds andcaged quail.Classification: UnacceptableAn additional volume was submitted that provided a summary, but no new information on birds.Classification: SupplementalThe OECD consensus document (OECD 2002) contains a review of information and states that effects on birds are not expectedfrom exposure to baculoviruses.Classification: Supplemental (acceptable for avian risk assessment)Avian inhalationtoxicity/pathogenicity(885.4100)N/A Not required: the nature of the microbial pesticide does not indicate potential toxicity or pathogenicity to birds.Wild mammaltoxicity/pathogenicity(885.4150)N/A Tests required by 40 CFR § 158.2140 are adequate and appropriate for assessment of hazards to wild mammals. The review ofinformation submitted to fulfill those data requirements does not indicate concerns for adverse effects on wild mammals.Classification: Acceptable forenvironmental risk assessmentFreshwater fish toxicity/pathogenicity (885.4200)48624720486247214862472548624718The LC50for rainbow trout (Oncorhynchus mykiss) in a 96-hour study was > 100 mgCydia pomonellaGV SC/L. At 100 mg/L, allfish showed a change in pigment to darker color, so the NOEC was determined to be 10 mg/L. The study is of too short duration todetermine pathogenicity.Classification: SupplementalA study withOrygia pseudotsugataNPV (Banowetz et al. 1976) showed no pathological or growth rate changes in salmonid fishcell lines exposed to this virus. Salmonid fingerlings and smolts showed no adverse effects with exposure through injection, inwater, and in food, and virus was inactivated within the fish within 24 hours after exposure. Rationale to bridge data on thisNPV toCydia pomonellaGV Isolate V22 is needed.Classification: SupplementalA study cited from the literature (Gröner et al. 1981) and the OECD consensus document (OECD 2002), provide information toshow that baculoviruses are generally understood not to be toxic or pathogenic to fish.Classification: SupplementalCydia pomonellaGranulovirusisolate V22DRAFT Biopesticides Registration Action DocumentPage 28 of 29Freshwater invertebratetesting(885.4240)486247224862472548624718A 48-hour study withCydia pomonellaGV SC demonstrated a LC50of > 100mg/L inDaphnia magna. The study is of too shortduration to assess pathogenicity.Classification: SupplementalA study cited from the literature (Gröner et al. 1981) and the OECD consensus document provide additional information thatbaculoviruses specific to Lepidoptera are not expected to cause adverse effects in freshwater invertebrates.Classification: SupplementalEstuarine/Marine fishtestingEstuarine/Marineinvertebrate testing(885.4280)486247244862472548624718A study was cited from the literature (Lightner et al.1973) that showed no effect and no infectivity ofAutographa californicaNPVon white (Penaeus setiferus) and brown (Penaeus azteca) shrimp. Rationale to bridge this NPV toCydia pomonellaGV IsolateV22 is needed.Classification: SupplementalA study cited from the literature (Gröner et al. 1981) providesadditional information that baculoviruses specific to Lepidoptera arenot expected to cause adverse effects in marine/estuarine fish or invertebrates. Additional discussion in the OECD consensusdocument supports this conclusion.Classification: SupplementalNontarget Plant Testing(885.4300)N/A Not required. The microbial pesticide is not taxonomically related to known plant pathogens.Nontarget Insect Testing(885.4340)48624727486247284862472648634723The 15-day LC50was > 900 ml/ha forCydia pomonellaGV SC in the ground beetle (Poecilus cupreusL.), which was 1.5X themaximum application rate for Madex Twin.Classification: AcceptableThe 7-day LC50forCydia pomonellaGV SC was > 900 ml/ha for the predatory mite (Typhlodromus pyriSCHEUTEN); the NOECfor reproduction was 900 mL/ha. Exposure route raises questions as to actual exposure level.Classification: SupplementalThe 48-hour LC50forCydia pomonellaGV SC was > 900 ml/ha for the parasitoid wasp (Aphidius rhopalosiphi); the NOEC forreproduction was 900 mL/ha. Exposure duration was too short to determine pathogenicity; exposure route raises questions as toactual exposure level.Classification: SupplementalA study (Gröner 1990) was cited that provided additional evidence thatCydia pomonellaGV Isolate V22 is unlikely to affectnontarget insect species.Classification: SupplementalHoneybee Testing(885.4380)4862472948624730The 72-hour acute oral LC50was > 3.6 x 107granula per bee, and the 48-hour acute contact LC50was >4.4 x 107granula per bee.This study is of too short duration to assess pathogenicity.Classification:SupplementalA study by Gröner et al. (1978) was cited, in which the authors tested an isolate ofCydia pomonellaGV and an isolate ofMamestra brassicaeNPB on bees via several routes. A 10-day pathogenicity test with honey bees at a daily dose of 5 x 1010granula/mL resulted in no significant mortality. Other results showing lack of effects of baculoviruses on bees were also discussed.Classification: SupplementalCydia pomonellaGranulovirusisolate V22DRAFT Biopesticides Registration Action DocumentPage 29 of 29APPENDIX B. PESTICIDE PRODUCTEPA FileSymbolRegistration NamePercentageActiveIngredientFormulation TypeUse Site(s)Method(s) ofApplicationApplicationRateTarget Pest(s)69553-1 Madex Twin 0.06%Contains aminimumof 3 x 1013viralocclusionbodies perliterDilutable Liquid Agricultural andresidential use.Spray 0.5 -3.0 fluidounces ofMadex Twinper acrediluted in 50-100 gallons ofwater peracre.Larvae of the Apple codling mothCydiapomonellaand the Oriental fruit mothGrapholita molesta.