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, 2012 

 
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TABLE OF CONTENTS 

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

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

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

A. Application for Pesticide Registration ................................................................................................................. 5

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

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

A. Product Analysis Assessment (40 CFR § 158.2120) ........................................................................................... 5

B. Human Health Risk Assessment (40 CFR 158.2140) .......................................................................................... 6

C. Environmental Assessment (40 CFR § 158.2150) ............................................................................................. 11

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

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

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

A. Final Printed Labeling ........................................................................................................................................ 19

B. Terms of Registration ......................................................................................................................................... 19

C. Reporting of Adverse Effects and Hypersensitivity Incidents ......................................................................... 19

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

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

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

B. Environmental Protection Agency Risk Assessment Memoranda .................................................................. 22

C. Other References ................................................................................................................................................. 22

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

APPENDIX B. PESTICIDE PRODUCT ................................................................................. 29

 
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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, MPB 
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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 granulovirus (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 butterflies. 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 multiplies rapidly within its host, causing an infection 

resulting in cell destruction and death, typically within 3-7 days. 

One microbial pesticide product containing CpGV-V22, Madex Twin Insecticidal Virus (EPA file symbol 

69553-R) is proposed for registration. Madex Twin Insecticidal Virus is a dilutable end-use product containing 

0.06% CpGV-V22. It is labeled for application to apple, quince, pear, peach, nectarine, apricot, plum, prunes, 

cherry, walnut, almond, pluot, and aprium in agricultural and residential settings to control apple codling moth 

and Oriental fruit moth. It is applied 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 harmful health effects to humans are expected 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 and 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 actions 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 product will result in the first outdoor and food uses for CpGV-V22. The 

following documents are being made available for comment in the docket, identification number EPA-HQOPP-2011-0984, and accessed through either http://www.regulations.gov or 

http://www.epa.gov/pesticides/regulating/registration-status.html: (1) draft Cydia pomonella granulovirus

isolate V22 (CpGV-V22) Biopesticides Registration Action Document (BRAD); (2) draft label for the end-use 

pesticide product, Madex Twin Insecticidal Virus. 

The public participation comment period is being initiated 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/.

 
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II. ACTIVE INGREDIENT OVERVIEW 

Biological Name: Cydia pomonella granulovirus isolate V22

Culture Collection: Deposited at the DSMZ Plant Virus Collection 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 this 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 microbial 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 apple 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 and aprium in agricultural and 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 announced receipt of an application for 

registration of this pesticide product for agricultural and residential 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 established for residues of the microbial pest control agent 

Occlusion Bodies of the 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) 
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Microbial pesticide product analysis data requirements1

 include product chemistry and composition, analysis 

and certified limits, and physical and chemical characteristics data. Product chemistry and composition data 

include information about the identity of the active ingredient, the manufacturing process, deposition of a 

sample of the microbial active ingredient in a nationally recognized culture collection, and discussion of the 

potential for formation of unintentional ingredients. Analysis and certified limits data include information on 

analysis of samples and certification of limits. Physical and chemical characteristics data describe basic 

characteristics of the registered pesticide products, including 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 new 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 proposed pesticide products. The majority of the required data were cited to studies 

conducted with CpGV-M, the Mexican isolate of CpGV, and 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 group that are closely related to CpGV-V22. 

The basis for citing these studies to fulfill the data requirements for CpGV-V22 is that all baculoviruses behave 

similarly (insect-specific) and present 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/Pathogenicity (OPPTS Harmonized Guidelines 885.3050 & 870.1100) (MRID No. 

42517202): In lieu of testing CpGV-V22, Andermatt cited data conducted with closely related CpGV (Mexican 

Isolate 1964). A distinct clearance pattern was evident in the feces and heart/lungs through day 7 when dosed 

with 2 x 1012 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 data conducted with closely related CpGV (Mexican 

Isolate 1964) were cited for Acute Pulmonary Toxicity/Pathogenicity in the Rat. A distinct clearance pattern 

was evident in the feces and heart/lungs through day 7 when dosed with 1 x 108 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. 
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CLASSIFICATION: Acceptable, Toxicity Category IV. 

Acute Injection Toxicity/Pathogenicity (Intravenous) – 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, transform 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. They also provided sequence alignment information 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 

accepted cell culture study (MRID 431612-02) that had been conducted with the CpGV-Mexican Isolate. 

Acute Oral Toxicity – Rat (OPPTS 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 indicate 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 indicate they do not present a toxicity hazard. 

Primary eye irritation (OPPTS Harmonized Guideline 870.2400) (MRID No. 48624716): 

This cited study was conducted in rabbits with Hoe083311 (the closely related Mexican isolate). There were no 

signs of ocular lesions or iritis during the test period. All animals showed slight to moderate discharge and 1of 

3 displayed diffuse crimson color at 1-2 hours after instillation. Diffuse crimson color was observed in 2 of 3 

animals at 24 and 48 hours. Positive signs of irritation 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 Harmonized Guideline 870.2500) (MRID No. 48624717): 

This cited study was conducted in rabbits with Granupom SC, which contains CpGV isolate Hoe083311 (the 

closely related Mexican isolate). No animals died during this study, and none showed any signs of erythema or 

edema. The study was terminated on day 10 after bandage removal. The lack of any signs of irritation indicates 

Granupom SC is Toxicity category IV for dermal irritation. 

CLASSIFICATION: Acceptable, Toxicity Category IV. 

Hypersensitivity Incidents (OPPTS Harmonized Guideline 885.3400): EPA regulations require 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, Tier II; Reproductive Fertility Effects, 

 Carcinogenicity, Immunotoxicity, and Infectivity/Pathogenicity Analysis, Tier 

 III 
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Tier II and Tier III studies were not required for Cydia pomonella Granulovirus Isolate V22, based on the lack 

of acute toxicity in the Tier I studies. For more information regarding these data requirements, 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 Disruptor Screening Program (EDSP) and has begun to implement the 

screening program that is to be used to test all pesticides to determine whether certain substances (including 

pesticide active and other ingredients) may have an effect in humans or wildlife similar to an effect produced by 

a “naturally occurring estrogen, or other such endocrine effects as the Administrator may designate.” FFDCA 

section 408(p)(4), authorizes the Administrator, by order, to exempt from the requirements of the Endocrine 

Disruptor Screening Program a biologic substance or other substance if a determination is made that the 

substance is anticipated not to produce 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 pesticide active ingredients and nine inert ingredients. 

CpGV-V22 is not among the group of 58 pesticide active ingredients on the initial list to be screened under the 

EDSP. 

The Agency believes that CpGV-V22 would not likely produce 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 screening 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 allows EPA to establish an exemption from the requirement for a tolerance 

(the legal limit for a pesticide chemical residue in or on a food) only if EPA determines that the exemption is 

“safe.” Section 408(c)(2)(A)(ii) of FFDCA defines “safe” to mean that “there is a reasonable certainty that no 

harm will result from aggregate exposure to the pesticide 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 settings but does not include 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 set 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 there 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 EPA consider “available information concerning the cumulative effects of 

[a particular pesticide’s] . . . residues and other substances that have a common mechanism of toxicity.” 

EPA performs a number of analyses to determine the risks from aggregate exposure to pesticide residues. First, 

EPA determines the toxicity of pesticides. Second, EPA examines exposure to the pesticide 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 scientific data and other relevant 

information, and considered its validity, completeness, and reliability, and the relationship of this information to 
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human risk. EPA also considered available information concerning the variability of the sensitivities of major 

identifiable subgroups of consumers, including infants and children. Based on the acute toxicity/pathogenicity 

data and information discussed previously 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 pesticide residue in food and all other non-occupational exposures, including 

drinking water from ground or surface water, pesticide use in gardens, residential and other lawns and 

landscapes, and buildings (residential and other indoor uses). This aggregate exposure potential 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 fruit crops grown in agricultural settings, and to fruit 

and nut trees in residential settings. Madex Twin Insecticidal 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 consumption of raw agricultural 

commodities and fruit and nuts grown in residential gardens that have been treated with Madex Twin 

Insecticidal Virus. This baculovirus is commonly found in soil and on plant undergrowth, 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 exposed to in the natural environment. Food exposure to 

pesticidal residues of the active ingredient 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 pesticide 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 pathogenicity 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 surface water (through runoff and spray 

drift), residues are expected to be very low since the product is diluted prior 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., chlorination, 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 unlikely event that residues of the pesticide occur in drinking water, no adverse 

effects would be expected, based upon the lack of toxicity and pathogenicity demonstrated in the submitted 

studies (refer to Section IV.B.1.a. of this BRAD). 

Non-occupational, Residential Exposure and Risk Characterization: Madex Twin Insecticidal Virus is labeled 

for use around the home, or in schools, day care facilities or other such settings, and its use may result in nonoccupational residential exposure. Non-occupational residential exposure may occur though contact with treated 

plants. EPA expects the aggregate exposure 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 processed at a water treatment facility, no adverse effects 

would be expected, based on the lack of toxicity and pathogenicity 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. 
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Section 408(b)(2)(D)(v) of FFDCA requires that, when considering whether to establish, modify, or revoke a 

tolerance exemption, EPA consider “available information 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 toxicity with other substances; therefore, cumulative 

effects with other substances 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 the U.S. Population, Infants and Children 

 

FFDCA section 408(b)(2)(C) provides that, in considering the establishment of a tolerance or tolerance 

exemption for a pesticide chemical residue, EPA shall assess the available information about consumption 

patterns among infants and children, special susceptibility of infants and children 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 that 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 and 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 retains the default value of 10X or uses a different additional 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 threshold effects of concern to infants, children, or adults when Madex Twin 

Insecticidal Virus is used as labeled in accordance with good agricultural practices. No toxicological effects or 

endpoints were identified in mammalian studies. The Agency 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 protect 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 directly above, the Agency concludes that there is a 

reasonable certainty that no harm will result to the U.S. population, including infants and children, from 

aggregate exposure to the residues of CpGV-V22 when it is used as labeled and in accordance with good 

agricultural practices as an insecticide. Such exposure includes all anticipated dietary exposures and all other 

exposures for which there is reliable information. EPA arrived at this conclusion because, considered 

collectively, the data available on CpGV-V22 do not demonstrate toxic potential to humans, including infants 

and children. 

3. Occupational Exposure and Risk Characterization

The available data (e.g., lack of toxicity noted for oral, dermal, and inhalation routes of exposure to CpGV-V22

do not demonstrate toxic potential to persons occupationally exposed to this microbial pest control agent. 

Standard personal protective equipment (PPE) on the label, including an N95 equivalent respirator, long-sleeve 

shirt, long pants, shoes, socks and waterproof gloves, further mitigates unintentional exposures to applicators. 

4. Human Health Risk Characterization 
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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 effects 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 purposes. Further testing of nontarget organisms at higher tier levels (i.e., 

Tiers II, III, and IV) is not required. EPA performed an environmental risk assessment based on data and other 

information provided by Andermatt, and has determined that adverse effects to nontarget 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 and 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 described in this section, refer to Table 4 in Appendix A. 

 a. Terrestrial Animals and Plants 

Avian Oral Toxicity (Harmonized Guideline 885.4050) (MRID Nos. 48624719 and 48624718): 

To meet this data requirement, Andermatt cited a study from the public literature (Lautenschlager et al. 1979; 

MRID 48624719) and an Organization for Economic Development (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 censuses, an experiment with caged birds, and collection of birds for necropsy 

and histopathological examination. The study area (location), NPV formulations, and treatment methods were 

described in a related paper that was not included with the submission. The bird census was carried out using 

“spot mapping” techniques on two NPV-treated plots (one plot treated with NPV mixed with an insecticide 

base, and one treated with NPV mixed with Sandoz Virus Adjuvant) and one was a control 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 that died before the end of the trial were sent to the 

laboratory for necropsy. In addition to the quail, free-ranging birds were collected weekly by shooting 

beginning one week after the second NPV treatment and continuing until August. These birds were also 

examined by necropsy, samples were taken for histopathology, and exposure to NPV was also confirmed via 

bioassay of alimentary canals and contents (methods were not described). The results showed no change in the 

population for most species, and the authors concluded that changes that did occur were the result of the change 

in habitat due to defoliation by the lepidopteran pest. For the caged quail, no mortality results were reported, 

even though it was apparent that mortality did occur based on the description of methods. For the 

histopathology and necropsy results, few 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, and results of the treatment were confounded with effects of defoliation by the 

pest. CLASSIFICATION: Unacceptable. 
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The OECD document (OECD 2002), provides support for a conclusion that adverse 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 provides 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 pesticide does not indicate potential toxicity or pathogenicity to birds. 

Wild Mammals Toxicity/Pathogenicity (Harmonized Guideline 885.4150): Information has been submitted 

to fulfill data requirements set forth in 40 CFR § 158.2140 for determining toxicity/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 mammals that could potentially be exposed to CpGV- V22. 

Nontarget Insects (Harmonized Guideline 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 exposure rate is 1.5X the proposed maximum 

application rate for Madex Twin Insecticidal Virus. Applications were made to moistened sand in the test 

chamber, and beetles were exposed to the granulovirus 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 survival or reproduction were taken. Based on mortality, the 

LC50 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). Adult 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 reductions in fecundity were not observed. The study also 

included a measurement of reduction in beneficial capacity; however, further details of a threshold of 

significance in this measure were not provided. While the study report stated that mortality was observed over 

the 14-day experimental period, results were provided only for the first seven days. Therefore, the 7-day LC50

(concentration at which 50% mortality 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 determine the certainty of oral exposure. The positive 

control substance likely affected the mites via exposure to their integument (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/hectare (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 period, so the 48-hour LC50 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 confirmed and the study is of short duration. As with the 
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other study described above, no evidence is provided to determine 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 Gröner (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 specific. 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 for nuclear polyhedral viruses than granuloviruses. 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 and virus are required for successful infection, some of which are controlled by a 

single gene. This study also discusses the results of tests with certain groups of nontarget 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 some studies with parasitoids did show a reduction in progeny emerging 

from infected lepidopteran larvae. The consensus between the results described in this paper and in the OECD 

document is that these effects are most likely indirect due to reduced food quality of the host insect that resulted 

from infection, rather than a direct 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 test with Granupom, containing Cydia pomonella 

granulovirus, (MRID 48624729). Honey bees were orally exposed 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 receiving Granupom and 2% mortality occurred in the control 

group. No mortality was observed in the group receiving Granupom via contact. 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 pathogenicity test was also performed, in which bees were fed 

either once at a rate of 5 x 1010 Cydia pomonella granulovirus, or daily at that rate. 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 fish was performed with rainbow 

trout (Oncorhynchus mykiss; MRID 48624720). This study was a 96-hour static 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 1013 occlusion bodies/L. No mortality related to the test material was demonstrated; however, 

the sublethal effect of a change in pigment coloration was observed in all fish at the highest test concentration. 
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Therefore, the LC50 was determined to be >100 mg/L; the LOEC (lowest 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 hours and this is not sufficient to test pathogenicity. 

CLASSIFICATION: Supplemental. 

 

Banowetz et al. (1976; MRID 48624721) was also cited to provide information 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, salmonid 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 water, and in food. Exposure in food and water occurred at 

100 acre-dosages, and lasted for 18 and 24 hours for water 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 fish 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, infected 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 sampled at time 0, 12, 24, 48, and 96 hours. Pooled kidneys, livers, and spleens 

from each sample group were assayed for baculovirus polyhedra. No pathological changes were observed in the 

cell lines exposed to the OpNPV, and no change occurred in the growth rates of the cells. Additionally, there 

was no evidence that the virus entered the cells. Histopathological examination showed no evidence of infection 

by the virus in salmonid fingerlings, and no virus was recovered 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 reject earthworms cut to the 

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

The Banowetz et al. (1976) study is helpful to understand that OpNPV (of which there are two types, both of 

which were tested) does not infect fish or cause adverse effects; however, 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 makes the point that high amounts of baculoviruses have been 

observed after natural epizootics, meaning that vertebrates, including fish, have been exposed under natural 

conditions without reported effects. The OECD consensus document (OECD 2002) also states that effects of 

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

of the data submitted shows that lethal effects are not expected in fish from exposure to Cydia pomonella

granulovirus Isolate V22. 

Freshwater Invertebrates (Harmonized 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/immobility 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 LC50 was determined to be >100 
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mg/L. Mortality occurred in positive controls 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 freshwater invertebrates is provided in 

Gröner et al. (1981). A certain type of baculovirus is known to occur in certain species of shrimp; however, no 

evidence is available to show that baculoviruses that are specific to Lepidopterans are pathogenic to 

crustaceans. Moreover, it is known that the host range of granuloviruses have narrower host ranges that are 

often restricted to a single species (OECD 2002). Therefore, it is unlikely that host range of CpGV-V22 would 

include aquatic invertebrates, and the information provided indicates that 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 virus is pathogenic to these 

marine invertebrates. White shrimp were wild caught from West Bay near Galveston, Texas; brown shrimp 

were obtained from artificially propagated laboratory stock. Exposure occurred 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 1011 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 102 to 2.5 x 108 nuclear polyhedral virus per shrimp. Dietary exposure occurred over 10 

days. In both experiments, animals were sampled for histopathological examination 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, control mortality for the injected white 

shrimp was high (46%). No explanation was provided, and the paper was unclear as to how the authors 

determined how much mortality was significant. Virus was not detected 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 that baculoviruses do affect certain species of shrimp within U.S. waters (this is 

also stated in Gröner [1990] described below); however, according to this review there is no knowledge that 

baculoviruses specific for Lepidoptera affect marine or estuarine fish and invertebrates. The OECD consensus 

document (OECD 2002) also states that granuloviruses generally have smaller host ranges than nuclear 

polyhedral viruses, so CpGV-V22 is not likely to include marine/estuarine organisms in its host range. 

Therefore, the weight of the evidence 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 the Tier I toxicity studies, and because of acceptable 

rationale to address certain 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 application to tree fruit and nut crops via spray at rates of 0.5 – 
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3.0 fluid ounces per acre (agricultural use) or 1⁄2 teaspoon per 5 gallons, applied to the point of wetting leaves 

without runoff (homeowner use). The proposed application instructions for agricultural 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 label directs reapplication after 6 – 8 sunny days if 

moths are still flying and laying eggs. Madex Twin Insecticidal Virus can be applied up to and including the day 

of harvest. The label for homeowner use is slightly different, and recommends Madex 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 generation. Additional applications (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. Application equipment is not described on either label. 

The proposed applications are expected to result in contact of the active ingredient with foliar and other above 

ground plant surfaces, and some degree of spray drift is expected to occur. The a.i. is expected to be deposited 

on soil at application and with rainfall, and some may move offsite due to runoff. Since reapplication is 

recommended after 6 – 8 sunny days, and because it is known that sunlight adversely affects the viability of 

CpGV (OECD 2002), CpGV-V22 is not expected to persist at high levels on most surfaces 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 predict the exact environmental fate of CpGV-V22, since environmental 

conditions can vary. Conditions that affect the fate of baculoviruses are discussed extensively in OECD (2002). 

Based on the type of application and expected environmental fate of CpGV-V22, exposure to terrestrial and 

aquatic nontarget organisms is possible as a result of the proposed labeled 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 result in significant exposure, though oral exposure may occur 

incidentally through grooming. The study cited from the literature is not useful for risk assessment for birds; 

however, other information that is available 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 indicate potential risk to wild mammals. Additionally, the 

OECD consensus document (OECD 2002) indicates that risk to wild mammals is unlikely. While birds and 

mammals may be exposed to CpGV-V22, there is sufficient evidence to show that this 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 other arthropods are not anticipated. 

Nontarget Insects and Honey Bees

Three studies were submitted with which 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 application 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 animals in two of the studies (with predatory mites and 

parasitoid wasp) exposure to treated surfaces) leaves uncertainty as to the adequacy of these studies for testing 

nontarget effects with a substance that must be ingested. Granuloviruses must be eaten to infect an insect, and it 
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is not clear from the study reports whether the test animals actually consumed the test material, or if they did, 

whether they consumed it at the dose tested. However, these 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 under 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 ground beetles is an exception, in that it appears from the 

study report that they and their initial dietary items were directly sprayed. 

A study was also submitted (MRID 48624729), and another was 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 whether the material tested is pathogenic to honey bees; however, the Gröner 

et al. (1978) paper tested 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 nontarget insects in honey bees. Granuloviruses generally have a very narrow host 

range, and no effects of baculoviruses have been observed 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 taxonomically 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 anticipated 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 and invertebrates. Based on 

these studies the LC50 for Cydia pomonella granulovirus SC is > 100 mg/L for 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 agricultural use application rate for Madex 

Twin Insecticidal Virus (3 fl. oz per acre), the expected 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. However, assuming a reasonable range of densities from 0.5 

g/mL to 2.0 g/mL, the LC50s determined in the freshwater studies amounts to 1.1 x 106

 – 4.4 x 106 OB/mL. 

Even if the test material density is outside the estimated range, the LC50s and NOECs are well above the 

estimated environmental concentration, and risk to freshwater fish and invertebrates is not expected. 

Considering these data, plus the additional information from the supplemental 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 effects to marine/estuarine 

fish and invertebrates are not expected. 
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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 anticipated for nontarget species exposed to CpGV-V22 as a 

result of the proposed labeled applications, effects to federally-listed threatened and endangered species and 

their designated critical habitats are also not expected. Therefore, a “No Effect” determination is made for direct 

and indirect effects to federally listed endangered and threatened species 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 meaningful 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 policies. Fair treatment means that no group of people, 

including racial, ethnic, or socioeconomic groups, should bear a disproportionate share of the negative 

environmental consequences resulting from industrial, municipal, and commercial operations or the execution 

of federal, state, local, and tribal environmental programs and policies. Meaningful involvement means that (1) 

potentially affected community residents have an appropriate opportunity to participate in decisions about a 

proposed activity that will affect their environment and/or health; (2) the public’s contribution can influence the 

regulatory agency’s decision; (3) the concerns of all participants involved will be considered in the decisionmaking process; and (4) the decision-makers seek out and facilitate the involvement of those potentially 

affected. EPA has this goal for all communities and persons across the U.S. 

To help address potential environmental justice issues, EPA seeks information on any groups or segments of the 

population who, as a result their location, cultural practices, or other factors, 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 atypical, unusually high exposure compared to the general population. 

For additional information regarding environmental justice issues, please visit EPA’s web site at 

http://www.epa.gov/compliance/environmentaljustice/index.html. 

VI. RISK MANAGEMENT DECISION 

Section 3(c)(5) of FIFRA provides for the registration 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 requirements of FIFRA; (3) it will perform its intended 

function without unreasonable adverse effects on the environment; 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 contradict 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 2 
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is satisfied by the current product labels, as well as the data and information presented in this document. It is 

believed that this end use product containing CpGV-V22 will not cause any unreasonable adverse effects on the 

environment and is likely to control the caterpillar pests described on the product labels, 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 label instructions. Therefore, Madex Twin Insecticidal 

Virus containing CpGV-V22 as a new active ingredient, is eligible for 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, Andermatt must submit the following: 

Results of storage stability (Harmonized Guideline 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 clearly 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 summary of these types of data 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, all incidents of hypersensitivity (including both 

suspected and confirmed incidents) must be reported to EPA under the provisions of 40 CFR § 

158.2140(d). 

 
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VIII. BIBLIOGRAPHY 

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

A. Studies Submitted to Support the Registration 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 (Tier 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 Formation 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. Unpublished study prepared by SciReg, Inc. 28p. 

48624703 Smith, F. (2011) Cydia pomonella Gravulovirus Isolate V22 Technical and MADEX Twin: 

Summary of Physical-Chemical Properties. Unpublished study prepared by SciReg, Inc. 5p. 

48624704 Andermatt, M. (2000) MADEX: Physical and 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 pomonella Granulosus Virus (CpGV) Hoe 083311 

Summary on Chemical and Physical Data. Project Number: A55428. Unpublished study prepared 

by 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. Project Number: 234/80. Unpublished study prepared 

by 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 LC08 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. 13 
Cydia 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/Corrosion: 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 Regulatory Oversight in Biotechnology 87p.

48624719 Lautenschlager, R.; Rothenbacher, H.; Podgwaite, 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, Salmonidae). Project Number: 96272/01/AAOM. 

Unpublished study prepared by GAB Biotechnologie Gmbh. 23p. 

48624721 Banowetz, G.; Fryer, J.; Iwai, P.; et al. (1976) Effects of the Douglas-Fir Tussock Moth 

Nucleopolyhedrosis Virus (Baculovirus) on Three 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 Invertebrates 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. Project 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 by Besana Consulting S.A.L. 15p.

48624733 Smith, F. (2011) Nontarget Organism 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 Formation 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. Unpublished study prepared by SciReg, Inc. 13p. 

48738103 Smith, F. (2012) Nontarget Organism Discussion (Madex Twin). Unpublished study 
Cydia 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 insect 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 MADEX 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 assessment 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. Nucleopolyhedroviruses and Granuloviruses (Insect Viruses) Preliminary Work Plan and 

Summary Document Registration Review: Initial Docket. September 26, 2011. 

C. Other References 

Banowetz, G.M., J.L. Fryer, P.J. Iwai, and M.E. Martignoni. 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 invertebrates 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: 25-27. (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. Podgwaite. 1979. Response of birds to aerial application 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.Adams, and A.M. Heimpel. 1973. Testing Penaeid shrimp for 

susceptibility to an insect nuclear polyhedrosis virus. Environmental Entomology 2: 611-613. 

Organization for Economic Cooperation and Development (OECD). 2002. Series on Harmonization of 

Regulatory Oversight in Biotechnology, No. 20: Consensus document on information used in the 

assessment of environmental applications 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 of Cydia pomonella Granulovirus Isolate V-2

 the End-Use Pesticide Product (EP) Madex Twin (40 CFR § 158.2120) 

Harmoniz

ed 

Guideline 

Number 

Data Requirement Results 

885.1100 Product Identity 

Supplemental data submitted by Andermatt were upgraded to acceptable based on additional info

provided by the Ccmpany. This sequence alignment information suggests that there has been no the polh/gran or lef 8 genes and that Isolate V22, along with the registered CpGV-Mexican isola

genome type A. 

885.1200 Manufacturing Process 

An in vivo production system was described. No details of how the laboratory colonies of Cydia 

and Grapholita molesta are maintained and assured to be disease-free. Confirmatory data will be 

term of registration. 

885.1250 

Deposition of a Sample 

in a Nationally 

Recognized Culture 

Collection 

The Andermatt has deposited Cydia pomonella Isolate V-22 in the Deutsche Sammlung von Mikroorganismen und Zellculturen GmbH under accession number DSMZ GV-0014. 

885.1300 

Discussion of Formation 

of Unintentional 

Ingredients 

The absence of microbial contaminants at levels appropriate for baculovirus based products is in with the OECD document on that topic except there is no mention of spore-forming bacterial conespecially Bacillus cereus. Isolates of Bacillus cereus can be an important cause of bacterial food 

and some measure should be added to the quality control procedures to characterize its presence 

product. Confirmatory data will be required as a term of registration. 

885.1400 Analysis of Samples Submitted data fulfill the requirement for analysis of samples. As a term of registration, EPA is r

submission of a storage stability and corrosion characteristics study within one year of registratio

885.1500 Certification of Limits Certified Limits are acceptable. 

TABLE 2. Physical and Chemical Characteristics for the Product Madex Twin, Containing Cydia pomonella Granulov

 (40 CFR § 158.2120)

Harmonized Guideline Number 

Data Requirement Results 
Cydia pomonella Granulovirus isolate V22 

DRAFT Biopesticides Registration Action Document 

TABLE 2. Physical and Chemical Characteristics for the Product Madex Twin, Containing Cydia pomonella Granulov

 (40 CFR § 158.2120)

Harmonized Guideline Number 

Data Requirement Results 

830.63021 Color GreyBrown830.63031 Physical State Liquid 830.63041 Odor Odorless 830.63131 

Stability to Normal and Elevated Temperatures, Metals, and Metal Ions 

The information that was submitted was supplemental, but does not need to be upgraded. As a registration, EPA is requiring submission of storage stability and corrosion characteristics data year of registration to confirm that the active ingredient maintains its activity against the OrienGrapholita molesta.

830.6317 Storage Stability 

A Results from a 3-year storage stability study for MADEX were submitted. Minimal contain

changes (<0.5%) over 24 months probably represent a loss due to evaporation of the contents. results were not provided. If there are data that demonstrate no decline in activity for 36 monthlonger storage stability statement could be considered. As a term of the registration, storage stcorrosion characteristics data within one year of registration to confirm that the active ingredieactivity against the Oriental fruit fly Grapholita molesta. 

830.63192 Miscibility Only required for emulsifiable liquid formulations of microbial pesticides.830.6320 Corrosion Characteristics Only required when microbial pesticides are packaged in metal, plastic, or paper containers. A

registration, EPA is requiring submission of a storage stability and corrosion characteristics styear of the date the product is registered. 

830.70001 pH The mean pH value of a 1%w/v aqueous dilution is 7.0 + 0.2.830.71003 Viscosity The viscosity is 1,200 mPascal-second. 830.7300

Density/Relative Density/Bulk Density (Specific Gravity) 

The relative density was determined to be ρ19.94.0= 1.16083 

 1 According to 40 CFR § 158.2120, these data are only required for the technical grade of the active ingredient. 
Cydia pomonella Granulovirus isolate V22 

DRAFT Biopesticides Registration Action Document 

 2 According to 40 CFR § 158.2120, these data are only required for emulsifiable liquid forms of microbial pesticides.

 3 According to 40 CFR § 158.2120, these data are only required for liquid forms of microbial pesticides. 

TABLE 3. Toxicology Data Requirements for Granulosis Virus of Cydia pomonella Isolate V-22, including 

 the End-Use Pesticide Product (EP) Madex Twin (40 CFR § 158.2140)

Harmonized Guideline Number 

Data Requirement Results 

Tier I 885.30501 Acute Oral 

Toxicity/ Pathogenicity 

Data conducted with closely related CpGV (Mexican Isolate 1964) were cited for Acute Oral Toxicity/Pathogenicity in 

distinct clearance pattern was evident in the feces and heart/lungs through day 7 when dosed with 2 x 1012 granulosis in

bodies/ml by oral gavage. Classification: Acceptable, Toxicity Category IV. 

885.31501

Acute Pulmonary Toxicity/ Pathogenicity 

Data conducted with closely related CpGV (Mexican Isolate 1964) were cited for Acute Pulmonary Toxicity/Pathogen

Rat. A distinct clearance pattern was evident in the feces and heart/lungs through day 7 when 1 x 108 granulosis inclusi

were administered intratracheally. Classification: Acceptable, Toxicity Category IV.

885.3200 

Acute Injection Toxicity/ Pathogenicity (Intravenous) 

 Not required for a virus (See 40 CFR § 158.2140, test note 2) 

885.3400 Hypersensitivity Incidents Hypersensitivity incidents, including immediate-type and delayed-type reactions of humans or domestic animals, that o

the testing or production of the TGAI or EP, or are otherwise known to Andermatt, must be reported if they occur (See 158.2140(d), test note 3). 

885.3500 Cell Culture 

Andermatt cited a study conducted with the CpGV-Mexican isolate that demonstrated that CpGV-Mexican does not in

in, transform or cause toxicity in mammalian cell lines. EPA requested additional information/data to confirm that CpG

similar to the CpGV-M1 (“Mexican”) strain. Andermatt supplied a statement of origin that Cydia pomonella granulovi

V22 is a selection derived from the Cydia pomonella granulovirus Mexico strain. They also provided sequence alignminformation for two genes (the granulin gene (polh/gran) and late expression factor gene (lef8)) and a Restriction Endo

(REN) profile DNA gel that support their statement of origin. This information permits bridging data from the previou

cell culture study (MRID 431612-02) that had been conducted with the CpGV-Mexican Isolate. Classification: Accep

870.1100 Acute Oral Toxicity Andermatt has provided information on the added inerts that indicate they are approved for food incorporation in other 

not present a toxicity hazard. (See 40 CFR 158.2040, test note 1). 

870.12003 Acute dermal toxicity Andermatt has provided information on the added inerts that indicate they are approved for food incorporation in other 

not present a toxicity hazard. (See 40 CFR 158.2040, test note 1). 

870.13003 Acute inhalation toxicity Andermatt has provided information on the added inerts that indicate they are approved for food incorporation in other 

not present a toxicity hazard. (See 40 CFR 158.2040, test note 1). 
Cydia pomonella Granulovirus isolate V22 

DRAFT Biopesticides Registration Action Document 

TABLE 3. Toxicology Data Requirements for Granulosis Virus of Cydia pomonella Isolate V-22, including 

 the End-Use Pesticide Product (EP) Madex Twin (40 CFR § 158.2140)

Harmonized Guideline Number 

Data Requirement Results 

870.2400 Eye Irritation 

This cited study was conducted in rabbits with Hoe083311 (the closely related Mexican isolate). There were no signs olesions or iritis during the test period. All animals showed slight to moderate discharge and 1of 3 displayed diffuse cri

1-2 hours after instillation. Diffuse crimson color was observed in 2 of 3 animals at 24 and 48 hours. Positive signs of 

were not significant by day 3 and cleared by day 7. Chemosis was visible on all animals at 1 hour, but was not significcleared by day 3. Classification: Acceptable. Toxicity category III. 

870.2500 Dermal Irritation 

This cited study was conducted in rabbits with Granupom SC, which contains CpGV isolate Hoe083311 (the closely re

Mexican isolate). No animals died during this study, and none showed any signs of erythema or edema. The study wa

on day 10 after bandage removal. The lack of any signs of irritation indicates Granupom SC is Toxicity category IV fo

irritation. Classification: Acceptable. Toxicity Category IV.

Tiers II and III Not required for Granulosis Virus of Cydia pomonella Isolate V-22, based on the lack of acute toxicity in the Tier I studies. 
Table 4. Non-target Organism Toxicity Requirements for Cydia pomonella Granulovirus Isolate V22 (40 CFR 158.2150) 

Requirement (OPPTS Guideline No.) 

MRID Toxicity and Classification 

Avian oral toxicity  (885.4050) 

48624719 48738103 48624718 

A study cited from literature (Lautenschlager et al. 1979) showed no effects of Lymantria dispar caged quail. Classification: Unacceptable An additional volume was submitted that provided a summary, but no new information on birds. Classification: Supplemental 

The OECD consensus document (OECD 2002) contains a review of information and states that efrom exposure to baculoviruses.Classification: Supplemental (acceptable for avian risk asses

Avian inhalation toxicity/ pathogenicity (885.4100) 

N/A Not required: the nature of the microbial pesticide does not indicate potential toxicity or pathoge

Wild mammal toxicity/ pathogenicity (885.4150) 

N/A Tests required by 40 CFR § 158.2140 are adequate and appropriate for assessment of hazards to 

information submitted to fulfill those data requirements does not indicate concerns for adverse efClassification: Acceptable for environmental risk assessment 

Freshwater fish toxicity/ pathogenicity (885.4200) 

48624720 48624721 48624725 48624718 

The LC50 for rainbow trout (Oncorhynchus mykiss) in a 96-hour study was > 100 mg Cydia pom

fish showed a change in pigment to darker color, so the NOEC was determined to be 10 mg/L. Tdetermine pathogenicity. Classification: Supplemental 

A study with Orygia pseudotsugata NPV (Banowetz et al. 1976) showed no pathological or growcell lines exposed to this virus. Salmonid fingerlings and smolts showed no adverse effects with water, and in food, and virus was inactivated within the fish within 24 hours after exposure. Rati

Cydia pomonella GV Isolate V22 is needed. Classification: Supplemental 

A study cited from the literature (Gröner et al. 1981) and the OECD consensus document (OECD show that baculoviruses are generally understood not to be toxic or pathogenic to fish. Classifica
Cydia pomonella Granulovirus isolate V22 

DRAFT Biopesticides Registration Action Document 

Freshwater invertebrate testing  (885.4240) 

48624722 48624725 48624718 

A 48-hour study with Cydia pomonella GV SC demonstrated a LC50 of > 100mg/L in Daphnia m

duration to assess pathogenicity. Classification: Supplemental A study cited from the literature (Gröner et al. 1981) and the OECD consensus document providbaculoviruses specific to Lepidoptera are not expected to cause adverse effects in freshwater inve

Classification: Supplemental

Estuarine/Marine fish testing Estuarine/Marine invertebrate testing (885.4280) 

48624724 48624725 48624718 

A study was cited from the literature (Lightner et al. 1973) that showed no effect and no infectiv

on white (Penaeus setiferus) and brown (Penaeus azteca) shrimp. Rationale to bridge this NPV t

V22 is needed. Classification: Supplemental 

A study cited from the literature (Gröner et al. 1981) provides additional information that baculonot expected to cause adverse effects in marine/estuarine fish or invertebrates. Additional discussdocument supports this conclusion. Classification: Supplemental

Nontarget Plant Testing (885.4300) 

N/A Not required. The microbial pesticide is not taxonomically related to known plant pathogens. 

Nontarget Insect Testing (885.4340) 

48624727 48624728 48624726 48634723 

The 15-day LC50 was > 900 ml/ha for Cydia pomonella GV SC in the ground beetle (Poecilus c

maximum application rate for Madex Twin. Classification: Acceptable The 7-day LC50 for Cydia pomonella GV SC was > 900 ml/ha for the predatory mite (Typhlodro

for reproduction was 900 mL/ha. Exposure route raises questions as to actual exposure level. Classification: Supplemental The 48-hour LC50 for Cydia pomonella GV SC was > 900 ml/ha for the parasitoid wasp (Aphidiu

reproduction was 900 mL/ha. Exposure duration was too short to determine pathogenicity; exposactual exposure level. Classification: Supplemental A study (Gröner 1990) was cited that provided additional evidence that Cydia pomonella GV Is

nontarget insect species. Classification: Supplemental

Honeybee Testing (885.4380) 

48624729 48624730 

The 72-hour acute oral LC50 was > 3.6 x 107 granula per bee, and the 48-hour acute contact LC50 

This study is of too short duration to assess pathogenicity. Classification: Supplemental 

A study by Gröner et al. (1978) was cited, in which the authors tested an isolate of Cydia pomon

Mamestra brassicae NPB on bees via several routes. A 10-day pathogenicity test with honey bee

granula/mL resulted in no significant mortality. Other results showing lack of effects of baculoviClassification: Supplemental
Cydia pomonella Granulovirus isolate V22 

DRAFT Biopesticides Registration Action Document 

APPENDIX B. PESTICIDE PRODUCT 

EPA File Symbol 

Registration Name Percentage 

Active Ingredient 

Formulation Type Use Site(s) Method(s) of 

Application 

Application Rate 

Target P

69553-1 Madex Twin 0.06% 

Contains a minimum of 3 x 1013

viral occlusion bodies per liter 

Dilutable Liquid Agricultural and 

residential use. 

Spray 0.5 -3.0 fluid 

ounces of Madex Twin per acre diluted in 50-100 gallons of water per acre. 

Larvae opomonelGraphol