BIOPESTICIDES REGISTRATION ACTION DOCUMENT



## Helicoverpa zea ABA Nucleopolyhedrovirus

- U

Pesticide Chemical (PC) Code: 107200

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

DRAFT

– February 4, 201 4Helicoverpa zea Nucleopolyhedrovirus Page 2 of 27

Biopesticides Registration Action Document

TABLE OF CONTENTS

I. EXECUTIVE SUMMARY

................................ ................................ ................................ ........................... 4

II. ACT IVE INGREDIENT OVERV IEW

................................ ................................ ................................ ....... 6

III. REGULATORY BACKGROUN D

................................ ................................ ................................ .............. 7

A. Applications for Pesticide Product Registration

................................ ................................ ............ 7

B. Food Tolerance Exemption

................................ ................................ ................................ .............. 7

IV. RISK ASSESSMENT

................................ ................................ ................................ ................................ .... 7

A. Product Analysis Assessment ( 40 CFR § 158.2120)

................................ ................................ ....... 8

B. Human Health Assessment (40 CFR § 158.2140)

................................ ................................ ........... 8

C. Environmental Assessment (40 CFR § 158.2150)

................................ ................................ ......... 14

V. ENVIRONMENTAL JUSTIC E

................................ ................................ ................................ ................. 16

VI. RISK MANAGEMENT DECI SION

................................ ................................ ................................ ......... 17

VII. A CTIONS REQUIRED OF T HE REGISTRANT

................................ ................................ ................... 17

VIII. GLOSSARY OF ACRONYMS AND ABBREVIATIONS

................................ ................................ ...... 18

IX. BIBLIOGRAPHY

................................ ................................ ................................ ................................ ....... 1 9

A. Studies Submitted to Support the Active Ingredient,

, Helicoverpa zea Nucleopolyhedrovirus

and Pesticide Product

, Heligen

, Registration

................................ ................................ ................................ ......... 1 9

B. Environmental Protection Agency Risk Assessme nt Memor anda

................................ ............. 21

C. Other References

................................ ................................ ................................ .......................... 21 2

APPENDIX A. MICROBI AL PESTICIDES DATA R EQUIREMENTS

................................ ........................... 23

APPENDIX B. PESTICI DE PRODUCT

................................ ................................ ................................ ................ 27Helicoverpa zea Nucleopolyhedrovirus Page 3 of 27

Biopesticides Registration Action Document

BIOPESTICIDES REGISTRATION ACTION DOCUMENT TEAM

Office of Pesticide Programs (OPP) Biopesticides and Pollution Prevention Division

Microbial Pestici des Branch

Science Reviews

Ibrahim S. Barsoum, Ph.D. Product Analysis, Human Health

John L. Kough, Ph.D. Product Analysis, Human Health Shannon Borges Environmental Effects

Regulations

Kimberly Nesci Chief, Microbial Pestici des Branch

Michael Glikes Regulatory Action LeaderHelicoverpa zea Nucleopolyhedrovirus Page 4 of 27

Biopesticides Registration Action Document

I. EXECUTIVE SUMMARY

Background

In September 201 2

, MacIntosh & Associates, Inc., on behalf of AgBiTech Pty Ltd., an Australian company, submitted application s for a new manufacturing use product (MP)

, Helicoverpa zea ABA Nucleopolyhedrovirus

- U (EPA File Symbol 87978

- R), and a new end

use product (EP)

, Heligen (EPA File Symbol 87978

- E), to the United States Environmental Protection Agency (EPA) under section 3 of the Federal Insecticide, Fungicide, and Rodenticide Act (FIFRA).

Helicoverpa zea Nucleopolyhedrovirus (Hz NPV) is a naturally occurring insect virus, originally isolated from Helothine Lepidoptera, specifically the c otton b ollworm ( Helicoverpa zea ) an d

t obacco budworm ( Heliothis virescens ) . In 1975, the EPA first registered Heliothis zea NPV

under the brand name Elcar for us e on cotton and tobacco to control the c otton b ollworm and

t obacco budworm. In 1984, the EPA issued a registration standard fo r the use of Heliothis zea

NPV on a variety of agricultural crops , and in 1990 the Agency issued a Registration Eligibility Decision (RED), which determined that the pesticide poses no unreasonable adverse effect to

human health and the environment. System atic entomologists changed the genus from "Heliothis" to "Helicoverpa" in the late 19 90s.

The proposed (MP) containing this active ingredient is an isolate of the registered strain, and it is

called Helicoverpa zea ABA Nucleopol yhedrovirus

- U (Hz ABA

- NPV

- U )

. T he proposed EP containing this active ingredient is Heligen

. AgBiTech’s first Hz NPV EP was commercialized in

Australia under the name ViVUS

. Th e proposed EP is for use in the United Sates in field

a pplications to control corn earworm, cotton bollworm, tomato fruitworm, and tobacco budworm

on food and non

- food crops , including sorghum, cereal grains, alfalfa, oilseed, potatoes, pulses, cotton, sweet corn, berry fruits, brassica, leafy vegetables, leg ume vegetables, pome fruits, celery, cucurbits, and ornamentals. It is proposed for application by ground or aerial spray or by chemigation.

EPA s cientists reviewed product analysis, toxicology, and nontarget organism data and information ( 40 CFR §§ 158.2 120, 158.2140, and 158.2150, respectively) submitted to su pport the registration of the proposed pesticide product s

. They found that, o verall, such data and information are adequate for risk assessment purposes, fulfill the current microbial pesticide data requirements, and support registration of the proposed product under FIFRA section 3(c)(5)

.

Product Analysis

For the purposes of FIFRA section 3(c)(5) registration, the product analysis data requirements

for Helicoverpa zea ABA Nucleopolyhedrovirus

- U (H z ABA

- NPV

- U)

, which is both the

tec h nical grade active ingredient (TGAI) and the MP, and the EP , Heligen , including product chemistry and composition, analysis of samples, physical and chemical characteristics, and additional data concerning genetic stability, were fulfilled by acceptable guideline studies.

Toxicology

The applicant submitted a dequate mammalian toxicology data and information to support the proposed pesticide products. A cute oral, pulmonary, and injection toxicity/pathogenic ity studiesHelicoverpa zea Nucleopolyhedrovirus Page 5 of 27

Biopesticides Registration Action Document

showed that, at a single high dose, Hz ABA

- NPV

- U is not toxic and/or p athogenic via these

route s of exposure

. Moreover, the applicant reported that no hypersensitivity incidents occurred during rese arch, dev elopment, or testing of Hz ABA

- NPV

- U

. In l ight of the results of the acute toxicity/pathogenicity data and the absence of hypersensitivity incidents, testing at higher tiers (i.e., Tiers II and III) was not required.

Tolerance Exemption

B ecause Helicove rpa zea Nucleopolyhedrovirus is already exempt from this requirement , t he applicant did not petition the EPA for an exemption fr om the requirement of a tolerance for

residues of the new isolate, H z ABA

- NPV

- U . In 1980 t he EPA granted an exemption from the requirement of a tolerance for residues of the active ingredient Hz NPV in or on all agricultural

commodities ( 7 CFR 180.1027) as long as the manufacturer meets cert ain specifications related to identity, contamination control, and safety. In its application to the EPA , the applicant provided data and other information to support coverage by the existing exemption from the

toler a nce requirement

. Hence, t he EPA conclud e s that there is reasonable certainty that no harm

will result to the U.S. population, including infants and children, from aggregate exposure to

residues of Hz ABA

- NPV

- U

. Data and information submitted by the applicant support the

conclusion that n o dieta ry risks are expected to occur as a result of the use of Hz ABA

- NPV

- U as an active ingredient in pesticide products. The EPA concludes that there will not be s ignificant

exposure to Hz ABA

- NPV

- U through food and drinking water

. Based on the review of acute oral toxicity and pathogenicity d ata by EPA scientists , the Agency determined that, even if Hz ABA

NPV

- U were present on food or in drinking water, human exposure to this insect virus would not

result in unreasonable adverse effects.

Occupational Exposur e

Despite the low toxicological profile of residues of the active ingredient, Hz ABA

- NPV

- U , in or on all agricultural commodities

, the EPA is requiring baseline personal protective equipment (PPE) for handlers that, due to their occupation

, may be subject to prolonged or repeated

exposure to the active ingredient

. The label on Heligen, the proposed end

- use product, directs

h andlers working with Hz NPV i n agricultural settings to wear a long

- sleeved shirt, long pants,

soc ks, and shoes. For future products, EPA may require additional PPE (e.g., protective eyewear

and/or a dust/mist filtering respirator meeting National Institute for Occupational Safety and Health (NIOSH) standards of at least N

- 95, R

- 95, or P

- 95 ) on a produ ct

- specific basis.

Nontarget Organisms

The applicant submitted rational e bridging studies previously submitted for a different isolate of Helicoverpa zea Nucleopolyhedrovirus t o support the Agency’s waiving nontarget organism testing for the new active ingredient (AI)

. T he EPA determined that this new AI is substantially

simil a r to the previously registered AI and that previously submitted data would provide an adequate measure of hazard for the new AI. Hence, the Agency accep ted th e bridging rational e and determined that the data 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. The EPA performed an environment al risk

assessment

, based on data and other information provided by the ap plicant and previously submitted data cited by the previous applicant , and determined that adverse effects to nontarget organisms are not anticipated from the proposed pesticidal uses of Heligen, which contains theHelicoverpa zea Nucleopolyhedrovirus Page 6 of 27

Biopesticides Registration Action Document

active ingredient H z ABA

- NPV

- U . Moreover, the EPA made a “No Effect” determination for

direct and indirect effects to federally listed threatened and endangered species an d their designated critical habitats resulting from these same proposed pesticidal uses.

Public Participation

On October 1, 2009, the EPA announced a new policy to provide a more meaningful opportunity for the public to participate i n major registr ation decisions before they occur. According to this policy, the EPA intends to provide a public comment period prior to making a registration decision for, at minimum, the following types of appli cations: new active ingredients; first food uses; first out door uses; first residential use s; or any other registration actions for which the Agency believes there may be significant public interest

.

Consistent with the policy of making registration actions more transparent, the proposed pesticide product conta ining Hz ABA NPV

- U , is subject to a 15

- day comment period. In addition to containing a new active ingredient, the registration of the pesticide product would

result in the first outdoor and food uses for H z NPV . The docket identification number associated with these registration actions

, and accessed through either http://www.regulations.gov or http://www.epa.gov/pesticides/regulating/registration

- status.html , is EPA

- HQ

- OPP

- 201 3

- 0 099

. The following documents are available for comment in EPA

- HQ

- OPP

- 201 3

- 0 099 : (1) draft Helicoverpa zea ABA Nucleopolyhedrovirus

- U Registration Acti on Document (BRAD); (2)

environmental risk assessment for Hz ABA NPV

- U ; (3) draft label for the manufacturing

- use product (File Symbol 87978

- R), and (4) draft label for the end

- use pesticide product, Heligen (EPA File Symbol 87978

- E ). While a final decisio n on registration is contingent upon review and consideration of public comments, EPA believes that, based upon the data and information submitted in support of Hz ABA NPV

- U and Heligen , it is appropriate to issue the re gistration. The basis for this preliminary decision can be found in the risk assessment for Hz NPV , which is characterized throughout this BRAD.

II. ACTIVE INGREDIENT OVERVIEW

Biological Name: Helicoverpa zea ABA Nucleopolyhedrovirus

- U

Culture D eposit: American Type Culture Collection : SD

- 6742

OPP Chemical Code: 107200

Type of Pesticide: Mi crobial Pesticide

– Insecticide

See Appendix B for specific information (e.g., use sites, application rate s , methods of application, formulation type s , and target pests) regarding the proposed pesticide products containing this active ingredient.Helicoverpa zea Nucleopolyhedrovirus Page 7 of 27

Biopesticides Registration Action Document

III. REGULATORY BACKGROUND

A. Applications for Pesticide Product Registration

In September 201 2

, MacIntosh & Associates , Inc. (address: 1203 Hartford Avenue, Saint Paul,

MN 55116 ) submitted an application on behalf of AgBiTech Pty. Ltd. (address: P.O. Box 537, Richmond, New South Wales 2753) to register a manufacturing

- use pesticide product, Helicoverpa zea ABA Nucleopolyhedrovirus

- U (EPA File Symbol 87978

- R), and a n end

- use product, Heligen (EPA File Symbol 87978

- E)

, under FIFRA section 3. On April 19 , 201 3 ( 76 FR

23558 ) , EPA announced receipt of the application to register a pesticide product containing a

ne w active ingredient for use in agriculture (outdoors)

. EPA opened a 3 0

- day public comment period pursuant to the provisions of FIFRA section 3(c)(4). No comme nts were received following this publication.

B. Food Tolerance Exemption

U nder Federal Food, Drug, and Cosmetic Act (FFDCA) section 408(d), the EPA in 1980 granted an exemption from the requirement of a tolerance for residues of the active ingredient Helicoverpa zea Nucleopolyhedrovirus in or on all agricultural commodities ( 40 CFR 180.1027)

as long as the manufacturer meets certain specifications related to identity, contamination control, and safety. In its application to the EPA in 2012

, MacIntosh & Associations, on behalf of the Australian company AgBiTech, provided data and other information to support coverage by the existing exemption from the requirement of a tolerance for residues of Hz ABA NPV

- U in or on all agricultural commodities.

IV. RISK ASSESSMENT

In the Federal Register of October 26, 2007 ( 72 FR 61002 )

, the EPA issued a Final Rule on the data requirements to support registration of microbial pesticides and updated the definition for microbial pesticides

. The rule became effective on December 26, 2007. Th e data and information evaluated for this BRAD were considered in light of these requirements.

The EPA classifies each data submission with an indication of the usefulness of the information contained in the documents for risk assessment. A rating of “ acc eptable ” indicates the study is scientifically sound and is useful for risk assessment. A “ supplemental ” rating indicates the data provide some information that can be useful for risk assessment. The studies may have certain aspects determined not to be sc ientifically acceptable (“ supplemental : upgradeable ”). If a study

is rated as “supplemental: upgradeable,” EPA always provides an indication of what is lacking or what can be provided to change the rating to “ acceptable .” If there is simply a “ supplemental ” rating, the reviewer will often state that t he study is not required by 40 CFR P art 158. Both

“ acceptable” and “supplemental ” studies may be used in the risk assessment process as appropriate. An “unacceptable ” rating indicates that new data must be subm itted.

For the acu te toxicity data requirements, Toxicity C ategories are assigned based on the hazard(s) identified from studies and/or other infor mation submitted to the EPA in support of a pesticide registration. The EPA classifies t he active ingredien t or particular product into Toxicity Category I, II, III, or IV, where Toxicity Category I indicates the highest toxicity and Toxicity Category IV indicates the lowest toxicity (see 40 CFR § 156.62 )

.Helicoverpa zea Nucleopolyhedrovirus Page 8 of 27

Biopesticides Registration Action Document

A. Product Analysis Assessment ( 40 CFR § 158.2120 )

For purposes of registration under FIFRA section 3(c)(5), the EPA determined that all product analysis data requirements for Helicoverpa zea ABA Nucleopolyhedrovirus

- U have been fulfilled. Refer to Tables 1, 2, 3, and 4 in Appendix A for a summary of the data requirements, including both generic and product

- specific information.

B. Human Health Assessment ( 40 CFR § 158.2140 )

1. Toxicity

All toxicology data requirements for Helicoverpa zea ABA Nucleopolyhedrovirus

– U have been

ful filled. Acceptable Tier I mammalian toxicology data and information support registration of

the pesticide product, Heligen

. Furthermore, Tier II and Tier III studies were not required for Hz

ABA NPV

- U because of the lack of acute toxicity/pathogenicity in the Tier I studies.

For a summary of the toxicology data for Hz ABA NPV

- U described in sections IV(B)(1)(a) and IV(B)(1)(b) (see pages 8

– 10 ), refer to Table 5 in Appendix A

.

a. Acute Toxicity/Pathogenicity

– Tier I

The appl icant evaluated the identi ty or similarity between NPV strains by extracting DNA, treating with restriction endonuclease and using electrophoresis to compare the gel patterns (MRID No. 490114

- 01) . The products, AgBiTech's Heligen and the previously registe red product containing Hz NPV

, Gemstar (EPA Registration Number 70051

- 45), were examined by light microscopy using Giemsa stain and X I000 magnification under phase contrast. This

process confirmed that the products contain a nucleopolyhedrovirus with occl usion bodies visually similar to each other. Researchers extracted t he virus from both products

, and they

extracted DNA from the virus occlusion bodies. Then, t his DNA was digested with restriction endonuclease (RENs) and the profile under electrophoresis on agarose was compared with that

of NPV in Hz NPV (Gemstar ). This analysis shows that the Hz ABA NPV

- U in the AgBiTech product is almost identical to th e Hz NPV found in Gemstar. Based on this, data previ ously submitted to support the registration of HzN PV (Gemstar) can be bridge d to this new strain.

Acute Oral Toxicity/Pathogenicity (Harmonized Guideline 885.3050; Master Record Identification Number (MRID No. 489417

- 06 ) : Th e EPA rated this scientific rationale for waiver

request as acceptable for risk assessment p urposes (U.S. EPA 2013; Barsoum and Kough). Hz NPV is a baculovirus with a long history of safe use as a microbial pesticide in the United States. Toxicology results from baculoviruses have raised no concerns for human or animal health.

Baculoviruses have narrow host ranges, and the mode of action is well understood and unique to insect hosts, with virtually no adverse effects in people, wildlife, or non

- target insects. Based on literature search for the period 1980

- 2012, there are no data demonstrating any toxic effects on humans or other mammals exposed to Hz NPV

.

Acute Pulmonary Toxicity/Pathogenicity(Harmonized Guideline 885.3150; MRID No. 4 89417

- 06 ) : Th e EPA rated this scientific rationale for waiver request as acceptable for risk assessment

purpos es (U.S. EPA 20 13 ; Barsoum and Kough ). Hz NPV is a baculovirus with a long history of safe use as a microbial pesticide in the United States. Toxicology results from baculoviruses

have raised no concerns for human or animal health. Baculoviruses have narro w host ranges, andHelicoverpa zea Nucleopolyhedrovirus Page 9 of 27

Biopesticides Registration Action Document

the mode of action is well understood and unique to insect hosts, with virtually no adverse effects in people, wildlife, or non

- target insects. Based on literature search for the period 1980

- 2012, there are no data demonstrating any t oxic effects on humans or other mammals exposed to

Hz NPV

.

Acute Injection Toxicity/Pathogenicity (Intra venous ) (Harmonized Guideline 885.3200; MRID

No 489 417

- 06 ) : Th e EPA rated this scientific rationale for waiver request as acceptable for risk

asses sment purposes (U.S. EPA 2013; Barsoum and Kough). Hz NPV is a baculovirus with a long history of safe use as a microbial pesticide in the United States. Toxicology results from

baculoviruses have raised no concerns for human or animal health. Baculovirus es have narrow host ranges, and the mode of action is well understood and unique to insect hosts, with virtually no adverse effects in people, wildlife, or non

- target insects. Based on literature search for the period 1980

- 2012, there are no data demons trating any toxic effects on humans or other mammals exposed to Hz NPV.

Hypersensitivity Incidents (Harmonized Guideline 885.3400; MRID 48 9417

- 06 ) : The applicant reported that n o hypersensitivity incidents, including immediate

- type or delayed

- type reactio ns of humans and domestic animals

, occurred during research, development, or testing of Hz ABA

NPV

- U

. Any future hypersensitivity incidents must be reported to EPA (refer to test note #3 of 40 CFR § 158.2140(d)).

Cell Culture (Harmonized Guideline 885.350 0) MRID 48941807 : Researchers tested for infectivity the following cells: human lung fibroblast WI38, primary human dermal fibroblast, primate continuous line CV

- 1, and primary Syrian hamster embryo cells. They incubated the four cells with Hz NPV (>1x10 v irus particles), commercial Hz NPV, a NPV control, and inactivated Hz NPV virus, sub

- cultured at days 7 and 14, and examined daily through day 7 and at days 14 and 21 after infection for cytopathic effect (CPE). Using Dot Blot techniques, the

researchers e xamined the cell cultures for the presence of viral nucleic acid at days 1, 2, 5, 7, 14 and 21, and then they resuspended the cultures in an appropriate volume based on the number of cells collected. Triplicate 25

- μ L samples were applied to positively cha rged nylon membrane at 300 to 22,250 cells per spot for most samples (inactivated virus samples had 0 to 4,250 cells per spot). To detect Hz NPV nucleic acids in the tested cells, researchers used purified enzyme restriction digested genomic viral DNA from the commercial virus Hz NPV. Helicoverpa zea cells infected with the commercial virus Hz NPV were used as positive control on the dot blot.

The researchers used mock

- infected H. zea cells (cells exposed to only cell culture media) as a negative control on the dot blot, photographed the blots, and stored samples below

- 60°C. They determined that the number of colonies per plate observed after 21 or 25 days for all cell lines with the virus infected cells was equivalent to the number of colonies per plate with no treatment. They observed no cytopathic effects or replication of Hz NPV exposed to any of the mammalian cell lines. Since no Hz NPV nucleic acid was detected, no further testing was necessary. The EPA rated this study as acceptable.

Acute Oral Tox icity (Harmonized Guideline 870.1100 ; Master Record Identification Number (MRID No. 489417

- 06 ) : The EPA rated this scientific rationale for waiver request as acceptable for risk assessment purposes (U.S. EPA 2013; Barsoum and Kough). Hz NPV is a baculovir us with a long history of safe use as a microbial pesticide in the United States. Toxicology results

from baculoviruses have raised no concerns for human or animal health. Baculoviruses have narrow host ranges, and the mode of action is well understood and unique to insect hosts, with virtually no adverse effects in people, wildlife, or non

- target insects. Based on literature searchHelicoverpa zea Nucleopolyhedrovirus Page 10 of 27

Biopesticides Registration Action Document

for the period 1980

- 2012, there are no data demonstrating any toxic effects on humans or other mammals exposed to Hz NPV. T he EPA classified the acute oral toxicity / pathogenicity as Toxicity Category IV

.

Acute Inhalation Toxicity (Harmonized Guideline 870

- 1300 ; MRID No. 489417

- 06 ) : The EPA rated this scientific rationale for waiver request as acceptable for risk assessment purposes (U.S. EPA 2013; Barsoum and Kough). Hz NPV is a baculovirus with a long history of safe use as a microbial pesticide in the United States. Toxicology results from baculoviruses have raised no

concerns for human or animal health. Baculoviruses have narrow host ranges, and the mode of action is well understood and unique to insect hosts, with virtually no adverse effects in people, wildlife, or non

- target insects. Based on literature search for the period 1980

- 2012, there are no data demonstrating any toxic effects on humans or other mammals exposed to Hz NPV. The

EPA classified the acute inhalation toxicity as Toxicity Category IV

.

Acute Dermal Toxicity

- Rat (Harmonized Guideline 870.1200; MRID No 489417

- 06 ) : The EPA

rated this scientific ratio nale for waiver request as acceptable for risk assessment purposes (U.S. EPA 2013; Barsoum and Kough). Hz NPV is a baculovirus with a long history of safe use as a microbial pesticide in the United States. Toxicology results from baculoviruses have raised no concerns for human or animal health. Baculoviruses have narrow host ranges, and the mode of action is well understood and unique to insect hosts, with virtually no adverse effects in people, wildlife, or non

- target insects. Based on literature search fo r the period 1980

- 2012, there are no data demonstrating any toxic effects on humans or other mammals exposed to Hz NPV. The EPA classified the dermal toxicity as Toxicity Category IV

.

Acute Eye Irritation Study (Harmonized Guideline 870.2400; MRID No 4 89417

- 06 ) : The EPA rated this scientific rationale for waiver request as acceptable for risk assessment purposes (U.S. EPA 2013; Barsoum and Kough). Hz NPV is a baculovirus with a long history of safe use as a microbial pesticide in the United States. Tox icology results from baculoviruses have raised no concerns for human or animal health. Baculoviruses have narrow host ranges, and the mode of action is well understood and unique to insect hosts, with virtually no adverse effects in people, wildlife, or no n

- target insects. Based on literature search for the period 1980

- 2012, there are no data demonstrating any toxic effects on humans or other mammals exposed to Hz NPV. T he EPA classified the eye irritation as Toxicity Category IV

.

Acute Dermal Irritatio n Study (Harmonized Guideline 870.2500; MRID No 489417

- 06 ) : The EPA rated this scientific rationale for waiver request as acceptable for risk assessment purposes (U.S. EPA 2013; Barsoum and Kough). Hz NPV is a baculovirus with a long history of safe use

as a microbial pesticide in the United States. Toxicology results from baculoviruses have raised no concerns for human or animal health. Baculoviruses have narrow host ranges, and the mode of action is well understood and unique to insect hosts, with virtual ly no adverse effects in people, wildlife, or non

- target insects. Based on literature search for the period 1980

- 2012, there

are no data demonstrating any toxic effects on humans or other mammals exposed to Hz NPV.

The EPA classified the primary dermal irritation as Toxicity Category IV

.

b. Acute Toxicology and Subchronic Toxicity/Pathogenicity

– Tier II; ReproductiveFertility Effects, Carcinogenicity, Immunotoxicity, and Infectivity/Pathogenicity Analysis

– Tier III

Tier II and Tier III studies we re not required for Helicoverpa zea ABA Nucleopolyhedrovirus

-Helicoverpa zea Nucleopolyhedrovirus Page 11 of 27

Biopesticides Registration Action Document

U based on the lack of acute toxicity/pathogenicity in the Tier I data requirements.

c. Endocrine Disruptors

As required by the Administrator und er the Federal Food, Drug, and Cosmetic Act ( FFDCA )

, the 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) author izes 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 to not 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 contains 58 pesticide active ingredients and 9 inert ingredients. Helicoverpa zea Nucleopolyhedrovirus (Hz NPV) is not among the group of 58 pesticide active ingredients on the initial list to be screened under the

EDSP. Hz ABA NPV

- U is substantially similar to Hz NPV.

The EPA believes that Hz ABA NPV

- U is a substance t hat would not 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), the EPA will determine in the future whether it can exempt Hz

ABA NPV

- U from the requiremen ts of the Section 408(p)(4) EDSP. In the event the EPA 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, the test guidel ines 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 the 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 the EPA determines that the exemption is “safe.” Section 408(c)(2)(A)(ii) of FFDCA defines

“safe” to mean that “there is a reasona ble 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 an d in residential settings

, but it 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, the EPA must take into account the factors set f orth in section 408(b)(2)(C) of FFDCA, which require the 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 the EPA consider “available information concerning the cumulative effects of [a particular pesticide's] . . . residues and other substances that have a common mechanism of toxicity.”

The EPA performs a number of analyses to determine the risks from aggregate exposure to pesticide residues. First, the Agency determines the toxicity of a pesticid e . Second, the EPAHelicoverpa zea Nucleopolyhedrovirus Page 12 of 27

Biopesticides Registration Action Document

examines exposure to the pesticide through food, drinking water, and through other exposures that occur as a result of pesticide use in residential settings.

Consistent with section 408(b)(2)(D) of FFDCA, the EPA has reviewed the availab le scientific data and other relevant information, and considered its validity, completeness, and reliability and the relationship of this information to human risk. The Agency also considered available information concerning the variability of the sensiti vities 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 5 in Appendix A , t he data required fo r a FFDCA risk assessment for Helicoverpa zea ABA Nucleopolyhedrovirus

- U 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 nonoccupational exposures, including drinking water from ground water or surface water and exposure through pesticide use in gardens, lawns, or buildings (residential and other indoor uses).

Food Exposure and Risk C haracterization : The EPA found that d ietary exposure to Helicoverpa

zea ABA Nucleopolyhedrovirus

- U

, a naturally occurring insect virus

, is anticipated to be negligible

. Due to the ubiquitous nature of the organism and the concentrations of Hz NPV and

oth er closely related viruses that already exist in the environment, the likelihood of increased risk

to humans or animals from the use of Heligen is low. The EPA concluded that t he risk posed to adults, infants, and children is likely to be minimal because o f the low acute oral toxicity/ p athogenicity potential of Hz ABA NPV

- U and any potential exposure to Hz ABA

NPV

- U also would be minimal.

Drinking Water Exposure and Risk Characterization : Exposure to residues of Helicoverpa zea

ABA Nucleopolyhedrovirus

- U i n consumed drinking water is unlikely. The proposed use patterns for Heligen are soil directed and/or soil incorporated , thereby limiting contact with surface water by drift and runoff. Furthermore, ground water is not expected to have significant

exposure to Hz NPV since, like other microorganisms, this microbial pesticide would likely be filtered out by the particulate nature of many soil types. If it were to be transferred to surface or ground waters that are intended for eventual human consumption (e.g. , through spray drift or runoff) and directed to wastewater treatment systems or drinking water facilities, it likely would not survive the conditions water is subjected to in such systems or facilities, including

chlor ination, pH adjustments, filtration

, and/or occasionally high temperatures (Centers for Disease Control and Prevention 2009; U.S. EPA 2004). In the remote likelihood that Hz NPV is

present in drinking water (e.g., water not subject to treatment systems or facilities), its target pest specific ity and available data indicate no toxicity and/or pathogenicity is likely to occur with any drinking water exposure to Hz ABA NPV

- U that results from pesticide application s made in accordance with good agricultural practice s (see section IV(B)(1)(a) on pa ges 8

– 10 and Table 5

in Appendix A ).

Non

- occupational, Residential Risk Characterization : Given that Hz ABA NPV

- U is naturally

occurring and usually not stable outside of its insect host

, EPA determined that non

- occupational

exp osure to the bacterium is unlikely

. Repeated exposure s to the microorganism from pesticidalHelicoverpa zea Nucleopolyhedrovirus Page 13 of 27

Biopesticides Registration Action Document

applications do not exceed EPA’s level of concern, particularly in light of available data that

demonstra te Hz ABA NPV

- U is not toxic (acute dermal toxicity and acute pulmonary toxicity/ pathogenicity), non

- irritating (primary dermal irritation), and not pathogenic (acute pulmonary toxicity/pathogenicity) (see section IV(B)(1)(a) on pages 8

– 10 and Table 5 in Appendix A ).

b. Cum ulative Effects from Substances with a Common Mechanism of Toxicity

Section 408(b)(2)(D)(v) of FFDCA requires that, when considering whether to establish, modify, or revoke a tolerance exemption, the EPA consider “available information concerning the

cumu lative effects of [a particular pesticide’s] . . . residues and other substances that have a

common mechanism of toxicity.”

The EPA has not found Helicoverpa zea ABA Nucleopolyhedrovirus

- U to share a common mechanism of toxicity with other substances, an d Hz ABA NPV

- U d oes not appear to produce a toxic metabolite against target pest s . For the purposes of the tolerance action, therefore, the EPA

has assumed that Hz ABA NPV

- U does not have a common mechanism of toxicity with other substances. Thus, section 408(b)(2)(D)(v) of the FFDCA does not apply. For information regarding the EPA’s efforts to determine which chemicals have a common mechanism of toxicity and to evaluate the cumulative effects of such chemicals, see the EPA’s website at http://www.epa.gov/pesticides/cumulative

.

c. Determination of Safety for the United States Population, Infants and Children

In considering the establishment of a tolerance or tolerance exemption for a pesticide c hemical residue, FFDCA section 408(b)(2)(C) provides that the 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 cumulativ e 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 the EPA shall apply an additional tenfold (10X) margin of exposure ( safety ) for infants and ch ildren in the case of threshold effects to account for prenatal and postnatal toxicity and the completeness of the database on toxicity and exposure

unless the EPA determines that a different margin of exposure ( safety ) will be safe for infants

and childre n. This additional margin of exposure ( safety ) is commonly referred to as the Food Quality Protection Act Safety Factor. In applying this provision, the EPA either retains the default value of 10X or uses a different additional safety factor when reliable data available to

the EPA support the choice of a different factor.

Based on the acute toxicity an d pathogenicity data/information discussed in section IV(B)(1)(a) (see pages 8

– 10 ) and Table 5 in Appendix A

, the EPA concludes th at there are no threshold effects of concern to infants, children, or adults when Helicoverpa zea ABA Nucleopolyhedrovirus

- U is used as labeled in accordance with good agricultural practices. As a result, the EPA concludes that no additional margin of ex posure (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.

Moreover, based on the same data /information and EPA analysis as presented directly above, t he Agency is able to conclude that there is a reasonable certainty that no harm will result to the

United States population, including infants and children, from aggregate exposure to the residues

of Helicoverpa zea ABA Nucleopolyhedrovirus

- U when it is used

— as labeled and inHelicoverpa zea Nucleopolyhedrovirus Page 14 of 2 7

Biopesticides Registration Action Document

accordance with good agricultural practices

— as a n insecticide . Such exposure includes all anticipated dietary exposures and all other exposures for which there is reliable information. The EPA has arrived at this conclusion because, considered collectively, the data and information

available on Hz ABA NPV

- U do not demons trate toxic and/ pathogenic potential to mammals, including infants and children.

3. Occupational Exposure and Risk Characterization

The EPA does not expect h andle r exposure to Helicoverpa zea ABA Nucleopolyhedrovirus

- U to pose any undue risk. Regardless, appropriate personal protective equipment (PPE) and precautionary statements are required on pesticide product labels to mitigate any potential risks to pesticid e handlers due to prolonged or numerous exposures. Handlers applying Heligen in agricultural settings must wear a dust/mist filtering respirator , waterproof gloves, long

- sleeved shirt, long pants, socks and shoes. For future products containing this insect virus as an active ingredient

, the EPA may require a dditional PPE (e.g., protective eyewear ) on a product

- specific

basis.

4. Human Health Risk Characterization

The EPA considered human exposure to Helicoverpa zea ABA Nucleopolyhedrovirus

- U in light of the standard for registration in FIFRA and the relevant safety factors in FFDCA. The Agency determin ed that no unrea sonable adverse effects to the United States population in general, and to infants and children in particular, will result when Hz ABA NPV

- U is used in

accordance with EPA

- approved labeling.

C. Environmental Assessment ( 40 CFR § 158.2150 )

Data and other information (e.g., scientific literature) submitted and cited by the applicant to

support requests to waive nontarget organism testing for Helicoverpa zea ABA Nucleopolyhedrovirus

– U are sufficient to fulfill the relevant microbial pest icide data requirements and for risk assessment purposes. In addition, the applicant submitted one non

guideline study on nontarget insects that partially fulfilled this data requirement. Further testing of nontarget organisms at higher tier levels (i.e., Tiers I I, III, and IV) is not required. The EPA performed an environmental risk assessment based on data and other information provided by the ap plicant, and the Agency determined that adverse effects to nontarget organisms are not anticipated from the pro posed pesticidal uses of Hz ABA NPV

- U . Moreover, the EPA has made

a “No Effect” determination for direct and indirect effects to federally listed threatened and endangered species and their designated critical habitats resulting from these same proposed

pe sticidal uses.

For a summary of the generic data requirements described in sections IV(C)(1) (see pages 14

– 1 5 ) , refer to Table 6 in Appendix A

.

1. Ecological Exposure and Risk Characterization

The applicant evaluated the identity or similarity between NPV strains by extracting DNA, treating with restriction endonuclease and using electrophoresis to compare the gel patterns. The products, AgBiTech's Heligen and the previously registered product containing Hz NPV, Gemstar ( EPA Registration Number 70051

- 45), were examined by light microscopy usingHelicoverpa zea Nucleopolyhedrovirus Page 15 of 27

Biopesticides Registration Action Document

Giemsa stain and X I000 magnification under phase contrast. This process confirmed that the products contain a nucleopolyhedrovirus with occlusion bodies visually similar to each ot her. Researchers extracted the virus from both products, and they extracted DNA from the virus occlusion bodies. Then, this DNA was digested with restriction endonuclease (RENs) and the profile under electrophoresis on agarose was compared with that of NPV in Hz NPV (Gemstar). This analysis shows that the Hz ABA NPV

- U in the AgBiTech product is almost identical to the

Hz NPV found in Gemstar. Based on this, data previ ously submitted to support the registration

of HzNPV (Gemstar) can be bridge d to this new strain.

a

. Terrestrial Animals and Plants

Wild Mammals (Harmonized Guideline 885.4150, MRID No. 489417

- 06), Birds (Harmonized

Guideline 885.4 10 0), Nontarget Insects (Harmonized Guideline 885.4340 , MRID No. 489418

- 10 ), Honey Bees (Harmonized Guideline 885.4380

, MRID No. 48 9 418

- 09 ) and Plants (Harmonized Guideline 885.4300) :

The proposed applications

— ground and aerial sprays and sprinkler chemigation

– would deposit

Hz ABA NPV

- U on foliar and other above ground plant surfaces. EPA expects deposition onto
these su rfaces to occur within the intended area of treatment

, as well as surrounding areas.

Persistence of Hz ABA NPV

- U in the environment, particularly in soil, is possible as long as conditions are favorable, but it does not pose risks to nontarget species because of its specificity. Testing on birds

, wild m ammals, nontarget insects, and honey bees were not required because the applicant instead submitted acceptable scientific rationale and /or cited previous studies. The

EPA additionally determined that, since Nz ABA NPV

- U is not taxonomically related to known plant pathogens, adverse effects to nontarget plants are not expected. Baculoviruses such as Hz NPV have a history of safe use as microbial pesticides, and they generally have narrow host ranges with no adverse effects on nontarget organisms (OEC D 2002). Typically, NPVs only infect the genus or perhaps family from which t hey w ere originally isolated. Searches of public

literature did not indicate reports of adverse effects of Hz NPV to these nontarget organisms.

An additional bioassay with a nontarget insec t

, Spodoptera frugiperda

, was submitted (MRID 48941809). T his study tested the toxicity of Hz ABA NPV

- U to this insect, and results indicated the potential for some effects in this nontarget lepidopteran

. However, effects to S

. f r u g i p e r d a would not be expected at field application rates, a n d s e n s i t i v i t y i s e x p e c t e d to be less in species that are not as closely related to the target pest

. See USEPA ( 2013d )

. Based on the information submitted and cited by the applicant, adverse effects to terrestrial nontarget organis ms are not expected to result from the proposed pesticidal uses of Hz ABA NPV

- U

.

b. Aquatic Animals and Plants

Freshwater Fish (Harmonized Guideline 885.4200 ), Freshwater Invertebrates (Harmonized Guideline 885.4240) , Estuarine/Marine Fish and Inverte brates (Harmonized Guideline 885.4280) (MRID No. 48 9418

- 10 ) :

The EPA determined that f reshwater and e stuarine fish and invertebrates are not likely to be exposed to Hz ABA NPV

- U from runoff and drift into aquatic areas and that Hz ABA NPV

- U is not pathoge nic to aquatic organisms. T he applicant submitted acceptable scientific rationale and/or cited previous studies. Baculoviruses such as Hz NPV have a history of safe use as microbial pesticides, and they generally have narrow host ranges with no adverse eff ects onHelicoverpa zea Nucleopolyhedrovirus Page 16 of 27

Biopesticides Registration Action Document

nontarget organisms (OECD 2002). Typically, NPVs only infect the genus or perhaps family from which they were originally isolated. Searches of public literature did not indicate reports of adverse effects of Hz NPV to these nontarget organisms.

Based on the information submitted and cited by the applicant, adverse effects to aquatic nontarget organisms are not expected to result from the proposed pesticidal uses of Hz ABA

NPV

- U

.

2. Environmental Fate Data

As t he data and information provided are sufficient to fulfill the Tier I nontarget organism data requirements and allow for nontarget organism risk asse ssment for Helicoverpa zea ABA Nucleopolyhedrovirus

- U

, f urther t esting at higher levels ( Tiers II, III, and IV) is not required.

3. Threatened and Endangered Species Assessment

Since the EPA has determined that no effects are ant icipated for any non target species exposed

to Helicoverpa zea ABA Nucleopol yhedrovirus

- U as a result of the proposed labeled uses

, effects to federally listed threatened and endangered species and their designated critical ha bitats

also are not expected. Therefore, a “No Effect” determination is made for direct and indirect

eff ects to listed species and their designated critical habitats resulting from the proposed uses of

Hz ABA NPV

- U

.

V. ENVIRONMENTAL JUSTICE

The EPA seeks to achieve environmental justice

— the fair treatment and meaningful involvement of all people regardles s 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 involveme nt 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 ag ency’s decision; (3) the concerns of all participants involved will

be considered in the decision

- making process; and (4) the decision

- makers seek out and facilitate the involvement of those potentially affected. The EPA has this goal for all communities a nd persons across the United States.

To help address potential environmental justice issues, t he EPA seeks information on any groups or segments of the population who, as a result of their location, cultural practices, or other factors, may have atypical , unusually high exposure to Helicoverpa zea ABA Nucleopolyhedrovirus

- U 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 add itional information regarding environmental justice issues, please visit the EPA’s web

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

.Helicoverpa zea Nucleopolyhedrovirus Page 17 of 27

Biopesticides Registration Action Document

V I

. RISK MANAGEMENT DECISION

Section 3(c)(5) of FIFRA permits for the registration of a pesticide provided that all the following determinations are made:

(1) Its composition is such as to warrant the proposed claims for it; (2) Its labeling and other material 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 Helicoverpa zea ABA Nucleopolyhedrovirus

- U product s, MP and EP,

ha ve well

- known properties. The EPA has no knowledge that would contradict t he claims made

on the label s
, and th e p esticide product s are not expected to cause unreasonable adverse effects

on the environment when used according to label instructions. Criterion 2 is satisfied by the

current product label s , as well as the data and information presented in this document. The Agency believe s that the Hz ABA NPV

- U p esticide product s will not cause any unreasonable

adverse effects on the environment . I n fact

, the EP, Heligen

, is likel y to provide protection against many infestations , satisfying criterion 3. Criterion 4 is satisfied in that the Hz ABA NPV

U pesticide product s are not expected to cause unreasonable adverse effects when used according to label instructions. Therefore, both the MP and the EP, Heligen

, which contain s Hz ABA NPV

U as a new active ingredient, are eligible for registration under FIFRA section 3(c)(5) for the

labeled uses.

V II

. ACTIONS REQUIRED OF THE REGISTRANT

A. Final Printed Labeling

Before the pro posed manufacturing

- use product, Helicoverpa zea ABA Nucleopolyhedrovirus

- U , and the proposed end

- use product, Heligen , may be released for shipment, the applicant is required to provide appropriate final printed labeling to the EPA

.

B. Terms of Registr ation

The EPA is not requiring additional information on the registration of these two products

.

C. Reporting of Adverse Effects and Hypersensitivity Incidents

Notwithstanding the information stated in the previous sections, it should be clearly unde rstood

that certain specific data must be reported to the EPA as a requirement for maintaining the federal registration for a pesticide product. Reports of all incidents of adverse effects to the

environment must be submitted to the EPA under the provision s stated in FIFRA section 6(a)(2). Additionally, all incidents of hypersensitivity (including both suspected and confirmed incidents) must be reported to the EPA under the provisions of 40 CFR § 158.2140(d).Helicoverpa zea Nucleopolyhedrovirus Page 18 of 27

Biopesticides Registration Action Document

VIII. GLOSSARY OF ACRONYMS AND ABBREVIATIONS

BRAD Biopesticides Registration Action Document

CFR Code of Federal Regulations

cSt centistokes

EDSP Endocrine Disruptor Screening Program

EP end

- use pesticide product

EPA United States Environmental Protection Agency (the “Agency”)

FFDCA Federal Food, Drug, and Cosmetic Act

FIFRA Federal Insecticide, Fungicide, and Rodenticide Act

FR Federal Register g/mL grams per milliliter

Hz NPV Helicoverpa zea Nucleopolyhedrovirus product lb/gal pounds per gallon

LD 50 median lethal dose. A statistically derived sin gle dose that can be expected

to cause death in 50% of the test animals when administered by the route indicated (oral, dermal, or inhalation). It is expressed as a weight of substance per unit weight of animal (e.g., mg/kg). mg/kg milligrams per kil ogram
MP manufacturing

- use pesticide product

MRID No. Master Record Identification Number

NIOSH National Institute for Occupational Safety and Health

OPP Office of Pesticide Programs

PC Code Pesticide Chemical Code

PP Pesticide Petition

PPE personal prote ctive equipment

TGAI technical grade of the active ingredient

U.S. United StatesHelicoverpa zea Nucleopolyhedrovirus Page 19 of 27

Biopesticides Registration Action Document

I X. BIBLIOGRAPHY

A. Studies Submitted to Support the Active Ingredient, Helicoverpa zea ABA Nucleopoly

hedrovirus

- U , and Pesticide Product Regi stration s for the MP and the EP

, Heligen

.

MRID No. Study Information

48941700 AgBiTech Pty Ltd. (2012) Submission of Product Chemistry and Toxicity Data in

Support of the Application for Registration of H eligen. Transmittal of 7 Studies

.

48941701 MacIntosh, S. (2012) Product Identity, Manufacturing Process, Product

Characterization: Viscosity & Discussion of Unintentional Ingredients: for Heligen (EP). Unpublished study prepared by MacIntosh & Associates, Inc. 17p.

48941702 MacIntosh, S. (2012) Analysis of Samples: for Heligen (EP). Unpublished study

prepared by MacIntosh & Associates, Inc. 15p.

48941703 Hawes, A. (2012) Certification of Limits: for Heligen (EP). Unpublished study

prepared by AgBiTech Pty Ltd. 8p.

48941704 Hawes, A. (2006) Storage Stability & Corrosion Characteristics for Heligen (EP).

Project Number: 06/001/2. Unpublished study prepared by Queensland Dept. of Primary Industries, Sonic Food and Water Te sting and AgBiTech Pty Ltd. 18p.

48941705 Hawes, A. (2012) Hypersensitive Incidents: Helifen (EP). Unpublished study

prepared by AGBITECH PTY LTD. 6p.

48941706 MacIntosh, S. (2012) Statisfaction of Data Requirements with Public Literature

and/ or Scientific Rationale: Acute Oral Toxicity: Acute Dermal Toxicity: Acute Inhalation Toxicity: Acute Eye Irritation Toxicity: Acute Dermal Irritation Toxicity: for Heligen. Unpublished study prepared by MacIntosh & Associates, Inc. 14p.

489417 07 MacIntosh, S. (2012) Satisfaction of Data Requirements with Public Literature and/or

Scientific Rationale: Freshwater Fish Toxicity/Pathogenicity: Freshwater Invertebrate Toxicity/Pathogenicity: for Heligen (EP). Unpublished study prepared by MacI ntosh & Associates, Inc. 9p

.

48941800 AgBiTech Pty Ltd. (2012) Submission of Product Chemistry and Toxicity Data in

Support of the Application for Registration of Helicoverpa zea Nucleopolyhedrovirus.

Transmittal of 10 Studies.

48941801 MacI ntosh, S. (2012) Product Identity, Manufacturing Process & Discussion of

Unintentional Ingredients: for Helicoverpa zea Nucleopolyhedrovirus. Unpublished study prepared by MacIntosh & Associates, Inc. 23p.

48941802 MacIntosh, S. (2012) Analysis of Samples: for Helicoverpa zea

Nucleopolyhedrovirus (TGAI/MP). Unpublished study prepared by MacIntosh & Associates, Inc. 16p.Helicoverpa zea Nucleopolyhedrovirus Page 20 of 27

Biopesticides Registration Action Document

MRID No. Study Information

48941803 Hawes, A. (2012) Certification of Limits: for Helicoverpa zea Nucleopolyhedrovirus.

Unpublished study prepared by AgBiTech Pty Ltd. 7p

.

48941804 Hawes, A. (2012) Product Chemistry: for Helicoverpa zea Nucleopolyhedrovirus.

Unpublished study prepared by AgBiTech Pty Ltd. 7p.

48941805 MacIntosh, S. (2012) Satisfaction of Data Requirement s with Public Literature

and/or Scientific Rationale: Storage Stability an d Corrosion Characteristics for Helicoverpa zea Nucleopolyhedrovi rus (TGAI/MP) . Unpublished study prepared by MacIntosh & Associates, Inc. 15p.

48941806 Hawes, A. (2012) Hypersensitive Incidents: Helicoverpa zea Nucleopolyhedrovirus

(TGAI/MP). Unpublished study prepared by AgBiTech Pty Ltd. 6p.

48941807 Pavloff, N. (2012) Cell Culture Tests for a Helicoverpa zea Nucleopolyhedrovirus.

Project Number: 173/4P/1R/CB/04/15/11. Unpublished study. 22p.

48941808 MacIntosh, S. (2012) Satisfaction of Data Requirements with Public Literature and/or

Scientific Rationale: for Helicoverpa zea Nucleopolyhedrovirus. Unpublished study prepared by MacIntosh & Associates, Inc. 18p.

48941809 Popham, H. (2012) Non

- Target Insect Bioassay: for Helicoverpa zea Nucleopoly

hedrovirus. Project Number: 12/016. Unpublished study prepared by USDA

- ARS BCIRL. 13p.

48941810 MacIntosh, S. (2012) Satisfaction of Data Requirements with Public Literature and/or

Scientific Rationale: for Helicoverpa zea nucleopolyhedrovirus (TGAI/MP). Unpublished study prepared by MacIntosh & Associates, Inc. 14p.

4901 1 400 AgBiTech Pty, Ltd. (2012) Submission of Product Chemistry and Toxicity Data in

Support of the Applications for Registration of Helicoverpa zea N ucleopoly

hedrovirus and Heligen. Tra nsmittal of 5 Studies.

4901 1 401 MacIntosh, S. (2012) Supplemental Information Product Identity: for Helicoverpa zea

Nucleopolyhedrovirus Supplemental to MRID No 489418

- 01. Unpublished study

p repared by MacIntosh & Associates, Inc. 22p

.

4901 1 402 MacIntosh, S. (2012) Satisfaction of Data Requirements with Public Literature and/or

Scientific Rationale for HzNPV for Toxicology Guideline Studies Supplement to:

M RID No 489418

- 08. U npublished study by MacIntosh & Associates, Inc. 13p

.

49011403 MacIntosh, S. (2012) Satisfaction of Data Requirements with Public Literature and/or

Scientific Rationale for HzNPV for Non

- target Organism Guideline Studies

S upplement to: MRID No 489418

- 10. Unpublished study prepared by MacIntosh &

Associates , Inc. 13p

.Helicoverpa zea Nucleopolyhedrovirus Page 21 of 27

Biopesticides Registration Action Document

MRID No. Study Information

49011404 MacIntosh, S. (2012) Satisfaction of Data Requirements with Public Literature and/or

Scientific Rationale for Heligen for Toxicology Guideline Studies Supple ment to:
MRID No 489417

- 06. Unpublished study prepared by MacIntosh & Associates, Inc

. 13p.

4901 1 405 MacIntosh, S. (2012) Publications in Support of Registration Applications for

H elicoverpa zea Nucleopolyhedrovirus and Heligen. Unpublis hed study prepared by MacIntosh & Associates, Inc. 322p.

49020400 AgBiTech Pty, Ltd. (2012) Submission of Product Chemistry Data in Support of the

Applications for Registration of Helicoverpa zea Nucleopolyhedrovirus and Heligen.

Transmitta l of 1 Study.

49020401 Popham, H.; Hawes, A. (2012) Supplemental Information: Product Identity: For

H elicoverpa zea Nucleopolyhedrovirus. Unpublished study prepared by Agbitech, Ltd and U.S. Dept. of Agriculture. 35p.

B. Environmental Protecti on Agency Risk Assessment Memoranda

U.S. EPA. 201 3 a. Review of Product Chemistry , Manufacturing Process, and Acute Toxicity

Studies of the TGAI Helicoverpa zea Nucleopolyhedrovirus (HZ NPV) for section 3 product registration (EPA Reg. no. 87978

- R ). Memora ndum from Ibrahim S. Barsoum,

Ph.D. and John L. Kough, Ph.D. to Michael Glikes

, dated Ju ne 4 , 201 3

.

U.S. EPA. 2013b. Review of Product Chemistry, Manufacturing Process, and Acute Toxicity Studies of the EP Heligen containing Helicoverpa zea Nucleopolyhedr ovirus (HZ NPV)

TGAI for section 3 product registration (EPA Reg. no. 87978

- E). Memorandum from Ibrahim S. Barsoum, Ph.D. and John L. Kough, Ph.D. to Michael Glikes, dated June 12,

2013.

U.S. EPA. 2013c. Review of the registrant’s response to the identifi cation and concentration of the surfactant used in the manufacturing process of the EP Heligen containing Helicoverpa zea Nucleopolyhedrovirus (HZ NPV) (EPA Reg. no. 87978

- E). Memorandum from Ibrahim S. Barsoum, Ph.D. and John L. Kough, Ph.D. to Michael

Gl ikes, dated June 10, 2013.

U.S. EPA. 201 3d

. Environmental Risk Assessment of Polyhedral occlusion bodies (OBs) of the nuclear polyhedrosis virus of Helicoverpa zea for FIFRA Section 3 Registrations of Helicoverpa zea Nucleopolyhedrovirus Manufacturing Us e product (EPA File Symbol

87978

- R) and Heligen End Use Product (EPA File Symbol 87978

- R) for Control of Helicoverpa zea and Heliothis virescens in Food and Nonfood Crops; PC Code: 107300; Decision No. 469835 and 469836; DP Barcodes: 408136 and 408138; Sub mission Nos. 928090 and 928232; MRIDs 48941809, 48941810, 49011403, and 48941707

.

U.S. EPA. Registration Eligibility Decision (RED) Facts: Heliothis Zea NPV, January 1991.Helicoverpa zea Nucleopolyhedrovirus Page 22 of 27

Biopesticides Registration Action Document

C. Other References

Annals of the Entomological Society of America. 2004. A Ne w Synonym of Helicoverpa zea (Boddie) and Differentiation of Adult Males of H. zea and H. armigera (Hubner). Vol.
97, no. 6.

Centers for Disease Control and Prevention. 2009. Drinking Water

- Water Treatment. Available

from http://www.cdc.gov/healthywater/drinking/public/water_treatment.html

.

DeFelice K, Wollenhaupt N, Buchholz D. 1993. Aquifers and Soil Filter Effect. Available from http://extension.missouri.edu/p/WQ24

.

U.S. EPA. 2004. Primer for Municipal Wastewater Treatment Systems. EPA 832

- R

- 04

- 001.

Available from http://www.epa.gov/npdes/pubs/primer.pdf

.Helicoverpa zea Nucleopolyhedrovirus Page 23 of 27

Biopesticides Registration Action Document

AP PENDIX A. MICROBIAL PESTICIDES DATA REQUIREMENTS

(40 CFR PART 158

– SUBPART V)

TABLE 1. Product Analysis Data Requirements for the Active Ingredient (TGAI), Helicoverpa zea AB Nucleopolyhedroviru

- U s (40 CFR § 158.2120)

Harmonized

G uideline

N umbe r Data Requirement Results MRID No.

885.1100 Product Identity Submitt ed data fulfill the requirement for product identity and composition. 48941801

885.1200 Manufacturing

Process Submitted data fulfill the requiremen t for manufacturing process

. 48941 801

N/A Deposition of a

Sample in a Nationally Recognized Culture

Collection Helicoverpa zea ABA Nucleopolyhedrovirus

- U is on deposit with the ATCC

. 48941801

885.1300 Discussion of

Formation of

Unintentional Ingredients Submitted data fulfill the req uirement for preventing and detecting any

microbial contaminants. 48941801

885.1400 Analysis of S amples Submitted data fulfill the requirement for analysis of samples. 4 8941802

8 85 .1 50 0 Certification of

L imits Submitted data fulfill the requirement fo r manufacturing process. 4 8941803

TABLE 2 . Product Analysis Data Requirements for the End

- Use Pesticide Product (EP), Heligen (40 CFR § 158.2120)

Harmonized

Guideline

Number Data Requirement Results MRID No.

885.1100 Product Identity Submitted data fulfill the requirement for product identity. 489 41701

885.1200 Manufacturing Process Submitted data fulfill the requirement for manufacturing

process. 48 941701

N/A Deposition of a Sample in a

Nationally Recognized

Culture Collection Submitted data fulfill the requirement for deposition of a sample

in a culture collection. N/A

885.1300 Discussion of Formation of

Unintentional Ingredients Submitted data fulfill the requirement for preventing and detecting any microbial contaminants. 48 941701

885.1 400 Analysis of Samples Submitted data fulfill the requirement for analysis of samples for purposes of FIFRA section 3(c)(5) registration. 4 8941702

885.1500 Certification of Limits Limits listed on the confidential statement of formula are adequate/ac ceptable. 48 941703Helicoverpa zea Nucleopolyhedrovirus Page 24 of 27

Biopesticides Registration Action Document

TABLE 3

. Physical and Chemical Characteristics for the Technical Grade of the Active Ingredient (TGAI)

, Helicoverpa zea ABA Nucleopolyhedrovirus

- U (40 CFR § 158.2120)

Harmonized

G uideline N umber Data Requirement Results MRID No.

830.6302 Color Grey

- brown translucent liquid

48 941804 830.6303 Physical State Liquid

830.630 4 Odor Earthy

830.6313 Stability to Normal and

E levated Temperatures, Metals,

and Metal I ons N/A N/A

830.6317 Storage Stability Submitted data fulfill the r equirement for storage stability. 48 941805

830.6319 Miscibility Non emulsifiable liquid. N/A

830.6320 Corrosion

Characteristics Submitted data fulfill the requirement for corrosion

characteristics. 48941805

830.7000 pH 5.0

– 6.0 48 941804

830.7100 Vis cosity N/A (not a homogenous solution) N/A

830.7300 Density/Relative

Density/Bulk Density (Specific Gravity) 1. 15

– 1.26 kg/L 48 941804

TABLE 4. Physical and Chemical Characteristics for the End

- Use Pesticide Product (EP), Heligen (40 CFR § 158.2120)

Harmonized

Guideline Number Data Requirement Results MRID No.

830.6302 1 Color Grey

- brown translucent liquid 48941704

830.6303 1 Physical State Liquid 48941704

830.6304 1 Odor Earthy 48941704

830.6313 1 Stability to Normal and

Elevated Temperatures,

Me tals, and Metal Ions N/A N/A

830.6317 Storage Stability Submitted data fulfill the requirement for storage stability. 48941704

830.6319 Miscibility Not required because the end

- use pesticide product, Serenade Soil DPZ, is not an emulsifiable liquid for m of microbial pesticides (refer to test note #2 of 40 CFR § 158.2120(d)). N/A

830.6320 Corrosion Characteristics Submitted data fulfill the requirement for corrosion

characteristics 48941704

830.7000 pH 5. 0

– 6.0 48941704

830.7100 Viscosity 6.05 4894 1704

830.7300 Density/Relative

Density/Bulk Density (Specific Gravity) 1. 15

– 1.26 kg/L 48941704

1 According to 40 CFR § 158.2120, these data are only required for the technical grade of the active ingredient. Since AgBiTech

LTD included this information with its application for Heligen , it is summarized appropriately in this table.Helicoverpa zea Nucleopolyhedrovirus Page 25 of 27

Biopesticides Registration Action Document

TABLE 5. Toxicology Data Requirements for the Active Ingredient Helicoverpa zea ABA Nucleopolyhedrovirus

- U

and the End Use Pesticide Product Heligen (40 CFR § 158.2140)

Harmonized

Guideline

Number Data Requirement Results MRID No.

Tier I

885.3050 Acute Oral

Toxicity/Pathogenicity Waived based on extensive list of toxicology studies on other strains of

NPV. Classification: Acceptable

TOXICITY CATEGORY IV 48941706

88 5.3150 Acute Pulmonary

Toxicity/Pathogenicity Waived based on extensive list of toxicology studies on other strains of

NPV. Classification: Acceptable 48941706

885.3200 Acute Injection

Toxicity/ Pathogenicity Waived based on extensive list of toxicology s tudies on other strains of

NPV. Classification: Acceptable 48941706

885.3200 Acute Injection

Toxicity/Pathogenicity (Intravenous) Waived based on extensive list of toxicology studies on other strains of

NPV. Classification: Acceptable 48941706

885.3400 H ypersensitivity

Incidents The applicant reported that n o hypersensitivity incidents, including

immediate

- type or delayed

- type reactions of humans and domestic animals,

occurred durin g research, development, or testing of the TGAI. Any future hypersensitivi ty incidents must be reported to EPA (refer to test note #3 of 40 CFR § 158.2140(d))

. 48941806

885.3500 Cell Culture No cytopathic effects or replication of Hz NPV exposed to any of the

mammalian cells were observed. Classification: Acceptable 48941807

870.1100 Acute Oral Toxicity Waived based on extensive list of toxicology studies on other strains of

NPV. Classification: Acceptable

TOXICITY CATEGORY I V 48941706

870.1200 Acute Dermal Toxicity Waived based on extensive list of toxicology studies on othe r strains of

NPV. Classification: Acceptable

TOXICITY CATEGORY IV 48941706

870.1300 Acute Inhalation

Toxicity Waived based on extensive list of toxicology studies on other strains of

NPV. Classification: Acceptable

TOXICITY CATEGORY I V 48941706

870.2400 Acute Eye Irritation Waived based on extensive list of toxicology studies on other strains of

NPV. Classification: Acceptable

TOXICITY CATEGORY IV 48941706

870.2500 Primary Dermal

Irritation Waived based on extensive list of toxicology studies on other st rains of

NPV. Classification: Acceptable

TOXICITY CATEGORY I V 48941706

NA NA Restriction Endonuclease Analysis Classification: Acceptable 49011402

Tiers II and III

Not required for Helicoverpa zea ABA Nucleopolyhedrovirus

- U based on the lack of acut e toxicity/pathogenicity in the Tier I studies.Helicoverpa zea Nucleopolyhedrovirus Page 26 of 27

Biopesticides Registration Action Document

TABLE 6

. Nontarget Organism Toxicity and Environmental Fate Data Requirements for the Technical Grade of the Active

Ingredient (TGAI), Helicoverpa zea ABA Nucleopolyhedrovirus

- U (40 CFR § 158.2150)

Harmonized Guideline

Number Data Requirement Results MRID No.

Tier I

885.4050 Avian Oral Toxicity Scientific rationale, in addition to citation of MRIDs 000814446 and 00081477, provide

sufficient information to determine that toxicity/pathogenicity to av ian wildlife is not expected from the proposed pesticidal uses. Classification: Acceptable 48 941810

885.4100 Avian Inhalation Toxicity/Pathogenicity Not required as the nature of the microbial pesticide does not indicate potential pathogenicity

to birds o r relatedness to any known bird pathogens (refer to test note #3 of 40 CFR § 158.2150(e)). N/A

885.4150 Wild Mammal Toxicity/Pathogenicity Scientific r ationale indicates that adverse effects to mammals would not be expected. Classification: Acceptable fo r wild mammal

risk assessment 48941810

885.4200 Freshwater Fish Toxicity/Pathogenicity Scientific rationale, in addition to citation of MRIDs 00081444 and 00081445, provide

sufficient information to determine that toxicity/pathogenicity to freshwater fish and invertebrates is not expected from the pesticide. Classification: Acceptable 48941810

885.4240 Freshwater Invertebrate Toxicity/Pathogenicity

885.4280 Estuarine/Marine Fish and Invertebrate Testing Not required because of the established toxicity/ pathogenicity profile, narrow host range, and history of Hz NPV use without apparent

adverse effects to estuarine/marine animals. N/A

885.4300 Nontarget Plant Testing Not required as the microbial pesticide is not taxonomically related to any known plant pathogens (refer to test note #7 of 40 CFR § 158.2150(e)). N/A

885.4340 Nontarget Insect Testing Scientific rationale, in addition to citation of
MRID 00049121, provide s sufficient information to determine that toxicity/pathogenicity to honey

bees and non target insects is not expected as a result of the proposed pesticidal uses. A nonguideline study also provided information on the potential for effects to Spodoptera frugiperda

. Classification: Acceptable 48941809

48 941810 885.4380 Honey Bee Testing

Tiers II, III, and IV Not required for Helicoverpa zea ABA Nucleopolyhedrovirus

- U based on the acceptability of the data and other information provided

for Tier I

.Helicoverpa zea Nucleopolyhedrovirus Page 27 of 27

Biopesticides Registration Action Document

APPENDIX B. PESTICIDE PRODUCT S

EPA File Symbol Registration

Name Percenta

ge Active Ingredie

nt Formulation

Type Use Site(s) Method(s) of Application Application Rate (s) Target Pest

87978

- R Helicoverpa

zea ABA

Nucleo poly

hedrovirus

- U 74.5% Manufacturing

use product N/A N/A N/A Cotton Bollworm ( Helicoverpa zea ) and

Tobacco bu dworm ( Heliothis virescens )

87978

- E Heligen 32% End

- use product Agricultural Ground application

Aerial Chemigation 0.7

– 2.4 fl. oz./ acre

depending on the crop Cotton Bollworm ( Helicoverpa zea ) and

Tobacco budworm ( Heliothis virescens )