Study Title

Tolerance Exemption Petition for Autographa californica multiple nucleopolyhedrovirus

(AcMNPV), strain FV #11, In or On All Raw and Processed Agricultural Commodities

When Used On Crops In Accordance With Good Agricultural Practices

Data Requirement

None

Completion Date

October 29, 2013

Author

SciReg, Inc.

12733 Director's Loop

Woodbridge,VA22192

Sponsor

Andermatt Biocontrol AG

Stahlermatten 6

CH-6146 Grossdietwil

Switzerland

Page 1 of 11
NO CLAIM OF CONFIDENTIALITY

No claim of confidentiality, on any basis whatsoever, is made for any information

contained in this document. I acknowledge that information not designated as within the

scope of FIFRA sec. 10(d)(l)(A), (B), or (C) and which pertains to a registered or

previously registered pesticide is not entitled to confidential treatment and may be

released to the public, subject to the provisions regarding disclosure to multinational

entities under FIFRA 10(g).

Submitter: 'WAAjW -p^^T^C^ Date: Z

Patricia Biggio

Agent (SciReg, Inc.)*

*SciReg, Inc. is the authorized agent for Andermatt Biocontrol AG

• • • • • • • ••

Page 2 of 11
TABLE OF CONTENTS

STATEMENT OF NO DATA CONFIDENTIALITY CLAIMS 2

TABLE OF CONTENTS 3

INTRODUCTION 4

PRODUCT NAME AND PROPOSED USE PRACTICES 4

PRODUCT IDENTITY/CHEMISTRY 4

TOXICOLIGAL PROFILE 5

AGGREGATE EXPOSURE 8

CUMULATIVE EXPOSURE 9

SAFETY DETERMINATION 9

EFFECTS ON THE IMMUNE AND ENDOCRINE SYSTEMS 10

EXISTING TOLERANCES 10

INTERNATIONAL TOLERANCES 10

SUMMARY/CONCLUSION 10

4

4

•

!

Page 3 of 11
INTRODUCTION

SciReg, Inc., 12733 Director's Loop, Woodbridge, VA, 22192, on behalf of Andermatt

Biocontrol AG, Stahlermatten 6, CH-6146 Grossdietwil, Switzerland, is proposing, pursuant

to section 408(d) of the Federal Food, Drug, and Cosmetic Act (FFDCA), 21 U.S.C.

346a, to establish an exemption from the requirement of a tolerance for the microbial

active ingredient, Autographa californica multiple nucleopolyhedrovirus (AcMNPV), strain

FV #11, applied in or on all raw agricultural commodities in accordance with good

agricultural practices.

SciReg, Inc., on behalf of Andermatt Biocontrol AG, is submitting the following

information, data, and arguments in support of their petition.

AcMNPV, strain FV #11, is being submitted for registration as a new active ingredient in

the end-use product, Loopex, for control of cabbage looper larvae (Trichoplusia ni) on

cole crops, green leafy vegetables, celery, beans, peas, peppers, cucumbers, tomatoes,

peppers, melons, ornamentals, and flowers.

PRODUCT NAME AND PROPOSED USE PRACTICES

Loopex will be applied pre-harvest in greenhouses and open fields to cole crops, green

leafy vegetables, celery, beans, peas, peppers, cucumbers, tomatoes, peppers, melons,

ornamentals, and flowers to protect these crops against infestation caused by cabbage

looper larvae (Trichoplusia ni). Application rates are projected to be up to 5 fluid

ounces/acre/application.

PRODUCT IDENTITY/CHEMISTRY

Identity:

Autographa californica MNPV strain FV #11

Physical/Chemical Properties

Property Result Reference

Color Light medium brown

Sylvar Technologies, Inc Report,

Loopex; Autographa californica

multiple nuclepolyhedrovirus

(AcMNPV) FV #11, batch #6 Physical

Properties

Physical state Liquid

Sylvar Technologies, Inc Report,

Loopex; Autographa californica

multiple nuclepolyhedrovirus

(AcMNPV) FV #11, batch #6 Physical

Properties

Odor Undetectable

Sylvar Technologies, Inc Report,

Loopex; Autographa californica

multiple nuclepolyhedrovirus

(AcMNPV) FV #11, batch #6 Physical

Properties

Page 4 of 11
Density

pH

Viscosity

1.154g/ml(9.63 Ibs/gal)

6.31

12.64cStat20°C.

5.86cStat40°C.

Stillmeadow Product Chemistry

Report, Study No. 17287-13

Stillmeadow Product Chemistry

Report, Study No. 17287-13

Stillmeadow Product Chemistry

Report, Study No. 17287-13

Magnitude of Residue and Method to Determine the Residue:

No magnitude of residue (MOR) studies have been conducted with Autographa californica

MNPV strain FV #11 based on the low toxicity of the active ingredient (see Toxicological

Profile, pages 6-12) and a total absence of any known infections to organisms other than

a narrow range of Lepidopteran insects. OPPTS Guideline 885.2000, section f,

Background for Residue Analysis of Microbial Pest Control Agents (MCPA), states:

"The Agency evaluates residue data for MCPAs used on food, feed, or raw

agricultural commodities only if toxic or other harmful properties were observed

in the maximum hazard toxicology tests (Tier I) prescribed in toxicology test

guidelines OPPTS 885.3050 through 885.3650 of this series. If Tier I toxicology

tests indicate no toxic or other harmful properties, then no residue data (with the

general exception of a monitoring method) would be indicated and thus a

recommendation for an exemption from the requirements of a tolerance can be

made."

SciReg, Inc., on behalf of Andermatt, is submitting a petition to establish an exemption

from the requirement of a tolerance and, therefore, an analytical method is not required.

TOXICOLIGAL PROFILE

The full reports that support this petition and which are summarized in this section were

previously submitted to, have been reviewed and accepted by the Agency, are cited

herein in support of the registration of another similar baculovirus product, Gusano, EPA

Reg. No. 70051-43.

Andermatt Biocontrol AG relied on toxicity/pathogenicity data cited from various

sources of public literature. Based on the data submitted and cited, it is established that

Autographa californica MNPV strain FV #11 is neither toxic, nor pathogenic to mammals,

birds, fish, aquatic invertebrates, plants, and nearly all non-Lepidopteran insects.

Acute Oral Toxicity/Pathogenicity:

Study 1 (MRID 74175): Twenty male and 20 female Sprague-Dawley rats were dosed

per os with Autographa californica MNPV at a rate of 4.35 x 107 occlusion bodies per

rat. Test animals exhibited body temperatures, mean body weights, and food consumption

similar to that of the controls. Four animals of each sex were sacrificed on the following

days: 0, 3, 7,14, and 21. Hematological and blood chemistry results appeared to be

Page 5 of 11
within normal limits. Histopathological examination revealed no microorganism related

lesions. There were no deaths and all animals appeared normal throughout the entire

study. It was concluded that A. californica MNPV was not injurious to rats.

Study 2 (MR1D 43912201): As cited per the National Pesticide Information Retrieval

System. *

Acute Dermal Toxicity:

Study 1(MRID 72638): Five male and five female albino guinea pigs were treated with a

single dermal application of 0.4 mL of suspended occlusion bodies (4.35 x 107 occlusion

bodies per rat.) on their shaved backs. Doses of 0.1 mL were applied to 2 abraded areas

and 2 areas of intact skin on each animal. There were no deaths and all A. californica

MNPV-treated animals appeared normal throughout the entire study. It was concluded

that A. californica was not injurious to rats and did not produce dermal toxicity.

Study 2(MRID 48624712): A dermal toxicity study was conducted with the test material

Baculovirus heliothis NPV. Rhesus monkeys were exposed to the organism. All monkeys

except for one survived the study and showed normal weight gains. One female monkey

in the subcutaneous injection group died of unknown causes at day 24 of the study. One

subcutaneous injected male had rapid sedimentation rate, elevated lecocytes and reduced

lymphocytes on day 181 which returned to normal on day 197. No test substance-related

effects were observed.

..

Acute Pulmonary Toxicity/Pathogenicity:

Study 1 (MRID 48624712): An acute pulmonary toxicity/pathogenicity study was

conducted with the test material Baculovirus heliothis NPV. Rhesus monkeys were

exposed to the organism at 1 mg/kg bw (1.2 x 10 PIB/kg bw), Ix weekly for 26 weeks.

All monkeys except for one survived the study and showed normal weight gains. One

female monkey in the inhalation treatment route had low platelet at day 181 which

returned to normal. No test substance-related effects were observed.

Acute Inhalation Toxicity:

Study 1 (MRID 74176) .Ten male and ten female Sprague-Dawley rats were exposed to a

single dose of 6 x 1011 polyhedral occlusion bodies of AcMNPV for one hour. The

nominal chamber concentration after one hour was 11.1 mg/L. All animals were observed

for mortality, signs of toxicity, and abnormal behavior throughout the one-hour exposure

period and daily for 14 days following exposure. Body weights, food consumption, and

organ weights were also observed. Inhalation exposure of rats to AcMNPV produces no

injurious results.

Page 6 of 11
Eye Irritation:

Study 1 (MRID 72639): A single application of 0.1 ml of each the test material was

administered into the conjunct! val sac of the left eye of each of five male and five female

albino (New Zealand White) rabbits weighing from 2.1 to 3.3 kg. The right eye of each

rabbit served as a control receiving a single application, 0.1 ml. of a 0.8% saline solution

containing 2% lactose. Initial fluorescein examination confirmed the absence of irritation and

corneal damage in all eyes. Observations for injury and irritation were recorded at 24, 48 ,

and 72 hours and seven and 14 days. No eye irritation was observed; the eye irritation score

was zero.

Dermal Irritation:

Study 1 (MRID 74177): Dermal irritation was assessed by applying 0.5 mL (2.17 x 107

polyhedral occlusion bodies) ofAutographa californica MNPV to intact and abraided

skin of six New Zealand White Rabbits. Assessment followed the Draize system. No

dermal irritation was observed and, therefore, the primary irritation score was zero.

Study 2 (MRID 48624717): A primary dermal irritation study was conducted using the

test substance Granupom SC (CpGV) containing 2.2 x 1013 granules/L. The test substance

was applied undiluted dermally in a single dose of 0.5 mL/animal onto the shaved dorsolumber region of 3 rabbits. The application site was covered with a patch and secured by

an occlusive bandage. After removal of the bandage at the end of the exposure period (4

h), the residual test substance was gently removed with water. Observations for mortality,

behavior, clinical signs, body weight, and skin reactions were made.

During the 10-day observation period, no mortalities and no systemic toxicity symptoms

were noted. Behavior was unaffected. The body weight gain was in the normal range. No

signs of skin irritation were found after the 4 h-exposure period (see Table).

Mean skin irritation scores" of 3 rabbits after dermal application of 0.5 ml Granupom

Time after bandage removal

1 hour

1 day

Further daily inspections up

to 1 0 days

Test substance

Erythema

0

0

0

Edema

0

0

0

Control2'

Erythema

0

0

0

Edema

0

0

0

"accordingto DRAIZE (0-4)

2)

 shaved area close to the exposure area

Granupom does not produce any dermal irritation in the rabbit. The lack of any signs of

irritation indicates Granupom is Toxicity category IV for dermal irritation.

Hypersensitivity Incidents: No reports of hypersensitivity have been associated with

Autographa californica MNPV strain FV #11.

•

Page 7 of 11
Cell Culture:

Study 1 (MRID 1431081: Thirty-five nontarget host cell lines, 23 of human and 12 of

nonhuman vertebrate origin, were exposed to Autographa californica multiple

nucleopolyhedrosis virus preparations derived from four different sources: polyhedra,

hemolymph,cell culture medium, and cultured cells. The virus and cells were incubated

together at two different temperatures, 28 or 37°C, for four different lengths of time, 16,

40, 64, or 168 h, and the cells were assayed for the presence of virus by a peroxidaseantiperoxidase detection method. The estimated sensitivity of the assay as routinely

conducted was 0.98 ng of alkali-liberated viral protein and 1.95 ng of budded viral

protein per mm2. No evidence offrank replication was obtained in any of the 35 cell lines

tested, although virus uptake appeared to be quite common. Virus uptake was confirmed

in some cases by electron microscopy. The degree of virus uptake appeared to be

dependent on cell type, time and temperature of incubation, and viral phenotype. Virus

purified from polyhedra was generally taken up more readily than were the other forms

tested.

Study 2 (MRID 143109): Autographa californica multiple nucleopolyhedrosis virus

(AcNPV) was examined to determine whether the virus or its DNA could replicate and/or

persitst in cultures of mammalian cells. Human HeLa cells or primary human embryonic

kidney cells, simian CV1 cells, hamster BHK21 (B3) cells or Muntiacus muntjak cells

growing in monolayer cultures were used in these studies. Cells were inoculated with

AcNPV at multiplicities ranging from 0.1 to 100 PFU/cell. Subsequently, the inoculated

cells were investigated for virus production and for the replication and the persistence of

viral DNA. Extracts of inoculated cells were also screened for the occurrence of AcNPVspecific RNA. AcNPV does not multiply in any of the cell lines studied. Viral DNA

replication or transcription could not be detected by blotting and nucleic acid

hybridization experiments using nick-translated, cloned viral probes. Furthermore, there

was not evidence for the persistence of viral DNA or of fragments o£ viral DNA in mass

cultures of mammalian cells.

Study 3 (MRID 147020): Mammalian cell lines from Chinese hamster, Indian muntjac

and mouse were inoculated with infectious supernatant of Autographa californica

(Speyer) multiple nucleopolyhedrosis virus (AcNPV) replicated in Mamestra brassicae

(L.) cell cultures (IZD-Mb-0503). There was no adverse effect on cell proliferation, nor

was a cytopathogenic effect (CPE) induced in such cultures. Cytogenetic data indicate

that uptake of Ac-NPVs into the cytoplasm of mammalian cells, as shown by an electron

microscopic study, induced neither numerical or structural chromosome aberrations nor

sister chromatid exchange (SCE) events.

AGGREGATE EXPOSURE

The potential for non-occupational, dietary, and non-dietary exposure to Autographa

californica MNPV strain FV #11 by the general population, including infants and children,

is likely through ingestion of treated crops.

Page 8 of 11
Dietary Exposure

1. Food: Dietary exposure to AcMNPV strain FV #11 should not be of concern due to

the low toxicity shown across the entire toxicological profile (see above). If

AcMNPV strain FV #11 is ingested, none of the studies have indicated any

toxicity/pathogenicity of the organism in mammals. In addition, Autographa

californica MNPV strain FV #11 does not replicate in mammalian cells.

2. Drinking water: AcMNPV strain FV #11 does not pose a risk to drinking water.

Anticipated agricultural practice prohibits direct application of the end-use product

directly to water. AcMNPV strain FV #11 is not likely to be present in aquatic

environments since it is not known to inhabit aquatic environments.

Non-Dietary Exposure

Non-dietary exposure to AcMNPV strain FV #11 will be much more likely through

handling by formulators and applicators, but in light of the subject product's

toxicological profile, should be of little concern.

CUMULATIVE EXPOSURE

Section 408(b)(2) of the FFDCA requires that, when considering whether to establish an

exemption from a tolerance, the Agency consider available information concerning the

cumulative effects of the particular ingredient's residues and other substances that have a

common mechanism of toxicity. These considerations include the possible cumulative

effects of such residues on infants and children.

Autographa californica MNPV strain FV #11 is effective by dissolving into the cabbage

looper larvae (Trichoplusia ni) midgut and releasing the occlusion-derived viral particles

(ODVs) that initially infect midgut tissues in order to systematically infect and replicate

in the nucleus of other tissues within the larvae. The infected larvae stop eating, become

sluggish and die of viral infection. Following death, the larvae melts and the progeny

virus is released in the surrounding environment and can in turn infect neighboring

cabbage looper larvae following ingestion.

Baculoviruses co-evolved with and have only been found in insects. Baculoviruses do not

infect Hymenopteran pollinators and parasitoids or insect predators such as praying

mantis and ladybug beetles. No member of the Baculoviridae family has been found in

vertebrates or plants, and AcMNPV does not replicate in mammalian cells. Baculoviruses

do not affect or infect fish, wildlife, humans, or beneficial organisms. This passive mode

of action does not have a common chemical-based mechanism of toxicity as can

sometimes be found with typical organic chemicals.

SAFETY DETERMINATION

U.S. Population

Page 9 of 11
Autographa californica MNPV strain FV #11 exhibits low mammalian toxicity as

demonstrated by the toxicological profile. When AcMNPV strain FV #11 is used on raw

agricultural commodities in accordance with good agricultural practice, it meets the

reasonable certainty of no harm requirement.

Infants and Children

It is believed that the potential exposure of infants and children to residues of AcMNPV

strain FV #11 through its use to protect plants and fruit against cabbage looper larvae is no

greater than that of the general population. Available information does not suggest that

infants and children are more susceptible to the effects of AcMNPV strain FV #11 than are

adults.

EFFECTS ON THE IMMUNE AND ENDOCRINE SYSTEMS

Extensive literature searches and review of Autographa californica MNPV strain FV #11

toxicity data indicate there is no evidence that it is an endocrine disrupter or that it has

any known effect on immune systems as AcMNPV does not replicate in mammalian cells.

EXISTING TOLERANCES

There are no known active agricultural tolerances or tolerance exemptions for Autographa

californica MNPV strain FV #11. A previous exemption from the requirement of a

tolerance for polyhedral occlusion bodies of Autographa californica MNPV (40 CFR

180.1125) was revoked in April 2004, based solely on non-payment of maintenance fees

and because there were no active FIFRA product registrations applicable to the

exemption.

INTERNATIONAL TOLERANCES

There are no known international tolerances for Autographa californica MNPV strain FV

#11 although experimental and commercial use of AcMNPV is reported from Central America

(1998).

SUMMARY/CONCLUSION

An end-use material containing Autographa californica MNPV strain FV #11 is being

submitted to EPA for registration. Loopex will be applied pre-harvest in greenhouses and

open fields to cole crops, green leafy vegetables, root and tuber vegetables, legume

vegetables, fruiting vegetables, cucurbit vegetables, watercress, cotton, tobacco, peanut,

ornamentals and flowers to protect these crops against infestation caused by cabbage

looper larvae (Trichoplusia ni). Application rates are projected to be up to 5 fluid

ounces/acre.

Page 10 of 11
Autographa californica MNPV strain FV #11 does not replicate in mammalian cells. As

such, AcMNPV would not be considered to be of pathological concern. Furthermore, the

organism shows no acute toxicity by the required routes of exposure.

The subject Autographa californica MNPV strain FV #11 has no potential aggregate

exposure through food or drinking water. The simple/rudimentary mode of action of the

organism against cabbage looper larvae also precludes any potential for cumulative

exposure. There is also no evidence that the organism affects the immune system or

causes endocrine disruption. When used in accordance with good agricultural practices,

Autographa californica MNPV strain FV #11 meets the reasonable certainty of no harm

requirement to the general US population and, specifically, the segment of the population

containing infants and children.

,

Page 11 of 11