EPA BIOPESTICIDES AND POLLUTION PREVENTION DIVISION

COMPANY NOTICE OF FILING FOR PESTICIDE PETITIONS PUBLISHED IN

THE FEDERAL REGISTER

EPA Biopesticides and Pollution Prevention Division contact: [Kimberly Nesci, 703-

308-8059]

SUBMISSION: E-mail the completed template to: reilly.sheryl@epa.gov.

TEMPLATE:

[Andermatt Biocontrol AG]

[Insert petition number]

EPA has received a pesticide petition ([insert petition number]) from

[Andermatt Biocontrol AG], [Stahlermatten 6 CH-6146 Grossdietwil Switzerland]

proposing, pursuant to section 408(d) of the Federal Food, Drug, and Cosmetic Act

(FFDCA). 21 U.S.C. 346a(d), to amend 40 CFR part 180.

(Options (pick one)

2. to establish an exemption from the requirement of a tolerance for Autographa

californica multiple nucleopolyhedrovirus (AcMNPV)

(Options (pick one)

1. microbial pesticide \Autographa californica multiple nucleopolyhedrovirus

(AcMNPV)]

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•

Pursuant to section 408(cl)(2)(A)(i) of FFDCA, as amended, [Andermatt ;...;.

Biocontrol AG] has submitted the following summary of information. daftf-MM

arguments in support of their pesticide petition. This summary was prepared Ry«[SciR«g, •

Inc.] and EPA has not fully evaluated the merits of the pesticide petition. Tffi? Summary ••

may have been edited by EPA if the terminology used was unclear, the summary ***• '

contained extraneous material, or the summary unintentionally made the reMJe? conclude,

that the findings reflected EPA's position and not the position of the petitioftftf .• • •

I, [Andermatt Biocontrol AG] Petition Summary •

I
A. Product Name and Proposed Use Practices

[An end-use material containing Autographa californica multiple

nucleopolyhedrovirus (AcMNPV) is being submitted to EPA for registration.

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.]

B. Product Identity/Chemistry

1. Identify of the pesticide and corresponding residues. Autographa californica

MNPV stain FV #11

Physical/Chemical Properties

Property

Color

Physical State

Odor

Density

PH

Viscosity

Result

Light medium brown

Liquid

Undetectable

1. 1 54 g/m 1(9.63 Ibs/gal)

6.31

12.64cStat20°C.

5.86 cSt at 40° C.

Reference

Sylvar Technologies, Inc Report,

Loopex; Autographa californica

multiple nuclepolyhedrovirus

(AcMNPV) FV #1 1, batch #6

Physical Properties

Sylvar Technologies, Inc Report,

Loopex; Autographa californica

multiple nuclepolyhcdrovirus

(AcMNPV) FV #11, batch #6

Physical Properties

Sylvar Technologies, Inc Report,

Loopex; Autographa californica

multiple nuclepolyhedrovirus

(AcMNPV) FV #1 1, batch #6

Physical Properties

Stillmeadow Product Chemistry

Report, Study No. 1 7287-13 1. . . I .

Stillmeadow Product Chemistry

Report, Study No. 17287-13 *•••*••

Stillmeadow Prod fie t'&ntmistry 9

Report, Study No. 3 72& 7*13 ". . .

••:••: : *..

••••••

2. Magnitude of residues til the time of harvest and method used to determine lmK

residue. [No magnitude of residue (MOR) studies have been conducted with

Autographa californica MNPVstrain FV #11 based on the low toxicity of the active
ingredient (see Toxicology Profile) and a total absence of any known infrections to

organisms other than a naroow 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 1) prescribed in toxicology test

guidelines OPPTS 885.3050 through 885.3650 of this series. If Tier 1 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."]

3. A statement of why an analytical method of detecting and measuring the levels

of the pesticide residue are not needed. [Andermatt Biocontrol AG is submitting a

petition to establish an exemption from the requirement of a tolerance and,

therefore, an analytical method is not required.)

C. Mammalian Toxicological 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, and cited herein in support of another similar baculovirus product,

Gusano, EPA REg. No 70051-43.

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

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

that Autograph a californica MNPV strain FV #11 is neither toxic, nor pathogenic

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-Daw ley rats were

dosed per os with Autographa californica MNPV at a rate of 4.35 x 10' 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 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 (MRID 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 w eight 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 monkgvs weft****

exposed to the organism at 1 mg/kg bw (1.2 x 108 PIB/kg bw), Ix weekjy fi>i*26 •

weeks. AH monkeys except for one survived the study and showed norjnaj}\eigh{*. *•

gains. One female monkey in the inhalation treatment route had low plaleki at dfav"*.

181 which returned to normal. No test substance-related effects were ojljs,e,r^'ed. *••

: : •• • • • Acute Inhalation Toxicity: I

• •

Study 1 (MRID 74l76):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.

Eye Irritation:

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

administered into the conjunctiva! sac of the left eye of each of five male and five

female albino (New Zeaand 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

in jury and irritation were recorded at 24, 48 , and 72 hours and seven and 14 days.

No eye irritation was observed; the eye irritiaton 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 101 granules/L. The test

substance was applied undiluted derm ally in a single dose of 0.5 mL/animal onto the

shaved dorso-Iumber 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 J...J.

symptoms were noted. Behavior was unaffected. The body weight gain was in thfe

normal range. No signs of skin irritation were found after the 4 h-exposurejf>eri<Jd?• • •

• • • *

Granupom does not produce any dermal irritation in the rabbit. The facj^o/anyj*. *.

signs of irritation indicates Granupom is Toxicity category' IV for dermal iiiritatHMi.*' *

Hypersensitiviry Incidents: No reports of hypersensitivity have been ass&ci&ed with*

Autographa californica MNPV strain FV #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 peroxidase-antiperoxidase 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

(AcMNPV) 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 AcMNPV 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 AcMNPV-specific RNA. AcMNPV 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 (AcMNPV) 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 culture^? • • • •

Cytogenetic data indicate that uptake of AcMNPV into the cytoplasm J>f J I •

mammalian cells, as shown by an electron microscopic study, induced'neither ;•. *.

numerical or structural chromosome aberrations nor sister chromatic! etch&nge"

(SCE) events.

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D. Aggregate Exposure §•••••

1 . Dietary exposure. [The potential for non-occupational, dietary, and non-«

dietary exposure to Autographa californica MNPV by the general population,

including infants and children, is likely through ingest ion of treated crops.]
1. i. Food. [Dietary exposure to Autographa calif arnica MNPV should not be of

concern due to the low toxicity shown across the entire toxicological profile (see

above). If Autographa californica MNPV is ingested, none of the studies have

indicated any pathogenicity of the organism in mammals. In addition, AcMNPV

does not replicate in mammalian cells.]

ii. Drinking water. [Autographa californica MNPV does not pose a risk to

drinking water. Anticipated agricultural practice prohibits direct application of the

end-use product directly to water. Autographa californica MNPV is not likely to be

present in aquatic environments since it is not known to inhabit aquatic

environments.]

2. Non-dietary exposure. [Non-dietary exposure to Autographa californica

multiple nucleopolyhedrovirus will be much more likely through handling by

formulators and applicators, but in light of the subjects toxicological profile, should

be of little concern.]

E. Cumulative Effects

[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 is effective by dissolving into the larval 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 decomposes and the progeny virus is released

in the surrounding environment and can in turn infect neighboring cabbage looper

larvae (Trichoplusia ni)_following ingestion.

Baculoviruses co-evolved with and have only been found in insects. Baculoviruses

not infect Hymenopteran pollinators and parasitoids or insect predators such as*

praying mantis and ladybug beetles. No member of the Baculoviridae family ha£

been found in vertebrates or plants, and AcMNPV does not replicate pi n}a}nmalian

cells. Baculoviruses do not affect or infect fish, wildlife, humans, or be'neficfal ;•. •.

organisms. This passive mode of action does not have a common chem icjj J-llased *

mechanism of toxicity as can sometimes be found with typical organicjch^icals.l *.*,

/'". Safely Determination ,•••••

i. U.S. population. [Autographa californica MNPV exhibits low mammalian *.,*.!

toxicity as demonstrated by the toxicological profile. When Autographa californica
MN.PV is used on raw agricultural commodities in accordance with good

agricultural practice, it meets the reasonable certainty of no harm requirement.]

2. Infants and children. [It is believed that the potential exposure of infants

and children to residues ofAutographa californica MNPV through their 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 ofAutographa californica MNPV) than are adults.]

G. Effects on the Immune and Endocrine Systems

[Extensive literature searches and review ofAutographa californica MNPV

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 current agricultural tolerances or tolerance exemptions

for Autographa californica MNPV. A previous exemption from the requirement of a

tolerance for polyhedral occlusion bodies ofAutographa 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 although experimental and commercial use of AcMNPV is reported from Central

America (1998).]

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