DRAFT

ADDENDUM TO BIOPESTICIDE REGISTRATION ACTION DOCUMENT

Metarhizium anisopliae strain F52 

### (PC Code 029056)

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

March 1 7 , 2011Metarhizum anisopliae F52 Page 2 of 31 Biopesticides Registration Action Document

– DRAFT Addendum March 1 7 , 2011

TABLE OF CONTENTS

I. BAC KGROUND 4

II. ACTIVE INGREDIENT OV ERVIEW 4

III. REGULATORY BACKGROUND 5

A. E xtensions of C onditional Registrations 5

B. Conditions of R egistration 6

C. Public Comment Period 7

D

. F ood C learances /T olerances 7

IV. RISK ASSESSMENT 8

A. P roduct A nalysis A ssessment 8

B

. Human Health Assessment 9

C. Environmental Assessment 1 2

D. P roduct P erformance D ata (E fficacy ) 1 5

V. RISK MANAGEMENT DECI SION 1 6

VI. ENVIRONMENTAL JUSTICE 1 7

VII. TERMS AND CONDITIONS OF THE REGISTRATION( S) 1 7

VIII. APPENDIX A. DATA REQUIREMENTS 1 8

IX. APPE NDIX B. MRID A ND CITED LITERATURE BIBLIOGRAPHY 24

X. GLOSSARY OF ACR ONYMS AND ABBREVIATI ONS 3 1Metarhizum anisopliae F52 Page 3 of 31 Biopesticides Registration Action Document

– DRAFT Addendum March 1 7 , 2011

BIOPESTICIDES REGIST RATION ACTION DOCUME NT TEAM

Office of Pesticide Programs:

Biopesticides and Pollution Prevention Division

Microbial Pesticides Branch ( M PB)

Ibrahim Barsoum Product Analysis, Human Health John Kough Product Analysis, Human Health

Shannon Borges Environmental Effects Zigfridas Vaituzis Environmental Effects

Gail Tomimatsu Environmental Effects

Sheryl Reilly Chief, M icrobial Pesticides Branch

Shanaz Bacchus Regulatory Action Leader

Leonard Cole Previous Regulatory Action LeaderMetarhizum anisopliae F52 Page 4 of 31 Biopesticides Registration Action Document

– DRAFT Addendum March 1 7 , 2011

I. BACKGROUND

Metarhizium anisopliae strain F52 (called MetF52 in this document ) is the active ingredient in

microbial insecticides that were conditionally registered between June 6, 2003

, and June 11, 2003

, to
Earth BioSciences under Section 3 of the Federal Insecticide, Fungicide and Rodenticide Act (FIFRA)

. The current registrant is Novozymes Biologicals, Inc

. The registered pesticide s contain living Met F5 2, a deuteromycetous ( or anamorphic ) fungus that is pathogenic to insects , mites and

ticks . The fungus is ubiquitous, and found in soil, river sediments, in associat ion with nematodes, and as a saprophyte on organic detritus. The kn own host range of M. anisopliae includes over 200

insect species. Most of the se belong to the Elateridae and Curculionidae families in the in sect Order Coleoptera, although it is known to infect insects found in several other O rders (Zimmerman 2007).

Met F 52 infects inse cts that come in to contact with its spores

. The spores attach to the surface of the insect, ge rminate and begin to grow. The growing fungus penetrate s the exoskeleton

of the insect host then grows very rapidly inside the insect , resulting in its death. Other

insects that come in to contact with infected insects also become inf ected, which leads to their

death.

The original Biopesticide Registration Action Document ( BRAD ) for Metar h izium anisopliae

F52 is available on the Biopesticides and Pollution Prevention Division’s (BPPD’s) website

at http://www.epa.gov/oppbppd1/biopesticides/ingredients/tech_docs/brad_ 029056.pdf

. The

BRAD summarizes the Agency’s exposure and risk assessments that were the basis of the conditional registration s of the MetF52 pesticide s between June 6, 2003, and June 11, 2003

. Th e assessments con tinue to support the currently registered uses of the pesticides . T his

addendum includes references to relevant

, previously evaluated data and other information

from the BRA D

, and provides new information and updated summari es of the Agency’s

current risk assessments for the proposed amendment to allow the appl ication of MetF52 to food crops.

II. ACTIVE INGREDIENT OVERVIEW

The MetF52 products conditionally registered are:

Common Name: Metarhizium anisopliae strain F52

Trade & Other Names: TAE

- 001 Technical Bioinsecticide

TAE

- 001 Granular Bioinsecticide;

TAENURE GRANULAR BIOINSECTICIDE [ Alternate Brand Name: Met52TM GRANULAR

BIOINSECTICIDE ]

TICK

- EX TM G

TICK

- EX TM EC

TICK

- EX EC BIOINSECTICIDEMetarhizum anisopliae F52 Page 5 of 31 Biopesticides Registration Action Document

– DRAFT Addendum March 1 7 , 2011

Culture Deposit: 1) American Type Culture Collection (ATCC ) Number is 90448 2) ARSEF (Agricultural Research Service Entomopatho genic Fungi) culture collection as ARSEF 7711 3) CABI (Centre for Agriculture and Biosciences International) collection as IMI 385045.

OPP Chemical Code : 029056

Type of Pesticide: Microbial pesticide, insecticide

Basic Manufacturer: Novozymes Bi ologicals, Inc.

Target Pests: M ites, thrips, ticks, whiteflies, weevils

III. REGULATORY BACKGROUND

Pesticides containing Met F52 were conditionally registered between June 6, 2003

, and June 11,

2003 to Earth BioSciences Inc. As condition s of regist ration, additional data were needed to :

**•** U pgrade the submitted product identity of the active ingredient;

**•** U pgrade the manufacturing process and disclosure of unintentional i ngredients stud ies ;

**•** R eassess the effects of Met F52 on aquatic vertebrates by rep eating the freshwater fish study ;

**•** D etermine the efficacy of Met F52 on the claimed public health pest s (e.g. blacklegged ticks, Ixodes scapularis ).

These data were to be submitted within one year from the date of the initial registration. The

additional data and information were submitted in 2004 , but were not sufficient to address the conditions of registration. In 2006

, the pesticide products were transferred to Novozymes Biologicals, Inc., (US EPA letter dated December 7, 2006 from Kathryn S. Bouve t o Mr. Theodore Melnik, Novozymes Biologicals, Inc. , referred to hereafter as Novozymes ).

A. Extension s of Conditional R egistrations

The conditional registrations for MetF52 pesticides were extended on February 12, 2007

, provided that the registrant submi tted acceptable product performance data for tick control by December 31, 2007

. On February 2, 2009, requests were granted to Novozymes to extend the conditional registrations to January 30

, , 2010 , pending submission of additional product chemistry data. On November 4, 2009, the conditional registrations were extended to January 30, 2011, pending review of the product chemistry data submitted to fulfill the conditions of registration , in addition to the proposed product amendment s for applications to food crops (see

Food Tolerances

, below) . The notice of receipt of the se amendment s was published in the Federal Register on March 5, 2010 (75FR10259). The current extension of the existing (non

food use) registrations to May 2, 2011 was granted to coincide w ith the Agency’s review of theMetarhizum anisopliae F52 Page 6 of 31 Biopesticides Registration Action Document

– DRAFT Addendum March 1 7 , 2011

product chemistry, dietary and ecological exposure and risk assessments for the proposed food

uses

.

B. Conditions of R egistration

1. Product chemistry

Novozymes submitted studies to satisfy the conditional data requirement s and obtain

unconditional registration s of the company’s MetF52 insecticides under FIFRA section 3(c) (5)

. The product chemistry data for the manufacturing and e nd

- use p roducts are a matter of

Confidential Business Information. The final submission inclu ded results from testing for fungal

and bacterial unintentional ingredients, and is a cceptable (US EPA BPPD DER MRID s 47123901 and 48177001, dated 03/04/10 and 1 2/30/2010 respectively , from Ibrahim Barsoum to Shanaz Bacchus)

. For more information

, see (see Table 1, Appendix A of this document )

.

2. Aquatic vertebrates

As a condition of registration on June 6, 2003 , the Agency required that the freshwater fish study be repeated. The registrant’s response to this condition was a rebuttal (dated December 09, 20 04)

and

, later

, a resubmission (dated January 14, 2008). The Agency accepted Novozymes’ rationale for not repeating the study based on the use patterns ( memorandum dated March 11, 2008, by Dr. Zigfridas Vaituzis), as discussed in Chapter 3C of this docume nt

. In the event the registrant seeks to add applications to aquatic sites to their products, additional data may be required.

3. Efficacy

The original conditional registrations (2003; extended February 12, 2007) were intended for non

food indoor and gr eenhouse use. One end

- use product, TICK

- EX TM G was intended for use on residential and institutional lawns and landscape perimeters to control ticks and certain grubs.

T he conditional registration notice (revised February 12, 2007 ) required that data mus t be

submitted by December 31, 2007 to demonstrate the efficacy of MetF52 against black

- legged

ticks . Novozymes submitted the required data on January 2008. The Agency accepted the explanation concerning the change of the products’ ownership for the late submission

, reviewed

the data and determined that it satisfied the product performance (efficacy) requirement for labeling ticks as target pests (see Product Performance, Chapter 3D in this document )

. Novozymes also submitted product performance data in A ugust 2010 to support control of mites, thrips , ticks, weevils and whiteflies in order to amend the labels to reflect these target pests. F or more information, see http://www.epa.gov/pesticides/pestlabels/

.

4. Other Label A mendments

L ab el a mendments to the non

- food use labels were submitted to include the Canadian and U

. S

. manufacturing establishments . These were approved December 15, 2010 , and the amended labels are available on EPA’s Pesticide Product Label System. For more information, see http://w ww.epa.gov/pesticides/pestlabels/ and search for EPA Reg. Nos. 70127

- 7, 70127

- 8,

70127

- 9, and 70127

- 10.Metarhizum anisopliae F52 Page 7 of 31 Biopesticides Registration Action Document

– DRAFT Addendum March 1 7 , 2011

C. Public C omment P eriod

This document and the associated risk assessments are posted in the docket ( EPA

- HQ

- OPP

20 10

- 0081 ) to allow the public 30 days to review and provide comments on the proposed

amendments to add food uses to registered products containing the insecticidal, microbial active ingredient, Met F52.

D. F ood Clearances/Tolerances

Novozymes submitted a petition to establish an exemption fr om the requirement of a tolerance for residues of the microbial pesticide, Metarhizium anisopliae strain F52 , in or on all food

commodities

. The Agency published a Notice of the filing of the petition ( [PP] 8F7508 ) i n the Federal Register on April 28, 2009 ( 74 FR 15969 ; EPA

- HQ

- OPP

- 2009

- 0194 ). Following a 30

- day comment period , the Agency determined that no comments w ere submitted in response to the published Notice

. This a ddendum provides the current asses sments of the data/information that were relied up on to support the request to exempt MetF52 from the requirement of a

tolerance.Metarhizum anisopliae F52 Page 8 of 31 Biopesticides Registration Action Document

– DRAFT Addendum March 1 7 , 2011

IV. RISK ASSESSMENT

On October 26, 2007, the Agency issued a Final Rule in the Federal Register on the data requirements to support registration of microbial pesticides and upd ated the definition for microbial pesticides ( 72 FR 61002 )

. The rule became effective on December 26, 2007. The data

and information evaluated for th e June 2003 BRAD

, published for the initial conditional registration

, and all subsequent data/information e valuated for registration amendments of

MetF52 pesticides

, were considered in light of these 2007 requirements.

The classifications that are found for each data submission are assigned by Agency science

reviewers and are an indication of the usefulness of the information contained in the documents for risk assessment. A rating of “ACCEPTABLE” 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 scientifically acceptable (“SUPPLEMENTAL: UPGRADABLE”). If a study is rated as “SUPPLEMENTAL: UPGRADABLE,” the Agency always provides an indication of what is lacking or what ca n be provided to change the rating to “ACCEPTABLE.” If there is simply a “SUPPLEMENTAL” rating, the reviewer will often state that the study is not required by the current 40 Code of Federal Regulations (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 need to be submitted.

For the acute toxicity data requirements, toxicity categories are assigned based on the hazard(s) identified from st udies and/or other infor mation submitted to the Agency in support of a pesticide registration. The active ingredient or particular product is classified into Toxicity Category I, II, III, or IV, where Toxicity Category I indicates the highest toxicity and Toxicity Category IV indicates the lowest toxicity.

A. Product Analysis Assessment ( 40 CFR § 158.2120 )

At the initial conditional registration of MetF52 products

, some product chemistry data were required to proceed to unconditional registration under FIFRA. These data were submitted and

evaluated

. Refer to revised Tables 1 and 2 in Appendix A of this document for a brief sum mary of the updated status of the se data requirements, including both generic and product

- specific

information. All product analysis data requirements (except for storage stability ) for MetF52 for

the labeled uses have now been satisfied

. Based on the existing usage of the pesticide for approximately 18 years without reports of adverse effects , and the registrant’s agreement to provide s torage stability data as a term of these registration amendments, these pesticides are

e ligible for registration under FIFRA Section 

### 3(c)(5). An analytical method (Guideline

885.1800 ) is not required for residues on food based on tolerance exemption. The analytical method for product identity is acceptable for product characterization. Add itionally, the Agency risk assessments that evaluate these data are available as “Supporting & Related Materials”

within Docket Number EPA

- HQ

- OPP

- 2010

- 0081 at http://www.regulations.gov

.Metarhizum anisopliae F52 Page 9 of 31 Biopesticides Registration Action Document

– DRAFT Addendum March 1 7 , 2011

B. Human Health Asse ssment ( 40 CFR § 158.2140 )

1. Toxicity

All Tier I mammalian toxicology data requirements for MetF52 have been satisfied by submitted

studies or acceptable waivers

. The acute mammalian oral study resulted in Toxicit y Category IV

classification. The a cute pulmonary toxicity/pathogenicity, acute dermal irritation and acute eye

irritation studies re sulted in Toxici ty Category III classifications. The acute inhalation study and primary dermal studies were waived due to the low potential exposure from the proposed uses of Met F52 as a pesticide, and the low toxicity potential demonstrated in the acute pulmonary and

acute dermal studies

. Tier II and Tier III studies were not required for MetF52 based on the lack of acute toxicity/pathogenicity in the Tier I studies. Refer to Table 2 in the original BRAD for a brief summary of the data evaluated for th ese data requirements

- see ( http://www.epa.gov/oppbppd1/biopesticides/ingredients/tech_docs/brad_029056.pdf ). Supporting

materials are available in this docket

, EPA

- HQ

- OPP

- 2010

- 0081

.

2. Effects on the Endocrine System

As required under FFDCA section 408(p), EPA has developed the Endocrine Disruptor Screening Program (EDSP) 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 occ urring estrogen, or other such endocrine effects as the Administrator may designate.” The EDSP employs a two

- tiered approach to making the statutorily required determinations. Tier 1 consists of a battery of 11 screening assays to identify the potential o f a chemical substance to interact with the estrogen, androgen, or thyroid (E, A, or T) hormonal

systems. Chemicals that go through Tier 1 screening and are found to have the potential to interact with E, A, or T hormonal systems will proceed to the next stage of the EDSP where EPA

will determine which, if any, of the Tier 2 tests are necessary based on the available data. Tier 2 testing is designed to identify any adverse endocrine related effects caused by the substance, and

establish a dose

- response rel ationship between the dose and the E, A, or T effect.

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.

This list of c hemicals was selected based on the potential for human exposure through pathways such as food and water, residential activity, and certain post

- application agricultural scenarios. This list should not be construed as a list of known or likely endocrine di sruptors.

Met F52 is not among the group of 58 pesticide active ingredients on the initial list to be screened under the EDSP. Under FFDCA sec. 408(p) the Agency must screen all pesticide chemicals. Accordingly, EPA anticipates issuing future EDSP test o rders/data call

- ins for all pesticide active ingredients

.

For further information on the status of the EDSP, the policies and procedures, the list of 67 chemicals, the test guidelines and the Tier 1 screening battery, please visit our website: http://ww w.epa.gov/endo/

.Metarhizum anisopliae F52 Page 10 of 31 Biopesticides Registration Action Document

– DRAFT Addendum March 1 7 , 2011

3. Food Quality Protection Act (FQPA) Considerations

Section 408(c)(2)(A)(i) of the Federal Food, Drug, and Cosmetic Act allows the Agency to establish an exemption from the requirement for a tolerance (the legal limit for a pesticide

chemical resi due in or on a food) only if the Agency determines that the exemption is “safe.” Section 408(c)(2)(A)(ii) of FFDCA defines “safe” to mean that “there is a reasonable certainty that no harm will result from aggregate exposure to the pesticide chemical residue, including all anticipated dietary exposures and all other exposures for which there is reliable information.” This includes exposure through drinking water and in residential settings, but does not include occupational exposure. Pursuant to section 408(c)(2)(B) of FFDCA, in establishing or maintaining in effect an exemption from the requirement of a tolerance, the Agency must take into account the factors set forth in section 408(b)(2 )(C) of FFDCA, which require the Agency to give special consideration to exposure of infants and children to the pesticide chemical residue in establishing a tolerance and to “ensure that there is a reasonable certainty that no harm will result to infants and children from aggregate exposure to the pesticide ch emical residue... ” Additionally, section 408(b)(2)(D) of FFDCA requires that the Agency consider “available information concerning the cumulative effects of [a particular pesticide's] residues and other substances that have a common mechanism of toxicity. ”

The Agency performs a number of analyses to determine the risks from aggregate exposure t o pesticide residues. First, the Agency determines the tox icity of pesticides. Second, the Agency examines exposure to the pesticide through food, drinking water, an d through other exposures that occur as a result of pesticide use in residential settings.

Consistent with s ection 408(b)(2)(D) of FFDCA, the Agency has reviewed the available scientific data and other relevant information, and considered its validity, com pleteness, and reliability and the relationship of thi s information to human risk. The Agency has also considered available information concerning the variability of the sensitivities of major identifiable subgroups of consumers, including infants and chil dren.

Based on the acute toxicity/pathogenicity data presented in Table 2 in the original BRAD t he data required for a FQPA risk assessment for Metarhizium anisopliae F52 have been satisfied

.

a. Aggregate Exposure

In examining aggregate exposure, sectio n 408 of FFDCA directs the Agency to consider available information concerning exposures from the pesticide residue in food and all other non

occupational exposures, including drinking water from ground water or surface water and exposure through pesticid e use in gardens, lawns, or buildings (residential and other indoor uses).

1. Dietary Exposure

Dietary exposure to residues of MetF52 from its proposed pesticidal use is anticipated to be negligible , as described below.Metarhizum anisopliae F52 Page 11 of 31 Biopesticides Registration Action Document

– DRAFT Addendum March 1 7 , 2011

a. Food

Pesticides containing Met F52 are intended to be applied at low application rates to row crops, transplants and dips, greenhouse foliar and root dips, or incorporation into soil

. T he spores,

which adhere to

, and infect insects, are not expected to grow on the crops. Though unlike ly, any spores remaining on food commodities at harvest would be removed by post

- harvest processes (washing, cooking, etc.).

b. Drinking Water

Exposure to Met F52 from drinking water is expected to be negligible because the microbial fungici de will not b e applied to water

. Further, t he spores are large and read i ly adhere to soil particles, and are unlikely to move through soils to reach underground water. Should any MetF52 spores reach underground water, the y would be removed during municipal water trea tment.

2. Other Non

- occupational Exposure

Deuteromycetous fungi , including Metarhizium anisopliae

, occur in nature around the world. The pesticidal uses of products containing Met F52 are not likely to increase non

- occupational human exposure signific antly above normal levels of these naturally occurring fungi

. Al so, if the proposed food uses are approved, pesticidal applications of Met F52 would occur in agricultural

and residential areas . However, t here have been no reports of pathogenicity or othe r adverse

effects in humans and animals from e xposure to the registered products or t o t he naturally occurring strain of this MetF52 fungus.

3

. Cumulative Effects from Substances with a Common Mechanism of Toxicity

Section 408(b)(2)(D)(v) of FFDCA requ ires that, when considering whether to establish, modify, or revoke a tolerance, the Agency consider “available information concerning the cumulative effects of [a particular pesticide’s] residues and other substances that have a common mechanism

of toxici ty.” Acute toxicity studies demonstrated that MetF52 is virtually non

- toxic to mammals

, and there is no known mechanism of toxicity that it share s with other insecticidal substances.

b

. Determination of Safety for the U.S. Population, Infants, and Childr en

FFDCA sectio n 408(b)(2)(C) provides that the Agency shall assess the available information about consumption patterns among infants and children, special susceptibility of infants and children to pesticide chemical residues, and the cumulative effects on infants and children of the residues and other substances with a common mechanism of toxicity. In addition, FFDCA

sectio n 408(b)(2)(C) provides that the Agency shall apply an additional tenfold margin of safety

for infants and children in the case of th reshold effects to account for prenatal and postnatal toxicity and the complet eness of the database unless the Agency determines that a different margin of safety will be safe for infants and children. Margins of exposure (safety), which are

often referre d to as uncertainty fa ctors, are incorporated into Agency risk assessments

, either directly or through the use of a margin of exposure analysis, or by using uncertainty (safety) factors in calculating a dose level that poses no appreciable risk.Metarhizum anisopliae F52 Page 12 of 31 Biopesticides Registration Action Document

– DRAFT Addendum March 1 7 , 2011

MetF52 has a low mammalian acute toxicity potential

, as discussed in Table 2 of the original

BRAD

, and Appendix A of this a ddendum . Registered pesticide products containing this active ingredient have been in use for almost 1 8 years without any reported adverse eff ects to humans

and/or the environment

. EPA concludes 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 o f MetF52

. This includes all antic ipated dietary exposures and all other exposures

for which there is reliable information. T he Agency has arrived at this conclusion because the

data and information available on MetF52 do not demonstrate toxic, pathogenic, and/or infective potential to ma mmals. Thus, there are no threshold effects of concern and, as a result, an additional margin of safety is not necessary.

c

. Occupational, Residential, School and Day Care Exposure and Risk Characterization

Tier I acute toxicity/pathogenicity study re sults demonstrated that Met F52 was not toxic or pathogenic in laboratory mammals via the oral, pulmonary, or intraperitoneal injection routes of exposure, and o ccupational handler exposure i s not expected to pose any unreasonable risks to agricultural or green house workers. Appropriate personal protective equipment and precautionary statements are required to be on product labels to mitigate the potential risks to pesticide handlers due to prolonged or repeated exposures. H andlers working with MetF52 mu st

wear a long

- sleeved shir t, long pants, socks, shoes, waterproof gloves, and 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 when mixing, loading, or applyin g the product.

The proposed agricultural and green house use sites are not expected to be in close proximity to or affect residential areas, schools

, and daycare centers . In the unlikely event exposure to the pesticide from the proposed uses occurs, t he lack of mammalian toxicity at high levels of

exposure to MetF52, in addition to the lack of adverse incident reports from the current pesticidal

uses
, s upport the Agency’s conclusions that no unreasonable adverse effect s would be expected

from the propose d food uses

.

C. Environmental Assessment ( 40 CFR § 1 58.2150 )

1. Nontarget Organism Toxicolog y

All nontarget toxicology data requirements for the proposed uses of Met F52 have been satisfied. Additionally, the Agency risk assessments that were evaluate d are available as “Supporting &

Related Materials” within Docket Number EPA

- HQ

- OPP

- 2010

- 0081 at http://www.regulations.gov

.

Acceptable Tier I nontarget toxicology data/information support the previous registrations, as

well as the proposed uses of MetF 52

. ( Refer to Table 3 in the original BRAD issued with the 6/6/03 and 6/11/03 conditional regis trations

. ) The s ubmitted studies demonstrated that Met F 52 poses no hazard to avian species, wild mammals and terrestrial and aquatic plant species from residential outdoor and institutional premise uses of the product. Data and other information

also s howed no potential adverse effects to beneficial non

- target organisms (lady beetles, green lacewings, parasitic wasps, honey bee larvae, honey bee adults and earthworms). For theMetarhizum anisopliae F52 Page 13 of 31 Biopesticides Registration Action Document

– DRAFT Addendum March 1 7 , 2011

original conditional registration, the data demonstrated that f reshwater and estuarine/marine fish, invertebrates and aquatic insects will not be affected if Met F52 enters the aquatic environment

from incidental drift and runoff from terrestrial uses. However, in light of laboratory studies reporting toxicity and pathogenicity t o immature aquatic vertebrate and invertebrate species, the Agency required additional hazard assessment prior to any registration for aquatic applications.

For the proposed food uses

, there is no direct application of the Met F52 pesticides to aquatic

sites. The Agency accepted the registrant’s rationale for not repeating the fish study. However, the rating of the fish study remains “Supplemental,” since a maxim um hazard dose was not obtained, and there are reports of laboratory induced pathological c hanges (silverside) in fish embryos and fry in the scientific literature. An adult fish study would not address advers e effects on developing embryos . T herefore, if in the future the use patterns are expanded to include aquatic use sites or otherwise inc rease exposure to freshwater fish, the Agency will require

additional studies on the effects of Met F52 TGAI (currently called TAE

- 001 Technical

Bioinsecticide ) on developing freshwater fish embryos.

See Appendix A of this document for a discussion of the rationale for this data requirement

Guideline 885. 4200: Freshwater Fish Toxicity/Pathogenicity

.

2. Environmental Fate and Ground Water Data

The environmental fate of microbial pest control agents (MPCAs) in general is difficult to predict with certaint y. The movement of the MPCA in the environment post application can be estimated based on the biology and ecology of the microorganism , but the population size in the treated and surrounding areas usually cannot be predicted beyond the time of application . Most microbial pesticides persist at initial population levels for a period of time, and then decrease to

ineffective levels. The biology of M. anisopliae has been characterized in the scientific literature, and sufficient information is available to e stimate where Met F52 would likely move and/or persist after applications (US EPA BPPD memorandum from Shannon Borges to Shanaz Bacchus, 02/17/11).

M. anisopliae is a soil borne fungus , though its persistence in soil is affected by environmental

condition s (e.g., temperature, soil moisture), bi otic factors, and can be strain

- dependent (Zimmerman 2007, Vanninen et al. 2000). Persistenc e may differ between laboratory, greenhouse

, and field conditions. M. anisopliae is not easily airborne, and is not an aqu atic fungus, though it can persist in water under some conditions. Little to no germination has occurred in water in laboratory tests (Zimmerman 2007).

Root exudates and decaying plant matter may provide a nutritive source for survival (Meyling

and Eil enberg 2007, Hu and St. Leger 2002) . H owever, insect s are believed to be the primary source of organic matter for population increase, and may account for some persistence in the

environment . T he fungus has been observed to be restricted to the insect ho st ’s cadaver in these cases (Meyling and Eilenberg 2007).

EPA has determined that soil applications will not result in movement off the treatment site or contamination of ground water. EPA has also determined that foliar sprays (both ground and

aerial applications) are likely to result in movement of Met F52 off the treatment site.Metarhizum anisopliae F52 Page 14 of 31 Biopesticides Registration Action Document

– DRAFT Addendum March 1 7 , 2011

However, based on the results of the acute non

- target toxicity testing, the need for environmental fate and groundwater data was not trigger ed.

3. Ecological Exposure and R isk Characterization

T he submitted non

- target organism effects information from publically available, scientific literature

indicated that the only ecological effects of some concern are to non

- target insects. Natural epizootic

infections caused by M. an isopliae occur among insect populations and the literature reports over 200 species of insects affected by this insect fungal pathogen. However, years of research have shown

that M. anisopliae adaptations result in strains that are more infectious for s om e insect species than

others. T he submitted studies show ed that M. anisopliae Strain F52 does not pose a hazard to the major beneficial insect s tested, including honey bees, but the potential risks to all non

- target insects

cannot be ruled out.

The Ag ency has performed a hazard assessment and determined that the proposed uses of Met F52

will have no adverse effects on avian species, wild mammal, and invertebrates, major beneficial insect species, including the honey bee, and terrestrial and aquatic pla nt species. The potential

adverse effect s to fish can be mitigated by limiting direct application to aquatic sites.

4. Endangered Species Assessment

T he BRAD for Met F52 states that an expansion of uses would trigger concerns for en dangered

and thre atened species, and that an extensive risk assessment would be necessary to determine all possible effects. These effects would include direct and indirect effects to threatened and endangered species and their habitat s

. Novozymes submitted an endangered species assessment (MRID 48175201) , which was reviewed by BPPD and rated “S upplemental. ” BPPD conducted

its own review , as discussed below (US EPA BPPD memorandum from Shannon Borges to Shanaz Bacchus, 02/17/11 ).

The known host range of M. anisopliae in cludes over 200 species , and most belong to the

Elateridae and Curculionidae families of Order Coleoptera, though infect ion of insects within

several other orders is documented (Zimmerman 2007).

The potential exposure level as a function of the proposed application methods of the pesticide was assessed, and BPPD ma de “no effect” determinations for direct effects, indirect effects, and affects to habitat (including designated critical habitat) for listed species for the following types of applications of MetF52 to the proposed food crops:

**•** Granular applications to growing medium

**•** All foliar and soil directed liquid applications to the proposed food crop plants treated within enclosed greenhouses,

**•** Outdoor soil drench treatments to the proposed food crop plan ts,

**•** R oot plant dip treatments.

Since the Agency has determined that no effects are anticipated for any nontarget species exposed to Met F52 as a result of the proposed applications to these sites, effects to threatenedMetarhizum anisopliae F52 Page 15 of 31 Biopesticides Registration Action Document

– DRAFT Addendum March 1 7 , 2011

and e ndangered species and their des ignated critical habitats are also not expected. There fore, a

“No Effect” determination is made for direct and indirect effects to listed species and their designated critical habitats resulting from the above proposed uses of Metarhizium anisopliae

F52.

BPPD concludes that the following application methods of Met F52 to the proposed food crops have the potential to cause direct effects, indirect effects, and effects to habitat (including designated critical habitat) to listed species :

**•** All foliage dips and outdoor foliar applications made by hand

- held sprayer, ground spray,

and aerial spray equipment.

BPPD considered the natural occurrence of Metarhizium species, the potential for the pesticidal fungal active ingredient to survive in the environment, and the alternatives that are intended for similar control of the target insect pests.

The natural occurrence of Metarhizium species is well

- documented. Growth of Met F52 was investigated in the laboratory at temperatures varying from 27 **°** C to 37 **°** C. Gr owth at 27 **°** C and

30 **°** C were similar in outcome, although the lower temperature required an additional 48 h of

incubation

. More time was required for growth at 34 **°** C in comparison to lower temperatures

, and no growth was observed when the fungus was grown at 37 **°** C for two weeks. Since there was substantial variance in the temperature of the incubator, conclusions or comparisons must be

made with caution (MRID 448447

- 17 , DER Chris Wozniak to Leonard Cole, 12/11/2001 )

. However, these data do suggest that tempe rature a ffects growth, and consequently, persistence of

Met F52 in the environment

. Moreover, MetF52 has a lower toxicity potential than t he

alternative insecticides (carbamates and pyrethrins ) for the propose d uses. Following application, the fungus ret urns to levels that cannot be distinguished from naturally occurring M. anisopliae

. Further analyses are needed to characterize the effects that are likely to occur and the species that could potentially be affected. This assessment will be condu cted d uring registration review for Met F52.

D. Product Performance Data (E fficacy )

Submission of product performance data (OPPTS 810.3000) is listed as a requirement for all pesticide products. Customarily, the Agency requires efficacy data to be submitt ed for review only in connection with the registration of products directly pertaining to the mitigation of disease bearing human health organisms and certain designated q uarantine p ests, i.e., ticks, mosquitoes, fleas, Mediterranean fruit flies, g ypsy m ot hs, Japanese b eetles

, etc. For a list of organisms considered by the Agency as “public health pests”, please refer to Pesticide Registration Notice 2002

- 1 ( http://www.epa.gov/PR_Notices/pr2002

- 1.p df ).

1. Ticks

Product performance data were provided for the use of MetF52 on ticks ( MRIDs 45723701, 47313401, 47313402 and 47313403; US EPA DERs dated October 2008 from Shannon Borges

and Gail Tomimatsu to Shanaz Bacchus )

. Some of the products con taining Met F52 claim ticksMetarhizum anisopliae F52 Page 16 of 31 Biopesticides Registration Action Document

– DRAFT Addendum March 1 7 , 2011

as pests

. The Agency evaluated the efficacy data that were submitted to satisfy condition s of registration, and determined that these data s upport the control of the tested ticks

, such as the lone star tick, black

- legged tick

, and American dog tick , but without efficacy data, a claim cannot be made to “ticks” in general for these products.

The pesticide products that claim ticks are only for use in greenhouses and residential or suburban terrestrial environments, and are not in cluded in the proposed amendments to add food uses to registered products containing Met F52. The products must not be used near critical habitats of

th reatened or endangered insects without data and/or information needed to support a “No Effect”

determin ation to endangered insects. The products are labeled only for urban and suburban residential terrestrial outdoor uses (not adjoining parks, forested areas) to mitigate the exposure and potential risks to endangered insects

.

2. Other Target P ests

The i nsect pathogenicity data submitted for the Environmental Risk Assessment of MetF52 Biological Insecticide, TICK

- EX TM , (formerly EPA Reg. No. 72098

- T ) performed in 2003 showed no adverse effects on nontarget insects. Submitted nontarget toxicity studies sh ow ed

that MetF52 poses no hazard to lady beetles, green lacewings, parasitic wasps, honey bee larvae, honey bee adults and earthworms

. The product performance data submitted by Novozymes ( cited above ) demonstrated that Met F52 provide s statistically signi ficant control of mites, thrips

, b lack v ine w eevils and white flies on agricultural crops when applied according to the label

directions (MRIDs 48150701, 4815070 2 , 4815070 3 , 4815070 4: US EPA BPPD memo and

DERs dated 02/15/11 from Zigfridas Vaituzis to Shan az Bacchus).

V. RISK MANAGEMENT DECI SION

Pesticides containing Metarhizium anisopliae F52 as the active ingredient were conditionally registered under FIFRA S e ction 3(c)7(C) on June 6 and 11, 2003. The current registrant, Novozymes Biologicals, Inc., has provided data/information to support an unconditional registration under Section 3(c)(5) of FIFRA active ingredients if it is determined that (A) its composition is such as to warrant the proposed claims for it; (B) its labeling and other materials

req uired to be submitted comply with the requirements of FIFRA; (C) it will perform its intended function without unreasonable adverse effects on the environment; and (D) when used in accordance with widespread and commonly recognized practice it will not gen erally cause

unreasonable adverse effects on the environment.

The four criteria of the Eligibility Determination for Pesticidal Active Ingredients are satisfied by

the science assessments supporting products containing Metarhizium anisopliae F52 . Such

p roducts are not expected to cause unreasonable adverse effects, and are likely to provide protection as claimed when used according to label instructions. Therefore, Metarhizium anisopliae F52 is eligible for FIFRA Section 3(c)(5) registration s for the lab eled uses and for the proposed new food uses

. Other considerations may apply to the labeled sites during registration

review of the Metarhizium case.Metarhizum anisopliae F52 Page 17 of 31 Biopesticides Registration Action Document

– DRAFT Addendum March 1 7 , 2011

The 2003 BRAD cited rationales in the public interest for the conditional registration of the pesticidal active ingredient. A public interest finding is not required for this Section 3(c)(5) registration of Metarhizium anisopliae F52

.

VI. ENVIRONMENTAL JUSTIC E

EPA seeks to achieve environmental justice

- the fair treatment and meaningful

involvement of all people, regardless of race, color, national origin, or income

- in the development, implementation, and enforcement of environmental laws, regulations, and policies. At this time EPA does not believe that use of pesticide products containing Metarhiz ium anisopliae F52 will cause harm or a disproportionate impact on at

- risk communities.

For additional information regarding environmental justice issues, please visit EPA’s website at: http://www.epa.gov/compliance/environmentaljustice/index.html

.

V I I

. TERMS AND CONDITIONS OF THE REGISTRATION(S)

As a term of registration, the registrant must su bmit one

- year storage stability data (40 CFR § 158.2120).Metarhizum anisopliae F52 Page 18 of 31 Biopesticides Registration Action Document

– DRAFT Addendum March 1 7 , 2011

VI I I

. APPENDIX A. DAT A REQUIREMENTS (40 CFR Part 158

- Subpart V )

On October 26, 2007, the Agency issued a Final Rule in the Federal Register on the data requirements to support registration of microbial pesticides and updated the definition for microbial pesticides ( 72 FR 61002 ). The rule became effective on December 26, 2007. The data and information evaluated for the BRAD published for the initial conditional registration and all subsequent data/information evaluated for registration amendments of MetF52 pesticides were

consid ered in light of these requirements. Data and info rmation reviewed for the original registration of the product s containing this active ingredient are summarized in the original BRAD (06/06/03). These tables are updated below to reflect recently reviewed submissions

.

*NOTE: MRID numbers listed in the following tables are representative of additional data evaluated to support the request to amend the registration to an unconditional registration under FIFRA Section 3(c)(5) and for the proposed food uses in PP 8F7508

. Subsequent to this registration amendment , there may be additional MRIDs that support registration of other products containing this active ingredient.

TABLE 1. Product Chemistry Data Requirements

Additional Data Reviewed for Metarhiziu m anisopliae F52 (40 CFR § 158. 2 120 )

OPPTS

Guideline

No. Study Results MRID &

references

885.1100 Product identity The data reviewed for the initial conditional registration s (June 6 and June 11, 2003) were not
sufficient to warrant the conclusion tha t the method could be used to separate or identify F52 from other commonly encountered strains of M. anisopliae

. The

stud y was classified as SUPPLEMENTAL (see BRAD, 06/06/03). Additional data reviewed for clonal

success of Metarhizium anisopliae F52 str ains ( DER

471239

- 01 dated 03/04/10 ) are SUPPLEMENTAL.

Another set of data (481770

- 01, DER 12/30/1 0 ) were considered ACCEPTABLE 456963

- 01 *

448547

- 01*

471239

- 01

481770

- 01

885.1200 Manufacturing

Process Additional data required. SUPPLEMENTAL 456 963

- 02 *

448547

- 02 *

456963

- 03 *

448547

- 03 *

885.1200 Additional data were submitted to satisfy the conditions for the initial conditional registration and

these studies were rated SUPPLEMENTAL (DER 03/04/10). More data were required. 478806

- 01**

478806

- 0 2***

480588

- 01**

480588

- 02***

Additional data reviewed are ACCEPTABLE. 481770

- 01

DER12/30/ 10Metarhizum anisopliae F52 Page 19 of 31 Biopesticides Registration Action Document

– DRAFT Addendum March 1 7 , 2011

TABLE 1. Product Chemistry Data Requirements

Additional Data Reviewed for Metarhiziu m anisopliae F52 (40 CFR § 158. 2 120 )

885.1300
Formation of

Unintentional Ingredients Additional data were required for the initial conditional registration and these studies were rated

SUPPLEME NTAL

Additional data reviewed were rated

SUPPLEMENTAL

. (DER 03/04/10). 448447

- 04 *

456963

- 03 *

478806

- 01**

478806

- 02***

480588

- 01**

480588

- 02***

885.1300
Additional data reviewed are ACCEPTABLE. 481770

- 01

DER12/30/1 0

885.1400 Analysis of Samples Approximately 48% of the extractable spores on the

solid matrix were recovered. The range of recovery was 40 to 59% and the viability of the recovered spores was highly consistent between batches. ACCEPTABLE. However, these data are not required

for resi dues on food based on tolerance exemption. Analytical method for product identity acceptable for product characterization. 456963

- 04*

456963

- 03*

448447

- 06*

448447

- 05*

885.1500 Certification

of Limits Ingredient limits were 1.7 to 2.3 % for the a.i. and 97.7 to 98.3% for the matrix of the biopesticide. Both the nominal concentration of F52 spores and the proposed range were within the scope of what was observed in production of this agent. ACCEPTABLE.

Additional data reviewed are ACCEPTABLE 456963

- 05 *

448447

- 02 *

448447

- 03 *

456963

- 03 *

481770

- 01

DER12/30/10

*Data reviewed for original conditional registration

* * TGAI

** * Taenure Granular BioinsecticideMetarhizum anisopliae F52 Page 20 of 31 Biopesticides Registration Action Document

– DRAFT Addendum March 1 7 , 2011

The following results were excerpted from an electronic file dated 12/11/01 ( DER from Chris Wozniak to Leonard Cole, USEPA/OPP/BPPD).

TABLE 2a . Physical and Chemical Properties of Metarhizium anisopliae F52 (40 CFR § 158. 2120)

OPPTS

Guideline

No. Property Description of Result MRID

830.6302 Color olive

- gray 448447

- 08

830.6303 P hysical State powder 448447

- 08

830.6304 Odor Odor

– halide; smell

– earthy. 448447

- 08

830.6313 Stability to Normal and elevated Temperatures,

Metals and Metal Ions Not reported or Not required N/A

830.6315 Flammability Not required N/A

830.6317 Storag e Stability Not reported in 2003

– term of unconditional registration 2011. N/A

830.6319 Miscibility Not required N/A

830.6320 Corrosion Characteristics Not reported N/A

830.7000 pH 1 % (w/v) suspension of spores was 5.66 at

23 **°** C. 448447

- 08

830.7050 UV /Visible Light Absorption Not required N/A

830.7100 Viscosity Not required N/A

830.7200 Melting Point/Range Not required N/A

830.7220 Boiling Point/Range Not required N/A

830.7300 Density Mean bulk density: 0.273 g / mL 448447

- 08

830.7520 Particle Siz e, Fiber Length

and Diameter Distribution Homogeneous consistency; no visible particulate foreign matter 448447

- 08

830.7550

830.7560

830.7570 Partition Coefficient (n

- Octanol/ Water) Not required N/A

830.7840 Water Solubility Not required N/A

830.7950 Vapor Pressure Not required N/AMetarhizum anisopliae F52 Page 21 of 31 Biopesticides Registration Action Document

– DRAFT Addendum March 1 7 , 2011

Th e following information was excerpted from an electronic file of a DER for EPA Reg. No.

70127

- 10 dated 12/11/01 ( from Chris Wozniak to Leonard Cole, USEPA/OPP/BPPD ). The asterisk (*) annotates that the data previous ly submitted and evaluated meet the c urrent data requirements in accordance with 40 CFR Part 158

- SubpartV (72 FR 61002)

.

Table 2b : Physical and Chemical Properties of Metarhizium anisopliae F52 End

- use Product (40 CFR § 158.2120)

*Guideline

No. Property Result MRID/

Reference

151

- 26

*830.6302 Color Not required (PR Notice 92

- 5) None

151

- 26

*830.6303 Physical state Not required

- liquid 46449614

151

- 26

*830.6304 Odor Not required

- (PR Notice 92

- 5) None

63

- 7

*830.7300 Specific gravity Not required

- 0 .92 46449614

63

- 12

*830.7000 pH Not required

- n ot dispersible in water; pH not measured 46449614

63

- 13

*830.6317 Stability (Shelf life) Greater than 15 months at 22 0 C

– 26 0 C 46580601

63

- 16

*830.6316 Explodability Not potentially explosive 46449614

63

- 18

*830.7100 Viscosity 40.6 Centipoises at 29.4 0 C (85 0 F) 46449614

63

- 20

*830.6320 Corrosion

Characteristics Not corrosive; does not react with

metals 46449614

Viability 5.6 x 10 9 cfu/g Letter 10/27/06;

46580601Metarhizum anisopliae F52 Page 22 of 31 Biopesticides Registration Action Document

– DRAFT Addendum March 1 7 , 2011

Acute Toxicity

– Health Effects

The Ag ency’s conclusions of the results of the acute toxicology tests , including the acute oral toxicity/pathogenicity test, for Metarhizium anisopliae F52

, previously reported as Table 2 of the

2003 BRAD

, are unchanged . Those assessments continue to support th e non

- food u ses, and support the proposed food uses of Met F52 pesticides (US EPA BPPD D ata E valuation R ecord

,

e lectronic file dated 12/11/01 from Chris Wozniak to Leonard Cole ) . They are summarized in the supporting materials in this docket.

Acute Toxic ity

– Ecological Effects

T he acute toxicity ecological effects exposure assessments for non

- target organisms continue to support the labeled non

- food uses and the proposed food uses of Metarhizium anisopliae F52

. These include effects on avian species, wildlife, non

- target and beneficial insects, freshwater vertebrates and invertebrates, honeybee and beneficial organisms).

However, additional product performance data were reported and summarized herein

. Ecological effects data support both the non

- fo od and proposed food uses of Metarhizium anisopliae F52 as labeled

. Additional studies of the developmental effects of Metarhizium anisopliae F52 on a quatic vertebrates (fish embryos ) will be required if amendments are requested for direct application to water.

Acceptable Tier I nontarget toxicology data/information support the previously labeled and , now, the currently proposed uses of Metarhizium anisopliae F52 . Refer to Table 3 in the original BRAD issued with the 6/6/03 and 6/11/03 conditional regist rations. Submitted studies summarized therein showed that MetF52 poses no hazard to avian species, wild mammals and terrestrial and aquatic plant species from residential outdoor and institutional premise uses of the product. Data/information also showed no potential adverse effects to beneficial non

- target organisms (lady beetles, green lacewings, parasitic wasps, honey bee larvae, honey bee adults and earthworms). Freshwater and estuarine/marine fish, invertebrates and aquatic insects will

not be affect ed by the quantities entering the aquatic environment from incidental drift and runoff from terrestrial uses. However, in light of laboratory studies reporting toxicity and pathogenicity to immature aquatic vertebrate and invertebrate species, the Agency r equired additional hazard assessment prior to any registration for aquatic applications.

Guideline 885

- 4200: Freshwater Fish Toxicity/Pathogenicity As a condition of registration, the Agency required that the freshwater fish study be repeated.

The 30

- d ay LC 50 for rainbow trout exposed to M. anisopliae was >53 mg/L, the highest concentration tested. All fish appeared normal and there were no mortalities in all treatment groups. The study was rated SUPPLEMENTAL since a maximum hazard dose was not obtained and laboratory induced pathological changes in fish (silverside) embryos and fry has been reported in the literature.

Novozymes’ response to this condition was a rebuttal (dated December 09, 2004) and a resubmission (dated January 14, 2008). The Agency accepted Novozymes’ rationale for not repeating the study based on the use patterns, as discussed immediately below (USEPA/BPPDMetarhizum anisopliae F52 Page 23 of 31 Biopesticides Registration Action Document

– DRAFT Addendum March 1 7 , 2011

memo dated March 11, 2008, by Dr. Zigfridas Vaituzis to Shanaz Bacchus).

The submitted study (“Toxicity and Pathogenicity Tes t with the Rainbow Trout”) was performed on adult fish, whereas previous published reports indicate adverse effects of some M. anisopliae strains to developing fish embryos. An adult fish study would not address adverse effects on developing embryos. The adult fish study cannot be upgraded from its supplemental rating, because analysis of the test vessel water indicated that the concentration of the active ingredient is below the 53 mg/L added to the test vessel. Therefore, the exposure to the test fish w as not at the required Maximum Hazard Dose concentration. Since the material is described as “hydrophobic,” this would further have limited the exposure of the active ingredient to the test

animals.

Considering that the product, TAE

- 001 Technical Bioins ecticide, is registered for terrestrial uses that do not result in significant aquatic exposure, the “Toxicity and Pathogenicity Test with the Rainbow Trout” does not have to be repeated. However, in the event that the use patterns are expanded for this pr oduct to include aquatic use sites or otherwise increase exposure to freshwater fish, the Agency will require additional studies on the effects of Metarhizium anisopliae F52 TGAI (currently called TAE

- 001 Technical BioI nsecticide ) on developing freshwater fish embryos.Metarhizum anisopliae F52 Page 24 of 31 Biopesticides Registration Action Document

– DRAFT Addendum March 1 7 , 2011

IX

. APPENDIX B. MRID S AND CIT ED LITERATURE BIBLIO GRAPHY

The following studies and references were relied upon as described in the original BRAD , and

some are referenced in this document.

MRID Citation Reference

44844700 Taensa, Inc. (1999) Submission of Product Chemistry, Toxicity, and Environmental Fate Data in Support of the Application for Registration of TAE

001. Transmittal of 28 Studies.

44844701 Butts, E. (1999) Product Identification and Disclosure of Ingredients
Metarhiziu m anisopliae Strain F52. Unpublished study prepared by Taensa, Inc. 21 p.

44844702 Butts, E. (1999) Description of Beginning Materials and Manufacturing Process

-

- 1 Metarhizium anisopliae Strain 52: TAE

- 001 Technical Bioinsecticide. Unpublished study pre pared by Taensa, Inc. 8 p.

44844703 Freudenthal, R. (1999) Description of Beginning Materials and Manufacturing

Process

-- 2 Metarhizium anisopliae Strain 52: TAE

- 001 Technical Bioinsecticide. Unpublished study prepared by Taensa, Inc. 9 p.

44844704 Fre udenthal, R.; Butts, E. (1999) Discussion of the Formation of Impurities During the Production of Metarhizium anisopliae Strain F52. Unpublished study prepared by Taensa, Inc. 4 p.

44844705 Gingras, B. (1999) Lot Analysis of Metarhizium anisopliae Strain F52: Lab Project Number: L08743SN24. Unpublished study prepared by IIT Research Institute. 23 p. {OPPTS 885.1400}

44844706 Butts, E. (1999) Analysis of Samples

-- Manufacturing Process 2 Metarhizium anisopliae Strain F52: TAE

- 001 Technical Bioinsecticide. Unpublished study prepared by Taensa, Inc. 9 p.

44844707 Butts, E. (1999) Certification of Ingredient Limits: Metarhizium anisopliae

Strain F52: TAE

- 001 Technical Bioinsecticide. Unpublished study prepared by Taensa, Inc. 6 p.

44844708 Jacobs, R.; Nix on, W. (1998) Determination of the Color, Physical State, Odor, pH and Density of Metarhizium anisopliae TGAI: Lab Project Number: 495C

109. Unpublished study prepared by Wildlife International Ltd. 23 p. {OPPTS 830.6302, 830.6303, 830.6304, 830.7000, 830. 7300}

44844709 Harrington, K. (1999) Toxicity/Pathogenicity Testing of Metarhizium anisopliae , Strain F52, Following Acute Oral Challenge in Rats: Final Report: Lab Project Number: L08743SN13. Unpublished study prepared by IIT Research Institute. 74 p. { OPPTS 885.3050}Metarhizum anisopliae F52 Page 25 of 31 Biopesticides Registration Action Document

– DRAFT Addendum March 1 7 , 2011

44844710 Findlay, J. (1999) Acute Dermal Toxicity/Pathology Study of Metarhizium anisopliae , Strain F52, in Rabbits: Lab Project Number: L08743SN16. Unpublished study prepared by IIT Research Institute. 28 p. {OPPTS 885.3100}

44844711 Harrington, K. (1999) Toxicity/Pathogenicity Testing of Metarhizium anisopliae , Strain F52, Following Acute Intratracheal Challenge in Rats: Lab Project Number: L08743SN14. Unpublished study prepared by IIT Research Institute. 111 p. {OPPTS 885.3150}

44 844712 Gingras, B. (1999) Toxicity/Pathogenicity Testing of Metarhizium anisopliae, Strain F52, Following Acute Intraperitoneal Challenge in Rats: Lab Project Number: L08743SN15. Unpublished study prepared by IIT Research Institute. 92 p. {OPPTS 885.3200}

44844713 Findlay, J. (1999) Primary Eye Irritation Study Metarhizium anisopliae , Strain F52, in Rabbits: Lab Project Number: L08743SN17. Unpublished study prepared by IIT Research Institute. 26 p.

44844714 Butts, E. (1999) Hypersensitivity Incidents Re port Metarhizium anisopliae, Strain F52. Unpublished study prepared by Taensa, Inc. 4 p.

44844715 Findlay, J. (1999) Dermal Sensitization Study of Metarhizium anisopliae , Strain F52 in Guinea Pigs Using the Buehler Method: Final Report: Lab Project

Numbe r: L08743 SN18. Unpublished study prepared by IIT Research Institute. 26 p. {OPPTS 870.2600}

44844716 Garthwaite, S. (1999) Measurement of Fungal Toxins Destruxin and Cytochalasin in Metarhizium anisopliae Growth Media: Lab Project Number: 8743SN32. Unpu blished study prepared by IIT Research Institute. 29 p.

44844717 Gingras, B. (1999) Growth Parameters of Metarhizium anisopliae Strain F52 at Various Temperatures: Lab Project Number: L08743 SN26. Unpublished study prepared by IIT Research Institute. 18 p.

44844718 Gingras, B. (1999) Sensitivity of Detection of Metarhizium anisopliae Strain F52 for Toxicity/Pathogenicity Testing in Rats: Lab Project Number: L08743 SN12. Unpublished study prepared by IIT Research Institute. 35 p. {OPPTS 885.0001, 885.315 0, 885.3200}

44844719 Foster, J.; Beavers, J. (1999) An Avian Oral Pathogenicity and Toxicity Study in

the Northern Bobwhite: Metarhizium anisopliae Strain F52: Lab Project

Number: 495

- 103. Unpublished study prepared by Wildlife International Ltd. 28

p. {OPPTS 885.4050}

44844720 Drottar, K.; Krueger, H. (1999) A 21

- Day Life

- Cycle Toxicity and Pathogenicity Test with the Cladoceran (Daphnia magna): Metarhizium anisopliae Strain F52: Final Report: Lab Project Number: 495A

- 104. Unpublished study prepared b y Wildlife International Ltd. 36 p.Metarhizum anisopliae F52 Page 26 of 31 Biopesticides Registration Action Document

– DRAFT Addendum March 1 7 , 2011

44844721 Drottar, K.; Krueger, H. (1999) A Five

- Concentration Toxicity and Pathogenicity Test with the Rainbow Trout (Oncorhynchus mykiss): Metarhizium anisopliae Strain F52: Final Report: Lab Project Number: 495A

10 5. Unpublished study prepared by Wildlife International Ltd. 30 p. {OPPTS 885.4200}

44844722 Palmer, S.; Krueger, H.; Hoxter, K. (1999) A Dietary Pathogenicity and Toxicity Study with Green Lacewing Larvae (Chrysoperla carnea): Metarhizium anisopliae Str ain F52: Final Report: Lab Project Number: 495

- 107. Unpublished study prepared by Wildlife International Ltd. 19 p. {OPPTS 885.4340}

44844723 Palmer, S.; Krueger, H.; Hoxter, K. (1999) A Dietary Pathogenicity and Toxicity Study with Ladybird Beetle (Hipp odamia convergens): Metarhizium anisopliae Strain F52: Final Report: Lab Project Number: 495

- 105. Unpublished study prepared by Wildlife International Ltd. 24 p. {OPPTS 885.4340}

44844724 Palmer, S.; Krueger, H.; Hoxter, K. (1999) A Dietary Pathogenicity and Toxicity Study with Parasitic Hymenoptera (Nasonia vitripennis): Metarhizium anisopliae Strain F52: Final Report: Lab Project Number: 495

- 106A. Unpublished study prepared by Wildlife International Ltd. 24 p. {OPPTS 885.4340}

44844725 Maggi, V. (1999 ) Evaluation of the Dietary Effect(s) of Metarhizium anisopliae

Strain F

- 52 on Honey Bee Larvae (Apis mellifera L.): Lab Project Number:

CAR 166

- 98. Unpublished study prepared by California Agricultural Research, Inc. 38 p. {OPPTS 885.4380}

44844726 Magg i, V. (1999) Evaluation of the Acute Contact Toxicity of Metarhizium anisopliae Strain F

- 52 on Adult Honey Bees (Apis mellifera L.): Lab Project

Number: CAR 168

- 98. Unpublished study prepared by California Agricultural Research, Inc. 81 p.

44844727 Hoxte r, K.; Palmer, S.; Krueger, H. (1999) An Acute Toxicity Study with the

Earthworm in an Artificial Soil Substrate: Metarhizium anisopliae Strain F

- 52: Final Report: Lab Project Number: 495

- 104. Unpublished study prepared by Wildlife International Ltd. 22 p.

44844728 Palmer, S.; Krueger, H. (1999) A 96

- Hour Toxicity Test with the Freshwater Alga (Selenastrum capricornutum): Metarhizium anisopliae Strain F52: Final Report: Lab Project Number: 495A

- 106. Unpublished study prepared by

Wildlife International Ltd . 37 p. {OPPTS 850.5400}

44854700 Taensa, Inc. (1999) Submission of Product Chemistry Data in Support of the Application for Registration of TAE

- 001 Granular Bioinsecticide. Transmittal of

3 Studies.

44854701 Butts, E. (1999) Product Identification and Di s closure of Ingredients Metarhizium anisopliae Strain F52 TAE

- 001 Granular Bioinsecticide. Unpublished study prepared by Taensa, Inc. 4 p.Metarhizum anisopliae F52 Page 27 of 31 Biopesticides Registration Action Document

– DRAFT Addendum March 1 7 , 2011

44854702 Freudenthal, R. (1999) Description of Beginning Materials and Manufacturing

Process Metarhizium anisopl iae Strain 52 TAE

- 001 Granular Bioinsecticide. Unpublished study prepared by Taensa, Inc. 9 p.

44854703 Butts, E. (1999) Analysis of Samples and Certification of Ingredient Limits

TAE

- 001 Granular Bioinsecticide. Unpublished study prepared by Taensa, Inc

. 4 p.

45696300 Taensa, Inc. (2002) Submission of Product Chemistry Data in Support of the Application for Registration of TAE

- 001 Technical Bioinsecticide and TAE

- 001

Granular Bioinsecticide. Transmittal of 5 Studies.

45696301 Butts, E. (2002) Product Identity and Disclosure of Ingredients

-- Metarhizium anisopliae Strain F52

-- TAE

- 001 Granular Bioinsecticide: Lab Project Number: 1. Unpublished study prepared by Taensa, Inc. 4 p.

45696302 Butts, E. (2002) Beginning Materials and Manufacturing Process 1

- - Metarhizium anisopliae Strain F52: Lab Project Number: 2. Unpublished study prepared by Taensa, Inc. 4 p.

45696303 Jenkins, N.; Butts, E. (2002) Response to EPA Letter Dated December 2001: Beginning Materials and Manufacturing Process 2

-- Metarhizium ani sopliae Strain F52: Discussion of Formation Impurities During the Production of Metarhizium anisopliae Strain F52: Lot Analysis of Metarhizium anisopliae Strain F52: Analysis of Samples

-- Manufacturing Process 2

-- Metarhizium anisopliae Strain F52

-- TAE

- 001 T echnical Bioinsecticide: Certification of Limits: Document Number: 3. Unpublished study prepared by CABI Bioscience and Taensa, Inc. 11 p.

45696304 Arzumanov, T.; Jenkins, N. (2002) Soft Extraction of Metarhizium anisopliae

100

- 1181 (Strain F52) from Ric e: Manufacturing Process and Analysis of Samples: Lab Project Number: 4. Unpublished study prepared by CABI Bioscience. 9 p.

45696305 Butts, E. (2002) Certification of Ingredient Limits Soft Extraction of Metarhizium anisopliae Strain F52 from Rice: Lab Project Number: 5. Unpublished study prepared by Taensa, Inc. 4 p.

45723700 Taensa Inc. (2002) Submission of Efficacy Data in Support of the Application for Registration of Tick

- Ex G. Transmittal of 1 Study.

45723701 Butts, E. (2002) Efficacy Data for Metarhizium anisopliae Strain F52 and Other Strains of Metarhizium anisopliae on Ticks. Unpublished study prepared by Taensa, Inc. 28 p.

45778800 Earth BioSciences (2002) Submission of Product Chemistry and Toxicity Data in Support of the Application for Registration of Tick

- Ex EC. Transmittal of 6

Studies.

45778801 Butts, E. (2002) Tick

- Ex EC: Manufacturing Process. Unpublished study prepared by Earth Biosciences Inc. 46 p.Metarhizum anisopliae F52 Page 28 of 31 Biopesticides Registration Action Document

– DRAFT Addendum March 1 7 , 2011

45778802 Lezotte, F.; Van Hoven, R.; Nixon, W. (2002) Determination of the Co lor, Physical State, Odor, pH, and Density (Specific Gravity) of TAE

- 001 Oil Experimental Bioinsecticide: Lab Project Number: 495C

- 113: 495/073102/234B/SUB4: 5/073102/234B/SUB495. Unpublished study prepared by Wildlife International, Ltd. 30 p. {OPPTS 830. 6302, 830.6303, 830.6304, 830.7000, 830.7300}

45778803 Findlay, J. (2002) Acute Oral Toxicity Study of TAE

- 001 Oil in Rats: Lab Project Number: RPTSN02

- 176: 02

- 176. Unpublished study prepared by Experimur. 28 p. {OPPTS 870.1100}

45778804 Findlay, J. (2 002) Acute Dermal Toxicity Study of TAE

- 001 Oil in Rabbits: Lab Project Number: 02

- 177: RPSN02

- 177. Unpublished study prepared by Experimur. 27 p. {OPPTS 870.1200}

45778805 Findlay, J. (2002) Acute Dermal Irritation Study of TAE

- 001 Oil in Rabbits:

Lab P roject Number: 02

- 175: RPTSN02

- 175. Unpublished study prepared by Experimur. 27 p. {OPPTS 870.2500}

45778806 Findlay, J. (2002) Acute Eye Irritation Study of TAE

- 001 Oil in Rabbits: Lab Project Number: 02

- 174: RPTSN02

- 174. Unpublished study prepared by

E xperimur. 24 p. {OPPTS 870.2400}

47123900 Novozymes Biologicals, Inc. (2007) Submission of Product Chemistry Data in Support of TAE

- 001 Technical Bioinsecticide, Taenure Granular Bioinsecticide, Tick Ex G and Tick Ex EC. Transmittal of 1 Study.

4712390 1 Turnbough, A. (2007) Clonal Success of Metarhizium a nisopliae Var. Anisopliae in Southwestern British Columbia is Independent of Habitat Association. Project Number: NOVOZYMES/0701. Unpublished Study Prepared by Agriculture Canada. 36 p.

47313400 Novozy mes Biologicals, Inc. (2007) Submission of Efficacy Data in Support of the Registrations of TAE

- 001 Technical Bioinsecticide, Taenure Granular Bioinsecticide, Tick

- Ex G, and Tick

- Ex EC. Transmittal of 3 Studies.

47313401 Gangloff

- Kaufmann, J.; Leland, J. (2007) Application of a Fungal Pathogen for the Management of Ticks in a County Park, Suffolk County, NY. Project

Number: METARHIZIUM /2007/001. Unpublished study prepared by Novozymes Biologicals, Inc. 8 p.

47313402 Shulze, T.; Leland, J. (2007) Field Eva luation of Metarhizium anisopliae Strain

F52 for Tick Control

- New Jersey. Project Number: METARHIZIUM/2007/002. Unpublished study prepared by Novozymes Biologicals, Inc. 11 p.

47313403 Bharadwaj, A.; Stafford, K.; Leland, J. (2007) Evaluation of Metarhi zium anisopliae Strain F52 for Tick Control. Project Number:

METARHIZIUM /2007/003. Unpublished study prepared by Novozymes Biologicals, Inc. 17 p.Metarhizum anisopliae F52 Page 29 of 31 Biopesticides Registration Action Document

– DRAFT Addendum March 1 7 , 2011

47880600 Novozymes Biologicals, Inc. (2009) Submission of Product Chemistry Data in Support of the Registrat ions of TAE

- 001 Technical Bioinsecticide, Taenure Granular Bioinsecticide, Tick

- Ex G and Tick

- Ex EC. Transmittal of 2 Studies.

47880601 Schreier, T.; Kauffman, J. (2009) Metarhizium anisopliae Strain F52: Three Batch Fungal Contamination Analysis of Ferme ntation Process and Five Batch Analysis of TGAI. Project Number: METARHIZIUM9/24/2009. Unpublished study prepared by Novozymes Biologicals, Inc. 30 p.

47880602 Schreier, T. (2009) Metarhizium anisopliae Strain F52: Five Batch Analysis of

Tick

- Ex EC. Proje ct Number: METARHIZIUM9/29/2009. Unpublished study prepared by Novozymes Biologicals, Inc. 30 p.

48058800 Exponent (2010) Submission of Product Chemistry in Support of the Registrations of TAE

- 001 Technical Bioinsecticide, Taenure Granular Bioinsecticide, Tick EX G, and Tick EX EC. Transmittal of 2 Studies.

48058801 Thome, D. (2010) Description of the Manufacturing Process for Taenure Granular Bioinsecticide and Tick EX G. Project Number: MET/52/10/01. Unpublished study prepared by Novozymes Biologicals, Inc. 8 p.

48058802 Thome, D. (2010) Description of the Manufacturing Process for Tick EX EC. Project Number: MET/52/10/02. Unpublished study prepared by Novozymes Biologicals, Inc. 8 p.

48150700 Novozymes Biologicals, Inc. (2010) Submission of Efficacy Data in Support of the Registrations of Tae

- 001 Technical Bioinsecticide, Taenure Granular Bioinsecticide, Tick

- EX G and Tick

- EX EC. Transmittal of 4 Studies.

48150701 Leland, J.; Thome, D. (2010) Report on Efficacy of Met52 EC Against Mites. Project Numb er: NOVOZYMES/10/02. Unpublished study prepared by Novozymes Biologicals, Inc. 100 p.

48150702 Leland, J.; Thome, D. (2010) Report on Efficacy of Met52 EC against Thrips. Project Number: NOVOZYMES/10/03. Unpublished study prepared by Novozymes Biologicals , Inc. 125 p.

48150703 Leland, J.; Thome, D. (2010) Report on Efficacy of Met52 EC against Whitefly. Project Number: NOVOZYMES/10/01. Unpublished study prepared by Novozymes Biologicals, Inc. 38 p.

48150704 Leland, J.; Thome, D. (2010) Report on Efficacy of Met52 G against Black Vine Weevils. Project Number: NOVOZUMES/10/04. Unpublished study prepared by Novozymes Biologicals, Inc. 17 p.

48175200 Novozymes Biologicals, Inc. (2010) Submission of Exposure and Risk Data in Support of the Amended Registratio ns of TAE

- 001 Technical Bioinsecticide, Taenure Granular Bioinsecticide, and Tick

- Ex EC and the Petition for Tolerance

of Metarhizium anisopliae Strain F52. Transmittal of 1 Study.Metarhizum anisopliae F52 Page 30 of 31 Biopesticides Registration Action Document

– DRAFT Addendum March 1 7 , 2011

48175201 Habig, C.; Costanzo, S.; Wiedling, R. (2010) Risk Assessment for Exposure of Endangered and Threatened Insects to Metarhizium anisopliae Strain F52. Project Number: MET52/10/07. Unpublished study prepared by Exponent, Inc. 45 p.

48177000 Novozymes Biologicals, Inc. (2010) Submission of Product Chemistry Data in Suppor t of the Registrations of TAE

- 001 Technical Bioinsecticide, Taenure Granular Bioinsecticide, Tick

- Ex G, and Tick

- Ex EC. Transmittal of 1 Study.

48177001 Thome, D. (2010) Report on Fungal Contaminants during Production of Metarhizium anisopliae Strain F52 based Products. Project Number: NOVOZYMES/10/CAN/01. Unpublished study prepared by Novozymes Biologicals, Inc. 20 p.

References

Hu, G. and R.J. St. Leger. 2002. Field studies using a recombinant mycoinsecticide ( Metarhizium anisopliae ) reveal that it is rhi zosphere competent. Appl. Env. Microbiol.,

68:6383

- 6387.

Meyling, N.V. and J. Eilenberg. 2007. Ecology of the entomopathogenic fungi Beauveria

bassiana and Metarhizium anisopliae in temperate agroecosystems: potential for conservation biolog ical control. Biol. Control, 43:145

- 155.

Vanninen, I., J. Tyni

- Juslin, and H. Hokkanen. 2000. Persistence of augmented Metarhizium anisopliae and Beauveria bassiana in Finnish agricultural soils. BioControl, 45:201

- 222.

Zimmerman, G. 2007. Review on safety of the entomopathogenic fungus Metarhizium anisopliae . Biocontrol Sci. and Technol., 17: 879

- 920.Metarhizum anisopliae F52 Page 31 of 31 Biopesticides Registration Action Document

– DRAFT Addendum March 1 7 , 2011

X

. GLOSSARY OF ACRONYMS AND ABBREVIATIONS

BPPD Biopesticides and Pollution Prevention Division

BRAD Biopesticides Registration Action Docu ment
CFR Code of Federal Regulations

cfu colony

- forming units

cfu/lb colony

- forming units per pound

°C degrees Celsius

DER Data Evaluation Record

EDSP Endocrine Disruptor Screening Program

EP end

- use product

EPA Environmental Protection Agency (the “Agen cy”)

FFDCA Federal Food, Drug, and Cosmetic Act

FIFRA Federal Insecticide, Fungicide, and Rodenticide Act FQPA Food Quality Protection Act

FR Federal Register mg/kg bw milligram per kilogram of body weight

MRID No. Master Record Identification Number

OCSP P Office of Chemical Safety and Pollution Prevention ; formerly, Office of Pesticides and Toxic Substances (OPPTS)

OPP Office of Pesticide Programs

TGAI t echnical grade of the active ingredient