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 17, 2011
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Biopesticides Registration Action Document – DRAFT Addendum March 17, 2011

TABLE OF CONTENTS 

I. BACKGROUND 4

II. ACTIVE INGREDIENT OVERVIEW 4

III. REGULATORY BACKGROUND 5

A. Extensions of Conditional Registrations 5

B. Conditions of Registration 6

C. Public Comment Period 7

D. Food Clearances/Tolerances 7

IV. RISK ASSESSMENT 8

A. Product Analysis Assessment 8

B. Human Health Assessment 9

C. Environmental Assessment 12

D. Product Performance Data (Efficacy) 15

V. RISK MANAGEMENT DECISION 16

VI. ENVIRONMENTAL JUSTICE 17

VII. TERMS AND CONDITIONS OF THE REGISTRATION(S) 17

VIII. APPENDIX A. DATA REQUIREMENTS 18

IX. APPENDIX B. MRID AND CITED LITERATURE BIBLIOGRAPHY 24

X. GLOSSARY OF ACRONYMS AND ABBREVIATIONS 31 
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BIOPESTICIDES REGISTRATION ACTION DOCUMENT TEAM

Office of Pesticide Programs:

Biopesticides and Pollution Prevention Division

Microbial Pesticides Branch (MPB)

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, Microbial Pesticides Branch

Shanaz Bacchus Regulatory Action Leader

Leonard Cole Previous Regulatory Action Leader
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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 pesticides contain 

living MetF52, a deuteromycetous (or anamorphic) fungus that is pathogenic to insects, mites and 

ticks. The fungus is ubiquitous, and found in soil, river sediments, in association with nematodes, 

and as a saprophyte on organic detritus. The known host range of M. anisopliae includes over 200 

insect species. Most of these belong to the Elateridae and Curculionidae families in the insect 

Order Coleoptera, although it is known to infect insects found in several other Orders 

(Zimmerman 2007). 

MetF52 infects insects that come into contact with its spores. The spores attach to the surface 

of the insect, germinate and begin to grow. The growing fungus penetrates the exoskeleton 

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

insects that come into contact with infected insects also become infected, which leads to their 

death.

The original Biopesticide Registration Action Document (BRAD) for Metarhizium 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 registrations of the MetF52 pesticides between June 6, 2003, and June 11, 2003. 

The assessments continue to support the currently registered uses of the pesticides. This 

addendum includes references to relevant, previously evaluated data and other information

from the BRAD, and provides new information and updated summaries of the Agency’s 

current risk assessments for the proposed amendment to allow the application 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-EXTM 

G

TICK-EXTM 

EC

TICK-EX EC BIOINSECTICIDE
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Culture Deposit: 1) American Type Culture Collection (ATCC) Number is 90448

2) ARSEF (Agricultural Research Service Entomopathogenic 

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 Biologicals, Inc.

Target Pests: Mites, 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 conditions of registration, additional data were needed to: 

• Upgrade the submitted product identity of the active ingredient;

• Upgrade the manufacturing process and disclosure of unintentional ingredients studies;

• Reassess the effects of Met F52 on aquatic vertebrates by repeating the freshwater fish 

study;

• Determine the efficacy of Met F52 on the claimed public health pests (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 to Mr. 

Theodore Melnik, Novozymes Biologicals, Inc., referred to hereafter as Novozymes). 

A. Extensions of Conditional Registrations

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

provided that the registrant submitted 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 amendments for applications to food crops (see 

Food Tolerances, below). The notice of receipt of these amendments was published in the 

Federal Register on March 5, 2010 (75FR10259). The current extension of the existing (nonfood use) registrations to May 2, 2011 was granted to coincide with the Agency’s review of the 
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product chemistry, dietary and ecological exposure and risk assessments for the proposed food 

uses.

 

B. Conditions of Registration

1. Product chemistry 

Novozymes submitted studies to satisfy the conditional data requirements and obtain 

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

The product chemistry data for the manufacturing and end-use products are a matter of 

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

and bacterial unintentional ingredients, and is acceptable (US EPA BPPD DER MRIDs

47123901 and 48177001, dated 03/04/10 and 12/30/2010 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, 2004) 

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 document. 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 nonfood indoor and greenhouse use. One end-use product, TICK-EXTM

G was intended for use on 

residential and institutional lawns and landscape perimeters to control ticks and certain grubs. 

The conditional registration notice (revised February 12, 2007) required that data must 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 August 2010 to support control of mites, 

thrips, ticks, weevils and whiteflies in order to amend the labels to reflect these target pests. For 

more information, see http://www.epa.gov/pesticides/pestlabels/. 

4. Other Label Amendments 

Label amendments 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://www.epa.gov/pesticides/pestlabels/ and search for EPA Reg. Nos. 70127-7, 70127-8, 

70127-9, and 70127-10.
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C. Public Comment Period

This document and the associated risk assessments are posted in the docket (EPA-HQ-OPP2010-0081) 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. Food Clearances/Tolerances 

Novozymes submitted a petition to establish an exemption from 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) in 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 were submitted in response to 

the published Notice. This addendum provides the current assessments of the data/information 

that were relied upon to support the request to exempt MetF52 from the requirement of a 

tolerance. 
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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 updated the definition for 

microbial pesticides (72 FR 61002). The rule became effective on December 26, 2007. The data 

and information evaluated for the June 2003 BRAD, published for the initial conditional 

registration, and all subsequent data/information evaluated 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 can 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) part 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 studies and/or other information 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 summary 

of the updated status of these 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 storage stability data as a term of these registration amendments, these pesticides are 

eligible 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. Additionally, 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.
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B. Human Health Assessment (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 Toxicity Category IV 

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

irritation studies resulted in Toxicity 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 these 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 occurring 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 of 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 relationship 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 chemicals 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 disruptors.

MetF52 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 orders/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://www.epa.gov/endo/.
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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 residue 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 chemical 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 to 

pesticide residues. First, the Agency determines the toxicity of pesticides. Second, the Agency

examines exposure to the pesticide through food, drinking water, and through other exposures 

that occur as a result of pesticide use in residential settings.

Consistent with section 408(b)(2)(D) of FFDCA, the Agency has reviewed the available 

scientific data and other relevant information, and considered its validity, completeness, and 

reliability and the relationship of this 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 children.

Based on the acute toxicity/pathogenicity data presented in Table 2 in the original BRAD the 

data required for a FQPA risk assessment for Metarhizium anisopliae F52 have been satisfied.

a. Aggregate Exposure

In examining aggregate exposure, section 408 of FFDCA directs the Agency to consider 

available information concerning exposures from the pesticide residue in food and all other nonoccupational exposures, including drinking water from ground water or surface water and 

exposure through pesticide use in gardens, lawns, or buildings (residential and other indoor 

uses).

1. Dietary Exposure 

Dietary exposure to residues of MetF52 from its proposed pesticidal use is anticipated to be 

negligible, as described below.
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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. The spores, 

which adhere to, and infect insects, are not expected to grow on the crops. Though unlikely, 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 

fungicide will not be applied to water. Further, the spores are large and readily adhere to soil 

particles, and are unlikely to move through soils to reach underground water. Should any

MetF52 spores reach underground water, they would be removed during municipal water treatment. 

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 significantly above normal levels of these naturally occurring fungi. Also, if the 

proposed food uses are approved, pesticidal applications of Met F52 would occur in agricultural 

and residential areas. However, there have been no reports of pathogenicity or other adverse 

effects in humans and animals from exposure to the registered products or to the 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 requires 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 toxicity.” Acute toxicity studies demonstrated that MetF52 is virtually non-toxic to mammals, 

and there is no known mechanism of toxicity that it shares with other insecticidal substances.

b. Determination of Safety for the U.S. Population, Infants, and Children

FFDCA section 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 

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

for infants and children in the case of threshold effects to account for prenatal and postnatal 

toxicity and the completeness 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 referred to as uncertainty factors, 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.
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MetF52 has a low mammalian acute toxicity potential, as discussed in Table 2 of the original 

BRAD, and Appendix A of this addendum. Registered pesticide products containing this active 

ingredient have been in use for almost 18 years without any reported adverse effects 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 of MetF52. This includes all anticipated dietary exposures and all other exposures 

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

data and information available on MetF52 do not demonstrate toxic, pathogenic, and/or infective 

potential to mammals. 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 results demonstrated that Met F52 was not toxic or 

pathogenic in laboratory mammals via the oral, pulmonary, or intraperitoneal injection routes of 

exposure, and occupational handler exposure is 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. Handlers working with MetF52 must 

wear a long-sleeved shirt, 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 applying 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, the 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, support the Agency’s conclusions that no unreasonable adverse effects would be expected 

from the proposed food uses. 

C. Environmental Assessment (40 CFR § 158.2150)

1. Nontarget Organism Toxicology 

All nontarget toxicology data requirements for the proposed uses of Met F52 have been satisfied. 

Additionally, the Agency risk assessments that were evaluated 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 MetF52. (Refer to Table 3 in the original BRAD issued with the 

6/6/03 and 6/11/03 conditional registrations.) The submitted studies demonstrated that Met F52 

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 showed no potential adverse effects to beneficial non-target organisms (lady beetles, green 

lacewings, parasitic wasps, honey bee larvae, honey bee adults and earthworms). For the 
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original conditional registration, the data demonstrated that freshwater 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 to 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 maximum hazard dose was not 

obtained, and there are reports of laboratory induced pathological changes (silverside) in fish 

embryos and fry in the scientific literature. An adult fish study would not address adverse effects 

on developing embryos. Therefore, if in the future the use patterns are expanded to include 

aquatic use sites or otherwise increase 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 certainty. 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 estimate 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 

conditions (e.g., temperature, soil moisture), biotic factors, and can be strain-dependent 

(Zimmerman 2007, Vanninen et al. 2000). Persistence may differ between laboratory, 

greenhouse, and field conditions. M. anisopliae is not easily airborne, and is not an aquatic 

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 Eilenberg 2007, Hu and St. Leger 2002). However, insects are believed to be the primary 

source of organic matter for population increase, and may account for some persistence in the 

environment. The fungus has been observed to be restricted to the insect host’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.
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However, based on the results of the acute non-target toxicity testing, the need for environmental 

fate and groundwater data was not triggered.

3. Ecological Exposure and Risk Characterization

The 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. anisopliae 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 some insect species than 

others. The submitted studies showed that M. anisopliae Strain F52 does not pose a hazard to the 

major beneficial insects tested, including honey bees, but the potential risks to all non-target insects 

cannot be ruled out. 

The Agency 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 plant species. The potential 

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

4. Endangered Species Assessment

The BRAD for Met F52 states that an expansion of uses would trigger concerns for endangered 

and threatened 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 habitats. Novozymes submitted an endangered species assessment

(MRID 48175201), which was reviewed by BPPD and rated “Supplemental.” 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 includes over 200 species, and most belong to the 

Elateridae and Curculionidae families of Order Coleoptera, though infection 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 made “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 plants,

• Root 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 threatened 
Metarhizum anisopliae F52 Page 15 of 31

Biopesticides Registration Action Document – DRAFT Addendum March 17, 2011

and endangered species and their designated critical habitats are also not expected. Therefore, a 

“No Effect” determination is made for direct and indirect effects to 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. Growth 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 temperature affects growth, and consequently, persistence of 

Met F52 in the environment. Moreover, MetF52 has a lower toxicity potential than the 

alternative insecticides (carbamates and pyrethrins) for the proposed uses. Following 

application, the fungus returns 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 conducted during 

registration review for Met F52. 

D. Product Performance Data (Efficacy)

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 submitted for review 

only in connection with the registration of products directly pertaining to the mitigation of 

disease bearing human health organisms and certain designated quarantine pests, i.e., ticks, 

mosquitoes, fleas, Mediterranean fruit flies, gypsy moths, Japanese beetles, 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.pdf). 

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 containing Met F52 claim ticks 
Metarhizum anisopliae F52 Page 16 of 31

Biopesticides Registration Action Document – DRAFT Addendum March 17, 2011

as pests. The Agency evaluated the efficacy data that were submitted to satisfy conditions of 

registration, and determined that these data support 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 included in the proposed amendments to add food uses to 

registered products containing Met F52. The products must not be used near critical habitats of 

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

determination 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 Pests

The insect pathogenicity data submitted for the Environmental Risk Assessment of MetF52 

Biological Insecticide, TICK-EXTM, (formerly EPA Reg. No. 72098-T) performed in 2003 

showed no adverse effects on nontarget insects. Submitted nontarget toxicity studies showed

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 provides statistically significant control of mites, thrips, 

black vine weevils and white flies on agricultural crops when applied according to the label 

directions (MRIDs 48150701, 48150702, 48150703, 48150704: US EPA BPPD memo and 

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

V. RISK MANAGEMENT DECISION

Pesticides containing Metarhizium anisopliae F52 as the active ingredient were conditionally 

registered under FIFRA Section 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 

required 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 generally 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 

products 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) registrations for the labeled 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 17, 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 JUSTICE

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 Metarhizium 

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.

VII. TERMS AND CONDITIONS OF THE REGISTRATION(S)

As a term of registration, the registrant must submit one-year storage stability data (40 CFR § 

158.2120).
Metarhizum anisopliae F52 Page 18 of 31

Biopesticides Registration Action Document – DRAFT Addendum March 17, 2011

VIII. APPENDIX A. DATA 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 

considered in light of these requirements. Data and information reviewed for the original 

registration of the products 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 PP8F7508. 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 Metarhizium anisopliae F52 (40 CFR § 158.2120)

OPPTS 

Guideline 

No. 

Study Results MRID & 

references

885.1100 Product 

identity

The data reviewed for the initial conditional

registrations (June 6 and June 11, 2003) were not 

sufficient to warrant the conclusion that the method 

could be used to separate or identify F52 from other 

commonly encountered strains of M. anisopliae. The 

study was classified as SUPPLEMENTAL (see BRAD, 

06/06/03). Additional data reviewed for clonal 

success of Metarhizium anisopliae F52 strains (DER 

471239-01 dated 03/04/10) are SUPPLEMENTAL. 

Another set of data (481770-01, DER 12/30/10) were 

considered ACCEPTABLE

456963-01*

448547-01*

471239-01

481770-01

885.1200

Manufacturing 

Process

Additional data required. SUPPLEMENTAL 

456963-02*

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**

478806-02***

480588-01**

480588-02***

Additional data reviewed are ACCEPTABLE.

481770-01

DER12/30/10
Metarhizum anisopliae F52 Page 19 of 31

Biopesticides Registration Action Document – DRAFT Addendum March 17, 2011

TABLE 1. Product Chemistry Data Requirements

Additional Data Reviewed for Metarhizium anisopliae F52 (40 CFR § 158.2120)

885.1300

Formation of 

Unintentional 

Ingredients

Additional data were required for the initial conditional 

registration and these studies were rated 

SUPPLEMENTAL

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

DER12/30/10

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 residues 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 Bioinsecticide 
Metarhizum anisopliae F52 Page 20 of 31

Biopesticides Registration Action Document – DRAFT Addendum March 17, 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 Physical 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 Storage 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 Size, 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/A
Metarhizum anisopliae F52 Page 21 of 31

Biopesticides Registration Action Document – DRAFT Addendum March 17, 2011

The 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 previously submitted and evaluated meet the current 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 - not dispersible in water; 

pH not measured

46449614

63-13

*830.6317

Stability (Shelf 

life)

Greater than 15 months at 220C – 260C 46580601

63-16

*830.6316

Explodability Not potentially explosive 46449614

63-18

*830.7100

Viscosity 40.6 Centipoises at 29.40C (850F) 46449614

63-20

*830.6320

Corrosion 

Characteristics

Not corrosive; does not react with 

metals

46449614

Viability 5.6 x 109 cfu/g Letter 

10/27/06; 

46580601
Metarhizum anisopliae F52 Page 22 of 31

Biopesticides Registration Action Document – DRAFT Addendum March 17, 2011

Acute Toxicity – Health Effects 

The Agency’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 the non-food uses, and 

support the proposed food uses of Met F52 pesticides (US EPA BPPD Data Evaluation Record, 

electronic file dated 12/11/01 from Chris Wozniak to Leonard Cole). They are summarized in the 

supporting materials in this docket.

Acute Toxicity – Ecological Effects 

The 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-food and proposed food uses of Metarhizium 

anisopliae F52 as labeled. Additional studies of the developmental effects of Metarhizium 

anisopliae F52 on aquatic 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 registrations. 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 affected 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 required 

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-day LC50 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/BPPD 
Metarhizum anisopliae F52 Page 23 of 31

Biopesticides Registration Action Document – DRAFT Addendum March 17, 2011

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

The submitted study (“Toxicity and Pathogenicity Test 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 was 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 Bioinsecticide, 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 product 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 BioInsecticide) on developing freshwater fish 

embryos.
Metarhizum anisopliae F52 Page 24 of 31

Biopesticides Registration Action Document – DRAFT Addendum March 17, 2011

IX. APPENDIX B. MRIDS AND CITED LITERATURE BIBLIOGRAPHY

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 TAE001. Transmittal of 28 Studies. 

44844701 Butts, E. (1999) Product Identification and Disclosure of Ingredients 

Metarhizium 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 prepared 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 Freudenthal, 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.; Nixon, W. (1998) Determination of the Color, Physical State, Odor, 

pH and Density of Metarhizium anisopliae TGAI: Lab Project Number: 495C109. 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 17, 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} 

44844712 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 Report 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 

Number: 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. Unpublished 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, 885.3150, 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 by Wildlife International Ltd. 36 p. 
Metarhizum anisopliae F52 Page 26 of 31

Biopesticides Registration Action Document – DRAFT Addendum March 17, 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: 495A105. 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 Strain 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 (Hippodamia 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 Maggi, 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 Hoxter, 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 Disclosure 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 17, 2011

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

Process Metarhizium anisopliae 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 anisopliae

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 Technical 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 Rice: 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 17, 2011

45778802 Lezotte, F.; Van Hoven, R.; Nixon, W. (2002) Determination of the Color, 

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. (2002) 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 Project 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 

Experimur. 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. 

47123901 Turnbough, A. (2007) Clonal Success of Metarhizium anisopliae Var. 

Anisopliae in Southwestern British Columbia is Independent of Habitat 

Association. Project Number: NOVOZYMES/0701. Unpublished Study 

Prepared by Agriculture Canada. 36 p.

47313400 Novozymes 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 Evaluation 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 Metarhizium 

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 17, 2011

47880600 Novozymes Biologicals, Inc. (2009) Submission of Product Chemistry Data in 

Support of the Registrations 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 Fermentation 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. Project 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 Number: 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 Registrations 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 17, 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 

Support 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 rhizosphere 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 biological 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 17, 2011

X. GLOSSARY OF ACRONYMS AND ABBREVIATIONS 

BPPD Biopesticides and Pollution Prevention Division

BRAD Biopesticides Registration Action Document

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 “Agency”)

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

OCSPP Office of Chemical Safety and Pollution Prevention; formerly, Office of Pesticides 

and Toxic Substances (OPPTS)

OPP Office of Pesticide Programs

TGAI technical grade of the active ingredient