Document ID: EPA-HQ-OPP-2010-0081-0009
Agency: epa
Document Type: Supporting & Related Material
Title: 
Posted Date: 2011-06-10T04:00Z

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

April 18, 2011

TABLE OF CONTENTS 

I.	BACKGROUND 									4

II.		ACTIVE INGREDIENT OVERVIEW 						4

III.	 	REGULATORY BACKGROUND 						5

  Extensions of Conditional Registrations 							5

  Conditions of Registration 								6

  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  

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

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_029
056.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. 

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: Met52™ GRANULAR BIOINSECTICIDE] 

							TICK-EXTM G

							TICK-EXTM EC

							TICK-EX EC BIOINSECTICIDE

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

      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).  

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
(non-food use) registrations to May 2, 2011 was granted to coincide with
the Agency’s review of the product chemistry, dietary and ecological
exposure and risk assessments for the proposed food uses. 

  

Conditions of Registration 

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).

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. 

 Efficacy 

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

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.

Public Comment Period

This document and the associated risk assessments were posted in the
docket (EPA-HQ-OPP-2010-0081) 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.  No comments were received to the documents posted
in this docket.

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.  

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 (  HYPERLINK
"http://ecfr.gpoaccess.gov/cgi/t/text/text-idx?c=ecfr&sid=f25d5426f66f63
ffeb9c986db92bf243&rgn=div8&view=text&node=40:23.0.1.1.9.16.1.3&idno=40"
 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  
HYPERLINK "http://www.regulations.gov"  http://www.regulations.gov .

	

	B.	Human Health Assessment (  HYPERLINK
"http://ecfr.gpoaccess.gov/cgi/t/text/text-idx?c=ecfr&sid=76805ba87ba7a2
c69b53feed9af452bb&rgn=div8&view=text&node=40:23.0.1.1.9.16.1.5&idno=40"
 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_02
9056.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/.

 

	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 non-occupational 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).

Dietary Exposure  

Dietary exposure to residues of MetF52 from its proposed pesticidal use
is anticipated to be negligible, as described below.

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.).

 	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.   

 

 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.

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 (  HYPERLINK
"http://ecfr.gpoaccess.gov/cgi/t/text/text-idx?c=ecfr&sid=c14a9a756506b0
93fafab6573b43e0fe&rgn=div8&view=text&node=40:23.0.1.1.9.16.1.6&idno=40"
 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 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.

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 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 (now called OCSPP)
Harmonized Guideline 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 (  HYPERLINK "http://www.epa.gov/PR_Notices/pr2002-1.pdf" 
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 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). 

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.

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:   HYPERLINK
"http://www.epa.gov/compliance/environmentaljustice/index.html" 
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).  These data have been submitted and
are under review.  However, the registrations are eligible for
unconditional registration based on its historical use for eight years
without any reports of adverse effects and the acceptable data evaluated
to satisfy the conditions of registration. 

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)

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

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  

The following results were excerpted from an electronic file dated
12/11/01 (DER from Chris Wozniak to Leonard Cole, USEPA/OPP/BPPD).  
These previously submitted and evaluated data meet the current data
requirements in accordance with 40 CFR Part 158-SubpartV (72 FR 61002).

TABLE 2. Physical and Chemical Properties of Metarhizium anisopliae F52
Technical – Data Requirements (40 CFR § 158.2120)

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

 

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 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.

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.

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 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: 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} 

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. 

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-105. 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. 

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. 

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.

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.

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.



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

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OPP	Office of Pesticide Programs

OPPTS	Office of Prevention, Pesticides and Toxic Substances (old name
for OCSPP)

TGAI 	Technical Grade of the Active Ingredient

Metarhizum anisopliae F52		Page   PAGE  22  of   NUMPAGES  30 

Biopesticides Registration Action Document –Addendum		April 18, 2011