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 1 8 , 2011Metarhizum anisopliae F52 Page 2 of 30

Biopestici des Registration Action Document

–Addendum April 18 , 2011 TABLE OF CONTENTS I. BAC KGROUND 4 II. ACTIVE INGREDIENT OVERVIEW 4 III. REGULATORY BACKGROUN D 5 A. E xtensions of C onditional Registrations 5 B. Conditions of Registration 6 C. Public Comment Period 7 D . F ood C learances /T olerances 7 IV. RISK ASSESSMENT 8 A. P roduct A nalysis A ssessment 8 B. Hum an Health Assessment 9 C. Environmental Assessment 1 2 D. P roduct P erform ance D ata (E fficacy) 1 5 V. RISK MANAGEMENT DECIS ION 1 6 VI. ENVIRONMENTAL JUSTICE 1 7 VII. TERMS AND CONDITIONS OF THE REGISTRATION( S) 1 7 VIII. APPENDIX A. DATA REQUIREMENTS 1 8 IX. APPENDIX B. MRID AND CITED LITER ATURE BIBLIOGRAPHY 24 X. GLOSSARY OF ACR ONYMS AND ABBREVIATIONS 3 1Metarhizum anisopliae F52 Page 3 of 30

Biopestici des Registration Action Document

–Addendum April 18 , 2011

BIOPESTICIDES REGIST RATION ACTION DOCUME NT TEAM

Office of Pesticide Programs: Biopesticides and Pollution Prevention Division Microbial Pesticides Branch (M PB) Ib rahim Barsoum Product Anal ys is, Human Health John Kough Product Anal ys is, Human Health Shannon Borges Environmental Effects Zi gfridas Vaituzis Environmental Effects Gail Tomimatsu Environmental Effects Sheryl Reill y Chief, Microbial Pesticides Branch Shanaz Bacchus Regulatory Action Leader Leonard C ole Previous Regulatory Action L eaderMetarhizum anisopliae F52 Page 4 of 30

Biopestici des Registration Action Document

–Addendum April 18 , 2011 I. BACKGROUND Metarhizium anisopliae strain F52 (called MetF52 in this document) is the active ingredient in microbial insecticides that were conditionally registered between June 6, 2003 , and June 11, 2003 , to E arth 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 MetF5 2, a deuteromycetous ( or anamorphic ) fungus that is pathogenic to insects , mites and ticks . The fungus is ubiquitous, and found in soil, river sediments, in associat ion with nematodes, and as a saprophyte on organic detritus. The known host range of M. anisopliae includes over 200 insect species. Most of th e se belong to the Elateridae and Curculionidae families in the in sect Order Coleoptera, although it is known to infect insects found in several other O rders (Zimmerman 2007). MetF 52 infects inse cts that come into contact with its spores
. The spores atta ch to the surface of the insect, germinate and begin to grow. The growing fungus penetrate s the exoskeleton of the insect host then grows very rapidly inside the insect , resulting in its death. Other insects that come into contact with infected insects also become inf ected, which leads to their death. The original Biopesticide Registration Action Document ( BRAD ) for Metarh izium anisopliae F52 is available on the Biopesticides and Pollution Prevention Division’s (BPPD’s) website at http://www.epa.gov/oppbp pd1/biopesticides/ingredients/tech_docs/brad_ 029056.pdf
. The BRAD summarizes the Agency ’s exposure and risk assessments that were the basis of the conditional registration s of the MetF52 pesticide s between June 6, 2003, and June 11, 2003
. Th e assessment s continue to support the currently registered uses of the pesticides . T his addendum includes references to relevant , previously evaluated data and other information from the BRAD , and provides new information and updated summari es of the Agency ’s current risk assessments for the proposed amendment to allow the appl ication of MetF52 to food crops. II. ACTIVE INGREDIENT OVERVIEW The MetF52 products conditionally registered are: Common Name: Metarhizium anisopliae strain F52 Trade & Other Names: TAE
- 001 Tec hnical Bioinsecticide TAE
- 001 Granular Bioinsecticide; TAENURE GRANULAR BIOINSECTICIDE [ Alternate Brand Name: Met52TM GRANULAR BIO INSECTI CIDE ] TICK
- EX TM G TICK
- EX TM EC TI CK
- EX EC BIOINSECTI CIDEMetarhizum anisopliae F52 Page 5 of 30

Biopestici des Registration Action Document

–Addendum April 18 , 2011 Culture Deposit: 1) American Ty pe Culture Collection (ATCC ) Number is 90448 2) ARSEF (Agricultural Research Service Entomopath ogenic 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: Novozy mes B iologicals, Inc. Target Pests: M ites, thrips, ticks, whiteflies, weevils III. REGULATORY BACKGROUND Pesticides containing Met F52 were conditionally registered between June 6, 2003 , and June 11, 2003 to Earth BioSciences Inc. As condition s of regis tration, additional data were needed to :

 Upgrade the submitted product identity of the active ingredient;  Upgrade the manufacturing process and disclosure of unintentional i ngredients studies ;  R eassess the effects of Met F52 on aquatic vertebrates by repeating the freshwater fish study ;  D etermine the efficacy of Met F52 on the claimed public health pest s (e.g. blacklegged ticks, Ixodes scapularis ). These data were to be submitted within one year from the date of the initial registration. The additional data and information were submitted in 2004 , but were not sufficient to address the conditions of registration. In 2006 , the pesticide products were transferred to Novozy mes Biologicals, Inc., (US EPA letter dated December 7, 2006 from Kathry n S. Bouve to Mr. Theodore Melnik, Novozy mes Biologicals, Inc. , referred to hereafter as Novozy mes ). A. Extension s of Conditional Registrations The conditional registrations for MetF52 pesticides were extended on February 12, 2007 , provided that the registrant submi tted acceptable product performance data for tick control by December 31, 2007
. On February 2, 2009, requests were granted to Novozy mes to extend the conditional registrations to January 30 , , 2010 , pending submission of additional product chemistry data. On November 4, 2009, the conditional registrations were extended to January 30, 2011, pending review of the product chemistry data submitted to fulfill the conditions of registration , in addition to the proposed product amendment s for applications to food crops (see Food Tolerances , below) . The notice of receipt of the se amendment s was published in the Federal Register on March 5, 2010 (75FR10259). The current extension of the existing (non
food use) registrations to May 2, 2011 was granted to coincide w ith the Agency ’s review of theMetarhizum anisopliae F52 Page 6 of 30

Biopestici des Registration Action Document

–Addendum April 18 , 2011 product chemistry , dietary and ecological exposure and risk assessments for the proposed food uses . B. Conditions of Registration 1. Product chemistry Novozy mes submitted studies to satisfy the conditional data requirements and obtain unconditional registration s of the company’s MetF52 insecticides under FIFRA section 3(c) (5)
. The product chemistry data for the manufacturing and e nd
- use p roducts are a matter of Confidential Business Information. The final submission includ ed results from testing for fungal and bacterial unintentional ingredients, and is a cceptable (US EPA BPPD DER MRID s 47123901 and 48177001, dated 03/04/10 and 1 2/30/2010 respectivel y , from Ibrahim Barsoum to Shanaz Bacchus)
. For more information , see (see Table 1, Appendix A of this document)
. 2. Aquatic vertebrates As a condition of registration on June 6, 2003 , the Agency required that the freshwater fish study be repeated. The registrant’s response to this condition was a rebuttal (dated December 09, 20 04) and , later , a resubmission (dated January 14, 2008). The Agency

accepted Novozy

mes’ rationale for not repeating the study based on the use patterns (memorandum dated March 11, 2008, b y Dr. Zi gfridas Vaituzis), as discussed in Chapter 3C of this document . Inthe event the registrant seeks to add applications to aquatic sites to their products, additional data may be required. 3. Efficacy The original conditional registrations (2003; extended February 12, 2007) were intended for non
food indoor and gr eenhouse use. One end
- use product, TI CK
- EX TM G was intended for use on residential and institutional lawns and landscape perimeters to control ticks and certain grubs. T he conditional registration notice (revised February 12, 2007 ) required that data mus t be submitted by

December 31, 2007 to demonstrate the efficacy of MetF52 against black
- legged ticks . Novozy

mes 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)
. Novozy mes 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. F or more information, see http://www.epa.gov/pesticides/pestlabels/
. 4. Other Label Amendments L ab el 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 Sy stem. 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.Metarhizum anisopliae F52 Page 7 of 30

Biopestici des Registration Action Document

–Addendum April 18 , 2011 C. Public Comment P eriod This document and the associated risk assessments we re posted in the docket ( EPA
- HQ
- OPP
20 10
- 0081 ) to allow the public 30 day s t o 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
. D. F ood Clearances/Toleranc es Novozy mes 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 ) i n the Federal Register on April 28, 2009 ( 74 FR 15969 ; EPA
- HQ
- OPP
- 2009
- 0194 ). Following a 30
- day comment period , the Agency determined that no comments w ere submitted in response to the published Notice
. This a ddendum provides the current asses sments of the data/information that were relied upon to support the request to exempt MetF52 from the requirement of a tolerance.Metarhizum anisopliae F52 Page 8 of 30

Biopestici des Registration Action Document

–Addendum April 18 , 2011 IV. RISK ASSESSMENT On October 26, 2007, the Agency issued a Final Rule in the Federal Register on the data requ irements 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 th e 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 b y Age ncy 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 Ag ency 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 b y the current 40 Code of Federal Regulations (CFR) p art 158. Both “ACCEPTABLE” and “SUPPLEMENTAL” studies may be used in the risk assessment process as appropriate. An “UNACCEPTABLE” rating indicates that new data need to be submitted. For the acute toxicity

data requirements, toxicity categories are assigned based on the hazard(s) identified from studies and/or other infor mation submitted to the Agency in support of a pesticide registration. The active ingredient or particular product is classified into Toxicity Category I , II, III, or IV, where Toxicity Category Iindicates 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 the se data requirements, including both generic and product
- specific information. All product anal ys is data requirements (except for storage stability ) for MetF52 for the labeled uses have now been satisfied
. Based on the existing usage of the pesticide for approximately

18 years without reports of adverse effects , and the registrant’s agreement to provide s torage stability data a s a term of these registration amendments, these pesticides are eligible for registration under FIFRA Section 

### 3(c)(5). An analy tical method (Guideline 885.1800 ) is not required for residues on food based on tolerance exemption. The anal yt ical 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
.Metarhizum anisopliae F52 Page 9 of 30

Biopestici des Registration Action Document

–Addendum April 18 , 2011 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 a cute pulmonary toxicity /patho genicity , acute dermal irritation and acute eye irritation studies resulted in Toxici ty Category I II 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 Sy stem 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 employ s a two - tiered approach to making the statutorily required determinations. Tier 1 consists of a battery of 11 screening assay s to identify the potential of a chemical substance to interact with the estrogen, androgen, or thyroid (E, A, or T) hormonal sy stems. Chemicals that go through Tier 1 screening and are found to have the potential to interact with E, A, or T hormonal sy stems 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 an y adverse endocrine re lated 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 pathway s such as food and water, residential activity , and certain post
- application agricultural scenarios. Thi s 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 pesti cide chemicals. Accordingl y, 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 guideline s and the Tier 1 screening battery , please visit our website: http://www.epa.gov/endo/
.Metarhizum anisopliae F52 Page 10 of 30

Biopestici des Registration Action Document

–Addendum April 18 , 2011 3. Food Quality Protection Act (FQPA) Considerations Section 408(c)(2)(A)(i) of the Federal Food, Drug, and Cosmetic Act allows the Agency to establish an exempt ion from the requirement for a tolerance (the legal limit for a pesticide chemical resi due in or on a food) onl y 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 cer tainty 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 re sidential 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 analy ses to determine the risks from aggregate exposure t o pesticide residues. First, the Agency determines the tox icity of pesticides. Second, the Ag ency 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 s ection 408(b)(2)(D) of FFDCA, the Agency has reviewed the available scientif ic data and other relevant information, and considered its validity , completeness, and reliability and the relationship of thi s information to human risk. The Agency has also considered available information concerning the variability of the sensitivities of major identifiable subgroups of consumers, including infants and 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 drinkin g 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 t o be negligible , as described below.Metarhizum anisopliae F52 Page 11 of 30

Biopestici des Registration Action Document

–Addendum April 18 , 2011
a. Food Pesticides containing Met F52 are intended to be applied at low application rates to row crops, transplants and dips, greenhouse foliar and root dips, or incorporation into soil
. T he spores, which adhere to , and infect insects, are not expected to grow on the crops. Though unlikel y, any spores remaining on food commodities at harvest would be removed b y post - harvest processes (washing, cooking, etc.). b. Drinking W

ater Exposure to Met F52 from drinking water is expected to be negligible because the microbial fungici de will not be applied to water
. Further, the spores are large and readi ly adhere to soil particles, and are unlikely to move through soils to reach underground water. Should any MetF52 spores re ach underground water, the y would be removed during municipal water treatment. 2. Other Non
- occupational Exposure Deuterom yc etous fungi , including Metarhizium anisopliae , occur in nature around the world. The pesticidal uses of products containing Met F52 are not likel y to increase non
- occupational human exposure signific antly above normal levels of these naturally occurring fungi
. Al so, if the proposed food uses are approved, pesticidal applications of Met F52 would occur in agricultural and residential areas . However, there have been no reports of pathogenicity or other adverse effects in humans and animals from e xposure to the registered products or t o the naturall y occurring strain of this MetF52 fungus. 3 . Cumulative Effects from Substances wi th 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 share s with other insecticidal substances. b . Determination of Safety for the U.S. Population, Infants, and Children FFDCA sectio n 408(b)(2)(C) provides that the Agency shall assess the available information about consumption patterns among infants and children, special susceptibility of infants and children to pesticide chemical residues, and the cumulative effects on infants and children of the residues and other substances with a common mechanism of toxicity . In addition, FFDCA sectio n 408(b)(2)(C) provides that the Agency shall apply an additi onal tenfold margin of safety for infants and children in the case of threshold effects to account for prenatal and postnatal toxicity and the complet eness of the database unless the Agency determines that a different margin of safety

will be safe for infants and children. Margins of exposure (safety), which are often referred to as uncertainty fa ctors, are incorporated into Agency risk assessments , either directly , or through the use of a margin of exposure anal ys is, or by using uncertainty (safety) facto rs in calculating a dose level that poses no appreciable risk.Metarhizum anisopliae F52 Page 12 of 30

Biopestici des Registration Action Document

–Addendum April 18 , 2011 MetF52 has a low mammalian acute toxicity potential , as discussed in Table 2 of the original BRAD , and Appendix A of this a ddendum . Registered pesticide products containing this active ingredi ent have been in use for almost 1 8 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 o f MetF52
. This includes all anticipated dietary exposures and all other exposures for which there is reliable information. T he Agency has arrived at this conclusion because the data and information available on MetF 52 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 o ccupational handler exp osure i s not expected to pose an y unreasonable risks to agricultural or green house workers. Appropriate personal protective equipment and precautionary

statements are required to be on product labels to mitigate the potential risks to pesticide handlers due to prolonged or repeated exposures. H andlers working with MetF52 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 (NI OSH ) st andards 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 unl ikel y 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 , s upport the Agency’s conclusions that no unreasonable adverse effect s would be expected from the proposed food uses
. C. Environmental Assessment ( 40 CFR § 1 58.2150 ) 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 evaluate d are available as “Supporting & Related Materials” within Docket Number EPA
- HQ
- OPP
- 2010
- 0081 at http://www.regulations.gov
. Acceptable Tier I nontarget toxicology data/information support the previous registrations, as well as the proposed uses of MetF 52
. ( Refer to Table 3 in the original BRAD issued with the 6/6/03 and 6/11/03 conditional registrations
. ) The s ubmitted studies demonstrated that Met F 52 poses no hazard to avian species, wild mammals and terrestrial and aquatic plant species from residential outdoor and ins titutional 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 theMetarhizum anisopliae F52 Page 13 of 30

Biopestici des Registration Action Document

–Addendum April 18 , 2011 o riginal conditional registration, the data demonstrated that f reshwater and estuarine/marine fish, invertebrates and aquatic insects will not be affected if Met F52 enters the aquatic environment from incidental drift and runoff from terrestrial uses. How ever, 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 w as 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 advers e effects on developing embryos . T herefore, 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 freshw ater 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 usuall y 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 characteriz ed 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), bi otic factors, and can be strain
- dependent (Zimmerman 2007, Vanninen et al. 2000). Persistenc e may differ between laboratory , greenhouse , and field conditions. M. anisopliae is not easily airborne, and is not an aquatic fungus, though it can persist in water under some conditions. L ittle 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. L eger 2002) . H owever, insect s are believed to be the primary source of organic matter for population increase, and may account for some persistence in the environment . T he fungus has been observed to be restricted to the insect 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 spray s (both ground and aerial applications) are likely to result in movement of Met F52 off the treatment site.Metarhizum anisopliae F52 Page 14 of 30

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–Addendum April 18 , 2011 However, based on the results of the acute non
- target toxicity testing, the need for environment al fate and groundwater data was not trigger ed. 3. Ecological Exposure and Risk Characterization T he submitted non - target organism effects information from publically available, scientific literature indicated that the only ecological effects of some c oncern 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 th at M. anisopliae adaptations result in strains that are more infectious for s ome insect species than others. T he submitted studies show ed that M. anisopliae Strain F52 does not pose a hazard to the major beneficial insect s tested, including honey bees, but the potential risks to all non - target insects cannot be ruled out. The 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 bene ficial insect species, including the honey bee, and terrestrial and aquatic plant species. The potential adverse effect s to fish can be mitigated by limiting direct application to aquatic sites. 4. Endangered Species Assessment T he BRAD for Met F52 states that an expansion of uses would trigger concerns for en dangered and 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 threatene d and endangered species and their habitat s . Novozy mes submitted an endangered species assessment (MRID 48175201) , which was reviewed b y BPPD and rated “S upplemental. ” BPPD conducted its own review , as discussed below (US EPA BPPD memorandum from Shannon Borges to Shanaz Bacchus, 02/17/11 ). The known host range of M. anisopliae includes over 200 species , and most belong to the Elateridae and Curculionidae families of Order Coleoptera, though infect ion of insects within several other orders is documented (Zimmerman 2007). The potential exposure level as a function of the proposed application methods of the pesticide was assessed, and BPPD ma de “no effect” determinations for direct effects, indirect effects, and affects to habitat (including designated c ritical habitat) for listed species for the following typ es 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 encl osed 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 threatenedMetarhizum anisopliae F52 Page 15 of 30

Biopestici des Registration Action Document

–Addendum April 18 , 2011 and e ndangered species and their designated critical habitats are also not expected. There fore, a “No Effect” determination is made for direct and indirect effects to listed species and their designated critical habitats resulting from the above proposed uses of Metarhizium anisopliae F52. BPPD concludes that the following application methods of Met F52 to the proposed food crops have the potential to cause direct effects, indirect effects, and effects to habita t (including designated critical habitat) to listed species :

 All foliage dips and outdoor foliar applications made b y 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 wa s 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 W ozniak to L eonard Cole, 12/11/2001 )
. However, these data do suggest that temperature a ffects growth, and consequently , persistence of Met F52 in the environment
. Moreover, MetF52 has a lower toxicity potential than the alternative insecticides (carbamate s and py rethrins ) for the propose d uses. Following application, the fungus returns to levels that cannot be distinguished from naturally occurring M. anisopliae
. Further analy ses are needed to characterize the effects that are likely to occur and the sp ecies that could potentially be affected. This assessment will be condu cted during registration review for Met F52. D. Product Performance Data (E fficacy) Submissi on of product performance data [OPPTS (now called OCSPP) Harmonized Guideline 810.30 00] 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 p ests, i.e., ticks, mosquitoes, fleas, Mediterranean fruit flies, g yps y m oths, Japanese b eetles , etc. For a list of organisms considered b y 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 473 13403; US EPA DERs dated October 2008 from Shannon BorgesMetarhizum anisopliae F52 Page 16 of 30

Biopestici des Registration Action Document

–Addendum April 18 , 2011 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 condition s of registratio n, and determined that these data s upport the control of the tested ticks , such as the lone star tick, black
- legged tick , and American dog tick , but without efficacy data, a claim cannot be made to “ticks” in general for these products. The pesticide prod

ucts that claim ticks are onl y 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 onl y for urban and suburban residential terrestrial outdoor uses (not adjoining parks, forested areas) to mitigate the exposure and potential risks to endangered insects
. 2. Other Target P ests The insect pathogenicity

data submitted for the Environmental Risk Assessment of MetF52 Biological Insecticide, TICK
- EX TM , (formerl y EPA R eg. No. 72098
- T ) performed in 2003 showed no adverse effects on nontarget insects. Submitted nontarget toxicity studies show ed that MetF52 poses no hazard to lady beetles, green lacewings, parasitic wasps, honey bee larvae, honey bee adults and earthworms
. The product performance data submitted by Novoz ym es ( cited above ) demonstrated that Met F52 provide s statistically significant control of mites, thrips , b lack v ine w eevils and white flies on agricultural crops when applied according to the label directi ons (MRIDs 48150701, 4815070 2 , 4815070 3 , 4815070 4: US EPA BPPD memo and DERs dated 02/15/11 from Zi gfridas Vaituzis to Shanaz Bacchus). V. RISK MANAGEMENT DECI SION Pesticides containing Metarhizium anisopliae F52 as the active ingredient were conditionall y registered under FIFRA S e ction 3(c)7(C) on June 6 and 11, 2003. The current registrant, Novozy mes Biologicals, Inc., has provided data/information to support an unconditional registration under Section 3(c)(5) of FIFRA active ingredients if it is determ ined 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 effe cts on the environment; and (D) when used in accordance with widespread and commonly recognized practice it will not generall y cause unreasonable adverse effects on the environment. The four criteria of the Eligibility Determination for Pesticidal Active In gredients are satisfied b y 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 l abel instructions. Therefore, Metarhizium anisopliae F52 is eligible for FIFRA Section 3(c)(5) registration s for the labeled uses and for the proposed new food uses
. Other considerations may appl y to the labeled sites during registration review of the Metarhizium case.Metarhizum anisopliae F52 Page 17 of 30

Biopestici des Registration Action Document

–Addendum April 18 , 2011 The 2003 BRAD cited rationales in the public interest for the conditional registration of the pesticidal active ingredient. A public interest finding is not required for this Section 3(c)(5) registration of Metarhizium anisopliae F52
. VI . ENVIRONMENTAL JUSTIC E EPA seeks to achieve environmental justice
- the fair treatment and meaningful involvement of all people, regardless of race, color, national origin, or income
- in the development, implementation, and enforcement of environment al 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
. VI I . TERMS AND CONDITIONS OF THE REGISTRATION(S) As a term of registration, the registrant must su bmit one
- ye ar 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 e ight years without any reports of adverse effects and the acceptable data evaluated to satisfy the conditions of registration
.Metarhizum anisopliae F52 Page 18 of 30

Biopestici des Registration Action Document

–Addendum April 18 , 2011 VI I I . APPENDIX A. DATA REQUIREMENTS (40 CFR Part 158 - Subpart V) On October 26, 2007, the Agency issued a Final Rule in the F ederal 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 publ ished 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 info rmation reviewed for the original registration of th e product s containing this active ingredient are summarized in the original BRAD (06/06/03). These tables are updated below to reflect recently reviewed submissions
. *NOTE: MRID numbers listed in the following tables are representative of additional dat a evaluated to support the request to amend the registration to an unconditional registration under FIFRA Section 3(c)(5) and for the proposed food uses in PP 8F7508
. Subsequent to this registration amendment , there may be additional MRIDs that support reg istration of other products containing this active ingredient. TABLE 1. Product Chemistry Data Requirements Additional Data Reviewed for Metarhizium anisopliae F52 (40 CFR § 158. 2 120 ) Guideline No. Study Results MRID & references 885.1100 Produc t

identity The data reviewed for the initial conditional

registration s (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. anisopli ae . 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 12/30/1 0 ) were con sidered 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 condition s 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/ 10Metarhizum anisopliae F52 Page 19 of 30

Biopestici des Registration Action Document

–Addendum April 18 , 2011 TABLE 1. Product Chemistry Data Requirements Additional Data Reviewed for Metarhizium anisopliae F52 (40 CFR § 158. 2 120 ) 885.1300 F ormation of

Unintentional In gredients 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 DER12/30/1 0 885.1400 Analysis of

Samples Approximately 48% of the extractable spores on the

solid matrix were recovered. The range of recovery was 40 to 59% and the viability of the recovered spores was highly consistent between batches. ACCEPTABLE. However, these data are not required for 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 In gredient 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 ** * Ta enure Granular BioinsecticideMetarhizum anisopliae F52 Page 20 of 30

Biopestici des Registration Action Document

–Addendum April 18 , 2011 The following results were excerpted from an electronic file dated 12/11/01 ( DER from Chris Wozniak to L eonard Cole, USEPA/OPP/BPPD). These previousl y 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) suspensio n 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 de nsity: 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 8 30.7840 Water Solubility Not required N/A 830.7950 Vapor Pressure Not required N/AMetarhizum anisopliae F52 Page 21 of 30

Biopestici des Registration Action Document

–Addendum April 18 , 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 M etarhizium anisopliae F52 , previousl y reported as Table 2 of the 2003 BRAD , are unchanged . Those assessments continue to support the non
- food u ses, and support the proposed food uses of Met F52 pesticides (US EPA BPPD D ata E valuation R ecord , e lectronic fi le dated 12/11/01 from Chris Wozniak to L eonard Cole ) . They are summarized in the supporting materials in this docket. Acute Toxicity

– Ecological Effects T he acute toxicity ecological effects exposure assessments for non
- target organisms continue to s upport 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, honey bee 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 Meta rhizium anisopliae F52 on a quatic vertebrates (fish embryos ) will be required if amendments are requested for direct application to water. Acceptable Tier I nontarget toxicology data/information support the previously labeled and , now, the currently

prop osed 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 l arvae, honey bee adults and earthworms). Freshwater and estuarine/marine fish, invertebrates and aquatic insects will not be affected b y the quantities entering the aquatic environment from incidental drift and runoff from terrestrial uses. However, in li ght 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 F ish Toxicity/Pathogenicity As a condition of registration, the Agency required that the freshwater fish study be repeated. The 30
- day

L C 50 for rainbow trout exposed to M. anisopliae was >53 mg/L, the highest concentration tested. All fish appeared normal and there were no mortalities in all treatment groups. The study

was rated SUPPLEMENTAL since a maximum hazard dose was not obtained and laboratory

induced pathological changes in fish (silverside) embryos and fry has been reported in the literature. Novo zy mes’ response to this condition was a rebuttal (dated December 09, 2004) and a resubmission (dated January 14, 2008). The Agency

accepted Novoz ym es’ rationale for notMetarhizum anisopliae F52 Page 22 of 30

Biopestici des Registration Action Document

–Addendum April 18 , 2011 repeating the study based on the use patterns, as discussed immediately below (USEPA/B PPD 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 anal ys is 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 Pathogeni city

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 stu dies on the effects of Metarhizium anisopliae F52 TGAI (currently

called TAE
- 001 Technical BioInsecticide ) on developing freshwater fish embryos.Metarhizum anisopliae F52 Page 23 of 30

Biopestici des Registration Action Document

–Addendum April 18 , 2011 IX . APPENDIX B. MRID S AND CIT ED LITERATURE BIBLIO GRAPHY The following studies and references were relied upon as described in the original BRAD , and some are referenced in this document. MRID Citation Reference 44844700 Taensa, Inc. (1999) Submission of Product Chemistry , Toxicity , and

Environmental Fate Data in Support of the Application for Registrati on 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, In c. 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 b y Taensa, Inc. 4 p. 44844705 Gingras, B. (1999) Lot Analy sis of Metarhizium anisopliae Strain F52: L ab

Project Number: L08743SN24. Unpublished study prepared by

IIT Research Institute. 23 p. {OPPTS 885.1400} 448447 06 Butts, E. (1999) Analy sis of Samples
-- Manufacturing Process 2 Metarhizium

anisopliae Strain F52: TAE
- 001 Technical Bioinsecticide. Unpublished study prepared b y Taensa, Inc. 9 p. 44844707 Butts, E. (1999) Certification of Ingredient Limits: Metarhiziu m 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, Phy sical State, Odor,

pH and Density of Metarhizium anisopliae TGAI : L ab 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 Acut e 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 24 of 30

Biopestici des Registration Action Document

–Addendum April 18 , 2011 44844710 Findlay , J. (1999) Acute Dermal Toxicity /Pathology

Study of Metarhizium anisopliae , Strain F52 , in Rabbits: Lab Project Number: L 08743SN16. 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 Intraperito neal 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: L ab Project Numb er: L08743SN17. Unpublished study prepared b y IIT Research Institute. 26 p. 44844714 Butts, E. (1999) Hy persensitivity

Incidents Report Metarhizium anisopliae, Strain F52. Unpublished study prepared by

Taensa, Inc. 4 p. 44844715 Findlay , J. (1999) Derm al 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. (199 9) Measurement of Fungal Toxins Destruxin and

Cy tochalasin 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 anis opliae Strain F52 at

Various Temperatures: Lab Project Number: L08743 SN26. Unpublished study prepared b y IIT Research Institute. 18 p. 44844718 Gingras, B. (1999) Sensitivity of Detection of Metarhizium anisopliae Strain F52 for Toxicity

/Pathogenicity

T esting 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 Bobwh ite: 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 L ife
- Cy cle Toxicity

and Pathogenicity Test with the Cl adoceran (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 25 of 30

Biopestici des Registration Action Document

–Addendum April 18 , 2011 44844721 Drottar, K.; Krueger, H. (1999) A Five
- Concentration Toxicity and Pathogen icity Test with the Rainbow Trout (Oncorhy nchus my kiss): Metarhizium anisopliae Strain F52: Final Report: L ab Project Number: 495A
105. Unpublished study

prepared by

Wildlife International Ltd. 30 p. {OPPTS 885.4200} 44844722 Palmer, S.; Krueger, H.; Hox ter, K. (1999) A Dietary Pathogenicity and Toxicity Study with Green L acewing Larvae (Chry soperla 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 Lady

bird Beetle (Hippodamia convergens): Metarhizium anisopliae Strain F52: Final Report: Lab Project Number: 495
- 105. Unpublished study prepare d 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 Hy

menoptera (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, In c. 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 melli fera 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: Metarhizi um 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): Me tarhizium anisopliae Strain F52: Final Report: L ab Project Number: 495A
- 106. Unpublished study prepared by Wildlife International Ltd. 37 p. {OPPTS 850.5400} 44854700 Taensa, Inc. (1999) Submission of Product Chemistry Data in Support of the Application for Registration of TAE
- 001 Granular Bioinsecticide. Transmittal of 3 Studies. 44854701 Butts, E. (1999) Product Identification and Di s closure of Ingredients

Metarhizium anisopliae Strain F52 TAE
- 001 Granular Bioinsecticide. Unpublished study

prepared by Taensa, Inc. 4 p.Metarhizum anisopliae F52 Page 26 of 30

Biopestici des Registration Action Document

–Addendum April 18 , 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) Analy sis 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 o f 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 b y Taensa, Inc. 4 p. 45696303 Jenki ns, 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: L ot Anal ys is of Metarhizium anisopliae Strain F52: Anal ys is 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 Analy sis of Samples: L ab 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 Dat a 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 Taens a, 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. Unpub lished study

prepared b y Earth Biosciences Inc. 46 p.Metarhizum anisopliae F52 Page 27 of 30

Biopestici des Registration Action Document

–Addendum April 18 , 2011 45778802 Lezotte, F.; Van Hoven, R.; Nixon, W. (2002) Determination of the Color,

Phy sical State, Odor, pH, and Density (Specific Gravity) of TAE
- 001 Oil Experimental Bioinsecticide: L ab Project Numbe r: 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 Rat s: L ab 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: L ab Project Number: 02
- 177: RPSN02
- 177. Unpublished study prepared b y 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 Novozy

mes Biologicals, Inc. (2007) Submission of Product Chemistry Data in Supp ort 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 a nisopliae Var.

Anisopliae in Southwestern British Columbia is Indepen dent of Habitat Association. Project Number: NOVOZYMES/0701. Unpublished Study Prepared b y Agriculture Canada. 36 p. 47313400 Novozy

mes Biologicals, Inc. (2007) Submission of Efficacy Data in Support of the Registrations of TAE
- 001 Technical Bioinsecticid e, 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: M

ETARHIZIUM

/2007/001. Unpublished study prepared by Novozy mes 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. Unpublish ed study prepared b y Novozy

mes Biologicals, Inc. 11 p. 47313403 Bharadwaj, A.; Stafford, K.; L eland, J. (2007) Evaluation of Metarhizium

anisopliae Strain F52 for Tick Control. Project Number: METARHIZIUM

/2007/003. Unpublished study prepared b y Novozy

mes Biologicals, Inc. 17 p.Metarhizum anisopliae F52 Page 28 of 30

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–Addendum April 18 , 2011 47880600 Novozy

mes 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 Studi es. 47880601 Schreier, T.; Kauffman, J. (2009) Metarhizium anisopliae Strain F52: Three

Batch Fungal Contamination Analy sis of Fermentation Process and Five Batch Analy sis of TGAI. Project Number: METARHIZIUM9/24/2009. Unpublished study prepared by

Novozy mes Biologicals, Inc. 30 p. 47880602 Schreier, T. (2009) Metarhizium anisopliae Strain F52: Five Batch Anal ys is of

Tick
- Ex EC. Project Number: METARHIZIUM9/29/2009. Unpublished study prepared b y 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

Novozy mes 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

Novozy mes Biologicals, Inc. 8 p. 48150700 Novozy

mes Biologicals, Inc. (2010) Submission of Efficacy Data in Support of the Registrations of Tae
- 001 Technical Bioinsecticide, Taenure Granular Bioinsecticide, Tick
- EX G and Ti ck
- 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 Novozy mes 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 Novozy mes 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 Novozy mes 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 Novoz ym es Biological s, Inc. 17 p. 48175200 Novozy

mes 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 M etarhizium anisopliae Strain F52. Transmittal of 1 Study
.Metarhizum anisopliae F52 Page 29 of 30

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–Addendum April 18 , 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. Unpublish ed study prepared b y Exponent, Inc. 45 p. 48177000 Novozy

mes 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 b y Novozy

mes Biologicals, Inc. 20 p. References Hu, G. and R.J. St. L eger. 2002. Field studies using a recombinant mycoinsecticide ( Metarhizium anisopliae ) reveal that it is rhi zosphere competent. Appl. Env. Microbiol., 68:6383
- 6387. Meyling, N.V. and J. Eilenberg. 2007. Ecology of the entomopathogenic fungi Beauveria bassiana and Metarhizium anisopliae in temperate agroecos ys tems: potential for conservation biological control. Biol. Control, 43:145
- 155. Vanninen, I., J. Ty

ni
- 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 30 of 30

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–Addendum April 18 , 2011 X . GLO SSARY 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 D ER 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 Qual ity 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 ; formerl y, Office of P revention,

P esticide s and Toxic Substances (OPPTS) OPP Office of Pesticide Programs OPPTS Office of P revention, P esticide s and Toxic Substances (old name for OCSPP ) TGAI Technical Grade of the Active Ingredient