                  BIOPESTICIDES REGISTRATION ACTION DOCUMENT

                                PC Code 115002

           Isaria fumosorosea (formerly Paecilomyces fumosoroseus)
                               Apopka Strain 97

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

                                August 10, 2011

                               TABLE OF CONTENTS
I.  	BACKGROUND	4
II.  	ACTIVE INGREDIENT OVERVIEW	4
III.	REGULATORY BACKGROUND	5
	A. Applications for Pesticide Registration	5
	B. Food Tolerance Exemptions	6
IV.	RISK ASSESSMENT	6
	A. Product Analysis Assessment (40 CFR § 158.2120)	7
	B.	Human Health Assessment (40 CFR § 158.2140)	7
	C. 	Environmental Assessment (40 CFR § 158.2150)	10
V.  	ENVIRONMENTAL JUSTICE	11
VI.  	RISK MANAGEMENT DECISION	12
VII. 	TERMS AND CONDITIONS OF THE REGISTRATION(S)	12
VIII.	GLOSSARY OF ACRONYMS AND ABBREVIATIONS	13
IX.  	BIBLIOGRAPHY	14
	A. Studies Submitted to Support the PFR-97 Pesticide Product	14
	B. Environmental Protection Agency Risk Assessment Memoranda	16
	C. Other References	16
Appendix A. Microbial Pesticides Data Requirements	18
                BIOPESTICIDES REGISTRATION ACTION DOCUMENT TEAM

                      Office of Pesticide Programs (OPP)
                Biopesticides and Pollution Prevention Division
                          Microbial Pesticides Branch

                                Science Reviews

Ibrahim Barsoum, Ph.D.				   Product Analysis, Human Health
John Kough, Ph.D.					   Product Analysis, Human Health
Shannon Borges					   Environmental Effects
Zigfridas Vaituzis, Ph.D.				   Environmental Effects
Gail Tomimatsu					   Environmental Effects

                                  Regulations

Sheryl Reilly, Ph.D.				               Chief, Microbial Pesticides Branch
Shanaz Bacchus	Regulatory Action Leader

I. BACKGROUND

The fungal active ingredient (ai), Paecilomyces fumosoroseus Apopka strain 97, (PC Code 115002) has been reported to be originally isolated in 1986 from a mealy bug in a greenhouse in Apopka, FL. It was recently reclassified as Isaria fumosorosea (ARSEF 2011; Zimmermann, 2008). In this and other Agency documents, it is referred to as Isaria fumosorosea (formerly Paecilomyces fumosoroseus) Apopka strain 97, or as Paecilomyces fumosoroseus Apopka strain 97 or as PFR-97. The pure culture was identified in 1988 and deposited at the American Type Culture Collection (ATCC # 20874). This fungal strain is a pathogen of many orders of insects. It occurs naturally in populations of infected and dead insects and randomly in soils around the world. It can be grown readily on various culture media and exists in several morphological stages, depending upon the culture medium. On solid medium, conidiophores, conidia, and mycelia are predominant, while submerged culture produces blastospores and mycelia.

PFR-97[TM] MUP is the manufacturing use product (MUP), registered under the Federal Insecticide, Fungicide and Rodenticide Act (FIFRA). It is formulated into the end-use product (EP), PFR-97[TM] 20% WDG that contains blastospores, mycelia and conidia. Under proper environmental conditions, spores of the fungus attach to and penetrate the cuticle of the target pest. The fungus grows inside the insect causing its death. The fungus then emerges from the dead insect to release more spores to infect other insects.

The product identity data submitted by the registrant were sufficient in 1998 to allow the proposed limited use patterns in greenhouses. The registrant submitted an amendment in 2009 to expand the uses to include food crops.

On October 1, 2009, the Environmental Protection Agency (EPA or the Agency) announced a new policy to provide a more meaningful opportunity for the public to participate in major registration decisions before they occur. According to this new policy, the Agency intends to provide a public comment period prior to making a registration decision for, at minimum, the following types of applications: new active ingredients; first food uses; first outdoor uses; first residential uses; and any other registration decisions for which the Agency believes there may be substantial public interest.

Consistent with the new policy of making registration actions more transparent, Paecilomyces fumosoroseus Apopka strain 97 is now subject to a 30-day comment period as a previously registered active ingredient whose registration amendment would result in a "first outdoor use," a "first food use," and possibly a "first residential use." The public comment period will conclude 30 days after the posting of these documents in the docket EPA-HQ-OPP-2010-0088. The risk assessments for Paecilomyces fumosoroseus Apopka strain 97 are characterized throughout this Biopesticides Registration Action Document (BRAD).

II. ACTIVE INGREDIENT OVERVIEW

    Biological Name:  	Isaria fumosorosea (formerly Paecilomyces fumosoroseus) Apopka strain 97

    Culture Deposit:		American Type Culture Collection in Manassas,
    		Virginia under Accession Number 20874

 	OPP Chemical Code: 		115002

	Type of Pesticide:				Microbial Pesticide/Biofungicide

III.	  REGULATORY BACKGROUND

A. Applications for Pesticide Registration

 On July 6, 1994, the Agency received an application from W.R.Grace to register PFR- 97[TM] MUP, a microbial insecticide, containing Paecilomyces fumosoroseus Apopka strain 97 as the active ingredient for manufacturing and end-use pesticide products. The registrant also requested the registration of two end-use products (EPs), PFR-97[TM] 20% WDG and PFR-97[TM] WDG, containing 20 percent (%) and
 12 % Paecilomyces fumosoroseus Apopka strain 97, respectively. The proposed pesticidal use was for control of whiteflies, aphids, thrips and spider mites on ornamentals in green houses, as well as food crops outdoors. A notice of receipt of the application for registration under FIFRA of Paecilomyces fumosoroseus Apopka strain 97 as a new active ingredient was published in the Federal Register (FR) on November 2, 1994 (59 FR 54093).

A pesticide petition for an exemption from tolerance (PP4F4372) was filed concurrently with the proposed Section 3 registration applications. The Notice of this Filing was published in the Federal Register simultaneously with the Notice above [59 FR 54092, Number 211 (Wednesday, November 2, 1994)]. No comments were received in response to either of the FR Notices within the 30-day comment period. On June 13, 1996, the pending products were transferred to Thermo Trilogy, which did not repropose the exemption from tolerance petition as required by the Food Quality Protection Act (FQPA) of 1996.  At that time, the Agency did not take any further action either on the request for the tolerance exemption or the pending food use registration application for PFR-97[TM] WDG. On April 22, 1998, the following PFR-97 products were registered to Thermo Trilogy for non-food uses:  (1) PFR-97[TM] MUP (EPA Reg. No. 70051-17), a manufacturing use product; and (2) PFR-97[TM] 20% WDG (EPA Reg. No. 70051-19), an end-use product.

The Agency published the notices of registration for these products in the FR on July 8, 1998 (63 FR 36899). Both products contained Paecilomyces fumosoroseus Apopka strain 97 as the active ingredient, and were intended for use on ornamentals in greenhouses and in interiorscapes. The company was informed that additional data would be required for more extensive use patterns.  On September 30, 1998, Thermo Trilogy formally withdrew the registration application for PFR-97[TM] WDG (12% ai), indicated that it would repropose the petition for food use as required by the FQPA and requested six months for providing non-target organism data required for additional uses.  The company did not follow through on these actions.

In 2004, the product registrations were amended to remove the interiorscape use from the labels. Thermo Trilogy has since changed its name to Certis USA, LLC (hereinafter referred to as "Certis"), currently located on 9145 Guilford Road, Suite 175, Columbia, MD 21046.

On December 17, 2009, Certis applied to amend these pesticide registrations to include new uses on all food commodities. The Agency announced receipt of the application in the FR on March 5, 2010 (EPA HQ-OPP-2010-0088; 75FR10259), and opened a 30-day public comment period pursuant to the provisions of FIFRA section 3(c)(4). No comments were received in response to this publication. This docket has been reopened to provide the public 30 days to review and submit comments on the posted documents that informed the Agency's proposed decision regarding Certis'proposed amendment to allow food uses to the product registrations.

B. Food Tolerance Exemptions

A numeric tolerance or exemption from the requirement of a tolerance was not needed when the first products were registered in 1998, since they were not intended for use on food or animal feed commodities.  Nevertheless, safety factors from the Food Quality Protection Act of 1996 (FQPA) were considered for potential secondary transfer of residues to drinking water during applications.

Included with the December 17, 2009, application to amend the product registrations was a petition to establish an exemption from the requirement of a tolerance on all food commodities for Paecilomyces fumosoroseus Apopka strain 97 (Pesticide Petition [PP] 9F7665) under the Federal Food, Drug, and Cosmetic Act (FFDCA) section 408(d). The Notice of Filing of the petition was published in the Federal Register of March 10, 2010 (EPA HQ-OPP-2010-0087, 75 FR11171), and opened a 30-day comment period. No comments were received in response to the FR publication.

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 this BRAD were considered in light of these requirements.

IV.	  RISK ASSESSMENT

The Agency assessed the data submitted by Certis in 2009 for the proposed pesticide registration amendments. The classifications assigned by Agency science reviewers to data that are submitted to support registration of a pesticide 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 must 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.

Refer to Tables 1, 2a, 2b, 3 and 4 in Appendix A for a brief summary of the data requirements, including both generic and product-specific information. Additionally, the comprehensive Agency risk assessments that evaluate these data are available either in the Appendix of this BRAD as summaries or as "Supporting & Related Materials" within Docket Number EPA-HQ-OPP-2010-0088 at http://www.regulations.gov

A. Product Analysis Assessment (40 CFR § 158.2120)

All product analysis data requirements for the proposed food uses of Isaria fumosorosea (formerly Paecilomyces fumosoroseus) Apopka strain 97 have been satisfied (Tables 1, 2a, 2b  -  Appendix A). A storage stability study shows that the pesticide product has a shelf life of 1 year at 4o C, but is less stable at 25o C. The sample stored at 4o C for 1 year was still effective at colonizing host insects, while that stored at 25o C was not. The product label must bear appropriate storage statements and a corresponding expiration date.

B.	Human Health Assessment (40 CFR § 158.2140)

1. Toxicity

All toxicology data requirements for Isaria fumosorosea (formerly Paecilomyces fumosoroseus) Apopka strain 97 have been satisfied. Acceptable Tier I mammalian toxicology data/information support the registration of this fungal active pesticide ingredient. Furthermore, Tier II and Tier III studies were not required for PFR-97 based on the lack of acute toxicity/pathogenicity in the Tier I studies. Refer to Table 3 in Appendix A for a brief summary of these data requirements, including both generic and product-specific information. Additionally, the summaries of the Agency risk assessments that evaluate these data are available as "Supporting & Related Materials" within Docket Number EPA-HQ-OPP-2010-0088 at http://www.regulations.gov.

2. Endocrine Disruptors

As required under FFDCA section 408(p), the Agency 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 the Agency 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 quantitative relationship between the dose and the E, A, or T effect.

Between October 2009 and February 2010, the Agency 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.

PFR-97 is not among the group of 58 pesticide active ingredients on the initial list to be screened under the EDSP. Under FFDCA section 408(p), the Agency must screen all pesticide chemicals. Accordingly, the Agency anticipates issuing future EDSP 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 3 in Appendix A, the data required for a FQPA risk assessment for Isaria fumosorosea (formerly Paecilomyces fumosoroseus) Apopka strain 97 have been satisfied. (See also, "Supporting & Related Materials" within Docket Number EPA-HQ-OPP-2010-0088 at http://www.regulations.gov.)

	a. Aggregate Exposure

In examining aggregate exposure, section 408 of FFDCA directs the Agency to consider
available information concerning exposures from the pesticide residue in food and all other nonoccupational exposures, including drinking water from ground water or surface water and exposure through pesticide use in gardens, lawns, or buildings (residential and other indoor uses).

Dietary Exposure:

Dietary exposure to the microbial pesticide is unlikely to occur, but if humans were exposed to Isaria fumosorosea (formerly Paecilomyces fumosoroseus) Apopka strain 97 through the diet, no adverse effects would be expected, based upon the lack of acute oral toxicity, infectivity, and/or pathogenicity in laboratory animals (results presented in Unit III.B. See additional discussion in Appendix A of this BRAD, and "Supporting & Related Materials" within Docket Number EPA-HQ-OPP-2010-0088 at http://www.regulations.gov). The following information is provided to demonstrate why residues of
Isaria fumosorosea (formerly Paecilomyces fumosoroseus) Apopka strain 97 are not expected in food or feed commodities, or drinking water.

	1. Food. Dietary exposure to the microbial active ingredient is expected to be minimal to non-existent. Isaria fumosorosea (formerly Paecilomyces fumosoroseus) Apopka strain 97 is intended for directed ground and spray application, or soil application, to food and feed commodities. The active ingredient rapidly degrades at room temperature, and as an insect pathogen, residues are not likely to accumulate on crops.

      2. Drinking Water: Drinking water exposure is expected to be negligible to non-existent, because the microbial fungicide will not be applied to water. Further, the insect pathogen Isaria fumosorosea (formerly Paecilomyces fumosoroseus) Apopka strain 97 would not proliferate in aquatic environments. The large fungal spores within the soil will not likely percolate into water due to their size and the fact that they readily adhere to soil particles.

      3. Other Nonoccupational Exposure:  PFR-97 is a naturally occurring microorganism and is ubiquitous in the environment. As a pesticidal active ingredient, PFR-97 will be applied to agricultural crops in the field and to crops grown in greenhouses and nurseries that are not expected to be in close proximity to residential areas. If applications take place in residential areas, there is no evidence for inhalation or dermal toxicity at exposure levels even several orders of magnitude higher than expected for a typical residential end-product user (see Appendix A of the BRAD and "Supporting & Related Materials" within Docket Number EPA-HQ-OPP-2010-0088 at http://www.regulations.gov). Additionally, there have been no reports of adverse effects to humans from inhalation or dermal exposure to this widespread fungus, which has been in use in greenhouses as a pesticide for the past thirteen years.

	b. 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 PFR-97, an insect pathogen, is practically non-toxic to mammals and no cumulative effects are anticipated.
	c. 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.
Based on the acute toxicity and pathogenicity data discussed in Table 3 in Appendix A of the BRAD and "Supporting & Related Materials" within Docket Number EPA-HQ-OPP-2010-0088 at http://www.regulations.gov , 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 PFR-97. 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 PFR-97 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.

3. Occupational Exposure and Risk Characterization

In light of the Tier I acute toxicity/pathogenicity studies, which did not show any toxicity or pathogenicity via the oral, pulmonary, or intraperitoneal injection routes of exposure, handler exposure to PFR-97 is not expected to pose any undue risk. Regardless, appropriate personal protective equipment and precautionary statements are required to be on the product label to mitigate any potential risks to pesticide handlers due to prolonged exposure. Handlers working with PFR-97 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.

C. Environmental Assessment (40 CFR § 158.2150)

1. Nontarget Organism Toxicology

All nontarget toxicology data requirements for the proposed uses of PFR-97 have been satisfied. Acceptable Tier I nontarget toxicology data/information support the registration of PFR-97. Refer to Table 4 in Appendix A for a brief summary of these data requirements. Additionally, the summaries of the Agency risk assessments that evaluate Table 4 in Appendix A of this BRAD are available as "Supporting & Related Materials" within Docket Number EPA-HQ-OPP-2010-0088 at http://www.regulations.gov.

2. Environmental Fate and Ground Water Data

As the information provided is sufficient to both satisfy the Tier I nontarget organism data requirements and for nontarget organism risk assessment for PFR-97, further testing at higher tier levels (i.e., Tiers II, III, and IV) is not required.

3. Ecological Exposure and Risk Characterization

The Agency has performed an environmental risk assessment based on the literature citations and studies provided by the registrant. PFR-97 is a naturally occurring strain of Isaria fumosorosea (formerly Paecilomyces fumosoroseus). Isaria fumosorosea has been isolated from many species of insects, particularly Lepidoptera, and has been collected from air, water, plants, other fungi, and often from soil. It is one of the most common entomopathogens known to affect nymphs and adults of Bemisia species of whiteflies, and has caused epizootics in this species and in the United States and other countries (Zimmerman, 2008). Data on natural concentrations of Paecilomyces fumosoroseus and persistence of this strain in the environment are not available.  Exposure on the field is expected to occur at levels that would affect sensitive insects.  Some offsite movement may occur with spray drift, but exposure is expected to occur at lower levels outside of treated areas (US EPA memorandum dated July 21, 2011, from Shannon Borges to Shanaz Bacchus)

The applicant has submitted data and rationales to waive testing to satisfy data requirements for nontarget organism risk assessment with the technical grade of the active ingredient (TGAI). Submitted data, data waiver rationales, and a search by the Agency of several literature databases provided sufficient information to determine that toxicity/pathogenicity to wild mammals, avian wildlife, freshwater and marine/estuarine vertebrates and invertebrates, and terrestrial and aquatic plants is not expected from the proposed label uses. There is uncertainty about the potential for adverse effects to certain non target insects and arachnids. While the honey bee study showed no toxicity to these beneficial insects, it was not of long enough duration to determine pathogenicity to them. Thus, the pesticide must not be applied while honey bees are actively foraging. Further testing of nontarget organisms at higher tier levels is not required at this time, but may be required in the future (US EPA Borges, July 21, 2011, Appendix A and "Supporting & Related Materials" within Docket Number EPA-HQ-OPP-2010-0088 at http://www.regulations.gov).

4. Threatened and Endangered Species Assessment

A complete Endangered Species (ES) has not been completed for this registration amendment and will be conducted during the registration review process. As indicated in the Environmental Risk Assessment (US EPA, Borges, July 21, 2011), included in this docket, EPA has determined that adverse effects to nontarget insects as a result of the proposed new uses of Paecilomyces fumosoroseus Apopka strain 97 are possible. EPA has determined that risk of adverse effects is minimal to birds, wild mammals, freshwater fish and invertebrates, estuarine/marine fish and invertebrates, and nontarget terrestrial and aquatic plants. Therefore, EPA makes conclusions as follows regarding federally threatened and endangered (listed) species for the proposed new uses of Paecilomyces fumosoroseus Apopka 97.

   * EPA makes "No Effect" determinations for direct effects to listed terrestrial vertebrates and plants and listed aquatic animals and plants for the proposed new uses.
   * EPA concludes that the proposed new uses of Paecilomyces fumosoroseus Apopka 97 have the potential to cause direct effects to listed terrestrial insects and arachnids, as well as indirect effects to listed species dependent on insects and arachnids (e.g., for food, pollination, habitat maintenance). Further analyses are needed to characterize the effects that may occur and the species potentially affected. Tier II environmental expression studies would allow for a more refined analysis and would likely reduce the number of species determined to be affected.

V. ENVIRONMENTAL JUSTICE

The Agency seeks to achieve environmental justice -- the fair treatment and meaningful involvement of all people regardless of race, color, national origin, or income -- with respect to the development, implementation, and enforcement of environmental laws, regulations, and policies. To help address potential environmental justice issues, the Agency seeks information on any groups or segments of the population who, as a result their location, cultural practices, or other factors, may have atypical, unusually high exposure to PFR-97, compared to the general population. Please comment if you are aware of any subpopulations that may have atypical, unusually high exposure compared to the general population.

For additional information regarding environmental justice issues, please visit the Agency's web site at http://www.epa.gov/compliance/environmentaljustice/index.html.

VI. RISK MANAGEMENT DECISION

Section 3(c)(5) of FIFRA provides for the registration of a new active ingredient 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 the product containing Isaria fumosorosea (formerly Paecilomyces fumosoroseus) Apopka 97. This product is not expected to cause unreasonable adverse effects and is likely to provide protection as claimed when used according to label instructions. The Agency believes, therefore, that Isaria fumosorosea (formerly Paecilomyces fumosoroseus) Apopka 97 is eligible for registration for the proposed uses.

VII. TERMS AND CONDITIONS OF THE REGISTRATION(S)

As a term of registration, the registrant must submit analyses of unintentional ingredients in five production batches (40 CFR § 158.2120).

VIII. GLOSSARY OF ACRONYMS AND ABBREVIATIONS

BRAD	Biopesticides Registration Action Document
CFR	Code of Federal Regulations
cfu	colony-forming units
cfu/g	colony-forming units per gram
°C 	degrees Celsius
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
g/mL	grams per milliliter
IR-4	Interregional Research Project Number 4
LLC 	limited liability company
MRID No.	Master Record Identification Number
NIOSH 	National Institute for Occupational Safety and Health
OCSPP	Office of Chemical Safety and Pollution Prevention
OPP	Office of Pesticide Programs
PP	Pesticide Petition
TGAI 	technical grade of the active ingredient

IX. BIBLIOGRAPHY

A. Studies Submitted to Support the Paecilomyces fumosoroseus Apopka strain 97 Pesticide Product
	Registration
                                 Bibliography

                                     MRID
                              Citation Reference

43139800
W.R. Grace & Co. (1994) Submittal of Product Chemistry, Toxicity/ Pathogenicity, and Environmental Fate Data in Support of Registration of PFR-MUP. Transmittal of 4 studies.
43139801
Eyal, J. (1994) PFR-MUP Product Chemistry: Lab Project Number: 4775-MUP-CHEM. Unpublished study prepared by W.R. Grace & Co.--Conn. 128 p.
43139802
Jones, M. (1993) Acute Pulmonary Toxicity/Pathogenicity Study of Paecilomyces fumosoroseus var. Apopka in Rats: Final Report: Lab Project Number: G-7444.228.001. Unpublished study prepared by Microbiological Associates, Inc. 83 p.
43139803
Jones, M. (1993) Acute Intraperitoneal Toxicity/Pathogenicity Study of Paecilomyces fumosoroseus var. Apopka in Rats: Final Report: Lab Project Number: G-7444.226.001. Unpublished study prepared by Microbiological Associates, Inc. 65 p.
43139804
Eyal, J. (1994) PFT-MUP Environmental Fate Data: Lab Project Number: 4775-MUP-ENVFATE. Unpublished study prepared by W.R. Grace & Co.--Comm. 19 p.
43139900
W.R. Grace & Co. (1994) Submittal of Product Chemistry Data in Support of Registration of PFR-97 WDG. Transmittal of 1 study.
43139901
Eyal, J. (1994) PFR-97 WDG Product Chemistry: Lab Project Number: 4775-PFR-97-CHEM. Unpublished study prepared by W.R. Grace & Co.--Conn. 89 p.
43146200
W. R. Grace & Co.-Conn. (1994) Submittal of Toxicity Data in Support of Registration
of PFR-MUP and PFR-97 WDG. Transmittal of 3 studies.
43146201
Wenk, M. (1994) Primary Eye Irritation Study of Paecilomyces fumosoroseus var. Apopka in Rabbits: Final Report: Lab Project Number: G-7473.230. Unpublished study prepared by Microbiological Associates, Inc. 32 p.
43146202
Wenk, M. (1994) Primary Dermal Irritation Test of Paecilomyces fumosoroseus var. Apopka in Rabbits: Final Report: Lab Project Number: G-7444.242. Unpublished study prepared by Microbiological Associates, Inc. 29 p.
43146203
Wenk, M. (1994) Dermal Sensitization Test of Paecilomyces fumosoroseus var. Apopka in Guinea Pigs: Final Report: Lab Project Number: G-7444.245. Unpublished study prepared by Microbiological Associates, Inc. 37 p.
43163900
W. R. Grace & Co. (1994) Submission of Toxicity Data for Paecilomyces fumosoroseus Apopka Strain 97 in Support of Registration. Transmittal of 1 study.
43163901
Jones, J. (1994) Acute Oral Toxicity/Pathogenicity Study of Paecilomyces fumosoroseus var. Apopka in Rats: Final Report: Lab Project Number: G-7444.222.006: 1602.222.006. Unpublished study prepared by Microbiological Associates Inc. 78 p.
43225500
W.R. Grace & Co. (1994) Submission of Toxicity Data for Paecilomyces fumosoroseus Apopka Strain 97 in Support of Registration. Transmittal of 1 study.
43225501
Wenk, M. (1994) Acute Dermal Toxicity Test of Paecilomyces fumosoroseus var. Apopka In Rabbits: Final Report: Lab Project Number: G-7444.232. Unpublished study prepared by Microbiological Associates, Inc. 36 p.
43292900
W.R. Grace & Co.-Conn. (1994) Submission of product chemistry data in support of registration for PFR-97 20% WDG. Transmittal of 1 study.
43292901
Eyal, J. (1994) PFR-97TM 20 (percent) WDG Product Chemistry: Lab Project Number: 4775/PFR/97/20/CHEM. Unpublished study prepared by W. R. Grace & Co. Conn. 89 p.
43534800
W. R. Grace and Co. (1995) Submission of toxicology data in support of registration of PFR-MUP, PFR-97, and PFR-97 20% WDG. Transmittal of 3 studies.
43534801
Wenk, M. (1994) Repeat Primary Eye Irritation Study of Paecilomyces fumosoroseus var. Apopka in Rabbits: Final Report: Lab Project Number: G/7373/230R. Unpublished study prepared by Microbiological Associates, Inc. 34 p.
43534802
Lawlor, T. (1994) Mutagenicity Test on Paecilomyces fumosoroseus Apopka Strain 97 in Salmonella/Mammalian-Microsome Reverse Mutation Assay (Ames Test) with a Confirmatory Assay: Final Report: Lab Project Number: 16203/0/401R. Unpublished study prepared by Hazleton Washington, Inc. 30 p.
43534803
Frey, L.; Grimes, J.; Beavers, J. (1994) Paecilomyces fumosoroseus Apopka Strain 97: An Avian Oral Pathogenicity and Toxicity Study in the Northern Bobwhite: Lab Project Number: 390/101. Unpublished study prepared by Wildlife International Ltd. 25 p.
43664400
W. R. Grace & Co.--Conn. (1995) Submission of Hazard to Non-Target Insects Data in Support of Applications for Registration of PFR-97 and PFR-97 20% WDG. Transmittal of 1 Study.
43664401
Eyal, J. (1995) PFR-97 20% WDG: Nontarget Insect Data: Effects on Beneficial Insects: Lab Project Number: 4775-20WDG-NTI. Unpublished study prepared by W. R. Grace & Co.--Conn. 101 p.
47939500
Certis USA, LLC (2009) Submission of Product Chemistry and Toxicity Data in Support of the Amended Registration of PFR-97 20% WDG and the Petition for Tolerance of Paecilomyces fumosoroseus Apopka Strain 97. Transmittal of 9 Studies.
47939501
Simpson, A. (2009) Discussion of Formation of Unintentional Ingredients: PFR-97 20% WDG. Unpublished study prepared by Certis USA, LLC. 38 p.
47939502
Simpson, A. (2009) PRF-97 20% WDG: Product Property Data Waivers: Physical and Chemical Characteristics. Unpublished study prepared by Certis USA, LLC. 6 p.
47939503
DeVane, W. (2009) PRF-97 20% WDG: Product Property Data Waivers: Storage Stability. Project Number: TTC0026/10. Unpublished study prepared by Certis USA, LLC. 20 p.
47939504
Dively, C. (2009) Paecilomyces fumosoroseus Apopka Strain 97: End Use Product: PFR-97 20%: Non-Target Plant Studies. Unpublished study prepared by Certis USA, LLC. 25 p.
47939505
Dively, C. (2009) Paecilomyces fumosoroseus Apopka Strain 97: End Use Product: PFR-97 20%: Non-Target Insect Studies. Unpublished study prepared by Certis USA, LLC. 45 p.
47939506
Dively, C. (2009) Paecilomyces fumosoroseus Apopka Strain 97: End Use Product: PFR-97 20%: Non-Target Freshwater Aquatic Invertebrate Testing, Tier I: Non-Target Freshwater Fish Testing, Tier I. Unpublished study prepared by Certis USA, LLC. 23 p.
47939507
Dively, C. (2009) Paecilomyces fumosoroseus Apopka Strain 97: End Use Product: PFR-97 20%: Non-Target Wild Mammal Studies. Unpublished study prepared by Certis USA, LLC. 16 p.
47939508
Dively, C. (2009) Paecilomyces fumosoroseus Apopka Strain 97: End Use Product: PFR-97 20%: Acute Toxicity Studies/End Product (Alternative Formulation). Unpublished study prepared by Certis USA, LLC. 10 p.
47939509
Nengel, S. (1998) PreFeRal WG: Acute Oral and Contact Toxicity to the Honey Bee, Apis mellifera L., in the Laboratory: Final Report. Project Number: 98338/01/BLEU. Unpublished study prepared by GAB Biotechnologie Gmbh. 23 p.

B. Environmental Protection Agency Risk Assessment Memoranda

U.S. EPA. 1995. Review of Product Chemistry, Manufacturing Process, and Acute Toxicity studies of the end use product (EP) PFR-97 (EPA Reg. Nos. 70051-17, 70051-19) containing the active ingredient (AI) Paecilomyces fumosoroseus Apopka strain 97. Memorandum from Cindy Schaffer and Roy Sjoblad, Ph.D. to Shanaz Bacchus dated September 8, 1995. Reviews of Product chemistry - MRIDs 43292901, 43139901, 43139801, acute toxicology - MRIDs 431363901, 43225501, 43139802, 43139803, 43146201, 43146202, 43146203, 43534802.

U.S. EPA. 2011. Review of Product Chemistry, Manufacturing Process, and Acute Toxicity studies of the end use product (EP) PFR-97 (EPA Reg. Nos. 70051-17, 70051-19) containing the active ingredient (AI) Paecilomyces fumosoroseus. Memorandum from I. Barsoum, Ph.D. and J. Kough, Ph.D. to Shanaz Bacchus, dated April 28, 2011 and July 20, 2011. See Tables 1, 2a, 2b, 3.

U.S. EPA. 2011. Environmental risk assessment for the FIFRA Section 3 new use registration of Paecilomyces fumosoroseus Apopka Strain 97 (EPA Reg. No. 70051-19). Memorandum from Shannon Borges and Zigfridas Vaituzis to Shanaz Bacchus, dated July 21, 2011.

C. Other References

ARS collection of Entompathogenic Fungal (ARSEF) Cultures. January 24, 2011. Isaria, plus Paecilomyces and Evlachovea USDA-ARS Biological Integrated Pest Management Research, Robert W. Holley Center for Agriculture and Health. 538 Tower Road, Ithaca, New York 14853-2901.

 Zimmermann, G. 2008. The entomopathogenic fungi Isaria farinosa (formerly Paecilmoyces farinosus) and the Isaria fumosorosea species complex (formerly Paecilomyces fumosoroseus):  biology, ecology and use in biological control. Biocontrol Science and Technology 18: 865-901.

Appendix A. Microbial Pesticides Data Requirements (40 CFR Part 158 - Subpart V)

*NOTE:  Master Record Identification Numbers listed in the following tables are representative
  of supporting data for the original registration of the product containing Isaria fumosorosea (formerly Paecilomyces fumosoroseus) Apopka strain 97.

1. Product Identity and Mode of Action

Product Identity

The Agency has classified Paecilomyces fumosoroseus Apopka strain 97 as an active ingredient for use in microbial pesticides. This fungal strain is an entomopathogenic fungus of many orders of insects. It occurs naturally in populations of infected and dead insects and randomly in soils around the world. It can be cultured readily on various culture media and exists in several morphological stages, depending upon the culture medium. On solid medium, conidiophores, conidia and mycelia are predominant while submerged culture produces blastospores and mycelia. PFR-97[TM] 20% WDG contains blastospores, mycelia and conidia.

The genus Paecilomyces was created in 1907 by Banier and was considered to be related to Penicillium. One taxonomic revision, done by Samson (1974),  redefined and monographed the genus as "those species with verticillate conidiophores bearing divergent whorls of branches and phialides". The latest taxonomic revision is reported in ARS collection of Entompathogenic Fungal (ARSEF) Cultures in January 24, 2011, which classifies the genus and species as Isaria fumosorosea.

When PFR-97 is grown on agar media or insects, the following observations have been noted. Colonies grow moderately fast on malt-agar, attaining a diameter of approximately 4 cm within 14 days at 25C. These colonies consist of a basal felt with raised floccose overgrowth, powdery when freshly isolated. In some isolates pink synnemata occur which may be branched. They are white at first, remaining so or changing to pink shades, especially when sporulating abundantly. Reverse is uncolored or yellow. Odor is absent. Vegetative hyphae are smooth-walled, hyaline, and 1.5 - 3.5 um wide. Conidial structures are mostly complex, consisting of erect conidiophores arising from submerged or laterally from aerial hyphae. Conidiophores are mono-or synnematous, up to 100um in length, 1.5 - 2 um wide, smooth-walled, hyaline, consisting of verticillate branches bearing whorls of 4 to 6 phialides. Phialides are 5.7-8 x 1-2 um, with a globose or ellipsoidal basal portion which tapers into a long distinct neck, about 0.5um wide. PFR Apopka strain 97 in the MUP has verticillate branches bearing whorls of 6 phialides. Conidia are cylindrical to fusiform, smooth-walled, hyaline to slightly pink, 3-4 x 1-2 um. Chlamydospores are absent.

Growth on insects. On insects the fungus produces monomenatous conidiophores or distinct but loose synnemata. Synnemata are erect, branched appearing pink and powdery due to the conidia, and can grow up to 3 cm in length and 400um in diameter. Conidiogenous elements are inflated. Condiophores are 2.5-4um wide. Verticillate branches are more or less globose or broadly cylindrical, 4-9 x 3-4.5 um. Phialides with a globose basal portion can be 2.3 - 3.5 um wide.

Mode of Action:

Conidia of the fungus attach to, and penetrate the cuticle of the host insect, where they germinate and grow. This leads to pathogenesis and eventual death of the diseased insect host.

      Product Chemistry

TABLE 1. Product Analysis Data Requirements for the Manufacturing Use Product (MUP), Isaria fumosorosea (formerly Paecilomyces fumosoroseus) Apopka strain 97, and Its Associated End-Use Product (EP)
                             (40 CFR § 158.2120)
                            OCSPP Guideline Number
                               Data Requirement
                                    Results
                                   MRID No.

                                      MUP
                                      EP

                                   885.1100

                               Product Identity
A sample of Paecilomyces fumosoroseus Apopka strain 97 is filed at American Type Tissue Collection # 20874.
(DER dated 09/08/1995, from Cindy Schaffer to Shanaz Bacchus,  MRIDs 43139901, 43139801). Change in taxonomic identification in 2011 identifies fungus as Isaria fumosorosea (Zimmerman, 2008) Apopka strain 97. Acceptable.
Submitted data satisfy the requirement for product identity. This product contains 20% w/w of the MUP Paecilomyces fumosoroseus Apopka strain 97 (ranged from 1.5 x 10[9] to 3.26 x 10[9] cfu/g).
(DER dated 09/08/1995 from Cindy Schaffer to Shanaz Bacchus, MRID [†]43292901).
Acceptable
[†]43292901
[†]43139901
[†]43139801

                                   885.1200

                             Manufacturing Process
The MUP is manufactured as liquid fermentation product at 10[8] cfu/ml onsite and is formulated into the End-use Product that is labeled at 10[9] cfu/gram (DER dated 09/08/95 from Cindy Schaffer to Shanaz Bacchus, MRID [†]43139801, Supplemental- required analysis of unintentional ingredients.
Manufacturing process for PFR-97 20% WDG was previously submitted in MRID 43292901. MSDS for the inert ingredients were provided in
in MRID 47939508
**US EPA DER I. Barsoum to S. Bacchus, 04/28/11. Acceptable
[†]43292901
[†]43139901
[†]43139801
47939508**
                                   885.1300
             Discussion of Formation of Unintentional Ingredients
`Beauveralides' were detected as secondary metabolites during the manufacturing process; significance of this was not determined, but secondary metabolites not a human health concern for the non-food greenhouse uses. Supplemental.
Levels of fungal and bacterial metabolites and MS-HPLC assay of negligible total beauverolides in broth acceptable; calculated beauverolides in sediment. Confirmation required in five production batches. ***US EPA DER 05/20/11 from I. Barsoum to S. Bacchus, MRIDs 48383001, 48357702.
Additional data provided for proposed food uses of the EP
in 2011 are acceptable for detection and Quality Assurance and Quality control of fungal, bacterial and other potential metabolites. **US EPA DER 04/28/11 from I. Barsoum to S. Bacchus; MRID 47939501, 47939508, dated 05/20/11; ***US EPA DER dated 05/20/11 from I. Barsoum to S. Bacchus, MRIDs 48383001, 48357702.
Acceptable
[†]43292901*
[†]43139901*
[†]43139801*
47939501**
47939501**
47939508**
48383001***
48357702***
                                Not applicable
     Deposition of a Sample in a Nationally Recognized Culture Collection
Paecilomyces fumosoroseus Apopka strain 97 is on deposit with ATCC Manassas, Virginia under Accession Number 20874.
                              Not required for EP
Certis' MSDS.
                                   885.1400
                              Analysis of Samples
Submitted data satisfy the requirement for analysis of samples.

The content of active ingredient ranged from 1.5 x 10[9] to 3.26 x 10[9] cfu/g, which meets the label claim. DER 04/28/11 from I. Barsoum to S. Bacchus, MRID 47939501.
†43139801
[†]43139901
†43292901
47939501

                                   885.1500

                            Certification of Limits
Limits listed on the Confidential Statement of Formula (CSF) are adequate/acceptable.

[†]43139801
[†]43139901
[†]43292901
47939501
                              Additional Studies
                                   830.1800
                         Enforcement Analytical Method
                                Not applicable
Acceptable
                                   47939501
[†]DER dated 09/08/95 from Cindy Schaffer to Shanaz Bacchus, MRIDs [†]43139801, [†]43292901*, [†]43139901*, [†]43139801
*DER Hanna Daoud, CA Dept. Pesticide Registration, April 27, 1995; †43139801, [†]43292901*, [†]43139901*, [†]43139801
For the initial registration of the active ingredient, US EPA evaluated the data submitted for this registration action in 1995, but considered and utilized the CA data assessments in order to streamline the review process.
**US EPA DERs from I. Barsoum to S. Bacchus; MRID 47939501 (04/28/11), 05/20/11;
***US EPA DER from I. Barsoum to S. Bacchus MRID 48357702, 48383001, dated 05/20/11.

TABLE 2a. Physical and Chemical Characteristics for the Manufacturing-Use Product (MUP), Isaria fumosorosea (formerly Paecilomyces fumosoroseus) Apopka strain 97 (40 CFR § 158.2120)
                            OCSPP Guideline Number
                               Data Requirement
                                    Results
                                  MRID Number
                                   830.6302
                                     Color
                                     Brown

43139801
                                   830.6303
                                Physical State
                  Solid and liquid (10% precipitate in broth)

                                   830.6304
                                     Odor
                                 Smoke flavor

                                   830.6313
                       Stability to Normal and Elevated
                             Temperatures, Metals,
                                and Metal Ions
Stable at 4o C up to 3 weeks; not stable at 25 o C. Stability to metals and metal ions not reported.
                                   43139801
                                   830.6317
                               Storage Stability
Up to three weeks @ 4ºC. (C. Schaffer, 1995) up to 6 months at 4C (BRAD 050405).
43139801

                                   830.6319
                                  Miscibility
Not required because PFR 97 is not an emulsifiable form of microbial pesticide [refer to test note #2 of 40 CFR § 158.2120(d)].
                                      N/A
                                   830.6320
                          Corrosion  Characteristics
                  Not corrosive in fermentation test vessels.

43139801

                                   830.7000
                                      pH
                                      5.7

                                   830.7100
                                   Viscosity
                                6.4 Centipoise

                                   830.7300
                                    Density
                                     1.06

                                   830.6315
                                 Flammability
                                 Not flammable

                                      N/A
                                   830.6314
                Oxidation/Reduction; Chemical Incompatibility
                                      N/A

                                   830.6316
                                 Explodability
                                     N/A.

                                   830.6321
                         Dielectric Breakdown Voltage
                                     N/A.

N/A = Not applicable

TABLE 2b. Physical and Chemical Characteristics for the End-Use Product (EP), Isaria fumosorosea (formerly Paecilomyces fumosoroseus) Apopka strain 97 (40 CFR § 158.2120)

                       Guideline Reference No./Property

                             Description of Result

                                    Methods

830.6302	Color
                                 Yellow/Brown

                                 MRID 47939502
                                Certis' MSDS

830.6303	Physical State
                                Granular solid

830.6304	Odor
                              Not required for EP

830.6313	Stability
                              Not required for EP

830.6314	Oxidation/Reduction:
        Chemical Incompatibility
  Not applicable, the product does not contain oxidizing or reducing agents.

830.6315	Flammability
      Not applicable, the product does not contain flammable ingredients.

830.6316	Explodability
      Not applicable, the product does not contain explosive ingredients.

830.6317	Storage Stability
Stable in product packaging at 4°C for 12 months. Less stable in product packaging at 25°C after 12 months. (Product label recommends storage at 4° to 6°C in a dry place. Sample stored at 4o C for 1 year was still effective at colonizing host insects while that stored at 25 o C was not).

                                 MRID 47939503

830.6319	Miscibility
          Not applicable, the product is not an emulsifiable liquid.

                                 MRID 47939502

830.6320	Corrosion Characteristics
                    Non-reactive to the product containers.

830.6321	Dielectric Breakdown Voltage
    Not applicable, the product is not for use around electrical equipment.

830.7000	pH
                                   6.4-6.5a
                                   6.0-7.00b

830.7100	Viscosity
                    Not applicable, the product is a solid.

830.7200	Melting Range
                              Not required for EP

830.7220	Boiling Range
                    Not applicable, the product is a solid.

830.7300	Density/Relative Density/Bulk Density
                                0.60-0.65 g/mLa
                                   600 g/Lb

830.7370	Dissociation Constant in Water
                              Not required for EP

830.7550	Partition Coefficient
                              Not required for EP

830.7840	Water Solubility
                              Not required for EP

830.7950	Vapor Pressure
                              Not required for EP

aData from CSF for basic formulation. Basic formulation withdrawn 07/28/11 (email from Chris Dively, Certis,USA, to Shanaz Bacchus, US EPA/OPP/BPPD).
bData from CSF for alternate formulation. This is new basic formulation.

TABLE 3. Toxicology Data Requirements for the Technical Grade of the Active Ingredient (TGAI), Isaria fumosorosea (formerly Paecilomyces fumosoroseus) Apopka strain 97 (40 CFR § 158.2140)
                            OCSPP Guideline Number
                               Data Requirement
                                    Results

                                  MRID Number
                                    Tier I

                                   885.3050

                      Acute Oral Toxicity/ Pathogenicity
An oral dose of 1.7 x 10[6] cfu/animal in a conidia spore suspension did not produce mortality or abnormal clinical effects. No signs of fungal contamination were reported for the brain, mesenteric lymph nodes, blood, kidney, spleen, liver, lung or cecum, and no infectivity or pathogenicity was recorded. [DERs to Shanaz Bacchus from Cindy Schaffer (09/08/95)* and from Ibrahim Barsoum (04/28/11)**]
Acceptable.     Toxicity Category IV
                                   43163901*
                                  47939508**

                                   885.3150

                    Acute Pulmonary Toxicity/ Pathogenicity
A single intratracheal dose (106 conidia spores/animal) resulted in no deaths and there were no signs of toxicity in any animal. No signs of infection were seen and no colonization of the lungs was observed. Total clearance of the fungus was attained by day eight after treatment.
Acceptable        Toxicity Category IV

                                   43139802

                                   885.3200

                    Acute Injection Toxicity/ Pathogenicity
Single intraperitoneal doses of suspensions containing 1.6 x 10[7] conidia spores per animal had no toxic or pathogenic effects and the spores were cleared from the body within two days.

                                   43139803

                                   870.1200
                             Acute Dermal Toxicity
2 grams of test substance applied to the skin of rabbits produced no deaths and there was no evidence of systemic toxicity.
Acceptable        Toxicity Category III.

                                   43225501

                                   870.2500
                           Primary dermal irritation
In a four-hour exposure, the test substance (2 g/animal) was found to be a slight dermal irritant.
Acceptable        Toxicity Category IV.

                                   870.2400
                             Acute eye Irritation
0.1 mL of diluted test substance containing >=107 cfu was instilled in the eye, which was examined at 1, 24, 48 and 72 hours, and at 4 and 7 days after treatment. The test substance produced slight irritation which was reversible by day 4 after treatment.  The product was classified as Acceptable         Toxicity Category IV.
                                   43146201

                             Dermal Sensitization
In guinea pig: The test substance (3.0 x 107  -  5.3 x 109 cfu) produced no irritation or sensitization reaction in treated animals. The product was not a dermal sensitizer.
Supplemental. Not required for registration
                                   43146202

                                   885.3400
                               Hypersensitivity
                                   Incidents
Published literature pertaining to incidents involving other strains of Paecilomyces have been evaluated and data submitted to demonstrate that those incidents do not involve PFR Apopka 97 strain. Incident data must be reported in a timely manner. None reported with regard to pesticide use during the time that the registered pesticide has been in use.

                                   885.3500
                                 Cell Culture
     Not required for a fungal active ingredient  -  required for viruses.
                                      NA

                              Mutagenicity assay
(Ames Assay): Sonicated blastospores and mycelium of Paecilomyces fumosoroseus Apopka strain 97 (100  -  5000 ug/plate) yielded negative results in a reverse mutation assay in both the presence and absence of microsomal enzymes (S9).
                                   43534802
                               Tiers II and III
Not required for  Paecilomyces fumosoroseus Apopka strain 97 based on the acceptability of the data and waiver rationales evaluated for Tier I.

Subchronic, Chronic Toxicity and Oncogenicity
Tier II tests (Guidelines 152-40 through 152-49) involving acute oral, acute inhalation, subchronic oral, acute intraperitoneal, intracerebral, primary dermal, primary eye, immune response, teratogenicity, virulence enhancement, and mammalian mutagenicity were (40 CFR 158.740(C)(2)(vi through xv) not required since survival, replication, infectivity, toxicity, or persistence of the microbial agent was not observed in the test animals treated in the Tier I acute oral infectivity test. Tier III tests (Guidelines 152-50 through 53) involving chronic testing, oncogenicity testing, mutagenicity, and teratogenicity were not required (40 CFR 158.740(C)(2)(xvi through xix) since Tier II tests were not required.

Acute toxicity  -  End-use Product
For the End-use Product (EP) PFR-97 20% WDG, the registrant requested that testing for data requirements be waived based on the results of the tests conducted on the TGAI and the nature of the inert ingredients (MRID 47939508). The rationales justifying these requests were acceptable to the Agency (US EPA DER dated 04/28/11 from I. Barsoum to Shanaz Bacchus).

Waiver Requests granted for EP:
 Acute Oral Toxicity (OPPTS 870.1100)
 Acute Dermal Toxicity (OPPTS 870.1200):
 Acute Inhalation Toxicity (OPPTS 870.1300)
 Acute Eye Irritation (OPPTS 870.2400)
 Acute Dermal Irritation (OPPTS 870.2500)

Ecological and Environmental Effects
See US EPA Borges July 21, 2011, for Nontarget Organism Toxicity and Environmental Fate Data Requirements  for the Technical Grade of the Active Ingredient (TGAI), for  Isaria fumosorosea (formerly Paecilomyces fumosoroseus) Apopka strain 97 (40 CFR § 158.2150) in this docket for summaries of the data reviewed and presented in the table below.

Table 4. Summary of data submitted to comply with nontarget organism data requirements published in 40 CFR § 158.2150 for support of the registration of Paecilomyces fumosoroseus Apopka strain 97.
Data Requirement
OPPTS Guideline No.
Results Summary and Classification
MRID No.
Avian oral toxicity/pathogenicity
885.4050
A study (supplemental) indicated that P. fumosoroseus Apopka Strain 97 is not toxic or pathogenic to birds. P. fumosoroseus Apopka Strain 97 does not grow at avian body temperatures and is not expected to pose a hazard to birds
43534803
Avian inhalation toxicity/pathogenicity
885.4100
Not required. P. fumosoroseus is not pathogenic to birds.
N/A
Wild mammal toxicity/pathogenicity

885.4150

Tests required by 40 CFR § 158.2140 are adequate/ appropriate for assessment of hazards to wild mammals. P. fumosoroseus Apopka Strain 97 was not toxic or pathogenic to albino rats at 1.7 x 10[6] cfu/animal. Scientific rationale also support the conclusion that adverse effects to wild mammals are not expected.
Acceptable
47939507

:: Freshwater fish toxicity/pathogenicity
:: Freshwater invertebrate toxicity/pathogenicity

885.4200

885.4240

Data waiver rationale provides sufficient information to determine that toxicity/pathogenicity to freshwater fish and invertebrates is not expected as a result of all applications except for aerial applications.
Supplemental for aerial applications.  Acceptable for all other applications
47939506
Estuarine/marine fish and invertebrate testing
885.4280
Not required. P. fumosoroseus is not to be applied directly to water and is not expected to reach estuarine or marine environments in significant quantities. P. fumosoroseus was also shown to be nonpathogenic to two species of estuarine invertebrates.
N/A
Nontarget plant testing
885.4300
Scientific rationale was sufficient to conclude that no adverse effects are expected to nontarget plants. Paecilomyces fumosoroseus Apopka strain 97 is not related to any known plant pathogen and testing is not required.
Acceptable
47939504
Nontarget insect testing

885.4340

Initial scientific rationale (MRID 47939505) was unacceptable. Additional rationale provides sufficient information to conclude that Paecilomyces fumosoroseus Apopka strain 97 will be pathogenic to some nontarget species of insects
Acceptable
47939505

48357703
Honey bee testing
885.4380
A honey bee study showed that Paecilomyces fumosoroseus Apopka strain 97 is not toxic to honey bees.
Supplemental
Additional rationale show that pathogenicity may not be likely, but a conclusion cannot be made without uncertainty.
Acceptable
47939509

48357704