# BIOPESTICIDES REGISTRATION

ACTION DOCUMENT

Paecilomyces fumosoroseus strain FE 9901



# DRAFT DOCUMENT

U.S. E NVIRONMENTAL 

## P ROTECTION 

## A GENCY 

## O FFICE O F 

## P ESTICIDE 

## P ROGRAMS


## B IOPESTICIDES A ND 

## P OLLUTION 

## P REVENTION 

## D IVISION



### October 5 , 2010ii



## T ABLE OF 

## C ONTENTS

I. REGULATORY BACKGROUND

................................ ................................ ................ 5 A. Application for Pesticide Registration

................................ ................................ ... 5 B. Petition to Establish an Exemption from Tolerance

................................ .............. 5

C . North American Free Trade Agreement Joint Review

................................ .......... 6

II. BIOLOGICAL CONSIDERATIONS

................................ ................................ ........... 6

III. ACTIVE INGREDIENT CHARACTERIZATION

................................ ........................ 6 A. Product Chemistry and Compositio n

................................ ................................ .... 7 1. Manufacturing Process

................................ ................................ .................... 7 2. Deposition of a Sample in a Nationally Recognized Culture Collection

........... 7 3. Discussion of Unintentional Ingredients

................................ ........................... 7 B. Physical and Chemical Properties

................................ ................................ ........ 7

IV. HUMAN HEALTH RISK ASSESSMENT

................................ ................................ .. 8 A. Toxicity

................................ ................................ ................................ ................. 8 1. Acute Toxicity

................................ ................................ ................................ .. 8 2. Hype rsensitivity I ncidents

................................ ................................ ................ 9 3. Cumulative Effects from... with a Common Mechanism of Toxicity

................. 9 4. Effects on the Endocrine System

................................ ................................ ... 10 B. Exposure

................................ ................................ ................................ ............ 10 1. Aggregate Exposure

................................ ................................ ...................... 10 2. Occupational Exposure

................................ ................................ ................. 11 C. Determination Of Safety For U.S. Population, Infants, and Children

.................. 11

D. Risk Characterization

................................ ................................ ......................... 12

V. ENVIRONMENTAL ASSESSMENT

................................ ................................ ........ 12

A. Risk Characterization

................................ ................................ ......................... 12 B. Endangered Species Assessment

................................ ................................ ...... 13

VI. ENVIRONMENTAL JUSTICE

................................ ................................ ................. 13

VII. RISK MANAGEMENT DECISION

................................ ................................ ......... 13

VIII. TERMS OF THE REGISTRATION

................................ ................................ ....... 14

BIBLIOGRAPHY

................................ ................................ ................................ ........... 15

APPENDIX A: Data Requirements and How the Applicant Me t Them

........................ A

- 1

APPENDIX B: Biology and Taxonomy of P. fumosoroseus

................................ ........ B

- 1

APPENDIX C: Summary of Toxicity Studies

................................ .............................. C

- 1



## List of Tables

Table 1. Select Physical and Chemical Properties of Paecilomyces fumosoroseus

strain FE 9901

................................ ................................ ................................ ... 7

Table 2 . Summary of Toxicology Data for Paecilomyces fumosoroseus strain FE 9901 9iii



## B IOPESTICIDES 

## R EGISTRATION 

## A C TION 

## D OCUMENT 

## (BRAD) T EAM



## Office of Pesticide Programs Biopesticides and Pollution Prevention Division

Microbial Pesticides Branch

S CIENCE 

## R EVIEW



### Ibrahim Barsoum Ph.D

. Product Chemistry and

Human Health Risk John Kough, Ph.D. Secondary Review of P roduct Chemistry and

Human Health Risk Gail Tomimatsu, Ph.D

. Environmental and Ecological Risk



## R EGULATORY



### Kathleen Martin, M

. S

. E

. S., M

. P

. A

. Regulatory Action Leader Sheryl Reilly, Ph.D. Chief, Microbial Pesticides Branchiv

ACRONYMS

Acro nym Extension

ARS Agricultural Research Service

BRAD Biopesticides Registration Action Document

CFU Colony forming units

DNA Deoxyribonucleic acid

EDSP Endocrine Disruptor Screening Program

EDSTAC Endocrine Disruptor Screening and Testing Advisory Co mmittee

EP End

- use product

EPA U.S. Environmental Protection Agency or

, the Agency

FFDCA Federal Food, Drug, and Cosmetic Act

FIFRA Federal Insecticide, Fungicide, and Rodenticide Act

FQPA The Food Quality Protection Act of 1996
g/cc Grams per cubi c centimeter

LC 50 Median Lethal Concentration

LD 50 Median Lethal Dose
mg/kg bw Milligrams per kilogram of body weight
mg/L Milligrams per liter

MPCA Microbial pesticide control agent

MRID Master Record IDentification (number)

NAFTA North American F ree Trade Agreement

NOF Notice of Filing

NR Not required

OCSPP Office of Chemical Safety and Pollution Prevention

OECD Organization for Economic Cooperation and Development

OPP EPA’s Office of Pesticide Programs

OPPTS EPA's Office of Prevention, Pes ticides and Toxic Substances;

n ow the Office of Chemical Safety and Pollution Prevention

P. fumosoroseus Paecilomyces fumosoroseus

PMRA Health Canada’s Pest Management Regulatory Agency

POTW Publicly owned treatment works

TGAI Technical grade active i ngredient

U.S. United States

UV Ultraviolet radiation

USDA U.S. Department of Agriculture

WP Wettable powderP. fumosoroseus strain FE 9901 page 5 Biopesticides Registration Action Document

Natural Industries Inc. has applied to the U.S. Environmental Protection Agency (EPA or the Agency) for registration of two products cont aining the new active ingredient Paecilomyces

fumosoroseus strain FE 9901 . The manufacturing

- use product (MUP) is NoFly TM Technical (73314

- T ); it contains 69.0% active ingredient. The end

- use product (EP) is NoFly TM WP ( 73314

- A ) and it contains 18% active ingredient. This new mycoinsecticide will be used in greenhouses ( on both food and nonfood sites ) and is intended to co ntrol whiteflies, aphids, thrips, psyllids, mealybugs, leafhoppers, plant bugs, weevils, grasshoppers, M ormon crickets, locust, and beetles

. Crops include ornamentals, vegetables, and herbs. The applicant has also petitioned for an exemption from the req uirement of tolerance for any P. fumosoroseus strain FE

9901 residues that may remain in or on food.

Public Participation

. On October 1, 2009, EPA announced a new policy to provide a more meaningful opportunity for the public to participate in major regi stration 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 fumosor oseus strain FE 9901 is subject to a 30

- day comment period as a “new active ingredient” whose registration would result in a “first food use. ” The public comment period runs from October 5 , 2010 to November 3 , 2010 . T o submit a comment, see the

instructi ons at EPA’s webpage titled, “ Instructions for Commenting During Open Comment

Periods ” ( www.epa.gov/pesticides/regulating/comment

- instruction.html ).

I. REGULATORY BACKGROUND

A. Application for Pesticide Re gistration

A Notice of Receipt

, announcing EPA’s receipt of Natural Industries Incorporated’s application s to register P. fumosoroseus strain FE 9901

, was published in the Federal Register

on March 10, 2010 ( 75 FR 11175 ) . This new mycoinsecticide is meant to control whiteflies, aphids, thrips, psyllids, mealybugs, leafhoppers , plant bugs, weevils, grasshoppers, M ormon crickets, locust, and beetles on all greenhouse and nursery crops including ornamentals, vegetables, and herbs

. You can find the Notice of Receipt and supporting materials (e.g., draft label) at regulations.gov; the docket number is EPA

- HQ

- OPP

- 2010

- 0093

. EPA received no

comments in response to the Notice of Receipt.

B. Petitio n to Establish an Exemption from the Requirement of Tolerance

Under section 408 of the Federal Food, Drug, and Cosmetic Act (FFDCA ), the applicant has petitioned ( PP 9F7587) the Ag ency to establish an exemption from the requirement of a

tolerance for residues of P. fumosoroseus s train FE 9901

. A Notice of Filing (NOF ) was published in the Federal Register on March 10, 2010 (75 FR 11171); it can be found at regulations.gov under dock et number EPA

- HQ

- OPP

- 2010

- 0092 . No comments were received in response to the NOF

.

EPA has made a preliminary conclusion that there is a reasonable certainty that no harm will result to the United States population, including infants and children, from a ggregate exposure to the residues of P. fumosoroseus s train FE 9901 , for the uses described on the pesticide product label.P. fumosoroseus strain FE 9901 page 6 Biopesticides Registration Action Document

C. North American Free Trade Agreement Joint Review

P. f umosoroseus s train FE 9901 was the subject of a North American Free Trade Agreement (NAFTA) joint registration review conducted by EPA’s Office of Pesticide Programs

and Health Canada ’s Pest Management Regulatory Agency (PMRA).

II. BIOLOGICAL CONSIDERA TIONS

P. fumosoroseus is a naturally occurring fungus that is found in soils around the world, including Europe ( Netherlands, Ireland, and Spain ); Asia ( China, Japan, Pakistan, and India ); Africa ( Ghana ); and the United States. In the environment, P. fumosoroseus is dispersed as conidia, either by air or water movement or by other insects and/or mites

. Conidia are spores and they are supported on specialized spore

- producing “ hyphae ” ( a filamentous stal k) known as conidiophores. As with other “ entomopathogenic fungi ,” 1

The infection cycle of the MPCA NoFly TM WP is particularly rapid under optimal

conditions. Initially, conidia attach to the cuticle or the dorsum of immature or adult whiteflies; first symptoms of infect ion typically are apparent within one to two days after contact. Conidia and hyphae are present in the host hemocoel (body cavity) within one day. The mycelium (a branching network of hyphae) is present on the dorsum of the insect body within two days. Sporulation occurs within three days and reaches a maximum within five to seven days. NoFly TM WP ’s target organism is the whitefly

— this MPCA is intended to control all stages of whitefly development P. fumosoroseus infects its target by conidial attachment followed by germination, penetration in to the host, vegetative growth within the organism, and then conidiogenesis (i.e., produc tion of more conidia). In laboratory testing, investigators determined that P. fumosoroseus strain FE 9901 grows best at (76.1 o F). At higher temperatures ( 79.7 o F to 86.0o F), growth is significantly reduced and at temperatures greater than 95.0 o F, P. fumosoroseus strain FE 9901 will no longer grow.

2

P. fumosoroseus s train FE 9901 does not exhibit any atypical morphological, physiological, biochemical, p esticidal, or resistance characteristics. For more information on the biology and taxonomy of P. fumosoroseus in general, and P. fumosoroseus strain FE

9901 specificall y, please see Appendix B : Biology and Taxonomy . Unless otherwise noted, the source of the information is from the joint review (U.S. EPA 2010) of the data supported to support the registration of this new active ingredient. (i.e., eggs, N1, N2, N3 and N4 larvae, and adults). Nontarget insect hosts include mealy bugs, beetles, caterpillars, and flies.

I I I. ACTIVE INGREDIENT CHA RACTERIZATION (Product A nalysis

— 40 CFR 158.2120 )

Most of the data requirements under 40 CFR 158.2120 (Product A nalysis 3

1 An entomopathogenic fungus is one that can act as a parasite of insects and kills or seriousl y disables them. ) have been satisfied. One of the requirements under Analysis and Certifi ed Limits

—Analysis of samples (OPPTS 885.1400)

— needs to be supplemented with additional information. Three of the Physical and Chemical Characteristics

— Stability to normal and elevated temperatures, metals and metal ions; Storage stability; and Corrosion characteristics

— have not been satisfied. Each of these will be a term of registration; for detailed information see, Section V II: Terms of t he

2 The ability to infect whitefly eggs is unique among entomopathogenic fungi.

3 Product Analysis data is the information EPA needs to identify the active and inert ingredient, and to guard against chemical and biological contamination.P. fumosoroseus strain FE 9901 page 7 Biopesticides Registration Action Document

Registration . F or a complete tabulation of all the Product Analysis data requirements

, see Table

A

- 1, Appendi x A. The Table also shows how the petitioner addressed each data requirement.

A. Product Chemistry and Composition

1

. Manufacturing Process (OPPTS 885.1200)

No Fly TM Technical (MUP) and NoFly TM WP (EP) are manufactured using a process typical for the growth of microbials. The applicant adequately described the manufacturing process (MRIDs 477919

- 01 and 477920

- 01).

2

. Deposition of a Sample in a Nationally Recognized Culture

Collection

A sample of P. fumosoroseus strain FE 9901 has been deposited in a nationally recognized culture collection . The sa mple is in the U.S. Department of Agriculture’s Agricultural Research Service Collection of Entomopathogenic Fungal Cultures ( USDA

ARS ) under the accession number ARSEF 4490.

3

. Discussion of Unintentional Ingredients (OPP TS 885.1300)

No substances (e.g., impurities, unintentional ingredients) of toxicological

concern have been identified for NoFly TM WP or NoFly TM Technical . This data requirement has been adequately satisfied.

B

. Physical a nd Chemical Properties

Provided in Table 1 is a summary of the physical and chemical properties of P.

fumosoroseus strain FE 9901.

Table 1 . Select Physical and Chemical Properties of P. fumosoroseus strain FE 9901

OPPTS

Guideline

Number Data Requirement VALUE

EP (NoFly TM WP) MUP (NoFly TM Technical)

830.6302 Color lilac lilac to brown

830.6303 Physical state powder slightly cohesive, dusty powder

830.6304 Odor yeast

- like yeast

- like

830.6313 Stability to normal and elevated temperatures,

metals and metal ions Term of registration Above 104  ̊ F (40  ̊ C) this MPCA will not survive for any length of time; NoFly TM Technical will not be exposed to metal or metal ions.

830.6317 Storage stability Term of registration

830.6320 Corrosion characteristics Term of registration

830. 7000 pH (suspension in water) 6.5 to 7.5 6.5 to 7.5

830.7300 Bulk density 0.65 to 0.75 g/cc 0.6 to 0.7 g/ccP. fumosoroseus strain FE 9901 page 8 Biopesticides Registration Action Document

I V

. HUMAN HEALTH RISK ASSESSMENT (U.S. EPA 2010)

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

in formation concerning the cumulative effects of [a particular pesticide's] residues and other

substances

The Agency performs a number of analyses to determine the risks from aggregate exposure to pesticide residues. First, the Agency determines the toxic ity of pesticides. Second, the Agency examines exposure to the pesticide through food, drinking water, and through other exposures that occur because 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 v ariability of the sensitivities of major identifiable subgroups of consumers, including infants

and children.

Following is the Agency’s human health risk assessment. It includes information on exposure, toxicity, and endocrine disruption. The cumulative effects aspect of FFDCA is discussed below under Toxicity ( section A.) ; aggregate exposure is under Exposure ( section B.) ; and s pecial consideration of infants and children is discussed under section C.

A

. Toxicity

1

. Acute Toxicity

Provided in Table 2 is a summary of the toxicity data for the end

- an d manufacturing

use products of P. fumosoroseus strain FE 9901 ; for brief descriptions of studies submitted, please see Appendix C: Summary of Toxicity Studies. Except for Acute injection toxicity/pathogenicity

, ( the applicant submitted a st udy but EPA requests additional information )

, all the toxicology data req uirements have been satisfied; see Section VII: Terms of the Regist ration for more information. Table A

- 2, Appendix A provides a complete tabulation of each data requirement along with how the applicant met the requirement.P. fumosoroseus strain FE 9901 page 9 Biopesticides Registration Action Document

Table 2

. Summary of Toxicology Data for Paecilomyces fumosoroseus

st rain FE 9901 (40 CFR 158.2140)

OPPTS

Guideline

Number Data Requirement Toxicity and Category 1

EP (NoFly TM WP) MUP (NoFly TM Technical)

870.1100 Acute oral toxicity >5,000 mg/kg bw
Category IV 600 mg/kg bw
Category I I I

870.1200 Acute dermal toxicit y 5,050 mg/kg bw
Category IV Data waived

870.1300 Acute inhalation toxicity >2.18 mg/L
Category III Data wa ived

870.2400 Acute eye irritation s ome redness
Category IV Data waived

870.2500 Primary dermal

irritation slightly irritating
Category IV Data waived

885.3050 Acute oral toxicity/ pathogenicity TGAI: Not infective or pathogenic to rats via the oral route

885.3150 Acute pulmonary toxicity/pathogenicity TGAI: not infective or pathogenic to ferrets via the inhalation route

885.3200 Acute inject ion toxicity/pathogenicity (intraperitoneal) TGAI: not infective or pathogenic to rats via injection, based on the

available results (more information is needed)

1 Category I has the highest toxicity while Category IV has the lowest. For a description of the Toxicity Categories along with their limits, see 40 CFR 1 56.62.

2

. Hypersensitivity I ncidents (OPPTS 885.3400 ; MRID 480514

- 02 )

The applicant reports that they have not seen any incidents of hype rsensitivity throughout the research, development, and testing of NoFly TM Technical and WP

. However, a literature search of the PubMed database ( www.ncbi.nlm.nih.gov/pubmed ) using the keywords “paecilomyces” and “allergy

, ” revealed 21 reports of hypersensi tivity

. Based on the assumption that most microorganisms contain substances that would elicit allergic reactions, EPA considers P. fumosoroseus strain FE 9901 to be a potential sensitizing agent.

3. Cumulative Effects from Substances with a

Common Mec hanism 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 i nformation concerning the cumulative effects of a particular pesticide’s residues and other substances that have a common mechanism of toxicity.’’ Based on the available toxicity information, EPA has not found P. fumosoroseus strain FE 9901 to share a com mon mechanism of toxicity with any other substances, and P. fumosoroseus strain FE

9901 does not appear to produce a toxic metabolite as its mode against its target pests.

For information regarding EPA’s efforts to determine which chemicals have a

common mechanism of toxicity and to evaluate the cumulative effects of such chemicals,

see EPA’s website at www.epa.gov/pesticides/cumulative

.P. fumosoroseus strain FE 9901 page 10 Biopesticides Registration Action Document

4

. Effects on the Endocrine System

As required under FF DCA 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.” 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 proc eed 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 estab lish 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 iner t 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

o f known or likely endocrine disruptors.

P. fumosoroseus strain FE 9901 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 Tie r 1 screening battery, please visit

our website: www.epa.gov/endo/



### .

B

. Exposure

Individuals may be exposed to P. fumosoroseus strain FE 9901 through the diet (i.e., consumption of vegetables and herbs that have been tr eated and drinking water) and exposure while applying the pesticide or working in a greenhouse where this pesticide has been applied.

1

. Aggregate Exposure

a. Dietary Exposure

Food . EPA expects that exp osure to P. fumosoroseus strain FE 9901 residues through consumption of treated vegetables and herbs to be low. P.

fumosoroseus strain FE 9901 does not grow well at very warm temperatures

— at

80 o F, growth is significantly reduced and at 95 o F, it does no t grow at all. Sunlight further inhibits the growth of P. fumosoroseus ( Fargues 1997) . Thus, EPA expects that once the treated food is out of the greenhouse, residues of P.

fumosoroseus strain FE 9901 would rapidly decrease.P. fumosoroseus strain FE 9901 page 11 Biopesticides Registration Action Document

Drinking water exposure

. EPA does not expect to find residues of P.

fumosoroseus strain FE 9901 in drinking water. Application of this MPCA is in greenhouses; wastewater from mixing or applying the pesticide is expected to go through a publicly owned treatment works (POTW) before being discharged back to the environment. Further, P. fumosoroseus does not thrive in the presence of sunlight. Fargues et al. (1997) studied the effects of ultraviolet (UV) radiation on the germinability, survival, and infectivity of P. fumosoroseus . T hey found the UVB range of ultraviolet light (sunlight includes light in the UVB, UVA, and visible ranges) was the most important factor “for the detrimental effects of sunlight on the persistence of conidia of entomopathogenic fungi in insolated environme nts ” (Fargues 1997) . Therefore, even if contaminated water is directly discharged to surface water, EPA expects that residues of P. fumosoroseus strain FE 9901

would not survive.

Residential, School, and Day Care Exposure

. I ndividuals in residential, schools, and day care settings will not be exposed to residues of P. fumosoroseus strain

FE 9901 because this MPCA is only applied in greenhouses.

2. Occupational Exposure

NoFlyTM WP, which is limited to greenhouse use, can be applied using conventional spraying equipment (but, use of manual sprayers is highly recommended). Occupational exposure via the inhalation and dermal is expected for persons who mix and/or applying t his pesticide. Dermal exposure is expected individuals who work in the greenhouse following application.

C

. Determination Of Safety For U.S. Population, Infants, a nd Children

FFDCA section 408(b)(2)(C) provides that EPA 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 e ffects 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 EPA shall apply an additional tenfold margin of safety for

infants and children in the case of t hreshold effects to account for prenatal and postnatal toxicity and the completeness of the database unless EPA determines that a different margin of safety

will be safe for infants and children. Margins of exposure (safety), which are often referred to a s uncertainty factors, are incorporated into EPA 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 avai lable toxicity and exposure information, as well as the natural

occurrence of P. fumosoroseus strain FE 9901 in the environment, EPA concludes that there is a reasonable certainty that no harm will result to the United States population, including infants and children, from aggregate exposure to the residues of P. fumosoroseus strain FE 9901 . This includes all anticipated dietary exposures and all other exposures for which there is reliable

information. The Agency has arrived at this conclusion because th e data and information

available on of P. fumosoroseus strain FE 9901 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 nec essary

.P. fumosoroseus strain FE 9901 page 12 Biopesticides Registration Action Document

D

. Risk Characterization

EPA does not expect harmful effects to humans from consuming food containing

residues of P. fumosoroseus strain FE 9901 (note: exposure is expected through the oral ro ute only; use of NoFly TM WP is limited to greenhouses). Based on the available testing, the acute oral toxicity P. fumosoroseus strain FE 9901 is low. The oral LD 50 is >5,000 mg/kg/ bw (which is the limit dose) and P. fumosoroseus strain FE 9901 is not infective or pathogenic t o rats via the

oral route . Further, the Agency expects any residues in food and drinking water to be low to

nonexistent because of the nature of P. fumosoroseus strain FE 9901 in the environment (i.e., it does not thrive in sunlight or warm temperatures). Therefore, even if some P. fumosoroseus

strain FE 9901 residues are present in food, the lack of acute oral toxicity, infectivity, and pathogenicity support s the establishment of an exemption from the requirement of a tolerance

for P. fumosoroseus strain FE 9901

.
Occupational exposure may occur through the dermal and inhalation routes. P.

fumosoroseus strain FE 9901 is not acutely toxic via the dermal and inhalation route s

— the

dermal LD 50 is 5,050 mg/kg bw (limit dose) and inhalation LC 50 is >2.18 mg/ L ( limit dose). There is concern, however, regarding sensitization. In searching the open literature, EPA noted a

number of hypersensitivity reports and a supplemental toxicity test (see Appendix C: Skin Sensitization) showed that NoFly TM WP is a skin sensitizer. Based on the assumption that most microorganisms contain substances that would elicit allergic reactions, EPA considers P.

fumosoroseus strain FE 9901 to be a potential sensitizing agent

. To mitigate potential

sensitization con cerns, the label requires protective eyewear ; chemical

- resistant gloves ; long

sleeved shirts and long pants and, for mixer/loaders, a dust/mist

- filtering respirator meeting

NIOSH standards of at least N

- 95, R

- 95, or P

- 95.

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 www.epa.gov/compliance/environmentaljustice/ index.html

.

V

. ENVIRONMENTAL ASSESSMENT

A. Risk Characterization

Because of the anticipated use pattern and expected low toxicity

, EPA has not required

enviro nmental effects data for this MPCA

. P. fumosoroseus strain FE 9901 will only be used in greenhouses; therefore, the Agency does not expect that birds, honeybees, and other nontarget organisms such as insects and fish, will be exposed. In a letter to the a pplicant (EPA 2008),

EPA pointed out that if “the intended use sites are restricted to indoors, then neither a honey bee study nor other nontarget assessments are likely to be needed .” For more details, including

a complete list of the Nontarget Organisms a nd Environmental Fate Data requirements under 40 CFR 158.2150, see Table A

- 3 of Appendix A.P. fumosoroseus strain FE 9901 page 13 Biopesticides Registration Action Document

B

. Endangered Species Assessment

EPA expects t hat nontarget organisms will not be exposed to P. fumoso roseus strain FE

9901 . Thus, the Agency has determined that it will have “No Effect ” 4 on any currently listed threatened or endangered species, or any designated critical habitat.

V I

. ENVIRONMENTAL JUSTICE

EPA seeks to achieve environmental justice

— the fair treatment and meaningful involvement of all people, regardless of race, color, national origin, or income

— with respect to the development, implementation, and enforcement of environmental laws, regul ations, 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 P. fumosoroseus strain FE 9901, compared to the general population.

V I I

. RISK MANAGEMENT DECISION

FIFRA Section 3(c)(5) provides for the registration of new active ingredients if it is

de termined that : (1) its composition is such as to warrant the proposed claims for it; (2) its labeling and other materials required to be submitted comply with the requirements of FIFRA ; (3) it will perform its intended function without unreasonable adverse effects on the environment; and (4) 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 for NoFlyTM WP (73314

- A) and NoFlyTM Technical (73314

- T). These products are not expected to cause unreasonable adverse effects, and are likely to provide protection as claimed when used according to label instructions. The refore, the new active ingredient P.

fumosoroseus strain FE 9901 is eligible for registration for the labeled uses. Please note, however, that certain terms of registration must be met.

4 The U.S. Fish and Wildlife Service define s the term “No Effect” to mean that a particular agency determines that “ its proposed

action will not affect a listed species or designated critical habitat” ( www.fws.gov/endangered/esa

- library/pdf/CH1

- 3.PDF )

.P. fumosoroseus strain FE 9901 page 14 Biopesticides Registration Action Document

VI I I

. TERMS OF THE REGISTRATION

EPA requires that the following data requirements be completed within the next several

months.



### Data Requirement Explanation Due

Date

40 CFR 158.2120 (Product Analysis)

Analysis and Certified Limits : Analysis of samples (OPPTS 885.1400):

EP and MP A five

- batch analysis of the EP and MP from the proposed manufacturing site must be submitted.

Physical and Chemical Characteristics

Stability to normal and elevated temperatures,

metals and metal ions (OPPTS 830 .6313): EP : This requirement has been satisfied for the MP ; however, it must be completed for the EP.

Storage stability (OPPTS 830.6317):

EP and MP Regarding the MP , MRID 480514

- 01 reports that this “will be conducted as a condition of registrat ion.” For the EP, the submitted storage stability study (477919

- 02) is inadequate, among other things, it only supports a storage period of three months at 4oC. A storage stability for the EP must be conducted using the proposed formulation of NoFly TM WP from the proposed manufacturing site.

Corrosion Characteristics (OPPTS 830.6320):

EP and MP For both the EP and MP , MRID 480514

- 01 reports that these two studies “will be conducted as a condition of registration.”

40 CFR 158.2140 (Toxicology data)

Tier 1 : Acute injection toxicity/pathogenicity

— intraperitoneal (OPPTS 885.3200): TGAI EPA is requiring that the applicant submit data showing microbial clearance from the body from matrices other than blood (e.g., organs, body fluids).15

BIBL IOGRAPHY

477919

- 00. Natural Industries, Inc. 2009. Submission of Product Chemistry and Toxicity Data in Support of the Application for Registration of NoFlyTM WP and the Petition for Tolerance of Paecilo myces fumosoroseus strain FE 9901, blastospores. Transmittal of 8 Studies. May 7, 2009. Studies, by MRID

477919

- 01. Curry, K.; Brookman, D. 2009. Product Chemistry of N o FLYTM WP. Project Number:

N o FLYTM/PC/002. Unpublished study prepared by Technology Sciences Group, I nc. 64 p. April 29, 2009.

477919

- 02. Bell, A. 2006. Storage and Stability of NoFlyTM WP. Project Number: CEMS/2836. Unpublished study prepared by MGS Laboratories, Ltd., and CEM Analytical Services, Ltd. 14 p. August 22,

2006.

477919

- 03. Kuhn, J. 2 009. Futureco NoFlyTM: Acute Oral Toxicity Study (UDP) in Rats: Final Report. Project Number: 12412/08. Unpublished study prepared by Stillmeadow, Inc. 12 p. January 13,

2009.

477919

- 04. Kuhn, J. 2009. Futureco NoFlyTM: Acute Dermal Toxicity Study in Rats: Final Report. Project Number: 12413/08. Unpublished study prepared by Stillmeadow, Inc. 13 p. January 13, 2009.

477919

- 05. Younger, C. 2009. Futureco NoFlyTM: Acute Inhalation Toxicity Study in Rats: Final Report. Project Number: 12414/08. Unp ublished study prepared by Stillmeadow, Inc. 19 p. January 29,

2009.

477919

- 06. Stahl, J. 2004. NoFlyTM WP Acute Eye Irritation Study (in Rabbits). Project Number: 04/831/005N. Unpublished study prepared by LAB International Research Centre Hungary, Ltd. 22 p. September 2, 2004.

477919

- 07. Stahl, J. 2004. NoFlyTM WP Primary Dermal Irritation Study (in Rabbits). Project Number: 04/831/006N. Unpublished study prepared by LAB International Research Centre Hungary, Ltd. 20 p. September 2, 2004.

477919

- 08. Stahl, J. 2004. NoFlyTM WP Skin Sensitization Study (in Guinea Pig). Project Number: 04/831/104T. Unpublished study prepared by LAB International Research Centre Hungary, Ltd. 34 p. September 27, 2004.

477920

- 00. Natural Industries, Inc. 2009. Submission of Product Chemistry and Toxicity Data in Support of the Application for Registration of NoFlyTM Technical and the Petition for Tolerance of Paecilomyces fumosoroseus Strain FE 9901, Blastospores. Transmittal of 7 Studies. May 7,

2009.

477920

- 01. Brookman, D.; Curry, K. 2009. Product Chemistry of NoFlyTM Technical. Project Number:

N o FLYTM/PC/001. Unpublished study prepared by Technology Sciences Group, Inc. 247 p. May 13, 2009.

477920

- 02. Cerna, Z. 2004. NoFlyTM Technical Acute Oral Toxicity (in Rats). Project Number: 006/04/C. Unpublished study prepared by BioTest, Ltd. 25 p. July 30, 2004.

477920

- 03. Mier, T.; Olivares

- Redonda, G.; Navarro

- Barranco, H.; et al. 2005. Acute Oral Intragastric Pathogenicity and Toxicity in Mice o f Paecilomyces fumosoroseus Isolated from Whiteflies.

Antonie van Leeuwenhoek 88:103

- 111. February 9, 2005.

477920

- 04. Marhan, J. 2004. NoFlyTM Technical Acute Pulmonary Toxicity/Pathogenicity (in Ferrets). Project Number: 004/04/C. Unpublished study prepared by BioTest, Ltd. 26 p. July 30, 2004.

477920

- 05. Cerna, Z. 2004. NoFlyTM Technical Acute Injection Toxicity/Pathogenicity (Intraperitoneal) (in Rats). Project Number: 005/04/C. Unpublished study prepared by BioTest, Ltd. 22 p. July 30,

2004.

477920

- 06. Even, M. 2009. Response to Tier 1 Microbial Pesticide Data Requirements for NoFlyTM

Technical. Unpublished study prepared by Technology Sciences Group, Inc. 41 p. May 7, 2009.16

477920

- 07. Vertesr, A. 2004. NoFlyTM Technical Bacterial Reverse Mutation Assay. Project Number: 04/831/007M. Unpublished study prepared by LAB International Research Cen tre Hungary, Ltd. 41 p. September 2, 2004.

479705

- 00. Natural Industries, Inc. 2010. Submission of Efficacy Data in Support of the Application for Registration of NoFlyTM WP. Transmittal of 1 Study. February 1, 2010.

479705

- 01. Corts, V.; Aucejo, S. 2007. Laboratory Trials to Test the Side Effect of NoFlyTM WP and Adults of Apis Mellifera: One Acute Oral and One Contact Toxicity Trial. Unpublished study prepared by recerca agricola/SynTech Research Spain S.L. 49 p. July 12, 2007.

480514

- 00. Natural Industries, Inc. 2010. Submission of Product Chemistry and Toxicity Data in Support of the Applications for Registration of NoFlyTM Technical and NoFlyTM WP. Transmittal of 3 Studies. April 2, 2010.

480514

- 01. Bjornson, H.; Lara, J. 2010. NoFlyTM Techn ical and NoFlyTM WP Response to PMRA and EPA Clarifications. Unpublished study prepared by Technology Sciences Group, Inc. 18 p. April 2,

2010.

480514

- 02. Bjornson, H. 2010. NoFlyTM Technical and NoFlyTM WP Response to Hypersensitivity Data Requirement. Unpublished study prepared by Technology Sciences Group, Inc. 4 p. April 1,

2010.

480514

- 03. Luangsa, J.; Hyel

- Jones, N.; Manoch, L.; et al. 2005. On the Relationships of Paecilomyces sect. Isarioidea Species. Mycol. Res. 109(5):581

- 589. May 2005.

Brown, A.H.S. and Smith, G. 1957. The genus Paecilomyces Bainier and its perfect stage Byssochlamys Westling. Transactions of the British Mycological Society. 40(1); 17

- 89. R eviews and other References

Catalogue of Life. 2010. Bisby et al., editors. Catalogue of Life: 2010 Annual Checklist. Integrated Taxonomic Information System (ITIS) and Species 2000. Reading, United Kingdom. Information retrieved on August 24,

2010. www.catalogueoflife.org/annual

- checklist/2010

.

1997. Fargues J. et.al. I nactivation of Conidia of Paecilomyces fumosoroseus by Near

- Ultraviolet (UVB and UVA) and Visible Radiation. Journal of Invertebrate Pathology 69, 70

- 78 (1997).

MycoBank. 2010. Species Paecilomyces fumosoroseus (Wize) A.H.S. Br. & G. Sm. 1957; MB30218 8. MycoBank Fungal Databases. Information Retrieved on August 25, 2010 (searching with the term "Paecilomyces

fumosoroseus ” ). www.mycobank.org

.

Padilla, M., Hernández, E., Carnero, A., Amador, S. 2004. Evaluation of the side effects of the Microbial Pest

Control Product FUTURECO NoF LYTM (BCA Paecilomyces fumosoroseus strain FE 9901) to whitefly natural enemies in the Canary Islands. Company Report No NF

- 04001.

PMRA. 2010a. Review of Acute Dermal Toxicity

— Rat for NoFly WP. Sub. No. 2009

- 1803 (WP). Pest Management Regulatory Agenc y (PMRA). Health Canada. Draft Document peer reviewed by Ibrahim Barsoum, U.S. EPA; August 30, 2010. Review of U.S. EPA MRID No. 477919

- 04.

PMRA. 2010b. Review of Acute Dermal Toxicity

—

Waiver Request for NoFly Technical. Sub. No. 2009

- 1802 (Technical) . Pest Management Regulatory Agency (PMRA). Health Canada. Draft Document peer reviewed by Ibrahim Barsoum, U.S. EPA; August 30, 2010. Partial Review of U.S. EPA MRID No. 477920

- 06.

PMRA. 2010c. Review of Acute Inhalation Toxicity

— Rat for NoFly WP. Sub. No. 2009

- 1803 (WP). Pest Management Regulatory Agency (PMRA). Health Canada. Draft Document peer reviewed by Ibrahim Barsoum, U.S. EPA; August 30, 2010. Review of U.S. EPA MRID No. 477919

- 05.

PMRA. 2010d. Review of Acute Inhalation Toxicity

— Wai ver Request for NoFly Technical. Sub. No. 2009

- 1802 (Technical). Pest Management Regulatory Agency (PMRA). Health Canada. Draft Document peer reviewed by Ibrahim Barsoum, U.S. EPA; August 30, 2010. Partial Review of U.S. EPA MRID No. 477920

- 06.17

PMRA. 2010e. Review of Acute Oral Infectivity and Toxicity

— Rat for NoFly Technical. Sub. No. 2009

- 1802 (Technical). Pest Management Regulatory Agency (PMRA). Health Canada. Draft Document peer reviewed by Ibrahim Barsoum, U.S. EPA; August 30, 2010. Review o f U.S. EPA MRID No. 477920

- 02.

PMRA. 2010f. Review of Acute Oral Toxicity

— Rat for NoFly WP. Sub. No. 2009

- 1803 (WP). Pest Management Regulatory Agency (PMRA). Health Canada. Draft Document peer reviewed by Ibrahim Barsoum, U.S. EPA; August 30, 2010. Review of U.S. EPA MRID No. 477919

- 03.

PMRA. 2010g. Review of Acute Pulmonary Infectivity and Toxicity

— Ferret for NoFly Technical. Sub. No. 2009

- 1802 (Technical). Pest Management Regulatory Agency (PMRA). Health Canada. Draft Document peer

reviewed b y Ibrahim Barsoum, U.S. EPA; August 30, 2010. Review of U.S. EPA MRID No. 477920

- 04.

PMRA. 2010h. Review of Acute Pulmonary Infectivity and Toxicity

— Waiver Request for NoFly Technical. Sub.

No. 2009

- 1802 (Technical). Pest Management Regulatory Agency ( PMRA). Health Canada. Draft Document peer reviewed by Ibrahim Barsoum, U.S. EPA; August 30, 2010. Partial Review of U.S. EPA MRID No. 477920

- 06.

PMRA. 2010i. Review of Dermal Irritation

— Rabbit for NoFly WP. Sub. No. 2009

- 1803 (WP). Pest Management

Re gulatory Agency (PMRA). Health Canada. Draft Document peer reviewed by Ibrahim Barsoum, U.S. EPA; August 30, 2010. Review of U.S. EPA MRID No. 477919

- 07.

PMRA. 2010j. Review of Dermal Irritation

— Waiver Request for NoFly Technical. Sub. No. 2009

- 1802 ( Technical). Pest Management Regulatory Agency (PMRA). Health Canada. Draft Document peer reviewed by Ibrahim Barsoum, U.S. EPA; August 30, 2010. Partial Review of U.S. EPA MRID No. 477920

- 06.

PMRA. 2010k. Review of Dermal Sensitization

— Guinea Pigs for NoFly WP. Sub. No. 2009

- 1803 (WP). Pest Management Regulatory Agency (PMRA). Health Canada. Draft Document peer reviewed by Ibrahim Barsoum, U.S. EPA; August 30, 2010. Review of U.S. EPA MRID No. 477919

- 08.

PMRA. 2010l. Review of Dermal Sensitizatio n

— Waiver Request for NoFly Technical. Sub. No. 2009

- 1802 (Technical). Pest Management Regulatory Agency (PMRA). Health Canada. Draft Document peer reviewed by Ibrahim Barsoum, U.S. EPA; August 30, 2010. Partial Review of U.S. EPA MRID No. 477920

- 06.

P MRA. 2010m. Review of Eye Irritation

— Rabbit for NoFly WP. Sub. No. 2009

- 1803 (WP). Pest Management Regulatory Agency (PMRA). Health Canada. Draft Document peer reviewed by Ibrahim Barsoum, U.S. EPA; August 30, 2010. Review of U.S. EPA MRID No. 477919

- 06.

PMRA. 2010n. Review of Eye Irritation

— Waiver Request for NoFly Technical. Sub. No. 2009

- 1802 (Technical). Pest Management Regulatory Agency (PMRA). Health Canada. Draft Document peer reviewed by Ibrahim Barsoum, U.S. EPA; August 30, 2010. Partia l Review of U.S. EPA MRID No. 477920

- 06.

PMRA. 2010o. Review of Genotoxicity

— Bacterial Reverse Mutation Assay for NoFly Technical. Sub. No. 2009

- 1802 (Technical). Pest Management Regulatory Agency (PMRA). Health Canada. Draft Document peer reviewed by Ibrahim Barsoum, U.S. EPA; August 30, 2010. Review of U.S. EPA MRID No. 477920

- 07.

PMRA. 2010p. Review of Intraperitoneal Infectivity

— Rat for NoFly Technical. Sub. No. 2009

- 1802 (Technical). Pest Management Regulatory Agency (PMRA). Health Canada. D raft Document peer reviewed by Ibrahim Barsoum, U.S. EPA; August 30, 2010. Review of U.S. EPA MRID No. 477920

- 05.

PMRA. 2010q. Review of Product Characterization and Analysis for NoFly WP and NoFly Technical. Sub. No.

2009

- 1803 (WP) and Sub. No. 2009

- 1 802 (Technical). Pest Management Regulator y Agency (PMRA). Health

Canada . Draft Document peer reviewed by Ibrahim Barsoum, U.S. EPA; August 30, 2010. Review of U.S. EPA

MRID No.’s 477919

- 01, 477919

- 02; 477920

- 01; 480514

- 03.

PMRA. 2010r. Review of Re porting of Hypersensitivity Incidence for NoFly Technical. Sub. No. 2009

- 1802 (Technical). Pest Management Regulatory Agency (PMRA). Health Canada. Draft Document peer reviewed by Ibrahim Barsoum, U.S. EPA; August 30, 2010. Partial Review of U.S. EPA MRID No. 477920

- 06.

PMRA. 2010s. Review of Tissue Culture

— Waiver Request for NoFly Technical. Sub.No.2009

- 1802 (Technical). Pest Management Regulatory Agency (PMRA). Health Canada. Draft Document peer reviewed by Ibrahim Barsoum, U.S. EPA; August 30, 2010. Partial Review of U.S. EPA MRID No. 477920

- 06

.18

TSG. 2009a. Letter from Heather Bjornson, Technology Sciences Group Inc. (TSG) on behalf of Natural Industries, Inc. to Sheryl Reilly, U.S. Environmental Protection Ag ency. "NoFlyTM Technical

— Application for new product registration...." May 7, 2009

.

TSG. 2009b. Letter from Heather Bjornson, Technology Sciences Group Inc. (TSG) on behalf of Natural Industries, Inc. to Sheryl Reilly, U.S. Environmental Protection Agency. "NoFlyTM WP

—Application for new product registration...." May 7, 2009

.
TSG. 2009c. Letter from Heather Bjornson, Technology Sciences Group Inc. (TSG) on behalf of Natural Industries, Inc. to Sheryl Reilly, U.S. Environmental Protection Agency. “Petition for an exemption from the requirement of a tolerance for residues of products containing the active ingredient “Paecilomyces fumosoroseus strain FE 9901” in or on all food commodities. May 7, 2009.

TSG. 2010. Letter from Heather R. Bjornson, Regulatory Consultant to Natural Industries to Sheryl Rei lly, U.S. EPA. “Re: Submission of Information to Support the Application to Register NoFlyTM Technical (EPA File Symbol:

73314

- L) and NoFlyTM WP (EPA File Symbol: 73314

- A).” March 2, 2010.

U.S. EPA. 2008. Letter from Sheryl Reilly, U.S. Environmental P rotection Agency to Heather Bjornson, Technology Sciences Group Inc. (TSG). Biopesticides and Pollution Prevention Division (BPPD) response to summary of the January 22, 2008 pre

- registration meeting to discuss registration of a new microbial active ingre dient, Paecilomyces fumosoroseus strain FE 9901 blastospores. September 3, 2008.

U.S. EPA. 2010. Memorandum from Ibrahim Barsoum to Kathleen Martin. Joint Review of data for the registration of a new TGAI, NoFly Technical, and a new end use product fo rmulation, NoFly Wettable Powder, containing the fungus Paecilomyces fumosoroseus strain FE 9001. September 15, 2010.A

- 1 

## A PPENDIX A: Data Requirements and How t he Applicant Met Them

In reviewing its data, EPA’s Office of Pesticide Programs (OPP) classifies each study as

either ‘Acceptable

, ’ Supplemental ,’ ‘Supplemental: Upgradable,’ or ‘Unacceptable.’ An Acceptable study is one that the Agency concludes is scientifically sound and is useful for risk

assessment. A Supplemental study is one that may provide some information that can be

useful in risk assessment ; o ften a study is classified as Supplemental because it was not required under 40 CFR 158. A ‘ Supplemental: Upgr adable ’ study is one that has the potential to be upgraded to Acceptable, provided certain addition information is submitted. Finally, an Unacceptable study is one that is not useful for risk assessment purposes.

Product Analysis

. Provided in Table A

- 1 is a summary of the Product Analysis d ata

r equirements for m icrobial pesticides (40 CFR 158.2120) with information regarding how the requirement was met. Note that four requirements (Analysis of samples ; Stability to normal and elevated temperatures ; meta ls and metal ions ; Storage stability ; and Corrosion characteristics) need to be completed as terms of registration

.

Table A

- 1

. Product Analysis Data Requirements for Paecilomyces fumosoroseus strain FE 9901

OPPTS

Guideline

Number Data Requirement How th e Requirement Was Met (U.S. EPA 2010)

End

- Use Product (73314

- A) Manufacturing

- Use Product (73314

- T)

MRID Conclusion, Review, Notes MRID Conclusion, Review, Notes

Product Chemistry and Composition

885.1100 Product Identity 477919

- 01 Accepta ble study 477920

- 01 Acceptable study

885.1200 Manufacturing Process 477919

- 01 Acceptable study 477920

- 01 Acceptable study

Deposition of a sample in a nationally recognized culture collection A pure culture of P. fumosoroseus strain FE 9901 was deposite d

.

885.1300 Discussion of formation of unintentional ingredients 477919

- 01 Acceptable study 477920

- 01 Acceptable study

Analysis and Certified Limits

885.1400 Analysis of samples 477919

- 01 Supplemental: pgradable 1 477920

- 01 Supplemental: Upgradable 1

88 5.1500 Certification of limits 477919

- 01 Acceptable study 477920

- 01 Acceptable study

Physical and Chemical Characteristics

830.6302 Color 477919

- 01 Acceptable study 477920

- 01 Acceptable study

830.6303 Physical state 477919

- 01 Acceptable study 477920

- 01 Acceptable study

830.6304 Odor 477919

- 01 Acceptable study 477920

- 01 Acceptable study

830.6313 Stability to normal and elevated temperatures, metals and metal ions Term of registration 480514

- 01 Acceptable study

830.6317 Storage stability 477919

- 02 Unacc eptable study 2 Term of registration

830.6319 Miscibility Only required for emulsifiable liquid forms of microbial pesticides. Because this product is a solid, this requirement does not apply.

830.6320 Corrosion characteristics Term of registration

830 .7000 pH 477919

- 01 Acceptable study 477920

- 01 Acceptable study

830.7100 Viscosity Only required for liquid forms of microbial pesticides. Because this product is a solid, this requirement does not apply.

830.7300 Bulk density 477919

- 01 Acceptable study 477920

- 01 Acceptable study

1 A five

- batch analysis of EP and MP from the proposed manufacturing site must be submitted as a term of registration.

2 As a term of registration, the applicant must conduct a storage stability study using the proposed formulat ion of NoFlyTM WP from the proposed manufacturing site.A

- 2 Toxicology

. Provided in Table A

- 2 is a summary of the Tier I Toxicology Data Requirements (40 CFR 158.2140) along with an explanation regarding how the requirement was met. Toxicology testing was limited to Tier I because the acute toxicity of P. fumosoroseus strain FE 9901 did not trigger the Tier II or III studies.

Table A

- 2 . Tier I Toxicology Data Requirements for Paecilomyces fumosoroseus strain FE 9901

OPPTS

Guideline

Number Data Requir ement How the Requirement Was Met (U.S. EPA 2010)

End

- Use Product (73314

- A) Manufacturing

- Use Product (73314

- T)

MRID Conclusion, Review, Notes MRID Conclusion, Review, Notes

870.1100 Acute oral toxicity 477919

- 03 Acceptable study 477920

- 02 Acceptabl e study

870.1200 Acute dermal toxicity 477919

- 04 Acceptable study 477920

- 06 Acceptable data waiver;

PMRA 2010b

870.1300 Acute inhalation toxicity 477919

- 05 Acceptable study 477920

- 06 Acceptable data waiver;

PMRA 2010d

870.2400 Acute eye irritation 477 919

- 06 Acceptable study 477920

- 06 Acceptable data waiver;

PMRA 2010n

870.2500 Primary dermal irritation 477919

- 07 Acceptable study 477920

- 06 Acceptable data waiver; PMRA 2010j

885.3050 Acute oral toxicity/ pathogenicity TGAI: 477920

- 02; 477920

- 03; 48 0514

- 01. Acceptable study

885.3150 Acute pulmonary toxicity/pathogenicity TGAI: 477920

- 04; 477920

- 06. Acceptable study.

885.3200 Acute injection toxicity/pathogenicity TGAI: 477920

- 05. Acceptable study; however, additional information is needed. 1

885.3400 Hypersensitivity

incidents 480514

- 02 Acceptable statement 480514

- 02 Acceptable statement

885.3500 Cell culture Data must be submitted only for products whose active ingredient is a virus; P. fumosoroseus is a fungus, therefore this requirement do es not apply.

1 As a term of registration, EPA requires that the applicant submit data showing microbial clearance from matrices other than blood (e.g., organs, body fluids).

Environmental Fate and Eco toxicity

. Provided in Table A

- 3 are explanation s for why environmental fate and ecotoxicity data were not required under 40 CFR 158.2150

.

Table A

- 3

. Tier I Nontarget Organisms and Enviro nmental Fate Data Requirements for Paecilomyces

fumosoroseus strain FE 9901

OPPTS

Guideline

Number Data Requirem ent How the Requirement Was Met (U.S. EPA 2010)

885.4050 Avian oral toxicity Tests for pesticides intended solely for indoor application would be required on

a case

- by

- case basis... In a letter to the applicant , EPA pointed out that if “the

intended use si tes are restricted to indoors, then neither a honey bee study nor

other non

- target assessments are likely to be needed” (EPA 2008). 885.4100 Avian inhalation toxicity/pathogenicity

885.4150 Wild mammal toxicity/pathogenicity NR 1 for greenhouse uses.

88 5.4200 Freshwater fish toxicity/ pathogenicity Tests for pesticides intended solely for indoor application would be required on

a case

- by

- case basis... In a letter to the applicant , EPA pointed out that if “the intended use sites are restricted to indoors, then neither a honey bee study nor

other non

- target assessments are likely to be needed” (EPA 2008). 885.4240 Freshwater invertebrate toxicity/ pathogenicity

885.4280 Estuarine/Marine fish testing; estuarine and marine invertebrate testing NR 1 for green house uses.

885.4300 Nontarget plant testing NR 1 for greenhouse uses.

885.4340 Nontarget insect testing Tests for pesticides intended solely for indoor application would be required on

a case

- by

- case basis... In a letter to the applicant , EPA pointed out that if “the intended use sites are restricted to indoors, then neither a honey bee study nor

other non

- target assessments are likely to be needed” (EPA 2008). 885.4380 Honey bee testing

1 NR=not required under 40 CFR 158B

- 1 

## APPENDIX B : Biology and Taxon omy of

P. fumosoroseus and strain FE 9901

Provided below is a summary of the biologic and taxonomic considerations for P.

fumosoroseus in general and strain FE 9901 in particular. Unless otherwise noted, the source of the information is from the joint review (U.S. EPA 2010) of the data supported to support the registration of this new active ingredient.

A. Origin, Derivation, and Identification of **Paecilomyces fumosoroseus** 

Syno nyms

Isaria fumosorosea (basionym; first described by Wize in 1904), . P. fumosoroseus , a mycoinsecticide, was first described by Wize in 1904 (Brown and Smith 1957). Synonyms (basionym, obligate, and taxonomic) for this fungus include (MycoBank 2010 , U.S. EPA 2010 ):

Spicaria fumosorosea (obligate synonym; Wize),

Spicaria aphodii (Vuillemin in 1910),

Penicillium aphodii (Vuillemin),

Monilia aquatilis (Malguth, in 1928),

Paecilomyces hibernicus (Kennelly & Grimes, in 1930), and

Paecilomyces isarioid es (Inagaki, in 1962).

Morphology and Growth

T he fungus produces blastospores in liquid media and conidia on solid media

. In synthetic media, growth is moderatel y rapid when incubated at 24 o C (75.2 oF) . In testing using

P. fumosoroseus strain FE 9901, investigators found that this MPCA grows best at 24.5 o C (76.1 o F). At higher temperatures ( 26.5oC and 30oC , or 79.7 o F and 86.0 o F) g rowth was significantly reduced an d at 35oC or 37oC (95.0 o F or 98.6 o F), it is unable to grow. In addition, P.

fumosoroseus strain FE 9901 was unable to resume growth after incubation at 37oC (98.6 o F) and subsequent incubation at room temperature, indicating that the organism is killed at 37oC (98.6 o F). Colonies reach a diameter of 25 to 30 mm after seven days and 50 to 65 mm after 14 days. P. fumosoroseus has an asexual life cycle

. However, exposure to ultraviolet light results in formation of auxotrophic mutants

, suggest ing that a para sexual life cycle also exists for the organism. P. fumosoroseus strain FE 9901 does not exhibit any atypical morphological, physiological, biochemical, p esticidal, or resistance characteristics. . When grown on potato dextrose agar, P. fumosoroseus strain FE 9901 is floccose (i.e., it is covered with tufts of soft hair) with a circular margin

. Colonies are white, often turning pale yellow from the margin out and changing to brownish

- gray when forming conidia. Conidia are spores and they are supported on specialized spore

- producing hyphae (stalk) known as conidiophores. The conidiophores are erect and can be about 100 μm in length and 1.5 to 2.0 μm in width (MycoBank 2010). The conidial structures consist of single phialides or in whorls occurring irregularly along aerial hyphae.

Taxonomy

Kingdom: Fungi . The taxonomic classification P. fumosoroseus i s provided below (Catalog of Life 2010). For a detailed explanation of the taxonomic designation, refer to the product

characterization data review (U.S. EPA 2010)

.

Phylum: Ascomycota

Class: Eurotiomycetes

Order: EurotialesB

- 2 F amily: Trichocomaceae

Genus: Paecilomyces
Species: fumosoroseus

Strain: FE 9901

The strain FE 9901 is a wild type

— it is neither a spontaneous nor an induced mutant and it has not been genetically modified. P. fumosoroseus strain FE 9901 was fi rst isolated from an adult

of Bemisia tabaci (a species of whitefly) in Padappai, India in 1992.

Identification and characterization of P. fumosoroseus strain FE 9901 has bee n made using BioLog FF MicroPlateTM. Growth was observed on a variety of solid media agar formulations (e.g., Sabouraud dextrose, MacConkey, and chitin) over a wide range of pH values. Molecular fingerprints of P. fumosoroseus strain FE 9901 were compared with those of

three other P. fumosoroseus isolates; two techniques were used: variable number tandem repeat and inter

- simple sequence repeat. Other technologies available to identify and

characterize the microbe include rDNA

- ITS sequencing and RAPD

- PCR (Random Amplification of Polymorphic DNA polymerase chain reaction).

B. Biological Properties

Natural Occurrence

. P. fumosoroseus is a naturally occurring fungus . It is found in most countries of the world and occurs in various soil types

, at very lo w densities

. The fungus has frequently been isolated from lepidopteran cadavers in Europe. Isolates occur in the Netherlands, Ireland, and Spain. The fungus has also been found in various non

- European countries such as China, Japan, Ghana, Pakistan, Ind ia, and the United States.

Target Organisms . NoFly TM WP is intended to control all stages of whitefly development (i.e., eggs, N1, N2, N3 and N4 larvae, and adults)

. The ability to infect whitefly eggs is unique among entomopathogenic fungi

.  Nontarget insect hosts include mealy bugs, beetles, caterpillar s, and flies.

Means of Infection

The infection cycle of P. fumosoroseus strain FE 9901 is particularly rapid under optimal conditions. Initially, conidia attach to th e cuticle or the dorsum of immature or adult whiteflies; first symptoms of infection typically are apparent within one to two days after contact. Conidia and hyphae are present in the host hemocoel (body cavity) within one day. Fungal multiplication take s place through formation of hyphal bodies or blastospores. The mycelium (a branching network of hyphae) is present on the dorsum of the insect body within two days. Sporulation occurs within three days and reaches a maximum within five to seven days. . In its natural environment, P. fumosoroseus strain FE 9901 is dispersed as conidia or blastospores, either by air or water movement or by other insects and/or

mites

. The typical mode of infection of the various entomo pathogenic fungi occurs via conidial attachment, germination, penetration, vegetative growth, and then conidiogenesis.

I nvestigators evaluated under laboratory conditions the ability of P. fumosoroseus strain FE 9901 to produce infections on eggs of two novel whitefly species. The strain produced infections on the N1 and N4 nymph stages and significantly reduced the hatchi ng capacity of eggs of Lecanoides floccissimus . It also produced infections on Aleurodicus dispersus N1 and N4 nymph stages and reduced the percent of hatching of eggs by 50 percent.

 An entomopathogenic fungus is one that can act as a parasite of insects and kills or seriously disables them.C

- 1 

## APPENDIX C : Summary of Submitted Toxicity Studies

Oral (OPPTS 870.1100 ; MRID 477919

- 03 ). F ollowing the Up

- and

- Down Method ( OECD Guideline 425), three female Sprague

- Dawley rats were gavaged with P. fumosoroseus strain

FE 9901 (containing containing 2×10 9 colon y forming units per gram, or CFU /g) at a dose of 5,000 mg/kg bw

. Investigators terminated the study following the stopping rules for this procedure. Each animal was then observed for a period of up to 14 days. None of the animals died or showed any symp toms of intoxication following treatment. The test material had no negative effect upon the body weight

, and no gross pathological changes were noted. Based on the results of this study, the oral LD 50 is > 5

, 000 mg/kg bw (w hich is the limit dose).

Derm al (OPPTS 870.1200 ; MRID 477919

- 04 ). A group of 7

- week

- old Sprague

- Dawley rats (five/sex) was exposed dermally to P. fumosoroseus strain FE 9901 ( containing 2×10 9

CFU /g ) at a limit dose of 5,050 mg/kg bw for 24 hours to an area of not less than 10% of the total body surface. Following exposure, the animals were observed for a period of 14 days. No

mortalities were observed

, and all the test animals appeared normal for the duration of the study. Very slight erythema and discoloration was observed in one f emale. At necropsy, two male rats had pale lungs at study termination. Based on these results, the dermal LD 50 is 5,050 mg/kg bw.

Inhalation (OPPTS 870.1300 ; MRID 477919

- 05 ). A group of 10

- week

- old Sprague

Dawley rats (five/sex) was exposed via the inh alation route to P. fumosoroseus strain FE 9901 (containing 2×10 9 CFU /g ) for four hours to nose only at a concentration of 2.18 mg/L . Animals then were observed for 14 days. No mortalities were observed; clinical observations included piloerection and de creases in activity which both resolved by Day 3. All test animals gained body weight throughout the study period

, and no observable abnormalities were observed at gross necropsy. Based on the findings of this study, the LC 50 is > 2.18 mg/L , which is the limit

dose.

Eye Irritation (OPPTS 870.2400 ; MRID 477919

- 06 ). In a primary eye irritation study, 0.1g of NoFly TM WP (containing 88% P. fumosoroseus strain FE 9901) was instilled into the conjunctival sac of the left eye of three adult male New Zealand W h ite rabbits. The treated eyes were not washed following application. The untreated right eye of each test animal served as a negative control

. Test animals then were observed for 72 hours. Irritation was scored by the Draize Method. After one hour, so me redness was observed in one rabbit, and slight to moderate increases in excretion discharge were noted in three rabbits. All test animals were

free of irritation at the 24

- , 48

- and 72

- hour observation timepoints. Based on the results of this study, N oFly TM WP is slightly irritating to the eyes of New Zealand White rabbits.

Primary Dermal Irritation (OPPTS 870.2500 ; MRID 477919

- 07 ). In a primary dermal irritation study, three adult male New Zealand W hite rabbit s were dermally exposed to 0.5g NoFly TM WP (containing 88% P. fumosoroseus strain FE 9901) in water for four hours to an area of skin of approximately six cm 2 . Animals then were observed for 72 hours. Irritation was scored by the method of Draize. Very slight erythema was noted in two test anim als one hour after patch removal. All test animals were free of irritation at the 24

- , 48

- and 72

- hour observation timepoints. Based on the results of this study, NoFly TM WP is slightly irritating to

the skin of male New Zealand White rabbits.C

- 2 Oral toxicity/pathogenicity (OPPTS 885.3050 ; MRID 47720

- 02 ). In an acute oral toxicity/ pathogenicity study, groups of fasted, young Wistar rats (15/sex) were given NoFly TM

Techn ical ( 1.0×10 9 CFU/g) in a single dose of 100 mg of test material per animal, each dose

contained 10 8 CFU. The animals were then observed for a period of up to 21 days with scheduled sacrifices 24 hours after application and on Days 3, 7, 14 and 21. None of the

animals died

, and no clinical observations were noted during the study period

. No gross pathological changes were noted in sacrificed test animals

. Based on the findings of this study,

P. fumosoroseus strain FE 9901 is not considered infective or pathogenic in the rat. In terms of

an LD 50 , the animals showed no signs of toxicity at a dose of 600 mg/kg bw. 5

Pulmonary toxicity/pathogenicity (OPPTS 885.3150 ; MRID 47720

- 04 ). In an acute pulmonary infectivity and toxicity study, groups of young adul t ferrets (14/sex) were exposed by

the intranasal route to NoFly TM Technical at a nominal dose of 1×10 8 CFU (in 0.5 mL) per animal. Animals were then observed for up to 21 days

. None of the animals died or showed any symptoms of intoxication following treatment. No gross pa thological changes were noted. Microbiologi cal examination of lungs, other prescribed organs, and body fluids proved minimal persistence and no infectivity of the test material. Based on the results of this study, P.

fumosoroseus strain FE 9901 is not considered infective or pathogenic in the ferr et. Please note that the rat is the preferred species for acute pulmonary toxicity.pathogenicity testing. For this MPCA, the applicant provided acceptable justification for using ferrets instead of rats.

Acute injection toxicity/pathogenicity

— intraper itoneal (OPPTS 885.3200; MIRD 47720

- 05). In an acute intraperitoneal infectivity study, a group of young adult Wistar rats (three/sex) were injected with NoFly TM Technical ( 1.0 × 10 9 CFU/g) in a single dose of 107 CFU per animal

. Animals were then observed for up to 21 days. None of the animals died or showed any symptoms of intoxication after treatment. The test item had no negative effect upon the body weight

, and no gross pathological changes were noted. Blood samples were collected during necropsy to evaluate clearance. Based on the available results from the study, P.

fumosoroseus strain FE 9901 is not considered infective or pathogenic in the rat via the intraperitoneal route of exposure. As a term of registration, EPA is requiring t hat the applica nt submit data showing microbial clearance from matrices other than blood (e.g., organs, body fluids).

Hypersensitivity incidents (OPPTS 885.3400 ; MRID 480514

- 02 ). The applicant stated reported that no hypersensitivity incidents have been reported thro ugh the research, development, and testing of NoFly TM Technical or WP . However, the EP has been found to be a dermal sensitizer (U.S. EPA 2010). In addition, a literature search of the PubMed database www.ncbi.nlm.nih.gov/pubmed ) using the keywords “paecilomyces” and “allergy” revealed 21 reports of hyper sensitivity

. Based on the assumption that most microorganisms contain substances that would elicit allergic reactions, P. fumosoroseus strain FE 9901 is considered a potential sensitizing agent.

Supplemental Toxicity Test

Skin Sensitization

5 . In a skin sensitization test ( 477 9 19

- 08 ) using the Magnusson and Kligman M ethod , female guinea pigs were exposed to NoFly TM WP that was administered as a
1% intradermal injection and a 25% topical application. Later, a challenge dose was applied ( 25% topical applic ation ). Study investigators observed a localized area of intense erythema and swelling in treated animals. EPA concluded that NoFly TM WP is a skin sensitizer.

Technical NoFlyTM mg 600 bw kg 0.163 Technical

NoFlyTM mg 100

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