BIOPESTICIDES REGISTRATION

ACTION DOCUMENT

Paecilomyces fumosoroseus strain FE 9901

DRAFT DOCUMENT

U.S. ENVIRONMENTAL PROTECTION AGENCY

OFFICE OF PESTICIDE PROGRAMS

BIOPESTICIDES AND POLLUTION PREVENTION DIVISION

October 5, 2010
ii

TABLE OF CONTENTS

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 Composition .................................................................... 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. Hypersensitivity Incidents ................................................................................ 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 Met 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 99019
iii

BIOPESTICIDES REGISTRATION ACTION DOCUMENT (BRAD) TEAM

Office of Pesticide Programs

Biopesticides and Pollution Prevention Division

Microbial Pesticides Branch

SCIENCE REVIEW

Ibrahim Barsoum Ph.D. Product Chemistry and 

Human Health Risk 

John Kough, Ph.D. Secondary Review of Product 

Chemistry and 

Human Health Risk

Gail Tomimatsu, Ph.D. Environmental and 

Ecological Risk 

REGULATORY

Kathleen Martin, M.S.E.S., M.P.A. Regulatory Action Leader

Sheryl Reilly, Ph.D. Chief, Microbial Pesticides Branch
iv

ACRONYMS

Acronym 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 Committee 

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 cubic centimeter

LC50 Median Lethal Concentration

LD50 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 Free 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, Pesticides and Toxic Substances; 

now 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 ingredient

U.S. United States

UV Ultraviolet radiation

USDA U.S. Department of Agriculture

WP Wettable powder
P. 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 containing the new active ingredient Paecilomyces 

fumosoroseus strain FE 9901. The manufacturing-use product (MUP) is NoFlyTM Technical 

(73314-T); it contains 69.0% active ingredient. The end-use product (EP) is NoFlyTM 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 control whiteflies, aphids, 

thrips, psyllids, mealybugs, leafhoppers, plant bugs, weevils, grasshoppers, Mormon crickets, 

locust, and beetles. Crops include ornamentals, vegetables, and herbs. The applicant has also 

petitioned for an exemption from the requirement of tolerance for any P.fumosoroseus strain FE 

9901residues 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 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 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. To submit a comment, see the 

instructions 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 Registration

A Notice of Receipt, announcing EPA’s receipt of Natural Industries Incorporated’s 

applications 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, Mormon 

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. Petition 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 Agency to establish an exemption from the requirement of a 

tolerance for residues of P.fumosoroseus strain 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 docket 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 aggregate 

exposure to the residues of P.fumosoroseus strain 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. fumosoroseus strain 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 CONSIDERATIONS

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 stalk) known as 

conidiophores. As with other “entomopathogenic fungi,”1

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

conditions. Initially, conidia attach to the 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. 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. 

NoFlyTM 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., production of more conidia). In laboratory testing, 

investigators determined that P. fumosoroseus strain FE 9901 grows best at (76.1o F). At 

higher temperatures (79.7o F to 86.0o F), growth is significantly reduced and at temperatures 

greater than 95.0o F, P. fumosoroseus strain FE 9901 will no longer grow. 

2

P. fumosoroseus strain FE 9901 does not exhibit any atypical morphological, 

physiological, biochemical, pesticidal, or resistance characteristics. For more information on the 

biology and taxonomy of P. fumosoroseus in general, and P. fumosoroseus strain FE 

9901specifically, 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.

III. ACTIVE INGREDIENT CHARACTERIZATION (Product Analysis—40 CFR 158.2120)

Most of the data requirements under 40 CFR 158.2120 (Product Analysis3

 1

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

) have been 

satisfied. One of the requirements under Analysis and Certified 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 VII: Terms of the 

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. For a complete tabulation of all the Product Analysis data requirements, see Table 

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

A. Product Chemistry and Composition

1. Manufacturing Process (OPPTS 885.1200)

No FlyTM Technical (MUP) and NoFlyTM 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 sample 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 (OPPTS 885.1300)

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

concern have been identified for NoFlyTM WP or NoFlyTM Technical. This data 

requirement has been adequately satisfied. 

B. Physical and 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 (NoFlyTM WP) MUP (NoFlyTM 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; 

NoFlyTM 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/cc
P. fumosoroseus strain FE 9901 page 8

Biopesticides Registration Action Document

IV. 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 

information 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 toxicity 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 variability 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 special 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- and manufacturinguse 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 study but EPA requests additional information),

all the toxicology data requirements have been satisfied; see Section VII: Terms of the 

Registration 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

strain FE 9901 (40 CFR 158.2140)

OPPTS 

Guideline 

Number

Data 

Requirement

Toxicity and Category1

EP (NoFlyTM WP) MUP (NoFlyTM Technical)

870.1100 Acute oral toxicity >5,000 mg/kg bw 

Category IV

600 mg/kg bw

Category III

870.1200 Acute dermal toxicity 5,050 mg/kg bw 

Category IV

Data waived

870.1300 Acute inhalation 

toxicity

>2.18 mg/L 

Category III

Data waived

870.2400 Acute eye irritation some 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 injection 

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 156.62. 

2. Hypersensitivity Incidents (OPPTS 885.3400; MRID 480514-02)

The applicant reports that they have not seen any incidents of hypersensitivity 

throughout the research, development, and testing of NoFlyTM 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 hypersensitivity. 

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 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.’’ Based on the available toxicity 

information, EPA has not found P. fumosoroseus strain FE 9901 to share a common 

mechanism of toxicity with any other substances, and P. fumosoroseus strain FE 

9901does 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 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.” 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 endocrinerelated 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.

P. fumosoroseus strain FE 9901is 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: 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 treated 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 exposure to P. fumosoroseus strain FE 9901 residues 

through consumption of treated vegetables and herbs to be low. P. 

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

80 o F, growth is significantly reduced and at 95 o F, it does not 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 9901in 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. They 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 environments” 

(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. Individuals in residential, schools, 

and day care settings will not be exposed to residues of P. fumosoroseus strain 

FE 9901because 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 this pesticide. Dermal exposure is expected individuals who work in the 

greenhouse following application. 

C. Determination Of Safety For U.S. Population, Infants, and 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 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 EPA 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 EPA 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 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 available 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 the 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 necessary. 
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 route 

only; use of NoFlyTM WP is limited to greenhouses). Based on the available testing, the acute 

oral toxicity P. fumosoroseus strain FE 9901 is low. The oral LD50 is >5,000 mg/kg/ bw (which is 

the limit dose) and P. fumosoroseus strain FE 9901is not infective or pathogenic to 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 supports 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 routes—the 

dermal LD50 is 5,050 mg/kg bw (limit dose) and inhalation LC50 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 NoFlyTM 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 concerns, the label requires protective eyewear; chemical-resistant gloves; longsleeved 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 

environmental effects data for this MPCA. P. fumosoroseus strain FE 9901will 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 applicant (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 and 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 that nontarget organisms will not be exposed to P. fumosoroseus 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. 

VI. ENVIRONMENTAL JUSTICE

EPA seeks to achieve environmental justice—the fair treatment and meaningful 

involvement of all people, regardless of race, color, national origin, or income— 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 P. fumosoroseus strain FE 9901, 

compared to the general population. 

VII. RISK MANAGEMENT DECISION

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

determined 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. Therefore, 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 defines 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

VIII. 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 registration.” 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 NoFlyTM 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

BIBLIOGRAPHY 

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 Paecilomyces 

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 NoFLYTM WP. Project Number: 

NoFLYTM/PC/002. Unpublished study prepared by Technology Sciences Group, Inc. 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. 2009. 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. Unpublished 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: 

NoFLYTM/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 of 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 Centre 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 Technical 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. 

Reviews 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. Inactivation 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; MB302188. 

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 NoFLYTM (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 Agency (PMRA). Health Canada. Draft Document peer reviewed by Ibrahim Barsoum, 

U.S. EPA; August 30, 2010. Review of U.S. EPA MRID No. 477919-04.

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—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.
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 of 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 by 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 

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. 477919-07.

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 Sensitization—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. 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. Partial 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. Draft 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. 2009-1802 (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.’s 477919-01, 477919-02; 477920-01; 480514-03. 

PMRA. 2010r. Review of Reporting 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 Agency. "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 Reilly, 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 Protection 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 

ingredient, 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 formulation, NoFly Wettable Powder, 

containing the fungus Paecilomyces fumosoroseus strain FE 9001. September 15, 2010.
A-1

APPENDIX A: Data Requirements and How the 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; often a study is classified as Supplemental because it was not 

required under 40 CFR 158. A ‘Supplemental: Upgradable’ 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 data 

requirements for microbial 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; metals 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 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

Product Chemistry and Composition

885.1100 Product Identity 477919-01 Acceptable 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 deposited.

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: pgradable1 477920-01 Supplemental: Upgradable1

885.1500 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 Unacceptable study2 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 formulation 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 

Requirement

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 Acceptable 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 477919-06 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; 480514-01. 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 does 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 Ecotoxicity. Provided in Table A-3 are explanations for why 

environmental fate and ecotoxicity data were not required under 40 CFR 158.2150. 

Table A-3. Tier I Nontarget Organisms and Environmental Fate Data Requirements for Paecilomyces 

fumosoroseus strain FE 9901 

OPPTS 

Guideline 

Number

Data Requirement 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 sites 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 NR1 for greenhouse uses.

885.4200 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

NR1 for greenhouse uses.

885.4300 Nontarget plant testing NR1 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 158
B-1

APPENDIX B: Biology and Taxonomy 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

Synonyms

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 isarioides (Inagaki, in 1962). 

Morphology and Growth

The fungus produces blastospores in liquid media and conidia on solid media. In 

synthetic media, growth is moderately rapid when incubated at 24oC (75.2oF). In testing using 

P. fumosoroseus strain FE 9901, investigators found that this MPCA grows best at 24.5oC

(76.1oF). At higher temperatures (26.5oC and 30oC, or 79.7oF and 86.0oF) growth was 

significantly reduced and at 35oC or 37oC (95.0oF or 98.6oF), it is unable to grow. In addition, P. 

fumosoroseus strain FE 9901 was unable to resume growth after incubation at 37oC (98.6oF) 

and subsequent incubation at room temperature, indicating that the organism is killed at 37oC

(98.6oF). 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, suggesting that a parasexual life cycle also exists for the 

organism. P. fumosoroseus strain FE 9901 does not exhibit any atypical morphological, 

physiological, biochemical, pesticidal, 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 is 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: Eurotiales
B-2

Family: 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 first 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 been 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 low 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, India, and the United States. 

Target Organisms. NoFlyTM 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, 

caterpillars, and flies. 

Means of Infection

The infection cycle of P. fumosoroseus strain FE 9901 is particularly rapid under optimal 

conditions. Initially, conidia attach to the 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 takes 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 entomopathogenic fungi occurs via conidial 

attachment, germination, penetration, vegetative growth, and then conidiogenesis. 

Investigators 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 hatching 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). Following the Up-and-Down Method (OECD

Guideline 425), three female Sprague-Dawley rats were gavaged with P. fumosoroseus strain 

FE 9901(containing containing 2×109 colony 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 symptoms 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 LD50 is >5,000 mg/kg bw (which is the limit dose). 

Dermal (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 2×109

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 female. At necropsy, two 

male rats had pale lungs at study termination. Based on these results, the dermal LD50 is 5,050 

mg/kg bw.

Inhalation (OPPTS 870.1300; MRID 477919-05). A group of 10-week-old SpragueDawley rats (five/sex) was exposed via the inhalation route to P. fumosoroseus strain FE 9901 

(containing 2×109 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 decreases 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 LC50 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 NoFlyTM WP (containing 88% P. fumosoroseus strain FE 9901) was instilled into the 

conjunctival sac of the left eye of three adult male New Zealand White 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, some 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, NoFlyTM 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 White rabbits were dermally exposed to 0.5g

NoFlyTM WP (containing 88% P. fumosoroseus strain FE 9901) in water for four hours to an 

area of skin of approximately six cm2

. Animals then were observed for 72 hours. Irritation was 

scored by the method of Draize. Very slight erythema was noted in two test animals 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, NoFlyTM 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 NoFlyTM

Technical (1.0×109 CFU/g) in a single dose of 100 mg of test material per animal, each dose 

contained 108 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 LD50, 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 adult ferrets (14/sex) were exposed by 

the intranasal route to NoFlyTM Technical at a nominal dose of 1×108 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 pathological changes were noted. 

Microbiological 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 ferret. 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—intraperitoneal (OPPTS 885.3200; MIRD 47720-

05). In an acute intraperitoneal infectivity study, a group of young adult Wistar rats (three/sex) 

were injected with NoFlyTM Technical (1.0×109 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 that the applicant

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 through the research, 

development, and testing of NoFlyTM 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 hypersensitivity. 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 (477919-08) using the Magnusson and 

Kligman Method, female guinea pigs were exposed to NoFlyTM WP that was administered as a 

1% intradermal injection and a 25% topical application. Later, a challenge dose was applied 

(25% topical application). Study investigators observed a localized area of intense erythema 

and swelling in treated animals. EPA concluded that NoFlyTM WP is a skin sensitizer. 

600 mg NoFlyTMTechnical

0.163 kg bw

100 mg NoFlyTMTechnical =