Isaria fumosoroseus strain FE 9901 – Petition to Establish a Tolerance Exemption 

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Petition for an exemption from the requirement of a tolerance 

for residues of products containing the active ingredient 

“Isaria fumosoroseus strain FE 9901” in and on all food 

commodities

Submitted for: Novozymes BioAg, Inc. (EPA Company No. 73314) 

13100 W. Lisbon Road, Suite 600 

Brookfield, Wisconsin 53005 

Submitted by: Technology Sciences Group Inc. 

 1150 18th Street, NW, Suite 1000 

 Washington, DC 20036 

 

SECTION A Name, Chemical identity, and Composition of the 
Isaria fumosoroseus strain FE 9901 – Petition to Establish a Tolerance Exemption 

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Pesticide Chemical 

Product Name: NoFLYTM Technical (EPA Reg. No. 73314-7) 

NoFLYTM WP (EPA Reg. No. 73314-6) 

Active Ingredient: Isaria fumosoroseus strain FE 9901 

Isaria fumosoroseus strain FE 9901 is a naturally occurring 

fungus found in most countries around the world and occurs in 

various soil types at very low densities. The fungus has 

frequently been isolated from Lepidoptera cadavers in Europe. 

Isolates occur in the Netherlands, Ireland, and Spain. The fungus 

has also been found in various non-European countries including 

China, Japan, Ghana, Pakistan, India, and the United States. 

Relevant information regarding the name, identity, and 

composition has been submitted to EPA and can be found in 

MRID Nos. 47792001, 48051401, and 48051403. 

Both the TGAI and end-use product formulation contain direct 

food additives as inert ingredients. The end-use product is 

currently used to control a variety of target insect pests (i.e. 

whiteflies, aphids, thrips, Psyllids, Mealybugs, and fungus gnats) 

on greenhouse nonfood plants but is proposed for use in/on 

growing agricultural crops, greenhouses, turf, nursery crops, 

commercial landscapes, and residential homes and gardens. 

Mode of Action: As a representative and primary target pest species, Isaria

fumosoroseus strain FE 9901 infects adult whiteflies, all immature 

whitefly stages, and whitefly eggs. The ability to infect whitefly 

eggs is unique among entomopathogenic fungi. I. fumosoroseus

strain FE 9901 is dispersed as conidia or blastospores, either by 

air or water movement or by other insects and/or mites. The mode 

of action of the various entomopathogenic fungi is functionally 

similar, and a typical infection cycle consists of: 1) conidial 

attachment; 2) germination; 3) penetration; 4) vegetative growth; 

and 5) conidiogenesis. 

 

 The infection cycle of I. fumosoroseus strain FE9901 is particularly 

rapid under optimal conditions. Initially, conidia are attached to 

cuticle or 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 

within one day. Fungal multiplication takes place through 

formation of hyphal bodies or blastospores. The mycelium 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. Relevant literature is summarized or 

referenced in Attachment A of MRID No. 47792001.
Isaria fumosoroseus strain FE 9901 – Petition to Establish a Tolerance Exemption 

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SECTION B Amount, Frequency, and Timing of Application of 

 the Pesticide Chemical 

The intended use of Isaria fumosoroseus strain FE 9901 in the end-use product, NOFLYTM WP 

(EPA Reg. No. 73314-6) is as an insecticide for use on greenhouse and outdoor ornamentals, 

trees and shrubs, nursery crops, turf, production agriculture crops, commercial landscapes, and 

residential/home and garden ornamentals, plants, and lawns by foliar spray or soil drench 

application, by chemigation, and/or by seed treatment. Target pests include whiteflies, aphids, 

thrips, Psyllids, Mealybugs, and fungus gnats. NOFLYTM WP can be applied at 5-15 day 

intervals at rates of 1⁄2 - 2 pounds per acre or 2 – 8 ounces per 10,000 square feet on 

ornamentals, trees and shrubs, nonfood crops, and food crops (corn, potatoes, cucurbit 

vegetables, fruiting vegetables, grains, cereals, soybeans, cotton, oil crops, herbs and spices, 

leafy and cole vegetables, legume vegetables, root and tuber/bulb vegetables, berry crops, 

citrus, grape, pome fruit, stone fruit, nuts, tropical fruit, asparagus, banana, plantain, watercress, 

mushroom, olive, and hops) up to and including the day of harvest. 

NOFLYTM WP can be applied to golf course tees, greens, fairways, and roughs, commercial and 

residential lawns, athletic fields, parks, cemeteries, sod farms, turf landscapes, herbaceous and 

woody ornamentals, trees and plants against whiteflies, aphids, thrips, Psyllids, Mealybugs, and 

fungus gnats at rates of 1⁄2 - 2 pounds per acre or 2 – 8 ounces per 10,000 square feet at the 

same application interval (5-15 days) as crop use. 

For residential/home and garden use, NOFLYTM WP can be applied as a foliar spray or soil 

drench to control whiteflies, aphids, thrips, Psyllids, Mealybugs, and fungus gnats on fruit and 

vegetable plants, annual and perennial bedding plants and flowers, roses, potted flowers, 

foliage plants, trees, shrubs, and lawns every 5-15 days at rates ranging from 2 – 8 tablespoons 

per gallon of water to 1⁄2 - 2 1⁄2 cups per 5 gallons of water. 
Isaria fumosoroseus strain FE 9901 – Petition to Establish a Tolerance Exemption 

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SECTION C Reports on Investigations Made with Respect to 

the Safety of the Pesticide Chemical

Toxicity of Isaria fumosoroseus strain FE 9901 

Studies to evaluate the safety to mammals were conducted on the technical grade active 

ingredient Isaria fumosoroseus strain FE 9901 and are summarized herein (see Table 1). The 

studies performed include toxicity/pathogenicity studies via the oral, pulmonary, and 

intraperitoneal injection pathways as well as the conventional acute toxicity (“6-pack”) studies to 

assess acute oral, dermal, and inhalation toxicity, eye and skin irritation, and dermal 

sensitization. The oral, dermal, and inhalation routes of exposure are the routes relevant for 

human health risk assessment. The results of these studies consistently demonstrated that 

Isaria fumosoroseus strain FE 9901 was non-toxic by all routes of exposure and only minimally 

irritating. Additionally, there was no evidence of pathogenically observed; the product was not 

infective or pathogenic to test animals when administered via the oral, pulmonary, or 

intraperitoneal routes. Even though positive for skin sensitization; no reports of hypersensitivity 

have been recorded from personnel working with this organism. 

Table 1. Summary of I. fumosoroseus strain FE 9901 Mammalian Toxicity Data 

Test Species Dose Tested Results 

Acute oral toxicity 

MRID No. 47791903 

Rat Gavage dose of 

5,000 mg/kg 

2.0x109

 CFU/g 

LD50 > 5,000 mg/kg 

Category IV 

Acute dermal toxicity 

MRID No. 47791904 

Rat Undiluted dose of 

5,050 mg/kg 

2.0x109

 CFU/g 

LD50 > 5,050 mg/kg 

Category III 

 

Acute inhalation 

toxicity 

MRID No. 47791905 

Rat 2.18 mg/L for 4 hours 

2.0x109

 CFU/g 

LC50 > 2.18 mg/L 

Category III 

Primary eye irritation 

MRID No. 47791906 

Rabbit 0.1 gram undiluted 

product for 24 hours 

Slightly irritating 

Category IV 

Dermal irritation 

MRID No. 47791907 

Rabbit 0.5 gram undiluted 

product for 4 hours 

Slightly irritating 

Category IV 

 

Dermal Sensitization 

MRID No. 47791908 

Guinea Pig 1% intradermal 

injection + 25% 

topical application 

Localized area of erythema 

and swelling 

Positive for skin sensitization 

Acute oral 

toxicity/pathogenicity 

MRID No. 47792002 

Rat 100mg of 108

CFU/rat 

Non-toxic, non-infective, nonpathogenic 

Acute pulmonary 

toxicity/pathogenicity 

MRID Nos. 47792004 

49044301 

Ferret 1.0x108

 (in 0.5mL) 

CFU/ferret 

Non-toxic, non-infective, nonpathogenic 

Acute intraperitoneal 

Toxicity/pathogenicity 

MRID Nos. 47792005 

49044301 

Rat 107

 CFU/rat Non-toxic, non-infective, nonpathogenic 
Isaria fumosoroseus strain FE 9901 – Petition to Establish a Tolerance Exemption 

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The results of toxicity testing on the organism show there is no risk to human health from the 

active ingredient. Isaria fumosoroseus strain FE 9901 is not toxic, not pathogenic, not infective, 

and only minimally irritating to mammals. Further, it is important to note that Isaria 

fumosoroseus strain FE 9901 does not grow or develop at temperatures in excess of 35°C 

(95°F) (MRID No. 47792001) and therefore is essentially inactivated in mammalian systems. 

 

Literature searches have been conducted against the Agency’s list of open literature database 

searches (July 26, 2010 edition). Those searches have demonstrated that there are no reports 

of ecological or human health hazards caused by Isaria fumosoroseus strain FE 9901. It does 

not produce recognized toxins, enzymes, or virulence factors normally associated with 

mammalian invasiveness or toxicity. 

A special analysis was conducted across five production batches of NOFLYTM Technical to 

determine the prevalence of beauverolide metabolites, beauvericin, and 2,6-pyridinedicarboxylic 

acid (PDCA) following the fermentation process. The results of this analysis (MRID No. 

49118301) concluded that there was no quantitation of beauvericin and 2,6-PDCA was only 

quantified at levels of 1-10μg/g. Across the fifteen beauverolide peptide species analyzed, 

Beauverolide I was determined to be the most prevalent (18% of the overall peptides 

combined), and 100% of the peptides combined were well below 0.1% of NOFLYTM Technical; 

therefore, no metabolite formation rises to a level of toxicological concern. This is further 

confirmed due to the low toxicity evidenced across the suite of mammalian studies on Isaria 

fumosoroseus strain FE 9901. 

 

As has been demonstrated, dietary exposure to Isaria fumosoroseus strain FE 9901 meets the 

FQPA 1996 safety standard. Studies submitted with this petition showed no evidence of toxicity, 

pathogenicity, or infectivity when this strain was tested in laboratory animals. 

Aggregate Exposure 

1) Dietary Exposure:

Dietary exposure from use of Isaria fumosoroseus strain FE 9901, as proposed, is 

minimal. The intended use of Isaria fumosoroseus strain FE 9901 is to agricultural 

crops, greenhouse and outdoor food and nonfood crops, nursery crops, trees and 

shrubs, commercial turf and landscapes, and residential lawns and gardens for the 

purposes of insect control. Isaria fumosoroseus strain FE 9901 does not grow or 

develop at temperatures in excess of 35°C (95°F), and residues are not expected due to 

its degradation in sunlight and in high temperatures (MRID No. 47792001). Any 

remaining residues would be removed during common post-harvest treatments including 

washing, packing, peeling, etc. 

 The results of toxicity testing indicate there is no risk to human health or the environment 

from Isaria fumosoroseus strain FE 9901. There are no reports of ecological or human 

health hazards caused by the microorganism Isaria fumosoroseus. It does not produce 

recognized toxins, enzymes, or virulence factors normally associated with mammalian 

invasiveness or toxicity. The absence of acute toxicity or pathogenicity in laboratory 

animals demonstrates the benign nature of this strain. Dietary exposures would not be 

expected to pose any quantifiable risk, due to a lack of residues of toxicological concern. 
Isaria fumosoroseus strain FE 9901 – Petition to Establish a Tolerance Exemption 

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2) Drinking Water Exposure:

Similarly, exposure to humans from residues of Isaria fumosoroseus strain FE 9901 in 

consumed drinking water would be unlikely. Potential exposure to surface water would 

be negligible and exposure to drinking water (well or ground water) would be impossible 

to measure. The intended use of Isaria fumosoroseus strain FE 9901 is to agricultural 

crops, greenhouse and outdoor food and nonfood crops, nursery crops, trees and 

shrubs, commercial turf and landscapes, and residential lawns and gardens for the 

purposes of insect control. The risk of the microorganism passing through the soil to 

ground water is minimal to unlikely. Additionally the fungi would not tolerate the 

conditions water is subjected to in a drinking-water treatment facility (including: 

chlorination, pH adjustments, high temperatures and/ or anaerobic conditions) (MRID 

No. 47792001). 

The results of toxicity testing indicate there is no risk to human health or the environment 

from Isaria fumosoroseus strain FE 9901. There are no reports of ecological or human 

health hazards caused by the microorganism Isaria fumosoroseus. It does not produce 

recognized toxins, enzymes, or virulence factors normally associated with mammalian 

invasiveness or toxicity. The absence of acute toxicity or pathogenicity in laboratory 

animals demonstrates the benign nature of this strain. Drinking water exposures would 

not be expected to pose any quantifiable risk, due to a lack of residues of toxicological 

concern. 

3) Non-Dietary Exposure:

The potential for non-dietary exposure to the general population, including infants and 

children, is unlikely. The intended use of Isaria fumosoroseus strain FE 9901 is to 

agricultural crops, greenhouse and outdoor food and nonfood crops, nursery crops, 

trees and shrubs, commercial turf and landscapes, and residential lawns and gardens 

for the purposes of insect control. The non-toxic and non-pathogenic nature of this 

product demonstrates that other incidental non-dietary exposures are not of concern. 

 The results of toxicity testing indicate there is no risk to human health or the environment 

from Isaria fumosoroseus strain FE 9901. There are no reports of ecological or human 

health hazards caused by the microorganism Isaria fumosoroseus. It does not produce 

recognized toxins, enzymes, or virulence factors normally associated with mammalian 

invasiveness or toxicity. The absence of acute toxicity or pathogenicity in laboratory 

animals demonstrates the benign nature of this strain. Non-dietary exposures would not 

be expected to pose any quantifiable risk due to a lack of residues of toxicological 

concern. 

Cumulative Effects 

It is not expected that, when used as proposed, Isaria fumosoroseus strain FE 9901 would 

result in residues that are of toxicological concern. The intended use of Isaria fumosoroseus

strain FE 9901 is to agricultural crops, greenhouse and outdoor food and nonfood crops, 
Isaria fumosoroseus strain FE 9901 – Petition to Establish a Tolerance Exemption 

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nursery crops, trees and shrubs, commercial turf and landscapes, and residential lawns and 

gardens for the purposes of insect control. The results of toxicity testing indicate there is no risk 

to human health or the environment from Isaria fumosoroseus strain FE 9901. There are no 

reports of ecological or human health hazards caused by Isaria fumosoroseus strain FE 9901. 

It does not produce recognized toxins, enzymes, or virulence factors normally associated with 

mammalian invasiveness or toxicity. The potential for toxic metabolite formation is diminutive in 

nature as demonstrated in the de minimus findings of beauverolide formation. The absence of 

acute toxicity or pathogenicity in laboratory animals demonstrates the benign nature of this 

strain. Any exposures would not be expected to pose any quantifiable risk due to a lack of 

residues of toxicological concern.

Safety Determination 

1) General US Population: 

 

Acute toxicity studies have demonstrated that Isaria fumosoroseus strain FE 9901 is not 

toxic, pathogenic, or infective to mammals. Other ingredients associated with the TGAI 

and end-use product are also direct food additives. The intended use of Isaria 

fumosoroseus strain FE 9901 is to agricultural crops, greenhouse and outdoor food and 

nonfood crops, nursery crops, trees and shrubs, commercial turf and landscapes, and 

residential lawns and gardens for the purposes of insect control. The results of toxicity 

testing indicate there is no risk to human health or the environment from Isaria 

fumosoroseus strain FE 9901, and there is a lack of residues on treated commodities. 

There are no reports of ecological or human hazards caused by the microorganism 

Isaria fumosoroseus. It does not produce recognized toxins, enzymes, or virulence 

factors normally associated with mammalian invasiveness or toxicity. The absence of 

acute toxicity or pathogenicity in laboratory animals demonstrates the benign nature of 

this strain. There is a reasonable certainty of no harm to the general US population from 

exposure to this active ingredient. 

2) Infants and Children:

As mentioned above, it is not expected that, when used as proposed, Isaria 

fumosoroseus strain FE 9901 would result in residues that are of toxicological concern. 

There is a reasonable certainty of no harm for infants and children from exposure to 

Isaria fumosoroseus strain FE 9901 from the proposed uses. 

Information on Endocrine Effects 

To date there is no evidence to suggest that Isaria fumosoroseus strain FE 9901 functions in a 

manner similar to any known hormone, or that it acts as an endocrine disrupter. 
Isaria fumosoroseus strain FE 9901 – Petition to Establish a Tolerance Exemption 

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Section D Tests on the Amount of Residue Remaining 

Not applicable, Novozymes BioAg, Inc. is petitioning for an exemption from the requirement of a 

tolerance for residues of Isaria fumosoroseus strain FE 9901. 

Analytical Method: An analytical method for residues is not applicable. It is expected 

that, when used as directed, Isaria fumosoroseus strain FE 9901will 

not result in residues that are of toxicological concern. 
Isaria fumosoroseus strain FE 9901 – Petition to Establish a Tolerance Exemption 

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SECTION E - Practical Methods for Removing Residues 

Not applicable, Novozymes BioAg, Inc. is petitioning for an exemption from the requirement of a 

tolerance for residues of Isaria fumosoroseus strain FE 9901. 
Isaria fumosoroseus strain FE 9901 – Petition to Establish a Tolerance Exemption 

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SECTION F – Proposed Tolerance Exemption 

Novozymes BioAg, Inc. is proposing that the following Exemption from the need of a tolerance be 

established for the pesticide, Isaria fumosoroseus strain FE 9901, similar to the current tolerance 

exemption in 40 CFR §180.1306 for a similar fungal strain, Isaria fumosorosea Apopka strain 97: 

“An exemption from the requirement of a tolerance is established for residues of Isaria 

fumosoroseus strain FE 9901 in or on all food commodities when applied as an insecticide or 

miticide and used in accordance with good agricultural practices.”
Isaria fumosoroseus strain FE 9901 – Petition to Establish a Tolerance Exemption 

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SECTION G Reasonable Grounds in Support of an Exemption 

from the Need of a Tolerance 

EXISTING TOLERANCES 

US EPA Tolerance -

While there is not a specific tolerance for Isaria fumosoroseus strain FE 9901, there is an 

established and existing tolerance exemption for a similar fungal strain (Isaria fumosorosea Apopka 

strain 97), codified under 40 CFR §180.1306, which states: 

“An exemption from the requirement of a tolerance is established for residues of Isaria 

fumosorosea Apopka strain 97 in or on all food commodities when applied as an insecticide 

or miticide and used in accordance with good agricultural practices.” 

International -

A Codex Alimentarium Commission Maximum Residue Level (MRL) is neither established nor 

required for Isaria fumosoroseus strain FE 9901. 

It is not expected that, when used as proposed, Isaria fumosoroseus strain FE 9901 would 

result in residues that are of toxicological concern. The intended use of Isaria fumosoroseus

strain FE 9901 is to agricultural crops, greenhouse and outdoor food and nonfood crops, 

nursery crops, trees and shrubs, commercial turf and landscapes, and residential lawns and 

gardens for the purposes of insect control. The results of toxicity testing indicate there is no risk 

to human health or the environment from Isaria fumosoroseus strain FE 9901. There are no 

reports of ecological or human health hazards caused by Isaria fumosoroseus strain FE 9901. It 

does not produce recognized toxins, enzymes, or virulence factors normally associated with 

mammalian invasiveness or toxicity. The absence of acute toxicity or pathogenicity in laboratory 

animals demonstrates the benign nature of this strain, and in conjunction with the established 

tolerance for similar fungal strains (Isaria fumosorosea Apopka strain 97), serves to 

demonstrate reasonable grounds in support of the petition for exemption from the requirement 

of a tolerance.