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UNITED STATES ENVIRONMENTAL PROTECTION AGENCY 

WASHINGTON, D.C. 20460 

MEMORANDUM ·sEP 1 Ji 2D1D 

SUBJECT: 

FROM: 

THROUGH: 

TO: 

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. I ·!... .. "" (; .. ~~ , 11rrfo 

Ibrahim S. Barsoum, Ph.D., Microbiologist ..JkW--

Microbial Pesticides Branch, Biopesticides and 

Pollution Prevention Division (751 lC) 

John Kough, Ph.D., Senior Scientist 

Microbial Pesticides Branch, Biope · cid s and 

Pollution Prevention Division (7511 

Kathleen Martin, Regulatory Action Leader 

Microbial Pesticides Branch, Biopesticides and 

Pollution Prevention Division (7511 C) 

ACTION REQUESTED: A joint review with PMRA, Canada, of product chemistry, acute 

toxicity/pathogenicity studies, and waiver requests 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 900lsubmitted by Natural Industries Inc., 

Houston, TX 

CONCLUSION: UPGRADABLE to ACCEPTABLE pending on the response of the registrant 

to the deficiencies reported in the recommendations. 

CONTAINS CONFIDENTIAL BUSINESS INFORMATION 

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DATA REVIEW ·RECORD 

Active Ingredient: 

Product Name: 

Company Name: 

Reg. No: 

Chemical Number: 

Decision Number: 

DP Barcode: 

MRIDNo: 

BACKGROUND: 

Paecilomyces fumosoroseus strain FE 900 I 

NoFly Technical, and NoFly Wettable Powder 

Natural Industries Inc., Houston, TX 

73314-T 

l I5003 

4I6429 

38I252 

47719-01, -02 Product Chemistry & Storage Stability ofNOFLY WP 

477I9-03 - 08 Acute Toxicity & Sensitization Studies ofNOFLY WP 

477920-0I Product Chemistry ofNOFL Y Technical 

477920-02 - 07 Acute Toxicity & Waiver requests ofNOFL Y Technical 

Paecilomyces fumosoroseus is a naturally-occurring fungus 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 USA. Target Organisms: NoFly WP is intended to control all stages of whitefly (eggs, 

NI, N2, N3 and N4 larvae, and adults). The ability to infect whitefly eggs is unique among 

entomopathogenic fungi. The target organisms are the whiteflies Trialeurodes vaporariorum, 

Bemisisa tabaci, Bemisia argentifolii, and Lecanoideus floccissimus. In its natural environment, P. 

fumosoroseus strain FE 990I is dispersed as conidia or blastospores, either by air or water 

movement or by other insects and/or mites. The mode of infection of the various entomopathogenic 

fungi is basically similar where a typical infection cycle proceeds as such: conidial attachment, 

germination, penetration, vegetative growth, and then conidiogenesis. The infection cycle of P. 

fumosoroseus strain FE 990I is particularly rapid under optimal conditions. Initially, conidia are 

attached 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 within one day. Fungal multiplication takes place through formation ofhyphal 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. 

Conidiospores generated from the mycelial growth on dead insects can infect additional live target 

pests. The ability of P. fumosoroseus strain FE 990 I to produce infections on eggs of two novel 

whitefly species was evaluated under laboratory conditions. The strain produced infections on the 

NI and N4 nymph stages and significantly reduced the hatching capacity of eggs of Lecanoides 

jloccissimus. It also produced infections on A/eurodicus disperse NI and N4 nymph stages and 

reduced the percent of hatching of eggs by 50%. 

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DISCUSSION: 

With the notable exception ofbeauverolides, no impurities of toxicological significance are 

associated with the TGAI or EP. NoFly Technical contains 

Test substances in all toxicity tests included 

these components in addition to the MPCA. Therefore, the potential toxicity of inert materials 

associated with the active ingredient in both TGAI and EP has been evaluated. 

Therefore, no side-products from chemical reactions are likely. The acute pulmonary 

infectivity and toxicity study was done in ferrets and not in rats. Although we usually prefer that this 

study be done in rats, the applicant has given enough justification for the use of ferrets that is 

acceptable. Also, in the acute intraperitoneal Infectivity study in rats, the applicant has studied 

microbial clearance only in blood samples and has not studied microbial clearance in other body 

organs. 

RECOMMENDATION: The following data are required for the unconditional registration of 

NoFly Technical; and NoFly Wettable Powder: 

1-A five batch analysis ofEP and TGAI from the proposed manufacturing site must be submitted. 

2- Microbial clearance from body organs in the acute intraperitoneal infectivity study in rats. 

SUMMARY OF DATA SUBMITTED: 

Product Chemistry & Storage Stability (MRID 47719-01, 47719-02, & 47720-01) 

NoFly Technical is a wettable powder consisting of

. NoFly WP is a wettable powder 

consisting of . NoFly Technical is a 

slightly cohesive, dusty powder with a bulk density of 0.6- 0.7 glee and 99% of the particles are 

less than 1 OOμm. Its color is lilac and has a yeast-like odor. A 1 % suspension of No Fly Technical in 

water has a pH of 6.5 - 7.5. NoFly WP is a powder with a bulk density of 0.65-0.75 glee. Its color 

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*Manufacturing process information may be entitled to confidential treatment*
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is lilac (Pantone 435M/662C) and has a yeast-like odor. A suspension of No Fly WP in water has a 

pH of 6.5 - 7.5. The potency estimation data for Nofly Technical supports a nominal guarantee of 

8.1 x l 09 CFU/g. A lower limit of for the TGAI is also supported by the data. The 

potency estimation data for NoFly WP supports a nominal guarantee of 2.0 x I 09 CFU/g. A lower 

limit of for the EP is also supported by the data. 

Manufacturing Process: 

The TGAI, NoFly Technical, and the formulated EP, NoFly WP, are produced

process of P.fumosoroseus strain FE 9901. Production ofNoFly Technical involves typical 

procedures for growth of microbial products and is summarized in Figure 1 

Figure 1. Flowchart for Manufacture of NoFfy•• T«lmlca/ 

Formation of Unintentional Ingredients: 

NoFly Technical 

Batch Number 

Parameterl'v1easured Pf Pf Pf 

0702 0706 0708 

Coliform count Abs Abs Abs 

Gram (-) Enteric Abs Abs Abs bacteria 

Salmonella/Shigella Abs Abs Abs 

4 

Pf Pf 

0712 0715 

Abs Abs 

Abs Abs 

Abs Abs 

Quality 

l'v1ean Control 

Specification 

Abs Absent in 

O.lg 

Abs Absent in 

O.lg 

Abs Absent in 

O.tg 

*Manufacturing process information may be entitled to confidential treatment*
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Lactobacilli 0 0 0 0 0 0 < lOJ CF 

Other fungi 0 0 0 0 0 0 < 1% 0 

f'Umosoro !!j f P. 

seus 

NoFly WP 

Batch Number Quality 

Parameter Measured FE07 FE07 FE07 FE07 FE08 Mean Control 

0920 0927 1010 0207 0409 Specification 

Coliform count Abs Abs Abs Abs Abs Abs Absent m 

O.lg 

Gram (-) Enteric Abs Abs Abs Abs Abs Abs Absent m 

bacteria O.lg 

Salmonella/Shigella Abs Abs Abs Abs Abs Abs Absent m 

0.lg 

Lactobacilli 0 0 0 0 0 0 < lOj CFIJ/g 

Other fungi 0 0 0 0 0 0 < 1% of P. 

fumosoroseus 

Analysis for Other Unintentional Ingredients demonstrated that known mammalian toxins are not 

produced or are not present in final products. 

Storage Stability Testing 

A storage stability study was conducted on FuturEco NoFly WP formulation; batch number FE 

051216 before and after storage for 6 months at 4°C. The viable count test (CFU/g) of the 

formulation showed a I 0-fold reduction over the storage period of six months set at 4°C (± 2 "C). 

There was no significant difference in the physical test results before and after storage. 

The reviewers agree that the details of the study support the conclusions drawn above. However, 

since the one batch of EP that was tested was not produced at the proposed manufacturing site, 

differs in formulation and the relative viability of EP after six months was mediocre, the proposed 

storage statement of six months at 4°C cannot be supported. A storage statement for a period of 3 

months at 4°C can be supported at this time with the condition that new data be produced usir_g the 

propo~ed formulation, manufactured at the proposed site. 

Deficiencies: A storage stability study must be conducted using the proposed formulation of NoFly 

WP from the proposed manufacturing site. 

Acute Oral Toxicity-Rat MRID # 47719-03 

In an acute oral toxicity study, Futureco No Fly (containing 2x 109 colony forming units of 

Paecilomycesfumosoroseus strain FE 9901 per gram) was administered to three female SpragueDawley rats as a gavage dose of 5000 mg/kg bw following the up and down procedme. The study 

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was terminated following the stopping rules for this procedure. Each animal was then observed for a 

period of up to 14 days. Oral LD50 is > 5000 mg/kg bw. 

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, Futureco NoFly is of LOW Toxicity (EPA Toxicity Category IV) 

to the rat via the oral route. This acute oral study is classified acceptable. This study satisfies the 

guideline requirement for an acute oral study (OPPTS 870.1100; OECD 425) in the rat. 

CLASSIFICATION: ACCEPTABLE 

Acute Dermal Toxicity - Rat MRID # 47719-04 

In an acute dermal toxicity study, a group of 7 week-old Sprague-Dawley rats (5/sex) was dermally 

exposed to Futureco NoFly (containing 2xl09 colony forming units of Paecilomycesfumosoroseus 

strain FE 9901 per gram) at a limit dose of 5050 mg/kg bodyweight 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. 

Dermal LD5o Males 

Females 

Combined 

> 5050 mg/kg bw 

> 5050 mg/kg bw 

> 5050 mg/kg bw 

There were no mortalities and all the test animals appeared normal for the duration of the study. All 

animals gained weight with the exception of one female which failed to gain weight between Days 7 

and 14. At necropsy, two male rats had pale lungs at study termination. 

Based on these results, Futureco NoFly is of LOW Toxicity to the rat via the dermal route and is 

minim'ally irritating (i.e., mean irritation score=O .1) following a 24-hour exposure. This acute 

dermal toxicity study is classified as acceptable. This study satisfies the guideline requirement for 

an acute dermal toxicity study (OPPTS 870.1200: OECD 402) in the rat. 

CLASSIFICATION: ACCEPTABLE, Toxicity Category IV 

Acute Inhalation Toxicity- Rat MRID # 47719-0S 

In an acute inhalation toxicity study, a group of 10-week old Sprague-Dawley rats (5/sex) were 

exposed by the inhalation route to undiluted Futureco No Fly (containing 2x109 colony forming units 

of Paecilomycesfumosoroseus strain FE 9901 per gram) for 4 hours to nose only at a concentration 

of2.18 mg/L. Animals then were observed for 14 days. 

Inhalation LC50 Males > 2.18 mg/L 

Females > 2.18 mg/L 

Combined > 2.18 mg/L 

Limit test 

There were no mortalities noted throughout the study. Clinical observations included piloerection 

and decreases in activity which 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 results of this study, Futureco NoFly is of LOW Toxicity in male and female SpragueDawley rats via the inhalation route. This acute inhalation study is classified as acceptable. It also 

satisfies the guideline requirement for an acute inhalation study (OPPTS 870.1300; OECD 403) in 

the rat. 

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CLASSIFICATION: ACCEPTABLE, Toxicity Category IV 

Primary Eye Irritation - Rabbit MRID # 47719-06 

In a primary eye irritation study, 0.1 g ofFutureco NoFly WP (containing 88% Paecilomyces 

fumosoroseus strain FE 9901) was instilled into the conjunctiva! sac of the left eye of 3 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 method of Draize. 

After I hour, some redness (grade 1) was observed in one rabbit, and slight to moderate increases in 

excretion discharge were noted in 3 rabbits. All test animals were free of irritation at the 24-, 48-

and 72-hour observation time points. In this study, Futureco NoFly WP (P. fumosoroseus strain FE 

990 l) is SLIGHTLY IRRITATING to the eyes of New Zealand white rabbits. 

This study is classified as acceptable. This study satisfies the guideline requirement for a primary 

eye irritation study (OPPTS 870.2400; OECD 405) in the rabbit. 

CLASSIFICATION: ACCEPTABLE, Toxicity Category IV 

Dermal Irritation -Rabbit MRID # 47719-07 

In a primary dermal irritation study, adult male New Zealand white rabbits (3) were dermally 

exposed to 0.5 g Futureco NoFly WP (containing 88% Paecilomycesfumosoroseus strain FE 9901 

[ARSEF]) in water for 4 hours to an area of skin of approximately 6 cm2. Animals then were 

observed for 72 hours. Irritation was scored by the method ofDraize. 

Very slight erythema was noted in two test animals I hour after patch removal. All test animals were 

free of irritation at the 24-, 48- and 72-hour observation time points. Based on the calculated 

maximum irritation score and an irritation index of 0.7/8 (1 h) and 0.17 (respectively), Futureco 

NoFly WP is slightly irritating to the skin of male New Zealand white rabbits. 

This study is classified as acceptable. This study satisfies the guideline requirement for a primary 

dermal irritation study (OPPTS 870.2500; OECD 404) in the rabbit. 

CLASSIFICATION: ACCEPTABLE, Toxicity Category IV 

Dermal Sensitization - Guinea Pig MRID # 47719-08 

In a skin sensitization study with Futureco NoFly WP (containing 88% Paecilomycesfumosoroseus 

strain FE9901), Dunkin Hartley guinea pigs (10 females) were tested using Magnusson and Kligman 

method. Guinea pigs were subjected to a two-stage induction procedure; a I% intradermal injection 

and a 25% topical application. Two weeks following the last induction exposure, a challenge dose 

was applied as a 25% topical application. Five additional female guinea pigs were exposed to the 

vehicle during induction then challenged with the test material. This group served as a negative 

control. The dermal reactions observed in test animals (left flank) following challenge includc:d 

confluent, intense erythema and swelling. No visible dermal reactions were observed to the vd1icles 

(right flank) or control animals. In this study, Futureco NoFly WP was a sensitizer in guinea pigs 

following the Magnusson and Kilgman procedure. This study satisfies the guideline requirerni~nt for 

a dermal sensitization study (OPPTS 870.2600; OECD 406) in the guinea pig. 

CLASSIFICATION: ACCEPTABLE, Toxicity Category IV 

Acute Oral lnfectivity and Toxicity- Rat MRID # 47720-02 

In an acute oral toxicity study, groups of fasted, young adult ( 6-7 weeks) Wistar rats (15 rats/sex) 

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were given a single oral dose of the MPCA, Paecilomycesfumosoroseus strain FE 9901 (100%; 

I .Ox 109 CFU/g), in 0.05% Tween 80 at a single dose of 108 CFU per animal. 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. Another group of rats (4 rats/sex) were untreated and served as a negative 

control. 

Oral LOSO Males 

Females 

Combined 

> 108 CFU/anirnal 

> I 08 CFU/animal 

> 108 CFU/animal 

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 results of this study, P. 

fumosoroseus strain FE 9901 is of LOW Toxicity and is not infective or pathogenic in the rat. 

CLASSIFICATION: ACCEPTABLE, Toxicity Category IV 

Acute Pulmonary Infectivity and Toxicity - Ferret 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 the microbial pest control agent (MPCA), Paecilomyces 

fumosoroseus strain FE 9901 (100%; I.OxI09 CFU/g), in 0.05% Tween 80 at a nominal dose of 

Ix 108 CFU (in 0.5 mL) per animal. Animals were then observed for up to 21 days. Another group 

of ferrets (4 ferrets/sex) were untreated and served as a negative control. 

Pulmonary LD50 Males > 0.9x 108 CFU/animal 

Females > 0.9x 108 CFU/anirnal 

Combined > 0.9x I 08 CFU/animal 

None of the animals died or showed any symptoms of intoxication following treatment. Some= 

animals lost weight during the first week of the study and it is not known if these effects were 

reversed by the end of the study since these data were not provided in the study report. No gross 

pathological changes were noted. Microbiological examination of lungs, other prescribed organs and 

body fluids proved minimal persistence and no infectivity of MPCA. 

Based on the results of this study, P. fumosoroseus strain FE 9901 is of SLIGHT Toxicity and the 

MPCA is not infective or pathogenic in the ferret. This acute pulmonary infectivity and toxicity 

study is classified as ACCEPTABLE. 

Ferrets are usually not used for acute pulmonary toxicity/pathogenicity testing, but is acceptable 

because of the justification given for their use 

CLASSIFICATION: ACCEPTABLE, Toxicity Category IV 

Acute Intraperitoneal lnfectivity MRID # 47720-05 

Jn an acute intraperitoneal infectivity study, a group of young adult Wistar rats (3/sex) were ir~ected 

with the MPCA, Paecilomycesfumosoroseus strain FE 9901 (100%; t.Oxl09 CFUig), in 0.05% 

Tween 80 at a dose of 107 CFU per animal. Animals were then observed for up to 21 days. Another 

group ofrats ( 4 rats/sex) were untreated and served as a negative control. Based on the resul~ of 

this study, P. fumosoroseus strain FE 9901 is not infective or pathogenic in the rat via the 

intraperitoneal route of exposure. 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. Microbial clearance was shown from blood samples but not from body organs. 

This intraperitoneal infectivity study is classified as supplemental. 

CLASSIFICATION: SUPPLEMENT AL 

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Hypersensitivity Incidence MRID # 4 7720-06 

The registrant submitted a statement indicating that there were no reports of hypersensitivity 

reactions with the microbial pest control agent (MPCA), Paecilomyces fumosoroseus strain FE 

9901, among researchers, product developers and manufacturers. An end-use product containing P. 

fumosoroseus strafo. FE 990 l was determined to be a sensitizer in a skin sensitization study and 21 

reports of hypersensitivity reactions were found in the PubMed database using the keywords 

''paecilomyces" and "allergy". Based on the assumption that most microorganisms contain 

substances that would elicit allergic reactions, the technical grade of the active ingredient, NoFly 

Technical, is considered a potential sensitizing agent. 

CLASSIFICATION: ACCEPTABLE 

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