EPA BIOPESTICIDES AND POLLUTION PREVENTION DIVISION 

COMPANY NOTICE OF FILING FOR PESTICIDE PETITIONS PUBLISHED IN 

THE FEDERAL REGISTER 

EPA Biopesticides and Pollution Prevention Division contact: ShaRon Carlisle; 

(703) 308-6427 

SUBMISSION: E-mail the completed template to: hollis.linda@epa.gov.

TEMPLATE: 

AFS009 Plant Protection, Inc. (EPA Co. No. 91197)

5F8410

 EPA has received a pesticide petition (5F8410) from AFS009 Plant Protection, 

Inc. (EPA Co. No. 91197), 104 T.W. Alexander Drive, Building 18, Research 

Triangle Park, NC 27709 proposing, pursuant to section 408(d) of the Federal Food, 

Drug, and Cosmetic Act (FFDCA), 21 U.S.C. 346a(d), to amend 40 CFR part 180 to 

establish an exemption from the requirement of a tolerance for microbial pesticide 

Pseudomonas chlororaphis subsp. aurantiaca strain AFS009.

 Pursuant to section 408(d)(2)(A)(i) of FFDCA, as amended, AFS009 Plant 

Protection, Inc. (EPA Co. No. 91197) has submitted the following summary of 

information, data, and arguments in support of their pesticide petition. This summary was 

prepared by AFS009 Plant Protection, Inc. (EPA Co. No. 91197) and EPA has not 

fully evaluated the merits of the pesticide petition. The summary may have been edited 

by EPA if the terminology used was unclear, the summary contained extraneous material, 

or the summary unintentionally made the reader conclude that the findings reflected 

EPA’s position and not the position of the petitioner. 

I. AFS009 Plant Protection, Inc. (EPA Co. No. 91197) Petition Summary

 5F8410 

A. Product Name and Proposed Use Practices

Product Name: HowlerTM Technical (100% ai; TGAI) – EPA File Symbol 

91197-R

 HowlerTM (50% ai; EP) – EPA File Symbol 91197-G 

Proposed Use Practice: HowlerTM Technical is a 100% ai Technical Grade 

Active Ingredient (TGAI) of Pseudomonas chlororaphis subsp. aurantiaca strain 
2

AFS009 and is proposed for manufacturing use only, for further formulation into 

registered end-use products. 

HowlerTM is a 50% ai is formulated end-use product for use on growing plants 

and crops to control plant diseases including Rhizoctonia, Pythium, Fusarium, 

Phytophthora and Botrytis. HowlerTM may be mixed with water and applied as a 

foliar spray, soil drench, in furrow spray, transplant spray or dip, cuttings or bare 

root dip, hydroponic or chemigation application in greenhouse, agricultural field, 

turf and ornamental and home and garden use sites. 

B. Product Identity/Chemistry 

 1. Identity of the pesticide and corresponding residues. Pseudomonas chlororaphis 

subsp. aurantiaca strain AFS009 (CAS No. Not applicable). 

 

Pseudomonas chlororaphis is a common bacterium identified primarily in the soil.

Pseudomonas chlororaphis subsp. aurantiaca is a Gram-negative, rod-shaped bacteria 

with one or more flagella for motility. Information regarding the name, identity and 

composition has been submitted to EPA and can be found in MRID No. 495680-01. 

Like other Pseudomonas chlororaphis strains Pseudomonas chlororaphis subsp. 

aurantiaca strain AFS009 is a plant-colonizing bacteria which controls fungal diseases 

by several modes of action, including competition and production of a variety of 

metabolites which are inhibitory to the fungi. (MRID No. 495680-01).

 2. Magnitude of residues at the time of harvest and method used to determine the 

residue. NA-Remove 

 3. A statement of why an analytical method of detecting and measuring the levels 

of the pesticide residue are not needed. An analytical method for residues is not 

applicable. It is expected that, when used as proposed, Pseudomonas chlororaphis 

subsp. aurantiaca strain AFS009, would not result in residues that are of toxicological 

concern.

C. Mammalian Toxicological Profile

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

grade active ingredient (TGAI) and are summarized as follows: 

1. Acute Oral Toxicity/Pathogenicity Study in Rats (OCSPP 885.3050): 

HowlerTM Technical (100% Pseudomonas chlororaphis subsp. aurantiaca strain 

AFS009) was not toxic or pathogenic in rats following acute oral exposure to a 

concentrate of 3.73 x 109 (MRID No. 495680-02). The MPCA test substance and 

the inactivated test substance were administered to rats by gavage in single high 

doses. The animals were observed frequently on the day of dosing (Day 0) for 

mortality and clinical signs of toxicity, and once daily thereafter for 21 days. An 

untreated control group was conducted concurrently. Tissue samples from 

treated rats were enumerated on days 0, 3, 7, 14, and 21. The test organism 

was not observed in plated blood, brain or liver. The urine from treated rats was 

completely clear of MPCA growth by 72 hours. The test organism cleared 
3

completely from the treated lungs, spleen, and kidney by Day 14. The mesenteric 

lymph nodes from treated rats did not achieve complete clearance, but declined 

to very low levels of growth found only in a single animal by Day 21. There were 

no signs of pharmacologic and/or toxicologic effects observed in any animal 

during the study, and no mortality occurred. The gross necropsy conducted at 

termination of the study revealed no internal abnormalities. The test substance, 

HowlerTM Technical, was determined to be non-toxic to rats and demonstrate a 

pattern of clearance when administered by oral gavage in a single dose of 3.73 x 

109 CFU/rat. 

2. Acute Oral Toxicity (OCSPP 870.1100): The TGAI, HowlerTM Technical 

(100% Pseudomonas chlororaphis subsp. aurantiaca strain AFS009), was not 

toxic following acute exposure by the oral route. An acute oral toxicity study was 

conducted on rats using the up-and-down procedure to determine the potential 

for HowlerTM Technical to produce toxicity from a single oral dose (MRID No. 

495680-03). An initial dose of 5,000 mg per kg body weight of the test substance 

was administered to a single female rat by gavage. This first rat survived and so 

two additional rats received a gavage dose of 5,000 mg/kg bw of the test 

material. All three animals survived, so no additional animals were tested. The 

rats were observed for mortality, clinical signs of toxicity and behavioral changes 

daily for 14 days. Body weights were recorded prior to exposure and again on 

Days 7 and 14. All animals survived test substance administration, gained body 

weight, and appeared active and healthy during the study. Necropsies were 

performed on all animals at terminal sacrifice. Under the study conditions, the 

acute oral LD50 of HowlerTM Technical was greater than 5,000 mg/kg body 

weight in female rats (Toxicity Category IV). 

3. Acute Dermal Toxicity (OCSPP 870.1200): The TGAI, HowlerTM 

Technical (100% Pseudomonas chlororaphis subsp. aurantiaca strain AFS009) 

was not toxic following acute exposure by the dermal route. An acute dermal 

toxicity study was conducted on rats to determine the potential for Howler 

Technical to produce toxicity from a single topical application (MRID No. 495680-

04). Five thousand milligrams (mg) of the test substance per kilogram (kg) of 

body weight was applied to the skin of 5 male and 5 female rats for 24 hours. 

The rats were observed for mortality, clinical signs of toxicity and behavioral 

changes daily for 14 days. Body weights were recorded prior to exposure and 

again on Days 7 and 14. Necropsies were performed on all animals at terminal 

sacrifice. Under the study conditions, the single dose acute dermal LD50 of 

HowlerTM Technical was greater than 5,000 mg/kg body weight in male and 

female rats (Toxicity Category IV). 

4. Acute Inhalation Toxicity Study in Rats (OCSPP 870.1300): An acute inhalation 

study demonstrates that the microbial pest control agent (MPCA), HowlerTM 

Technical is not toxic by the inhalation route (MRID No. 495680-05). The TGAI, 

HowlerTM Technical was not toxic to rats following an acute inhalation exposure. 

An acute inhalation toxicity study was conducted on rats to determine the 

potential for HowlerTM Technical to produce toxicity from a single 4-hour 

inhalation exposure. Ten healthy rats (5/sex) were exposed to the test 

atmosphere in a nose-only chamber for 4 hours. Chamber concentration and 

particle size distributions of the test atmosphere were determined periodically 

during the exposure period. The gravimetric chamber concentration was 5.04 
4

mg/L and the average mass median aerodynamic diameter was estimated to be 

2.39 μm. The animals were observed for mortality, clinical signs of toxicity, and 

behavioral changes daily for 14 days following exposure. Body weights were 

recorded prior to exposure and again on Days 1, 3, 7, and 14. Necropsies were 

performed on all animals at terminal sacrifice. Under the study conditions, the 

single dose acute inhalation LC50 of Howler Technical for a 4-hour exposure was 

greater than 5.05 mg/L (Toxicity Category IV). 

 5. Primary Eye Irritation (OCSPP 870.2400): The TGAI, Howler Technical 

(100% Pseudomonas chlororaphis subsp. aurantiaca strain AFS009), was not an 

eye irritant following a 24-hour ocular exposure to rabbits. One-tenth of a milliliter 

of Howler Technical was instilled into the right eyes of three healthy rabbits 

(MRID No. 495680-06). The left eyes remained untreated and served as 

controls. Ocular irritation was evaluated using the Draize scoring method. No 

ocular irritation was observed in any treated eye during the study, and the test 

substance was classified as non-irritating to the eye (Toxicity Category IV). 

 

6. Primary Dermal Irritation (OCSPP 870.2500): The TGAI, HowlerTM 

Technical (100% Pseudomonas chlororaphis subsp. aurantiaca strain AFS009), 

was not a dermal irritant following a 4-hour dermal exposure to rabbits. Fivetenths of a milliliter of Howler Technical was applied to the skin of three healthy 

rabbits and covered for four hours with a gauze pad and semi-occlusive tape 

(MRID No. 495680-07). Following exposure, dermal irritation was evaluated 

using the Draize scoring method. No dermal irritation was observed in any rabbit 

during the study, and the test substance was classified as non-irritating to the 

skin (Toxicity Category IV). 

7. Hypersensitivity Incidents (OCSPP 885.3400): The registrant has noted that no 

incidents of hypersensitivity or any other adverse effects have occurred through 

the research, development or testing of the active ingredient and its related enduse product. Should any incidents occur, they will be reported per FIFRA 

Section 6(a)(2) (MRID No. 495680-16). 

Literature searches have demonstrated that there are no reports of ecological or human 

health hazards caused by Pseudomonas chlororaphis strains. It does not produce 

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

invasiveness or toxicity. Pseudomonas chlororaphis is an entomopathogenic fungus 

and a search of the literature demonstrates it is not reported to be pathogenic to 

humans. A search of the National Library of Medicine, PubMed, using the terms 

“Pseudomonas chlororaphis” AND “mammal” AND “pathogenicity” resulted in “No items 

found” (MRID No. 495680-16). 

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

ingredient. Pseudomonas chlororaphis subsp. aurantiaca strain AFS009 is not toxic, 

pathogenic, infective or irritating to mammals.

D. Aggregate Exposure

 1. Dietary exposure. 
5

 i. Food. Dietary exposure from use of Pseudomonas chlororaphis subsp. 

aurantiaca strain AFS009, as proposed, is minimal. The intended use of 

Pseudomonas chlororaphis subsp. aurantiaca strain AFS009 is as a biological 

fungicide to growing plants in greenhouses, agricultural fields and home gardens 

for the purposes of disease control. 

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

environment from Pseudomonas chlororaphis subsp. aurantiaca strain AFS009. 

There are no reports of ecological or human health hazards caused by this strain 

of Pseudomonas chlororaphis. 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.

 ii. Drinking water. Similarly, exposure to humans from residues of 

Pseudomonas chlororaphis subsp. aurantiaca strain AFS009 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. Pseudomonas chlororaphis is a common bacterium 

found in soils (MRID No. 495680-16). 

The intended use of Pseudomonas chlororaphis subsp. aurantiaca strain 

AFS009 is to growing plants in greenhouses, agricultural fields and home 

gardens for the purposes of disease control. The risk of the microorganism 

passing through the soil to ground water is minimal to unlikely. Additionally the 

fungus would not tolerate the conditions water is subjected to in a drinking water 

facility (including: chlorination, pH adjustments, high temperatures and/or 

anaerobic conditions). 

 

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

environment from Pseudomonas chlororaphis subsp. aurantiaca strain AFS009. 

There are no reports of ecological or human health hazards caused by this 

microorganism. 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.

 2. Non-dietary exposure. The intended use of Pseudomonas chlororaphis subsp. 

aurantiaca strain AFS009 is to growing plants in greenhouses, agricultural fields 

and home gardens for the purposes of disease control. Personal Protective 

Equipment (PPE) mitigates the potential for exposure to applicators and handlers 

of the proposed products, when used in agricultural settings. Pseudomonas 

chlororaphis is a common bacterium found in soils (MRID No. 495680-16). 

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

environment from Pseudomonas chlororaphis subsp. aurantiaca strain AFS009. There 

are no reports of ecological or human health hazards caused by this microorganism. 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 
6

exposures would not be expected to pose any quantifiable risk due to a lack of residues 

of toxicological concern.

E. Cumulative Effects

It is not expected that, when used as proposed, Pseudomonas chlororaphis 

subsp. aurantiaca strain AFS009 would result in residues that are of toxicological 

concern. The intended use of Pseudomonas chlororaphis subsp. aurantiaca strain 

AFS009 is to growing plants in greenhouses, agricultural fields and home gardens for 

the purposes of disease control. The results of toxicity testing indicate there is no risk to 

human health or the environment from Pseudomonas chlororaphis subsp. aurantiaca 

strain AFS009. There are no reports of ecological or human health hazards caused by 

this microorganism. 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.

F. Safety Determination

 1. U.S. population. Acute toxicity studies have shown that Pseudomonas 

chlororaphis subsp. aurantiaca strain AFS009 is not toxic, pathogenic, infective or 

irritating to mammals. The intended use of Pseudomonas chlororaphis subsp. 

aurantiaca strain AFS009 is to growing plants in greenhouses, agricultural fields and 

home gardens for the purposes of disease control. The results of toxicity testing indicate 

there is no risk to human health or the environment from Pseudomonas chlororaphis 

subsp. aurantiaca strain AFS009. There are no reports of ecological or human health 

hazards caused by this microorganism. 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, Pseudomonas chlororaphis subsp. aurantiaca strain AFS009 would result 

in residues that are of toxicological concern. There is a reasonable certainty of no harm 

for infants and children from exposure to Pseudomonas chlororaphis subsp. aurantiaca 

strain AFS009 from the proposed uses.

G. Effects on the Immune and Endocrine Systems 

To date there is no evidence to suggest that Pseudomonas chlororaphis subsp. 

aurantiaca strain AFS009 functions in a manner similar to any known hormone, or that it 

acts as an endocrine disrupter.

H. Existing Tolerances

There is no US EPA tolerance or tolerance exemption for Pseudomonas 

chlororaphis subsp. aurantiaca strain AFS009. 
7

I. International Tolerances

A Codex Alimentarium Commission Maximum Residue Level (MRL) is not 

established for Pseudomonas chlororaphis subsp. aurantiaca strain AFS009.