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TABLE OF CONTENTS

I. EXECUTIVE SUMMARY.....................................................................................................................................4

II. ACTIVE INGREDIENT OVERVIEW ...............................................................................................................6

III. REGULATORY BACKGROUND......................................................................................................................6

A. Applications for Pesticide Product Registration .................................................................................................6

B. Food Tolerance Exemption ...................................................................................................................................6

IV. RISK ASSESSMENT...........................................................................................................................................7

A. Product Analysis Assessment (40 CFR § 158.2120) ............................................................................................7

B. Human Health Assessment (40 CFR § 158.2140) ................................................................................................8

C. Environmental Assessment (40 CFR § 158.2150)..............................................................................................14

V. ENVIRONMENTAL JUSTICE .........................................................................................................................17

VI. RISK MANAGEMENT DECISION ................................................................................................................17

A. Final Printed Labeling.........................................................................................................................................18

B. Terms of Registration ..........................................................................................................................................18

C. Reporting of Adverse Effects and Hypersensitivity Incidents .........................................................................18

VIII. GLOSSARY OF ACRONYMS AND ABBREVIATIONS..........................................................................19

IX. BIBLIOGRAPHY ..............................................................................................................................................20

A. Studies Submitted to Support the Active Ingredient, Pseudomonas fluorescens strain D7 and Pesticide 

Product Registrations for the Fluorescens Technical and the EP, D7 ..................................................................20

B. Environmental Protection Agency Risk Assessment Memoranda...................................................................22

C. Other References .................................................................................................................................................23

APPENDIX A. MICROBIAL PESTICIDES DATA REQUIREMENTS ...........................................................24

APPENDIX B. PESTICIDE PRODUCTS..............................................................................................................29
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BIOPESTICIDES REGISTRATION ACTION DOCUMENT TEAM

Office of Pesticide Programs (OPP)

Biopesticides and Pollution Prevention Division

Microbial Pesticides Branch

Science Reviews

Ibrahim S. Barsoum, Ph.D. Product Analysis, Human Health

John L. Kough, Ph.D. Product Analysis, Human Health

In-Soon You, Ph.D. Environmental Effects

Gail Tomimatsu, Ph.D. Environmental Effects

Shannon Borges Environmental Effects

Regulatory

Kimberly Nesci Chief, Microbial Pesticides Branch

Susanne Cerrelli Regulatory Action Leader
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I. EXECUTIVE SUMMARY

Background

On October 5, 2012, Ag-Chem Consulting on behalf of Northwest Agricultural Products 

submitted applications for a new technical grade active ingredient (TGAI), Fluorescens 

Technical (EPA File Symbol 71975-G), and a new end-use product (EP), D7 (EPA File Symbol

71975-U), to the United States Environmental Protection Agency (EPA) under section 3 of the 

Federal Insecticide, Fungicide, and Rodenticide Act (FIFRA). These products both contain the 

new active ingredient, Pseudomonas fluorescens strain D7.

Pseudomonas fluorescens strain D7 is a naturally occurring bacterium originally isolated from 

winter wheat roots. On February 14, 2012, an Experimental Use Permit no. 71975-EUP-1 was 

issued to Northwest Agricultural products for experimental use on a total of 1,020 acres in 

Oregon and Washington states. This permit is effective from March 1, 2012 to March 1, 2015. 

In September 2009, EPA completed a Final Registration Review Decision for Pseudomonas 

fluorescens, Registration Review Case 6006 

(http://www.regulations.gov/#!documentDetail;D=EPA-HQ-OPP-2007-0567-0014). In its 

decision, EPA concluded that the standards of Registration Review were met for the

Pseudomonas fluorescens and identified no general risk concerns. This strain will now be 

considered as part of the Pseudomonas fluorescens Registration Review Case 6006.

The EP containing Pseudomonas fluorescens strain D7 is proposed for use in field applications

for suppression of downy brome (cheatgrass), medusahead, Japanese brome, and jointed 

goatgrass on fields of turf and grasses grown for seed, alfalfa, wheat, barley, triticale, oat, and 

rangeland. The proposed EP label contains applications as a seed treatment and also by ground or 

aerial spray. The label directs the user to apply the EP before weed seeds germinate in the spring 

or autumn when daytime temperatures are below 50oF and just prior to expected rainfall. It can 

be applied up to 4 times per year for a maximum annual application of 8g/acre. 

EPA scientists reviewed product analysis, toxicology, and nontarget organism data and 

information (40 CFR §§ 158.2120, 158.2140, and 158.2150, respectively) submitted to support 

the registration of the proposed pesticide products. They found that, overall, such data and 

information are adequate for risk assessment purposes, fulfill the current microbial pesticide data 

requirements, and support registration of the product under FIFRA section 3(c)(5).

Product Analysis

For the purposes of FIFRA section 3(c)(5) registration, the product analysis data requirements 

for the TGAI (also the manufacturing use product) and EP were fulfilled by acceptable guideline 

studies. These data requirements include product chemistry and composition, analysis of 

samples, physical and chemical characteristics.

Toxicology

The applicant submitted adequate mammalian toxicology data and information to support the 

pesticide products. An acute injection toxicity/pathogenicity study submitted with the original 

application and conducted with high dosages of Pseudomonas fluorescens strain D7, showed 
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adverse effects including mortality, abnormal clinical signs, transient weight loss with resultant 

decreased absolute body weight, abnormal gross necropsy findings, and altered organ weights.

This injection study was conducted at an inappropriate excessive dose. The applicant repeated 

the acute injection study at an appropriate lower dose with additional controls to account for 

observed results. This repeated injection toxicity/pathogenicity study along with the acute oral

and pulmonary toxicity/pathogenicity studies, showed that Pseudomonas fluorescens strain D7 is 

not toxic or pathogenic via these routes of exposure. In light of the results of the acute 

toxicity/pathogenicity data, EPA did not require testing at higher tiers (i.e., Tiers II and III).

Tolerance Exemption

In its application to the EPA in 2012, Ag-Chem Consulting on behalf of Northwest Agricultural 

Products provided a petition, data and other information to support an exemption from the 

requirement for a tolerance for residues of Pseudomonas fluorescens strain D7 when used in or 

on growing crops and rangeland. 

Occupational Exposure

Despite the low toxicological profile of residues of the active ingredient, Pseudomonas 

fluorescens strain D7, in or on all agricultural commodities, the EPA is requiring baseline 

personal protective equipment (PPE) for handlers that, due to their occupation, may be subject to 

prolonged or repeated exposure to the active ingredient. The label on D7, the end-use product, 

directs handlers working with Pseudomonas fluorescens strain D7 in agricultural settings to wear 

a long-sleeved shirt, long pants, socks, shoes, waterproof gloves, and a dust/mist filtering 

respirator meeting National Institute for Occupational Safety and Health (NIOSH) standards of at 

least N-95, R-95, or P-95. 

Nontarget Organisms

P. fluorescens occurs naturally in soil and water (Pallerroni, 2005), and strain D7 is a rootassociated bacterium that was isolated originally from the West Coast region of the United 

States. Soil-dwelling nontarget organisms in those areas are expected to currently have some 

exposure to this bacterium. The nontarget organism data requirements have been adequately 

addressed for the purposes of FIFRA section 3(c)(5) registration, based on the submitted data, 

rationales, and limited anticipated exposure to nontarget organisms. The specific findings for 

each nontarget organism are addressed in Section IV below.

Public Participation

On October 1, 2009, the 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 

policy, the EPA 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; or any other registration actions for which the 

Agency believes there may be significant public interest.

Consistent with the policy of making registration actions more transparent, the pesticide products

containing Pseudomonas fluorescens strain D7 were subject to a 15-day comment period. In 

addition to containing a new active ingredient, the registration of the pesticide product will result 
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in the first outdoor and food uses for Pseudomonas fluorescens strain D7. The docket 

identification number associated with these registration actions, and accessed through either 

http://www.regulations.gov or http://www.epa.gov/pesticides/regulating/registration-status.html, 

is EPA-HQ-OPP-2013-0570. During this comment period, no comments were received. 

Registration Decision

Therefore EPA maintains that based upon the risk assessment and information submitted in 

support of registration of the Fluorescens Technical (TGAI) and the D7 EP, it is appropriate to 

issue these registrations. The basis for this decision can be found in the risk assessment for 

Pseudomonas fluorescens strain D7 which is characterized throughout this BRAD. 

II. ACTIVE INGREDIENT OVERVIEW

Biological Name: Pseudomonas fluorescens strain D7 

Culture Deposit: Agricultural Research Service Culture Collection NRRL B-18293

OPP Chemical Code: 016418

Type of Pesticide: Microbial Pesticide –Herbicide

See Appendix B for specific information (e.g., use sites, application rates, methods of 

application, formulation types, and target pests) regarding the pesticide products containing this

active ingredient.

III. REGULATORY BACKGROUND

A. Applications for Pesticide Product Registration

On October 5, 2012, Ag-Chem Consulting (address: 12208 Quinque Lane, Clifton, VA 20124)

on behalf of Northwest Agricultural Products (address: 821 South Chestnut Avenue, Pasco, WA 

99301) submitted applications for a new TGAI, Fluorescens Technical (EPA File Symbol 71975-

G), and a new EP, D7 (EPA File Symbol 71975-U), to the EPA under section 3 of FIFRA.

On October 30, 2013 (78 FR 64937), EPA announced receipt of the application to register a 

pesticide product containing a new active ingredient for use in agriculture (outdoors). EPA

opened a 30-day public comment period pursuant to the provisions of FIFRA section 3(c)(4). No 

comments were received following this publication. 

B. Food Tolerance Exemption

Concurrent with its registration applications and under Federal Food, Drug, and Cosmetic Act 

(FFDCA) section 408(d), Northwest Agricultural Products submitted a petition to establish an 

exemption from the requirement for a tolerance for residues of the pesticide active ingredient, 

Pseudomonas fluorescens strain D7 [Pesticide Petition (PP) 2F8103] in or on growing crops and 

rangeland. EPA published a Notice that Northwest Agricultural Products filed a petition to 

establish an exemption of the requirement for a tolerance for residues of Pseudomonas 

fluorescens strain D7 in or on growing crops and rangeland in the Federal Register of September 

12, 2013 (78 FR 56185) and opened a 30-day comment period. While rangeland is not a food 
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commodity and “growing crops” is not a recognized commodity term used by the Agency in 

tolerance actions, the Agency is interpreting the petitioner’s request as seeking a tolerance 

exemption for “all food commodities.” The term “growing crops” is quite broad and does not 

limit that types of food commodities that it covers; therefore, the Agency believes a reasonable 

interpretation of that term allows for establishment of an exemption from the requirement of a 

tolerance for “all food commodities.” 

One comment was received in the docket for the petition for Pseudomonas fluorescens strain D7

tolerance exemption that expressed general opposition to the sale of pesticides, and to their 

negative effects on bees and human beings. The comment was not specific to Pseudomonas 

fluorescens strain D7 and was not found to be relevant to the proposed exemption of the 

requirement for a tolerance. For Pseudomonas fluorescens strain D7, no hazard to bees was 

identified and the risk posed to adults, infants, and children is likely to be minimal because of the 

low acute oral toxicity/pathogenicity potential of Pseudomonas fluorescens strain D7. The 

exposure to bees to this pesticide is expected to be low based on the label application 

instructions. 

IV. RISK ASSESSMENT

In the Federal Register of October 26, 2007 (72 FR 61002), the EPA issued a Final Rule on the 

data requirements to support registration of microbial pesticides and updated the definition for 

microbial pesticides. The rule became effective on December 26, 2007. The data and information 

evaluated for this BRAD were considered in light of these requirements.

The EPA classifies each data submission with an indication of the usefulness of the information 

contained in the documents for risk assessment. A rating of “acceptable” indicates the study is 

scientifically sound and is useful for risk assessment. A “supplemental” rating indicates the data 

provide some information that can be useful for risk assessment. The studies may have certain 

aspects determined not to be scientifically acceptable (“supplemental: upgradeable”). If a study 

is rated as “supplemental: upgradeable,” EPA always provides an indication of what is lacking or 

what can be provided to change the rating to “acceptable.” If there is simply a “supplemental” 

rating, the reviewer will often state that the study is not required by 40 CFR Part 158. Both 

“acceptable” and “supplemental” studies may be used in the risk assessment process as 

appropriate. An “unacceptable” rating indicates that new data must be submitted.

For the acute toxicity data requirements, Toxicity Categories are assigned based on the 

hazard(s) identified from studies and/or other information submitted to the EPA in support of a 

pesticide registration. The EPA classifies the active ingredient or particular product into Toxicity 

Category I, II, III, or IV, where Toxicity Category I indicates the highest toxicity and Toxicity 

Category IV indicates the lowest toxicity (see 40 CFR § 156.62). 

A. Product Analysis Assessment (40 CFR § 158.2120)

For purposes of registration under FIFRA section 3(c)(5), the EPA determined that all product 

analysis data requirements for Pseudomonas fluorescens strain D7 have been fulfilled. Refer to 

Tables 1, and 2, in Appendix A for a summary of the data requirements, including both generic 

and product-specific information. 
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B. Human Health Assessment (40 CFR § 158.2140)

1. Toxicity

All applicable mammalian toxicology data requirements supporting the request for an exemption 

of the requirement for a tolerance for residues of Pseudomonas fluorescens strain D7 in or on all 

food commodities have been fulfilled with data submitted by the applicant or scientific rationale. 

Furthermore, Tier II and Tier III studies were not required for Pseudomonas fluorescens strain

D7 because of the lack of acute toxicity/pathogenicity in the Tier I studies.

The toxicity and pathogenicity tests (dermal, toxicity) and irritation tests (acute eye and 

primary dermal irritation) that address potential routes of exposure to the active ingredient are

all classified in Toxicity Category IV (see below or U.S. EPA 2014a and b) and revealed little

to no toxicity attributed to Pseudomonas fluorescens strain D7. Finally, the applicant reported 

that no hypersensitivity incidents occurred during the research, development, and testing of this 

active ingredient.

The overall conclusions from all toxicological information submitted by the applicant are briefly

described below, in sections IV(B)(1)(a) and IV(B)(1)(b) (see pages 8–11), and summarized in

Table 3 in Appendix A. 

a. Acute Toxicity/Pathogenicity – Tier I

Acute Injection Toxicity/Pathogenicity – Rat (OCSPP Guideline Number 885.3200) 

(MRID 49349701) 

A study was repeated to re-evaluate the acute intravenous toxicity infectivity and pathogenicity, 

of a microbial pest control agent (MPCA), Pseudomonas fluorescens D7, at a single exposure of 

106and 107 CFU/rat doses. The MPCA test substance, inactivated MPCA, or sterile filtrate was 

injected into the tail vein of one of four groups of rats. A control group (untreated) was 

conducted concurrently. The animals were observed frequently on day of dosing for mortality 

and signs of pharmacological and/or toxicological effects and once daily thereafter for 21 days. 

Tissue and blood samples taken at interim sacrifices from the group receiving the active test 

substance were cultured to provide quantitative measurements of the test microbe's clearance 

pattern. There was no mortality in any group during the study. During observations for clinical 

signs, all animals appeared normal for the duration of the study. The gross necropsy conducted at 

termination of the study revealed no observable abnormalities. The test organism had cleared 

completely from Groups IV and V (MPCA) blood, kidneys, mesenteric lymph nodes, lungs, and 

brain, liver, spleen and cecum contents by Day 7. After two consecutive interim sacrifices 

showed no growth of test organism in the organs plated, no further tissue samples were taken. 

The test substance Pseudomonas fluorescens D7 was determined to be non-toxic and not 

pathogenic or infective to rats when injected intravenously at a dose of 3.4 x 107 CFU/rat or at 

3.7 x 106 CFU/rat. EPA determined this study to be acceptable.

This study (MRID 49349701) addresses EPA recommendations to repeat an acute injection 

toxicity study using a lower dose of the microorganisms that includes additional controls such as 

filtered culture medium, and autoclaved medium to account for the observed injection results. 

This study supersedes the previously submitted acute injection toxicity/pathogenicity in rat study 

(MRID 48966402) that tested Pseudomonas fluorescens strain D7 and showed adverse effects on 

rats following a single intravenous injection administration of 1.3 x 108 CFU/rat, including 
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mortality, abnormal clinical signs; transient weight loss, abnormal gross necropsy findings, and 

altered organ weights. 

Acute Oral Toxicity/Pathogenicity – Rat (OCSPP Guideline Number 885.3050) 

(MRID 48966403)

Sprague-Dawley rats, 8 week old, (12/sex) were given a single oral dose of MPCA,

Pseudomonas fluorescens strain D7, of 1.3 X 109 CFU/animal. The animals were observed three 

times on day 0 after dosing and daily through day 21 with interim scheduled sacrifices on Days 

3, 7, and 14. Five males and five females were treated with autoclaved Pseudomonas 

fluorescens strain D7 as inactive MPCA controls, two untreated animals per sex were used as 

“shelf controls,” and two animals per sex were designated as untreated controls.

No observable abnormalities were found during observations for clinical signs or at necropsy, 

and there were no toxicologically relevant differences between the body weights of the treated 

animals and those of controls. Relative to their respective controls, MPCA-treated females had 

increased relative liver weights (+16.0%; p<0.05), and MPCA-treated males had increased 

relative spleen weights (+16.7%; p<0.05), and these differences correlated with increased mean 

absolute weights of these same organs (+11.7% and +27.1% for MPCA-treated female absolute 

liver and MPCA-treated male absolute spleen weights, respectively). No CFUs (or in one 

instance a single individual count of <100 CFU/g) were recovered from the brain, lungs, liver, 

kidneys, or lymph nodes of any active-treated animal. Pseudomonas fluorescens strain D7 was 

found in feces, urine, and cecum contents of MCPA-treated animals and appeared to be 

completely cleared by day 14 following oral administration to rats. Based on the results of this 

study, Pseudomonas fluorescens strain D7 does not appear to be toxic, infective, and/or 

pathogenic in rats when dosed at 1.3 X 109 CFU/animal. The EPA rated this study as acceptable.

Acute Pulmonary Toxicity/Pathogenicity – Rat (OCSPP Guideline Number 885.3150)

(MRID 489664-04)

Groups of 12 week old Sprague Dawley rats (3/sex/Group) were exposed by the intratracheal 

route to Pseudomonas fluorescens strain D7 in sterile PBS at a constant dose volume of 0.1 mL/ 

animal and a dose of 4.6 x 108cfu/animal. Eight males and eight females were treated with 

autoclaved Pseudomonas fluorescens strain D7 as autoclaved controls; five males and five 

females were not treated and used as untreated control; and four males and four females were not 

treated and used as shelf controls. The animals were then observed for up to 21 days, with 

interim sacrifices on day 0 (all groups) and days 3, 7, and 14 (active-MCPA group). 

There were no test substance related clinical signs, gross necropsy findings, or differences in 

organ weights. Two males in the active test material treated group sacrificed on day 3 lost 

weight and one female in autoclaved test material treated group sacrificed on day 21 did not gain 

weight during the first week but gained weight by day 14. All other animals gained weight prior 

to scheduled sacrifice. The test organisms were not seen or were present at < 100 cfu/g in lungs 

from the animals sacrificed after dosing on day 0. The test organisms were not seen or were 

present at < 100 cfu/g in blood, brain, lungs, spleen, liver, kidneys, mesenteric lymph nodes, and 

cecum content removed from animals sacrificed on days 3 and 7. Based on these results, 

Pseudomonas fluorescens strain D7 does not appear to be toxic, infective, and/or pathogenic in 

rat when dosed at 4.6 x 108cfu /animal. The EPA rated this study as acceptable.
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Acute Dermal Toxicity/Pathology – Rabbit (OCSPP Guideline Number 870.1200)

(MRID 489664-05)

Five male and five female New Zealand White rabbits, 16-18 weeks old, were dermally exposed 

to a 2000 mg/kg bw dose of Pseudomonas fluorescens strain D7 moistened with 2.0 mL of 

deionized water/g test substance for 24 hours to an area of approximately 10% of body surface 

area. Following exposure, the animals were observed for a period of 14 days. All animals 

survived and had no abnormal systemic clinical signs during the study. Very slight to welldefined Erythema was noted on all dose sites on day 1 and resolved by day 4. With the 

exception of one female that lost weight during the second week, all animals gained weight 

throughout the study. No observable abnormalities were found at necropsy. The dermal LD50

for males was greater than 2000 mg/kg bw; for females was greater than 2000 mg/kg bw; and for 

both sexes combined was greater than 2000 mg/kg bw. Based on the results of this study, 

Pseudomonas fluorescens strain D7 is of low toxicity. EPA classified the acute dermal toxicity 

of Pseudomonas fluorescens strain D7 as Toxicity Category IV and rated this study as 

acceptable.

Primary Eye Irritation – Rabbit (OCSPP Guideline Number 870.2400) (MRID 49100801)

100 mg of undiluted Pseudomonas fluorescens strain D7 (Batch No. 201109210301; purity 95%; 

pH not reported) was instilled as supplied into the conjunctival sac of the right eye of three male 

and three female New Zealand White rabbits. Untreated left eyes served as controls. Animals 

were observed at 1, 24, 48, and 72 hours after test material instillation. Irritation was scored by 

the method of Draize and classified by the system of Kay and Calandra.

No corneal opacity, iritis, or conjunctival irritation was noted on any rabbit throughout the study. 

Pseudomonas fluorescens strain D7 was not irritating to the eye and EPA classified the acute eye 

irritation as Toxicity Category IV for primary eye irritation. The EPA rated this study as 

acceptable.

Primary Dermal Irritation – Rabbit (OCSPP Guideline Number 870.2500)

(MRID 49100802)

Three male and three female New Zealand White rabbits were dermally exposed to 500 mg of 

Pseudomonas fluorescens strain D7 moistened with 1.0 mL of DI water for 4 hours on an 

approximately 2.5 x 2.5 cm area of the body surface. The animals were observed at 1, 24, 48, 

and 72 hours after patch removal. Irritation was scored by the method of Draize. No dermal 

irritation was noted on any animal during the study. The primary irritation index was 0.0. 

Pseudomonas fluorescens strain D7 was not irritating and EPA classified the dermal irritation 

toxicity as Toxicity Category IV. The EPA rated this study as acceptable.

Hypersensitivity incidents (OCSPP Guideline Number 885.3400)

The applicant reported that no hypersensitivity incidents, including immediate-type or delayedtype reactions of humans and domestic animals, occurred during research, development, or

testing of Pseudomonas fluorescens strain D7

b. Acute Toxicology and Subchronic Toxicity/Pathogenicity – Tier II; 

Reproductive, Fertility Effects, Carcinogenicity, Immunotoxicity, and Infectivity/Pathogenicity

Analysis – Tier III

Tier II and Tier III studies were not required for Pseudomonas fluorescens strain D7 based on the 

lack of acute toxicity/pathogenicity in the Tier I data requirements.
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c. Endocrine Disruptors

As required by the Administrator under the Federal Food, Drug, and Cosmetic Act (FFDCA), the 

EPA has developed the Endocrine Disruptor Screening Program (EDSP) and has begun to 

implement the screening program that is to be used to test all pesticides 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.” FFDCA section 408(p)(4) authorizes the 

Administrator, by order, to exempt from the requirements of the Endocrine Disruptor Screening 

Program a biologic substance or other substance if a determination is made that the substance is 

anticipated to not produce any effect in humans similar to an effect produced by a naturally 

occurring estrogenic substance. Between October 2009 and February 2010, EPA issued test 

orders/data call-ins for the first group of 67 chemicals, which contains 58 pesticide active 

ingredients and 9 inert ingredients. Pseudomonas fluorescens strain D7 is not among the group 

of 58 pesticide active ingredients on the initial list to be screened under the EDSP. 

The EPA believes that Pseudomonas fluorescens strain D7 is a substance that would not produce 

any effect in humans similar to an effect produced by a naturally occurring estrogenic substance. 

As such, and pursuant to Section 408(p)(4), the EPA will determine in the future whether it can 

exempt Pseudomonas fluorescens strain D7 from the requirements of the Section 408(p)(4) 

EDSP. In the event the EPA does determine to exempt this substance from the EDSP, an order 

will be issued. 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: 

http://www.epa.gov/endo/.

2. Federal Food, Drug, and Cosmetic Act (FFDCA) Considerations

Section 408(c)(2)(A)(i) of FFDCA allows the EPA to establish an exemption of the requirement 

of a tolerance (the legal limit for a pesticide chemical residue in or on a food) only if the EPA 

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 it 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 for a tolerance, the EPA must take into account the factors set forth in section 

408(b)(2)(C) of FFDCA, which require the EPA to give special consideration to exposure of 

infants and children to the pesticide chemical residue in establishing a tolerance exemption, 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 EPA consider “available information concerning the cumulative effects 

of [a particular pesticide's] . . . residues and other substances that have a common mechanism of 

toxicity.”

The EPA performs a number of analyses to determine the risks from aggregate exposure to 

pesticide residues. First, the Agency determines the toxicity of a pesticide. Second, the EPA 

examines exposure to the pesticide through food, drinking water, and through other exposures 

that occur as a result of pesticide use in residential settings. Consistent with section 408(b)(2)(D) 
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of FFDCA, the EPA 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 also considered available information concerning the variability of 

the sensitivities of major identifiable subgroups of consumers, including infants and children.

Based on the acute toxicity/pathogenicity data and information discussed previously and 

presented in Table 3 in Appendix A, the data required for a FFDCA risk assessment for 

Pseudomonas fluorescens strain D7 have been fulfilled.

a. Aggregate Exposure

In examining aggregate exposure, section 408 of FFDCA directs EPA to consider 

available information concerning exposures from the pesticide residue in food and all other 

nonoccupational exposures, including drinking water from ground water or surface water and 

exposure through pesticide use in gardens, lawns, or buildings (residential and other indoor 

uses).

Food Exposure and Risk Characterization: The EPA found that increased dietary exposure to

Pseudomonas fluorescens strain D7, a naturally occurring bacterium, is anticipated to be 

negligible. For the proposed use of Pseudomonas fluorescens strain D7 as an herbicide, the 

applications are made before crop plants emerge, or as seed treatment, and consequently oral 

exposure to residues from such use is expected to be minimal. Pseudomonas fluorescens strain 

D7 and other closely related Pseudomonas fluorescens bacteria already exist in the soil 

environment. The EPA concluded that the risk posed to adults, infants, and children is likely to 

be minimal because of the low acute oral toxicity/pathogenicity potential of Pseudomonas 

fluorescens strain D7.

Drinking Water Exposure and Risk Characterization: Exposure to residues of Pseudomonas 

fluorescens strain D7 in consumed drinking water is not likely to increase because there are no 

use sites for the pesticide with direct applications to water. There is a possibility of spray drift 

from aerial and ground applications or runoff of prepared fields and rangelands into surface 

waters. Ground water is not expected to have significant exposure to Pseudomonas fluorescens

strain D7 since, like other microorganisms, this microbial pesticide would likely be filtered out 

by the particulate nature of many soil types. If it were to be transferred to surface or ground 

waters that are intended for eventual human consumption (e.g., through spray drift or runoff) and 

directed to wastewater treatment systems or drinking water facilities, it likely would not survive 

the conditions water is subjected to in such systems or facilities, including chlorination, pH 

adjustments, filtration, and/or occasionally high temperatures (Centers for Disease Control and 

Prevention 2009; U.S. EPA 2004, and DeFelice et.al., 1993). In the remote likelihood that

Pseudomonas fluorescens strain D7 is present in drinking water (e.g., water not subject to 

treatment systems or facilities), its target pest specificity and available data indicate no toxicity

and/or pathogenicity is likely to occur with any drinking water exposure to Pseudomonas 

fluorescens strain D7 that results from pesticide applications made in accordance with good 

agricultural practices (see section IV(B)(1)(a) on pages 8–10 and Table 3 in Appendix A).

Non-occupational, Residential Risk Characterization: Given that Pseudomonas fluorescens

strain D7 use sites do not include residential settings and because the bacterium is naturallyoccurring, EPA determined that non-occupational exposure to the bacterium is unlikely.

Repeated exposures to the Pseudomonas fluorescens strain D7 microorganism from pesticidal 

applications do not exceed EPA’s level of concern, particularly in light of available data that 
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demonstrate Pseudomonas fluorescens strain D7 is not toxic (acute dermal toxicity and acute 

pulmonary toxicity/ pathogenicity), is non-irritating (primary dermal irritation), and is not 

pathogenic when used as labeled in accordance with good agricultural practices (acute 

pulmonary toxicity/pathogenicity, and acute injection toxicity/pathogenicity) (see section 

IV(B)(1)(a) on pages 8–11 and Table 3 in Appendix A).

b. 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 exemption, the EPA consider “available information concerning the 

cumulative effects of [a particular pesticide’s] . . . residues and other substances that have a 

common mechanism of toxicity.” The EPA has not found Pseudomonas fluorescens strain D7 to 

share a common mechanism of toxicity with other substances. Pseudomonas fluorescens strain 

D7 does not appear to be toxic to humans via dietary, dermal and pulmonary exposure. For the 

purposes of the tolerance action, therefore, the EPA has assumed that Pseudomonas fluorescens

strain D7 does not have a common mechanism of toxicity with other substances. Thus, section 

408(b)(2)(D)(v) of the FFDCA does not apply. For information regarding the EPA’s efforts to 

determine which chemicals have a common mechanism of toxicity and to evaluate the 

cumulative effects of such chemicals, see the EPA’s website at 

http://www.epa.gov/pesticides/cumulative.

c. Determination of Safety for the United States Population, Infants and Children

In considering the establishment of a tolerance or tolerance exemption for a pesticide chemical 

residue, FFDCA section 408 (b)(2)(C) provides that the 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 the EPA shall apply an additional tenfold (10X) 

margin of exposure (safety) for infants and children in the case of threshold effects to account for 

prenatal and postnatal toxicity and the completeness of the database on toxicity and exposure 

unless the EPA determines that a different margin of exposure (safety) will be safe for infants 

and children. This additional margin of exposure (safety) is commonly referred to as the Food 

Quality Protection Act Safety Factor. In applying this provision, the EPA either retains the 

default value of 10X or uses a different additional safety factor when reliable data available to 

the EPA support the choice of a different factor.

Based on the acute toxicity and pathogenicity data/information discussed in section IV(B)(1)(a) 

(see pages 8–11) and Table 3 in Appendix A, the EPA concludes that there are no threshold 

effects of concern to infants, children, or adults when Pseudomonas fluorescens strain D7 is used 

as labeled in accordance with good agricultural practices. As a result, the EPA concludes that no 

additional margin of exposure (safety) is necessary to protect infants and children and that not 

adding any additional margin of exposure (safety) will be safe for infants and children.

Moreover, based on the same data/information and EPA analysis as presented directly above, the 

Agency is able to conclude 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 Pseudomonas fluorescens strain D7 when it is used—as labeled and in accordance with good
Pseudomonas fluorescens strain D7 Page 14 of 29

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agricultural practices—as an herbicide. Such exposure includes all anticipated dietary exposures 

and all other exposures for which there is reliable information. The EPA has arrived at this 

conclusion because, considered collectively, the data and information available on Pseudomonas 

fluorescens strain D7 do not demonstrate toxic or pathogenic potential to mammals, including 

infants and children. 

3. Occupational Exposure and Risk Characterization

The EPA does not expect handler exposure to Pseudomonas fluorescens strain D7 to pose any 

undue risk. Regardless, appropriate personal protective equipment (PPE) and precautionary 

statements are required on pesticide product labels to mitigate any potential risks to pesticide 

handlers due to prolonged or numerous exposures. Handlers applying D7 in agricultural settings 

must wear a dust/mist filtering respirator, waterproof gloves, long-sleeved shirt, long pants, 

socks and shoes. 

4. Human Health Risk Characterization

The EPA considered human exposure to Pseudomonas fluorescens strain D7 in light of the 

standard for registration in FIFRA and the relevant safety factors in FFDCA. The Agency 

determined that no unreasonable adverse effects to the United States population in general, and 

to infants and children in particular, will result when products containing Pseudomonas 

fluorescens strain D7 are used in accordance with EPA-approved labeling.

C. Environmental Assessment (40 CFR § 158.2150)

Four original guideline studies with the TGAI and three sets of data waiver rationale were 

submitted to meet data requirements for nontarget organisms per 40 CFR Part 158.2150 for 

Pseudomonas fluorescens strain D7, which are sufficient to fulfill the relevant microbial 

pesticide data requirements and for risk assessment purposes. Further testing of nontarget 

organisms at higher tier levels (i.e., Tiers II, III, and IV) is not required. Adverse effects to 

nontarget organisms, including federally listed threatened and endangered species, are not 

expected to result from the proposed registration of P. fluorescens strain D7 when applied in 

accordance with the directions on the proposed label. 

For a summary of the generic data requirements described in sections IV(C)(1) (see pages 14–

15), refer to Table 4 in Appendix A.

1. Ecological Exposure and Risk Characterization 

According to the proposed label for the EP, P. fluorescens strain D7 is intended for use in 

suppression of downy brome, medusahead, Japanese brome, and jointed goatgrass on wheat, 

barley, triticale, oats, and rangeland. The EP containing 95% P. fluorescens strain D7 is a freezedried powder that is to be mixed in water and applied as a direct spray to the soil surface at a rate 

of 2 g /acre (4 x 1011 cells/acre). The label directs the user to apply the EP before weed seeds 

germinate in the spring or autumn when daytime temperatures are below 50oF. The EP is to be 

applied just prior to expected rainfall, and can be applied up to 4 times per year for a maximum 

annual application of 8g/acre. The EP may also be used as a seed treatment to control the listed 

weeds at planting.
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The proposed EP label indicates that the product is applied by spray (except for seed treatments), 

and may be applied using aerial equipment. Aerial sprays are generally expected to have greater 

potential for nontarget exposure compared to other application methods (e.g., ground sprays, 

sprays using hand-held equipment). However, because the EP is to be applied directly to the soil 

surface, in autumn or spring when temperatures are cooler, and at preemergence, the potential 

exposure of nontarget organisms to the EP is expected to be more limited. Spray drift may result 

in some exposure to nearby areas, including aquatic environments. However, based on the Tier I 

aerial agricultural application scenario in the AgDRIFT spray drift model, assuming fine to 

medium droplet size, a maximum of 12.5% of the amount applied is expected to reach aquatic 

areas. P. fluorescens also occurs naturally in soil and water (Pallerroni, 2005). As discussed 

above, strain D7 is a root-associated bacterium which was isolated originally from the West 

Coast region of the United States, so soil-dwelling nontarget organisms in those areas currently 

have some exposure to this bacterium.

Birds and Mammals 

The guideline study submitted for the avian oral toxicity/pathogenicity testing requirement 

showed no adverse effects in the bird species tested (Northern bobwhite, Colinus virginianus). P. 

fluorescens strain D7 also is not likely to grow at a bird’s high body temperatures (see McNab, 

1966). Based on expected relatively low exposure and lack of adverse effects observed in birds 

exposed to P. fluorescens strain D7, risk to nontarget birds resulting from the proposed 

registration is minimal.

An acute oral toxicity/pathogenicity study with laboratory rats indicated no adverse effects of P. 

fluorescens strain D7 when administered at a dose of 1.3 X 109 CFU/animal (MRID No. 

48966403). As discussed above, other studies with laboratory mammals also showed no adverse 

effects upon exposure to P. fluorescens strain D7 (USEPA, 2014a; 2014b). Some wild mammals 

may be exposed to P. fluorescens strain D7 in treated areas, but the exposure of most wild 

mammals is expected to be limited because of the root-borne nature of P. fluorescens strain D7

(i.e., it is a rhizobacterium), application timing (e.g., pre-emergence) and type of use (e.g., 

applied to soil surface). Since adverse effects were not observed in the mammalian studies, risk 

to wild mammals is expected to be low from the proposed registration of the new a.i.

Nontarget Insects and Honey Bees

The guideline study submitted to meet the requirement for nontarget insect testing did not show 

toxicity or pathogenicity to ladybird beetles (Hippodamia convergens) upon a 7-day dietary 

exposure to corn earworm (Helicoverpa zea) sprayed with P. fluorescens strain D7 at 1 x 106

cfu/ml. The applicant also provided a study with honey bees to support a previous Experimental 

Use Permit for P. fluorescens D7 (MRID No. 48668301) which showed no adverse effects at a 

dose of 1 x 107cfu/ml over seven days. This study was classified as supplemental due to the 

short duration (USEPA, 2012), so the applicant submitted rationale (MRID No. 49177701) to 

address the honey bee data requirement for the proposed section 3 registration of P. fluorescens

strain D7. The rationale referenced the former study and also cited a literature study (Johnson et 

al., 1993) that involved honey bees as a vector for the dispersal of another strain of this 

microorganism, Pseudomonas fluorescens strain A506, to apple and pear blossoms without 

observable impact on honey bees.
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Exposure to nontarget insects and honey bees may occur in the treated areas. However, under 

the recommended application conditions (cool, below 50oF, and wet), insects including honey 

bees are not likely active, thus no significant exposure of these insects to P. fluorescens strain D7 

is expected. 

Based on the above information (no hazard and low exposure), adverse effects to nontarget 

honey bees and insects are not likely from the proposed registration of the new a.i.

Nontarget Plants

P. fluorescens occurs naturally in soil and water. P. fluorescens strain D7, the a.i., was isolated 

initially from the West Coast region of the United States. No adverse effects of the a.i. on tested 

dicots and most monocots were shown in greenhouse, growth chamber or field studies. 

Exposure to nontarget plants is expected with the proposed registration of P. fluorescens strain 

D7. Scientific rationale submitted for a waiver of the requirement for Nontarget Plant Testing is 

based on its selective mode of action on monocotyledonous weeds (particularly to Bromus spp.) 

and limited number of seasonal applications to soil surfaces, and the microorganism’s reported 

ecology. The rationale provided was sufficient to conclude that the proposed uses of P. 

fluorescens strain D7 to suppress certain weeds (downy brome, medusa head, Japanese brome, 

and jointed goat grass) is not expected to result in increased exposure to, or adverse effects in 

nontarget plants of economic importance.

Freshwater Fish and Invertebrates

The studies submitted for the nontarget freshwater fish and invertebrate testing requirements 

showed no toxicity/pathogenicity of P. fluorescens strain D7 to Oncorhynchus mykiss (rainbow 

trout) and Daphnia magna when these aquatic animals were exposed to P. fluorescens strain D7. 

Some exposure to freshwater fish and invertebrates in nearby aquatic environments may occur 

with proposed applications of P. fluorescens D7, especially if the proposed treatments are 

applied in aerial sprays. However, as described above, the exposure of freshwater fish and 

invertebrates would be somewhat limited because of the type of use (e.g., applied directly to soil 

surfaces and before the rain) of P. fluorescens D7 and limited number of applications in autumn 

and spring, respectively, when it is cool (below 50oC) and wet. Based on the studies submitted 

and anticipated low exposure to strain D7 sprayed onto soil surfaces, adverse effects on 

nontarget freshwater organisms are not likely. 

Marine/Estuarine Fish and Invertebrates

The applicant did not submit a guideline study to meet the requirement for nontarget 

marine/estuarine fish and invertebrate testing, but submitted scientific rationale in lieu of testing. 

This testing is conditionally required if significant exposure of nontarget marine/estuarine fish 

and invertebrates to P. fluorescens strain D7 is expected. Similar to the nontarget organisms in 

fresh water environments, P. fluorescens strain D7 is not expected to reach marine/estuarine 

environments in significant quantities, thus significant exposure of the estuarine/marine fish and 

invertebrates to stain D7 is not likely as a result of the proposed registration of the a.i. Therefore, 

adverse effects to marine/estuarine fish and invertebrates are not anticipated. 
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2. Environmental Fate Data 

As the data and information provided are sufficient to fulfill the Tier I nontarget organism data 

requirements and allow for nontarget organism risk assessment for Pseudomonas fluorescens

strain D7, further testing at higher levels (Tiers II, III, and IV) is not required. 

3. Threatened and Endangered Species Assessment

Since EPA has determined that no effects are anticipated for any nontarget species exposed to 

Pseudomonas fluorescens strain D7 as a result of the proposed labeled applications, effects to 

federally-listed threatened and endangered species and their designated critical habitats are also 

not expected. Therefore, a “No Effect” determination is made for direct and indirect effects to 

listed species and their designated critical habitats resulting from the proposed uses of 

Pseudomonas fluorescens strain D7, as labeled.

V. ENVIRONMENTAL JUSTICE

The 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. 

Fair treatment means that no group of people, including racial, ethnic, or socioeconomic groups, 

should bear a disproportionate share of the negative environmental consequences resulting from 

industrial, municipal, and commercial operations or the execution of federal, state, local, and 

tribal environmental programs and policies. Meaningful involvement means that (1) potentially 

affected community residents have an appropriate opportunity to participate in decisions about a 

proposed activity that will affect their environment and/or health; (2) the public’s contribution 

can influence the regulatory agency’s decision; (3) the concerns of all participants involved will 

be considered in the decision-making process; and (4) the decision-makers seek out and facilitate 

the involvement of those potentially affected. The EPA has this goal for all communities and 

persons across the United States. 

To help address potential environmental justice issues, during the 15-day public participation 

comment period, the EPA sought 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 Pseudomonas fluorescens strain D7, compared to the general population. The EPA 

received no public comments on this particular matter.

For additional information regarding environmental justice issues, please visit the EPA’s website 

at http://www.epa.gov/compliance/environmentaljustice/index.html.

VI. RISK MANAGEMENT DECISION

Section 3(c)(5) of FIFRA permits for the registration of a pesticide provided that all the 

following determinations are made:

(1) Its composition is such as to warrant the claims for it;

(2) Its labeling and other material required to be submitted comply with the
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 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.

To satisfy criterion 1, the Pseudomonas fluorescens strain D7 products, the TGAI and EP, have

well-known properties. The EPA has no knowledge that would contradict the claims made on the 

labels, and the pesticide products are not expected to cause unreasonable adverse effects on the 

environment when used according to label instructions. Criterion 2 is satisfied by the product 

labels, as well as the data and information presented in this document. The Agency believes that

the Pseudomonas fluorescens strain D7 pesticide products will not cause any unreasonable 

adverse effects on the environment and based on cited field testing publications these products

offer biocontrol of certain weed species, satisfying criterion 3. Criterion 4 is satisfied in that the

Pseudomonas fluorescens strain D7 pesticide products are not expected to cause unreasonable 

adverse effects when used according to label instructions. Therefore, both the MP and the EP, 

D7, which contains Pseudomonas fluorescens strain D7 as a new active ingredient, are eligible 

for registration under FIFRA section 3(c)(5) for the labeled uses. 

VII. ACTIONS REQUIRED OF THE REGISTRANT

A. Final Printed Labeling

Before the manufacturing-use product, Pseudomonas fluorescens strain D7 TGAI and the enduse product, D7, may be released for shipment, the applicant is required to provide appropriate 

final printed labeling to the EPA.

B. Terms of Registration

The EPA is not requiring additional information on the registration of these two products.

C. Reporting of Adverse Effects and Hypersensitivity Incidents

Notwithstanding the information stated in the previous sections, it should be clearly understood 

that certain specific data must be reported to the EPA as a requirement for maintaining the 

federal registration for a pesticide product. Reports of all incidents of adverse effects to the 

environment must be submitted to the EPA under the provisions stated in FIFRA section 6(a)(2). 

Additionally, all incidents of hypersensitivity (including both suspected and confirmed incidents) 

must be reported to the EPA under the provisions of 40 CFR § 158.2140(d).
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VIII. GLOSSARY OF ACRONYMS AND ABBREVIATIONS

BRAD Biopesticides Registration Action Document

CFR Code of Federal Regulations

CFU colony forming units

cSt centistokes

EDSP Endocrine Disruptor Screening Program

EP end-use pesticide product

EPA United States Environmental Protection Agency (the “Agency”)

FFDCA Federal Food, Drug, and Cosmetic Act

FIFRA Federal Insecticide, Fungicide, and Rodenticide Act

FR Federal Register

g/mL grams per milliliter

lb/gal pounds per gallon

LD50 median lethal dose. A statistically derived single dose that can be expected 

to cause death in 50% of the test animals when administered by the route 

indicated (oral, dermal, or inhalation). It is expressed as a weight of 

substance per unit weight of animal (e.g., mg/kg).

mg/kg milligrams per kilogram

MP manufacturing-use pesticide product

MPCA microbial pest control agent 

MRID No. Master Record Identification Number

NIOSH National Institute for Occupational Safety and Health

OPP Office of Pesticide Programs

PC Code Pesticide Chemical Code

PP Pesticide Petition

PPE personal protective equipment

TGAI technical grade of the active ingredient

U.S. United States
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IX. BIBLIOGRAPHY

A. Studies Submitted to Support the Active Ingredient, Pseudomonas fluorescens strain D7 

and Pesticide Product Registrations for the Fluorescens Technical and the EP, D7

MRID No. Study Information

48497800 Northwest Agricultural Products (2011) Submission of Product Chemistry Data in 

Support of the Experimental Use of Bacterium Pseudomonas fluorescens Strain D7 

on Aggressive Weeds. Transmittal of 1 Study.

48497801 Brooks, M. (2011) D7, Product Chemistry, Group: Product Identity, Composition 

and Analytical Test Guidelines and Group B: Physical and Chemical Properties 

Test Guidelines. Project Number: NAP01. Unpublished study prepared by 

NorthWest Agricultural Products. 21 p.

48524500 Northwest Agricultural Products (2011) Submission of Product Chemistry Data in 

Support of the Experimental Use of D7. Transmittal of 1 Study.

48524501 Brooks, M. (2011) D7, Product Chemistry, Deposition of a Sample in a Nationally 

Recognized Cullture Collection. Discussion of Formation of Unintentional 

Impurities. Analysis of Samples. Project Number: NAP02. Unpublished study 

prepared by Ag-Chem Consulting. 7 p.

48548901 Kennedy, A. (2011) Effect of Pseudomonas Strain D7 on Nontarget Vegetation. 

Project Number: NAP003. Unpublished study prepared by Washington State 

University. 13p.

48668300 Northwest Agricultural Products (2011) Submission of Toxicity Data in Support of 

the Experimental Use of Pseudomonas fluorescens Strain D7. Transmittal of 1 

Study.

48668301 Younger, C. (2011) Pseudomonas fluorescens D7: Honey Bee, Apis mellifera, Nontarget Insect Microbial Testing: Final Report. Project Number: 14619/10. 

Unpublished study prepared by Stillmeadow, Inc. 12p.

48724301 Brooks, M. (2012) Metabolite Analysis of Pseudomonas Strain D7. Project 

Number: NAP04. Unpublished study prepared by Ag-Chem Consulting, LLC. 33p.

48966400 Northwest Agricultural Products (2012) Submission of Product Chemistry and 

Toxicity Data in Support of the Application for Registration of D7 and Fluorescens 

Technical. Transmittal of 10 Studies.

48966401 Wend, C. (2012) Identification of Pseudomonas fluorescens Strain D7. Project 

Number: NAP20121004. Unpublished study prepared by Northwest Ag. Products. 

27p.

48966402 Monds, K. (2011) Pseudomonas fluorescens D7: Intravenous Toxicity / 

Pathogenicity Study in Rats: Final Report. Project Number: 14620/10. Unpublished 

study prepared by Stillmeadow, Inc. 34p.
Pseudomonas fluorescens strain D7 Page 21 of 29

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48966403 Doig, A. (2012) Pseudomonas fluorescens Strain D7: Acute Oral Toxicity / 

Pathogenicity Study in Rats: Final Report. Project Number: 15730/11. Unpublished 

study prepared by Stillmeadow, Inc. 29p.

48966404 Monds, K. (2012) Pseudomonas fluorescens Strain D7: Acute Pulmonary Toxicity / 

Pathogenicity Study in Rats: Final Report. Project Number: 15728/11. Unpublished 

study prepared by Stillmeadow, Inc. 30p.

48966405 Monds, K. (2012) Pseudomonas fluorescens Strain D7: Acute Dermal Toxicity / 

Pathogenicity Study in Rabbits: Final Report. Project Number: 15729/11. 

Unpublished study prepared by Stillmeadow, Inc. 12p.

48966406 Brooks, M. (2012) D7, Product Chemistry, Stability to Normal and Elevated 

Temperatures, Metals and Metal Ions, Analysis of Samples, Storage Stability and 

Corrosion Characteristics. Project Number: NAP20121002. Unpublished study 

prepared by Northwest AG Product. 17p.

48966407 Younger, C. (2012) Pseudomonas fluorescens Strain D7: Ladybird Beetle Nontarget Insect Microbial Testing. Project Number: 16338/12. Unpublished study 

prepared by Stillmeadow, Inc. 11p.

48966408 Hartwell, T. (2011) Pseudomonas fluorescens Strain D7: Microbial Pest Control 

Agent (MPCA) Freshwater Fish Test with Oncorhynchus mykiss (Rainbow Trout): 

Final Report. Project Number: 14622/10. Unpublished study prepared by

Stillmeadow, Inc. 17p.

48966409 Younger, C. (2011) Pseudomonas fluorescens D7: Acute Oral Toxicity Study in 

Bobwhite Quail: Final Report. Project Number: 14716/10. Unpublished study 

prepared by Stillmeadow, Inc. 17p.

48966410 Hartwell, T. (2011) Pseudomonas fluorescens D7: Microbial Pest Control Agent 

(MPCA) Freshwater Aquatic Invertebrate Test with Daphnia magna: Final Report. 

Project Number: 14623/10. Unpublished study prepared by Stillmeadow, Inc. 27p.

49021100 Northwest Agricultural Products (2012) Submission of Fate Data in Support of the 

Application for Registration of Fluorescens Technical and D7. Transmittal of 1 

Study.

49021101 Brooks, M. (2012) Effect of Pseudomonas Strain D7 on Nontarget Vegetation -

Supplemental Report. Project Number: NAP003A. Unpublished study prepared by 

U.S. Dept. of Agriculture/Pacific West Area. 165p.

49100800 Northwest Agricultural Products (2013) Submission of Toxicity Data in Support of 

the Applications for Registration of D7 and Fluorescens Technical. Transmittal of 2 

Studies.

49100801 Kuhn, J. (2013) Pseudomonas fluorescens Strain D7 Acute Eye Irritation in 

Rabbits. Project Number: 17085/12. Unpublished study prepared by Stillmeadow, 

Inc. 16p.

49100802 Kuhn, J. (2013) Pseudomonas fluorecens Strain D7: Acute Dermal Irritation in 

Rabbits. Project Number: 17085/12. Unpublished study prepared by Stillmeadow, 

Inc. 10p.
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49157600 Northwest Agricultural Products (2013) Submission of Toxicity Data in Support of 

the Applications for Registration of Fluorescens Technical and D7. Transmittal of 1 

Study.

49157601 Younger, C. (2013) Pseudomonas fluorescens Strain D7: Ladybird Beetle NonTarget Insect Microbial Testing. Project Number: 16338/12. Unpublished study 

prepared by Stillmeadow, Inc. 12p.

49177700 Northwest Agricultural Products (2013) Submission of Product Chemistry, 

Toxicity, and Fate Data in Support of the Applications for Registration of 

Fluorescens Technical and D7. Transmittal of 3 Studies. 

49177701 Brooks, M. (2013) Pseudomonas Strain D7 Nontarget Insect Testing/ Supplemental 

Report. Project Number: NAP7162012A1. Unpublished study prepared by AgChem Consulting. 71p.

49177702 Brooks, M. (2013) Effect of Pseudomonas Strain D7 on Nontarget VegetationSupplemental Report 2. Project Number: NAP003B. Unpublished study prepared 

by Washington State University and U.S. Dept. of Agriculture. 36p.

49177703 Brooks, M. (2013) Pseudomonas Strain D7 Discussion of Formation of 

Unintentional Ingredients: Supplemental Report. Project Number: NAP71612013. 

Unpublished study prepared by Northwest Agricultural Products. 10p.

49349700 Northwest Agricultural Products (2014) Submission of Toxicity Data in Support of 

the Applications for Registration of Fluorescens Technical and D7. Transmittal of 1 

Study.

49349701 Doig, A. (2014) Pseudomonas fluorescens D7: Intravenous Toxicity/Pathogenicity 

Study in Rats. Project Number: 17873/13. Unpublished study prepared by 

Stillmeadow, Inc. 38p.

B. Environmental Protection Agency Risk Assessment Memoranda

US Environmental Protection Agency (USEPA). 2012. Memorandum from S. Borges 

(OPP/BPPD) to S. Cerrelli, Re: Environmental risk assessment for Pseudomonas fluorescens

strain D7 in response to an application for a FIFRA Section 5 Experimental Use Permit. Dated 

February 6, 2012.

USEPA. 2014a. Memorandum from I. Barsoum (OPP/BPPD) to S. Cerrelli, Re: Review of 

Acute Toxicity/Pathogenicity study for section 3 registration of the TGAI, Fluorescens 

Technical, and the EP, D7, containing the active ingredient Pseudomonas fluorescens strain D7. 

Dated April 15, 2014.

USEPA. 2014b. Memorandum from I. Barsoum (OPP/BPPD) to S. Cerrelli, Re: Review of 

Acute Toxicity/Pathogenicity study for section 3 registration of the TGAI, Fluorescens 

Technical, and the EP, D7, containing the active ingredient Pseudomonas fluorescens strain D7. 

Dated April 15, 2014.

USEPA. 2014c. Memorandum from I. You, Ph.D. through S. Borges (OPP/BPPD) to S. Cerrelli 

Environmental risk assessment for the FIFRA Section 3 registration of the TGAI, Fluorescens
Pseudomonas fluorescens strain D7 Page 23 of 29

Biopesticides Registration Action Document

Technical (EPA Reg. No. 71975-G), and the EP, D7 (EPA Reg. No. 71975-U), containing the 

active ingredient Pseudomonas fluorescens Strain D7. Dated July 18, 2014.

C. Other References

Centers for Disease Control and Prevention. 2009. Drinking Water - Water Treatment. Available 

from http://www.cdc.gov/healthywater/drinking/public/water_treatment.html.

DeFelice K, Wollenhaupt N, Buchholz D. 1993. Aquifers and Soil Filter Effect. Available from 

http://extension.missouri.edu/p/WQ24.

U.S. EPA. 2004. Primer for Municipal Wastewater Treatment Systems. EPA 832-R-04-001. 

Available from http://www.epa.gov/npdes/pubs/primer.pdf

Johnson, K. B., V. O. Stockwell, D. M. Burgett, D. Sugar, and J. E. Loper. 1993. Dispersal of 

Erwinia amylovora and Pseudomonas fluorescens by Honey Bees from Hives to Apple and Pear 

Blossoms. Phytopathology 83:478-484. 

Kremer, R. J. and A. C. Kennedy. 1996. Rhizobacteria as Biocontrol Agents for Weeds. Weed 

Technology 10:601-609.

McNab, B. K. 1966. An Analysis of the Body Temperatures of Birds. The Condor 68:47-55.

Palleroni, N. J. 2005. Pseudomonas. Pp. 323 – 379. In: Bergey’s Manual of Systematic 

Bacteriology, Vol. 2. D. J. Brenner, N. R. Krieg, and J. T. Staley (Eds.) Springer, New York, 

New York

Skipper, H. D. A. G. Ogg, and A. C. Kennedy. 1996. Root Biology of Grasses and Ecology of 

Rhizobacteria for Biological Control. Weed Technology 10: 610-620. 
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APPENDIX A. MICROBIAL PESTICIDES DATA REQUIREMENTS 

(40 CFR PART 158 – SUBPART V)

TABLE 1. Product Analysis Data Requirements for the Active Ingredient (TGAI), Pseudomonas fluorescens strain D7 and the 

End-Use Pesticide Product (EP), D7 (40 CFR § 158.2120) 

Harmonized 

Guideline 

Number

Data Requirement Results MRID No.

885.1100 Product Identity Submitted data fulfill the requirement for product identity and composition. 48966401

885.1200 Manufacturing 

Process Submitted data fulfill the requirement for manufacturing process. 48497801

885.1250

Deposition of a 

Sample in a 

Nationally 

Recognized Culture 

Collection

Pseudomonas fluorescens strain D7 is on deposit at Agricultural Research 

Service Culture Collection as NRRL B-18293

885.1300

Discussion of 

Formation of 

Unintentional 

Ingredients

Submitted data fulfill the requirement for preventing and detecting any 

microbial contaminants. 4917703

885.1400 Analysis of Samples Submitted data fulfill the requirement for analysis of samples.

48966406

885.1500 Certification of 

Limits Submitted data fulfill the requirement for manufacturing process. CSFs dated 9/25/12
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TABLE 2. Physical and Chemical Characteristics for the Technical Grade of the Active Ingredient (TGAI), Pseudomonas 

fluorescens strain D7 and the End-Use Pesticide Product (EP),D7 (40 CFR § 158.2120)

Harmonized 

Guideline Number Data Requirement Results MRID No.

830.6302 Color Brown

48497801 830.6303 Physical State Granular

830.6304 Odor Sweet sugar smell

830.6313

Stability to Normal and 

Elevated

Temperatures, Metals,

and Metal Ions

Stability to Metals: N/A

Stability to Elevated Temperatures: This product is not stable 

at elevated temperatures and must be stored refrigerated or 

frozen. This product will be stored in paper and used 

immediately when tank mixed.

N/A

830.6317 Storage Stability Submitted data fulfill the requirement for storage stability and 

demonstrate stabilty for s test period of 361 days.

48966406

830.6319 Miscibility Not applicable (chemical is solid).

830.6320 Corrosion 

Characteristics

Submitted data fulfill the requirement for corrosion 

characteristics

48966406

830.7000 pH The pH of a 1% solution is 5.1 48497801

830.7100 Viscosity Not applicable to a solid

830.7300

Density/Relative 

Density/Bulk Density 

(Specific Gravity)

12.48 lbs/ ft3 48497801
Pseudomonas fluorescens strain D7 Page 26 of 29

Biopesticides Registration Action Document

TABLE 3. Toxicology Data Requirements for the Active Ingredient Pseudomonas fluorescens strain D7 

and the End Use Pesticide Product D7 (40 CFR § 158.2140)

Harmonized 

Guideline 

Number

Data Requirement Results MRID No.

Tier I

885.3050 Acute Oral 

Toxicity/Pathogenicity

Pseudomonas fluorescens strain D7 does not appear to be toxic, 

infective, and/or pathogenic in rats, when dosed at 1.3 X 109

CFU/animal.

Classification: Acceptable

48966403

885.3150 Acute Pulmonary 

Toxicity/Pathogenicity

Pseudomonas fluorescens strain D7 does not appear to be toxic, 

infective, and/or pathogenic in rat, when dosed at 4.6 x 108cfu / 

animal.

Classification: Acceptable

48966404

885.3200

Acute Injection 

Toxicity/

Pathogenicity

Average CFU counts of the cecum contents were 8.6 x 104, 5.5 x 105, 

2.9 x 105, and 2.1 x 102 CFU/g on days 3, 7, 14, and 21, respectively. 

Pseudomonas fluorescens strain D7 was TOXIC and/or 

PATHOGENIC as tested in this study.

Classification: Acceptable, however, injection with the active MPCA 

at a dose of 1.3 x 108 CFU/rat resulted in mortality, abnormal clinical 

signs; transient weight loss with resultant decreased absolute body 

weight, abnormal gross necropsy findings, and altered organ weights.

48966402

This study supersedes MRID 48966402 and was conducted to evaluate 

the acute intravenous toxicity and infectivity of the microbial pest 

control agent (MPCA), Pseudomonas fluorescens D7, at two 

concentration exposures and at adequate post-exposure observation 

period. No abnormalities were seen during daily observations or at 

necropsy. There was no mortality in any group during the study. The 

test organism had cleared completely from the active 106 CFU/rat and 

active 107 CFU/rat groups blood, kidneys, mesenteric lymph nodes, 

lungs, brain, liver, spleen and cecum contents by Day 7. The observed 

differences in spleen weights for groups treated with live Pseudomonas 

fluorescens D7 are considered a normal immune response during the 

clearance phenomenon. The test substance Pseudomonas fluorescens 

D7 was determined to be non-toxic to rats when injected intravenously 

at a dose of 3.4 x 107 CFU/rat or at 3 .7 x 106 CFU/rat .

Classification: Acceptable

49349701
Pseudomonas fluorescens strain D7 Page 27 of 29

Biopesticides Registration Action Document

TABLE 3. Toxicology Data Requirements for the Active Ingredient Pseudomonas fluorescens strain D7 

and the End Use Pesticide Product D7 (40 CFR § 158.2140)

Harmonized 

Guideline 

Number

Data Requirement Results MRID No.

885.3400 Hypersensitivity

Incidents

The applicant reported that no hypersensitivity incidents, including 

immediate-type or delayed-type reactions of humans and domestic 

animals, occurred during research, development, or testing of the 

TGAI. Any future hypersensitivity incidents must be reported to EPA 

(refer to test note #3 of 40 CFR § 158.2140(d)). 

885.3500 Cell Culture N/A

870.1100 Acute Oral Toxicity

This requirement addressed by Acute Oral Toxicity/pathogenicity 

study described above. 

Classification: Acceptable

48966403

870.1200 Acute Dermal Toxicity

Dermal LD50 Males >2000 mg/kg bw; Females > 2000 mg/kg bw; 

Combined > 2000 mg/kg bw. Based on the results of this study, 

Pseudomonas fluorescens strain D7 is of low toxicity.

Classification: Acceptable

TOXICITY CATEGORY IV

48966405

870.1300 Acute Inhalation 

Toxicity

This requirement addressed by Acute pulmonary 

Toxicity/pathogenicity study described above

Classification: Acceptable

48966404

870.2400 Acute Eye Irritation

No corneal opacity, iritis, or conjunctival irritation was noted on any 

rabbit throughout the study. Pseudomonas fluorescens strain D7 was 

not irritating to the eye.

Classification: Acceptable

TOXICITY CATEGORY IV

49100801

870.2500 Primary Dermal 

Irritation

No dermal irritation was noted on any animal during the study. The 

primary irritation index was 0.0.

Classification: Acceptable

TOXICITY CATEGORY IV

49100802

Tiers II and III

Not required for Pseudomonas fluorescens strain D7 based on the lack of acute toxicity/pathogenicity in the Tier I studies.
Pseudomonas fluorescens strain D7 Page 28 of 29

Biopesticides Registration Action Document

TABLE 4. Nontarget Organism Toxicity and Environmental Fate Data Requirements for the Technical Grade of the Active 

Ingredient TGAI, Pseudomonas fluorescens strain D7. (40 CFR § 158.2150)

Harmonized 

Guideline 

Number

Data Requirement Results MRID No.

Tier I

885.4050 Avian Oral Toxicity

An avian oral test indicated that P. fluorescens strain D7 is not toxic or 

pathogenic to Northern bobwhite (Colinus virginianus) dosed at 2.1 x 

109cfu/kg body wt/day. 

Classification: Acceptable

48966409

885.4100 Avian Inhalation 

Toxicity/Pathogenicity

Not required as the nature of the microbial pesticide does not indicate 

potential pathogenicity to birds or relatedness to any known bird 

pathogens (refer to test note #3 of 40 CFR § 158.2150(e)).

N/A

885.4150 Wild Mammal 

Toxicity/Pathogenicity

Wild mammal testing was not conducted nor was a waiver requested. 

Studies conducted with P. fluorescens strain D7 on laboratory animals 

are appropriate for use in risk assessment. Acceptable studies with 

laboratory rats indicated no concerns for toxicity/pathogenicity in 

mammals.

Classification: Acceptable 

48966403,

48966402

885.4200 Freshwater Fish 

Toxicity/Pathogenicity

A freshwater fish test with P. fluorescens strain D7 showed no toxic or 

pathogenic effect on Rainbow trout (Oncorhynchus mykiss) dosed at 1 x 

106cfu/ml. 

Classification: Acceptable 

48966408

885.4240 Freshwater Invertebrate 

Toxicity/Pathogenicity

A freshwater invertebrate study with P. fluorescens strain D7 showed 

no adverse effects on mobility or reproduction of Daphnia magna at 

exposure to 1 x 106cfu/ml of test medium. 

Classification: Acceptable

48966410

885.4280

Estuarine/Marine Fish 

and Invertebrate 

Testing

Scientific rationale was insufficient to conclude that no adverse effects 

are expected on nontarget fish and invertebrates for all applications. P. 

fluorescens strain D7 is not to be applied directly to water and thus is 

not expected to reach estuarine or marine environments in significant 

amounts. However, aerial applications over large areas may lead to 

some exposure of P. fluorescens D7 in nearby estuarine/marine 

environments. 

Classification: Supplemental

No MRID, but a 

letter (dated 

June 3, 2013) 

requesting a 

waiver for 

Estuarine/

Marine Fish and 

Invertebrate 

Testing was 

submitted. 

885.4300 Nontarget Plant Testing

Additional data and published literature submitted were sufficient to 

support the nontarget plant testing requirement for P. fluorescens strain 

D7.

Classification: Acceptable

49021101,

48966401

885.4340 Nontarget Insect 

Testing

A laboratory bioassay with adult ladybird beetles (Hippodamia 

convergens) at 1X the field application rate showed no adverse effects 

of P. fluorescens strain D7 to the test organism (Helicoverpa zea) within 

7 days of testing. The study is not of sufficient duration and was not 

carried out at the maximum hazard dose of 10X. 

Classification: Supplemental 

49157601

49177701

885.4380 Honey Bee Testing

Scientific rationale was sufficient to conclude that no adverse effects of 

P. fluorescens strain D7 on honey bees are likely to occur. Because of 

recommended preemergence uses in cool and wet (< 50oF) conditions, 

the exposure of nontarget honey bees to D7, even when it occurs, is 

expected to be low. 

Classification: Acceptable

49177701

Tiers II, III, and IV

Not required for based on the acceptability of the data and other information provided for Tier I.
Pseudomonas fluorescens strain D7 Page 29 of 29

Biopesticides Registration Action Document

APPENDIX B. PESTICIDE PRODUCTS

EPA 

File 

Symbol 

Registration 

Name

Percentage 

Active 

Ingredient

Formulation 

Type

Use Site(s) Method(s) of 

Application

Application Rate(s) Target Pest

71975-G

Fluorescens 

Technical 95% Technical Manufacturing use only N/A N/A

Downy Brome 

(cheatgrass), 

Medusahead, 

Japanese Brome, 

and Jointed 

Goatgrass

71975-U D7 95% End-use 

product

Agricultural

turf and grasses 

grown for seed, 

alfalfa; wheat, 

barley, triticale, 

oat, and 

rangeland

Ground 

application,

Aerial 

2g/acre (see label for 

dilution instructions)

Seed 

treatments 2-4 g/100 lbs. of seed