- 2

- TABLE OF CONTENTS

ACRONYMS

................................ ................................ ................................ ................... 3

REGISTRATION REVIEW TEAM

................................ ................................ ................... 4

PRELIMINARY WORK PLAN

................................ ................................ ........................ 5

A. Introduction

................................ ................................ ................................ ..... 5

B. Background and Regulatory Information

................................ ..................... 5

C. Anticipated Risk Assessment and Data Needs

................................ ............ 6

1. Product Analysis (40 CFR 158.2120)

................................ ..................... 6

2. Human Health Risk Assessment Status (40 CFR 158.2140)

................. 8

3. Environmental Fate and Ecological Risk Assessment Status

........... 10

4

. Risk to Threatened and Endangered Species

................................ ..... 12

5. Endocrine Effects

................................ ................................ .................. 12

6. Incidents

................................ ................................ ................................ . 13

7. Summary of Data Gaps

................................ ................................ ......... 13

D. Timeline

................................ ................................ ................................ ......... 13

E. Guidance for Commenters

................................ ................................ ........... 14

F. Environmental Justice

................................ ................................ .................. 14

H. Trade Irritants

................................ ................................ ................................ 15

I. Additional Information

................................ ................................ .................. 15

J. N ext Steps

................................ ................................ ................................ ..... 16

BIBLIOGRAPHY: Pseudomonas aureofaciens strain Tx

- 1

................................ ............. 17- 3

-



### ACRONYMS

Acronym Extension

μg microgram

a.i. Active ingredient
ATTC American Type Tissue Collection

B. cereus Ba cillus cereus
CFR Code of Federal Regulations

CFU Colony forming units

DCI Data Call

- in
EDSP Endocrine Disruptor Screening Program

EP End

- use product

EPA U.S. Environmental Protection Agency or, the Agency EPA Reg. No. EPA Registration Number: a two

- part number assigned by EPA to identify each

pesticide product registration.

ETE Emetic Toxin
FFDCA Federal Food, Drug, and Cosmetic Act

FIFRA Federal Insecticide, Fungicide, and Rodenticide Act
FQPA Food Quality Protection Act of 1996

FR F ederal Register

g gram HBL Hemolytic enterotoxin
IDS Incident Data System

L liter

LD 50 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 indic ated (oral, dermal, inhalation) mg/kg bw Milligrams per kilogram of body weight mg/L Milligrams per liter (approximately equivalent to ppm)

mL Milliliter

MP Manufacturing

- use product
MPCA Microbial pesticide control agent

MRID Master Record I d entification (number): EPA's system of recording and tracking

studies submitted to the Office of Pesticide Programs.

MRL Maximum residue limits

Nhe Nonhemolytic enterotoxin
NPIC National Pesticides Information Center

OPP EPA’s Office of Pesticide Pr ograms OPPTS Office of Prevention, Pesticides and Toxic Substances; now the Office of

Chemical Safety and Pollution Prevention (OCSPP)

PC Code Pesticide Chemical Code: a six

- digit number assigned by OPP to identify

pesticide chemicals.

Ps

. Pseudomonas Sy n Synonym

TMDL Total M aximum Daily L oad- 4

- 

### REGISTRATION REVIEW TEAM
Office of Pesticide Programs Biopesticides and Pollution Prevention Division

Microbial Pesticides Branch

Human Health

John Kough, Ph.D.

Environmental Effects

Gail Tomimatsu , Ph.D.

Risk Management

Susanne Cerrelli Sheryl Reilly, Ph.D.

Office of General Counsel

Philip Ross, Esq.- 5

-



### PRELIMINARY WORK PLAN

A. Introduction

The Food Quality Protection Act (FQPA) of 1996 mandated the continuous review of existing pesticides. All pesticides distributed or sold in the United States must generally be registered by the U.S. Environmental Protection Agency (EPA or the Agency), based on scientific data showing that they will not cause unreasonable risks to human health

o r the environment when used as directed on the product labeling. The Registration Review program is intended to make sure that, as the ability to assess risk evolves and as policies and practices change, all registered pesticides continue to meet the statu tory
standard of no unreasonable adverse effects. Changes in science, public policy, and pesticide use practices will occur over time. Through the Registration Review program, the Agency periodically reevaluates pesticides to make sure that as change occur s, products in the marketplace can continue to be used safely. Information on this program is provided at: http://www.epa.gov/oppsrrd1/registration_review/.

The Agency has begun to implement the Registration Review program pursuant to Federal Insecticide, Fungicide, and Rodenticide Act (FIFRA) section 3(g) and will review

each registered pesticide every 15 years to determine whether it continues to meet the

FIFRA standard for registration. The Agency will consider benefits information and data as required by FIFRA. The public phase of Registration Review begins when the initial docket is opened for each case. The docket is the Agency’s opportunity to state what it

knows about the pesticide and what additional risk analyses and data or information it

believe s are needed to make a Registration Review decision. After reviewing and responding to comments and data received in the docket during this comment period, the Agency will develop and commit to a final work plan and schedule for the

r egistration r eview of Pseudomonas aureofaciens

.

This Registration Review Preliminary Work Plan and Summary Document evaluates the microbial pest control agent ( MPCA) classified under Case 6009 : P seudomonas

aureofaciens

. More information about this case is available in the publ ic docket at regulations.gov. To access it, enter the docket number, which is E PA

- HQ

- OPP

- 2012

- 0421 in the blue Search box.

B. Background and Regulatory Information

Pseud omonas aureofaciens strain Tx

- 1 (PC Code 006473 ) was isolated from turf grass soil i n 1956 at Michigan S tate University. Pseudomonas aureofaciens ( synonym: Pseudomonas chlororaphis subspecies aureofaciens ) is a naturally occurring bacterium that is ubiquitous in soil and plant rhizospheres. Strains of this species have been used

for bio logical control of several fungal plant diseases that attack the root zone of various plant species. A n antibiotic (phenazine) produced by Pseudomonas aureofaciens is known to have antifungal properties that are useful in limiting the spread of fungal phy topathogens.- 6

-

The only registered pesticide product containing Pseudomonas aureofaciens strain Tx

- 1

as a pesticide active ingredient is Spot

- less Biofungicide, EPA Reg. No 75801

- 1

, w hich is registered for use for management of turf diseases on golf course s and other turf

sites

. This product was conditionally registered o n February 11, 1999

. This product is unusual in that a trained operator ferments the pesticide product on site in a Bioject fermentation system , and performs quality control prior to night

- time application s of the product to turf grass (including golf courses). T he product is applied to turf through chemigation system s or with conventional sprayers. The Spot

- less product label contains a use restriction that states that the product is not to be applied to turf or sod grown for sale, commercial seed production , and/or r esearch.

T he Pseudomonas aureofaciens species ha ve been reclassified in scientific literature as Pseudomonas chlororaphis subspecies aureofaciens based on the level of DNA

si milarity of Pseudomonas aureofaciens and Pseudomonas chlororaphis ( Johnson and Palleroni, 1989 ; and Palleroni, N.J., 2005 )

, b ut the registrat ion has not been amended to reflect the change in nomenclature for this microorganism. The Registration Review

case name Pseudomonas aureofaciens reflects the substance name provided when the Spot

- less product was initially registered.

Table 1 . Summary of Pesticide Product Containing Pseudomonas aureofaciens strain

Tx

- 1 (PC Code 006473 ) as an Active Ingredient

Regi stration

No Product Name Registrant First Registered

75801

- 1 Spot

- less

Biofungicide Turf Science

Laboratories, Inc. February 11, 1999

C

. Anticipated Risk Assessment and Data Needs

The following summaries describe how the current data requirements for m icrobial

p esticides have been addressed for the registration of Pseudomonas aureofaciens

strain Tx

- 1

. The pertinent data areas include: p roduct a nalysis

, r esidue d ata

, t oxicology (human health)

, n ontarget o rganisms

, and e nvironmental f ate (ecotoxicol ogy). D ata

gaps are also discussed

, and are summarized in Table 4

.

1. Product Analysis (40 CFR 158.2120)

At the time of submission, the data were adequate to support the registration of

Pseudomonas aureofaciens strain Tx

- 1. A dditional data are anticipate d to be needed, however, to ensure that the organism is currently d eposit ed in a nationally recognized culture collection ( 40 CFR 158.2120 ) . The Agency also expects to require the registrant

t o confirm the product identity (including the new species design ation, strain, and isolate) of the organism if it has not been previously deposited in a nationally recognized

culture collection. S torage stability

, miscibility

, and viscosity ( 40 CFR 158.2120 ) are data requirement s that remain outstanding and are expecte d to b e in need of be ing

addressed during the registration review of this case

.- 7

-

This assessment of the product analysis data submitted to support registration is based upon adequate quality control as provided at the time of culture of Ps. aureofaciens T x

1 (i.e., when the product is fermented on

- site in the BioJect System). The pesticide product label and the BioJect® Fermentation and Injection System Operator’s manual include q uality control measures necessary to assure the integrity of the product

.

Ta ble 1

. Summary of Product Analysis Data for Pseudomonas aureofaciens strain Tx

- 1

40 CFR 158.2120 Data Requirement (OPPTS Guideline) How it Has Been Addressed

MRID (S) EPA Review , Conclusion s , Comments)

Product Identity (885.110 0) 44337101

44422401 Acceptable

U .S. EPA 1999 a I n conjunction with redepositing the Tx

1 strain culture in a nationally recognized culture collection

, the Agency anticipates requiring confirmatory product identity data to

insure that the characteristics ar e identical to the originally registered product.
Manufacturing process (885.1200) 44337101

44437501

44461701 Acceptable

U .S. EPA 1999 a

Deposition of a sample in a nationally recognized culture collection (885.1200) 44337101 Additional information is exp ected to

be required to confirm that this requirement is currently addressed.

Discussion of formation of unintentional ingredients (885.1300) 44337101

44461701 Acceptable

U .S. EPA 1999 a

Analysis of samples (885.1400) 44337102

45048501

45077601 Acceptab le

U .S. EPA 2003
Certification of limits (885.1500) CSF dated

44337102 Additional data are expected to be

required. If the nomenclature of the active ingredient is amended or a cell deposit number is submitted, a new
CSF will need to be submitted that reflects the change(s).

Color (830.6302) 44337103 Orange Acceptable
Physical state (830.6303) 44337103 Liquid Acceptable
Odor (830.6304) 44337103 Slightly musty Acceptable- 8

-

40 CFR 158.2120 Data Requirement (OPPTS Guideline) How it Has Been Addressed

MRID (S) EPA Review , Conclusion s , Comments)

pH (830.7000) 44337103 7.5 Acceptable
Density/relative density/bulk

dens ity (specific gravity) (830.7300) 44337103 9.1 lbs . / gal. Acceptable

Corrosion characteristics (830.6320) 44337103 Noncorrosive to plastic Acceptable
Storage stability (830.6317) T hese data are expected to be required.
Stability to normal and elevate d temperatures, metals and metal ions (830.6313) 44337103 Stable in water at normal and elevated temperature (85 ̊F max). Not stable if stored in direct sunlight. Acceptable
Miscibility (830.6319) Th ese data are expected to be required for Spot

- less Biofungicide

. Viscosity (830.7100) Th ese data are expected t o be required for Spot

- less Biofungicide

. NON

- GUIDELINE

- Literature

Review For Pseudomonas produced antibiotics 45077601 Acceptable

. U .S EPA 2003

2. Human Health Risk Assessment Status (40 CFR 158.2140)

All mammalian toxicology data requirements hav e been satisfied. During initial registration

, EPA determined that there would be no human health concerns arising

from the use of Pseudomonas aureofaciens strain Tx

- 1 in pesticide products, and provided a public interest finding in support of pesticides containing Pseudomonas

aureofaciens strain Tx

- 1 as an alternative to more toxic conventional pesticides for turf disease control ( U.S. EPA, 1999a. )

The acute oral toxicity and dermal irritation studies submitted for the active ingredient

Ps. aureofaciens Tx

- 1 demonstrate a lack of toxicity or adverse effects following dosage with amounts ( expressed as colony forming units or “ cfu ”) surpassing those that are present following application of this bacterium through irrigation lines and sprinklers on golf cour ses. T his determination is for the active ingredient as applied through the BioJect Automated Microbial Fermentation and Injection System as currently designed; any changes in the identity of the active ingredient or alterations to the BioJect System

will require a full assessment for potential adverse effects.

The human health toxicity and pathogenicity data are summarized in the following table :- 9

-

Table 2

. Summary of Toxicology Data for Pseudomonas aureofaciens strain Tx

- 1

40 C FR 158.2140 Data Requirement (OPPTS Guideline) How it Has Been Addressed

MRID EPA Review , Conclusions , Comments

Acute oral toxicity/ pathogenicity (885.3050) 40384802 Data were bridged from test material Ps

. fluorescens

EG

- 1053

- TGAI.

There was no si gnificant effect on mean body weight gain, and no mortality associated with treatment . Not pathogenic at 2.6 x 10 9 cfu/kg of body weight. Toxicity
Ca tegory IV. Classification: Acceptable . U .S. EPA

1999 a Acute pulmonary toxicity/pathogenicity (885.3150) 40384806 Data were bridged from test material Ps

. fluorescens

EG

- 1053

- TGAI

. Ps

. fluorescens was not infective

, toxic , n or pathogenic to rats when administered at 1.5 x 10 5
cfu/animal. There was no effect on mean body weight gain , no mortality, and no trea tment

- related lesions or other adverse effects

. Toxicity Ca tegory IV.
Classification: Acceptable U .S. EPA 1999 a Acute injection toxicity/ pathogenicity

— intravenous (885.3200) 40384808 Data were bridged from test material Pseud omonas fluorescens EG

- 1053

- TGAI. There was no significant effect on mean body weight gain, no mortality, and no treatment related lesions

when administered at 2.2 x 10 8 cfu/kg. Toxicity Ca tegory IV. Classification: Acceptable

U .S. EPA 1999 a

Cell culture (885.3500) Not Applicable

. Ps

. aureofaciens is not a virus (refer to test note #4 of 40 CFR § 158.2140(d)).

Hypersensitivity incidents (885.3400) Not Applicable No hypersensitivity incidents occurring during the

testing, production, or use of Ps

. aureofaciens were reported by the registrant. Any future hypersensitivity incidents must be reported ( refer to

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

Acute oral toxicity (870.1100) 44337104 One female rat displayed weight loss from day 7 to day 14 ; and dilated pupils were noted in 3 fe male

rats on days 1 to 4. No Abnormalities noted at necropsy. LD 50 > 5000 mg/kg of BW. Toxicity Category IV

. U .S. EPA 1999 a

Acute dermal toxicity (870.1200) 40384804 Data were bridged from test material Ps

. fluorescens EG

- 1053

- TGAI

. Mean body weight s ( rats ) increased during the 14 day study; Ps.

fluorescens was considered non

- toxic when applied

to rabbit skin

. Toxicity Category IV

U .S. EPA 1999 a- 10

-

40 C FR 158.2140 Data Requirement (OPPTS Guideline) How it Has Been Addressed

MRID EPA Review , Conclusions , Comments

Acute inhalation toxicity (870.1300) Not applicable to the TGAI test substance.

Acute eye irritation (8 70.2400) Not applicable to the TGAI test substance.

Primary dermal irritation (870.2500) 44337105 Slight dermal irritation and erythema at 1 hour; resolved by 72 hours. Tan discoloration at test site in all rabbits. Primary irritation index=0.63

Toxici ty Ca tegory IV. Classification: Acceptable

U .S. EPA 1999 a

3. Environmental Fate and Ecological Risk Assessment Status (40 CFR 158.2150)

The ecotoxicity data which satisfy Tier I data requirements are summarized in Table 3 below. No data gaps have b een identified.

Table 3. Ecological Toxicity Data for Pseudomonas aureofaciens strain Tx

- 1

40 CFR 158.2150 Data Requirement (OPPTS Guideline) How it Has Been Addressed

MRID EPA Review , Conclusions, Comments

Avian oral toxicity ( 885.4050) 45040804

45 040805 Pseudomonas aureofaciens strain Tx

- 1 is practically

nontoxic tand is not pathogenic to Mallard Duck and Bob White Quail (NOEL= 5 x10 9 CFU/kg/day ) Classification: Acceptable

U .S. EPA 2008b and c

Avian inhalation toxicity/pathogenicity ( 885.4100) Not applicable No testing expected to be required . Results of

acceptable oral toxicity studies determined that

Pseudomonas aureofaciens strain Tx

- 1 is practically nontoxic and not pathogenic to avian species.

Wild mammal toxicity/ pathogenicity ( 885.4150 ) Not applicable No testing expected to be required

. R esults of the

Tier 1 toxicity tests required by 40 CFR § 158.2140 (Table 2) are adequate and appropriate for

assessment of hazards

Freshwater fish toxicity/ pathogenicity ( 885.4200) 45040801

45040803 Pseudomonas aureofaciens strain Tx

- 1 is not

infectious and practically nontoxic to both coldwater and warm water fish species. The 30 day LC 50 is > 2.2 x10 7 CFU/ml for rainbow trout and

>3.4 x 10 5 for blue gill. The NOEC for rainbow trout and blue gill is 9.3 x10 5 CFU/ml. Classification: Acceptable- 11

-

40 CFR 158.2150 Data Requirement (OPPTS Guideline) How it Has Been Addressed

MRID EPA Review , Conclusions, Comments

U.S. EPA 2007a. and U.S. EPA 2008a.

Freshwater invertebrate toxicity/ pathogenicity ( 885.4240) 45040802 Pseudomonas aureofaciens strain Tx

- 1 is not infectious and practically nontoxic to Daphnia magna at 1. 7 x 10 4 CFU/ml. Classification: Acceptable

U .S. EPA 2007b.

Estuarine/Marine fish testing

Estuarine and marine invertebrate testing ( 885.4280) Not applicable No testing expected to be required. Significant

exposures to estuarine/marine organisms are not expected organisms are not expected as a result of application to golf courses. Also, Pseud omonas aureofaciens strain Tx

- 1 is not infectious and practically nontoxic to

freshwater fish and aquatic invertebrate specie s Nontarget plant testing ( 885.4300) Not applicable No testing expected to be required, if exposures to

Nontarget plants are limited to those species on turf and golf courses.

U.S. EPA 1998.

Nontarget insect testing ( 885.4340) 45265201 For green lace wings the LC 50 is >8.5 x 10 6 CFU/g

and t he NOEC is 8.5 x 10 6 CFU/g. Classification: Acceptable

U .S. EPA 2002.

Nontarget insect testing ( 885.4340) 45225201 For Ladybird Beetles the dietary LC 50 is >4 .28 x10 7
CFU/g and the NOEC is 4.28 x10 7 CFU/g. Classification: Acceptable

U .S. EPA 200 1.

Nont arget insect testing ( 885.4340) 45225202 For parasitic Hymenoptera the dietary LC 50 is >4. 28 x

10 7 CFU/g and the NOEC is 4.28 x10 7 CFU/g. Classification: Acceptable

U .S. EPA 200 1.

Honey bee testing ( 885.4380) 45077602 The reviewed data indicate Pseudomon as aureofaciens strain Tx

- 1 is not toxic at 10 6

- 10 7

CFU/ml, or pathogenic at 10 3 CFU/ml. In addition,

Pseudomonas aureofaciens has never been reported as pathogenic to bees. Classification: Acceptable

US EPA 2000

.- 12

- 4. Risk to Threatened and Endang ered Species

Based on the specific use patterns and the absence of data gaps in ecological data, the Agency does not anticipate the need for new data for Pseudomonas aureofaciens strain

Tx

- 1

. B ecause this a.i. ha s a product that is registered for outdoo r use, the Agency is required to consider any effects on nontarget species, particularly those with endangered or threatened status.

EPA has not conducted a risk assessment that supports a complete endangered species determination for Pseudomonas aureof aciens strain Tx

- 1

. The ecological risk assessment planned during Registration Review will allow the Agency to determine

whether use of Pseudomonas aureofaciens strain Tx

- 1 will have “no effect” or “may affect” federally

- listed threatened or endangered sp ecies (listed species) or their designated critical habitat. When an assessment concludes that a pesticide’s use “may affect” a listed species or its designated critical habitat, the Agency will consult with the

U.S. Fish and Wildlife Service and/or Natio nal Marine Fisheries Service (the Services), as appropriate.

5. Endocrine Effects

As required by the Administrator under the Federal Food, Drug, and Cosmetic Act (FFDCA) section 408(p), EPA has developed the Endocrine Disruptor Screening Program (EDSP) a nd has begun to implement the screening program that is to be used to test all pesticides in order to determine whether certain substances (including pesticide active and other ingredients) may have an effect in humans or wildlife similar to an effect prod uced 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 Estrogenic Substances Screening Program a biologic substance or

other substance if a determination is made that the substance is not anticipated to 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 nine inert ingredients. Pseudomonas aureofaciens strain Tx

- 1 is not among the group of 58 pesticide active ingredients on the init ial list to be screened under the EDSP.

The Agency believes that the active ingredient involved in this Registration Review case likely is a substance that would not produce any effect in humans similar to an effect produced by a naturally occurring est rogenic substance. As such, pursuant to Section 408(p)(4), EPA will determine if it can exempt Pseudomonas aureofaciens strain Tx

1 from the requirements of the Section 408(p) EDSP. If the Agency determines that it is appropriate to exempt th is active ingr edient from EDSP screening, an order will be

issued.- 13

-

For further information on the status of the EDSP, the policies and procedures, the list of 67 chemicals, future lists, the test guidelines and the Tier 1 screening battery, please

visit our website: http://www.epa.gov/endo/

.



### 6. Incidents

The National Pesticides Information Center (NPIC) database reported no incidents associated with products containing Pseudomonas aureofaciens strain Tx

- 1 use

. The

Office of Pest icide Programs’ Incident Data System (IDS) indicates that there have been

no reports of adverse human, domestic animal , or environmental incidents for

Pseudomonas aureofaciens strain Tx

- 1 products

.

7. Summary of 

### Anticipated Data Needs

Table 4 include s anticipated data needs for Pseudomonas aureofaciens

. For additional discussion of the anticipated data needs, see section C. 1. Product Analysis

above.

Anticipated Data Needs for Registration Review of Pseudomonas aureofaci ens

Guideline

Number Study Title Test Material Estimated

Timeframe

(Months from receipt of DCI)

885.1100 Product Identity 1 E P 12

Deposition of a sample in a nationally recognized culture collection TGAI 12

885.1500 Certification of limits EP 1 2

830.6317 Storage stability TGAI and EP 12

830.6319 Miscibility EP 12

830.7100 Viscosity EP 12

TGAI = tech nical grade active ingredient; EP = end

- use product

1 In conjunction with redepositing the Tx

- 1 strain culture in a nationally recognized culture collection, the Agency anticipates requiring confirmatory product identity data to insure that the characteristics are identical to the originally registered product.

D

. Timeline

The projected timeline for Case 6009

— Pseudomonas aureofaciens strain Tx

- 1 is listed in table 5, below. This may be revised, if more time is needed (e.g., issuance of a DCI ESA Consultation with Fish and Wildlife Services, etc.) , and included in the Final Work

Plan.- 14

-

Table 5

. Projected Timeline for the Registration Review of Case 60 09

Activity Estimated Month

and Year

Phase 1 : Open the Public Docket

Open Comment Period for Docket J une 30, 201 2

Close Comment Period August 30, 201 2

Phase 2 : Case Development

Develop Final Work Plan January 2013

Issue Data C all

- in (DCI) January 2014

Data Submission January 2016

Open Public Comment Period for Preliminary Risk Assessments July 2017

Close Public Comment Period August 2017

Phase 3 : Registration Review Decision

Open Comment Period for Proposed Registra tion Review Decision January 2018

Close Comment Period March 2018

Final Decision and Begin Post

- Decision Follow

- up 2018

Estimated Total (years) 6

E

. Guidance for Commenters

The public is invited to comment on EPA’s preliminary work plan and ration ale

. The Agency will consider all comments as well as any additional information or data provided in a timely manner prior to issuing a final work plan for th is case.

F

. Environmental Justice

EPA seeks to achieve environmental justice, the fair treatm ent and meaningful involvement of all people, regardless of race, color, national origin, or income, in the development, implementation, and enforcement of environmental laws, regulations, and policies. To help address potential environmental justice issue s, the Agency seeks

information on any groups or segments of the population who, as a result of their location, cultural practices, or other factors, may have atypical, unusually high exposure

to Pseudomonas aureofaciens strain Tx

- 1 compared to the general population. Please comment if you are aware of any subpopulations that may have atypical, unusually high exposure compared to the general population.

G

. Water Quality

Pseudomonas aureofaciens strain Tx

- 1 is not identified as a cause of impairment for

an y water bodies listed as impaired under section 303(d) of the Clean Water Act, based

on information provided at : http://www.epa.gov/owow/tmdl/. In addition, no Total- 15

-

Maximum Daily Loads (TMDL) have been developed for Pseudomonas aureofaciens

strain Tx

- 1 based on information provided at :

http://iaspub.epa.gov/tmdl_waters10/text_search.tmdl_search_form

More information on impaired water bodies and TMDL ’ s can be found at:

http://www.epa.gov/owow/tmdl/

The Agency invites submission of water quality data for this pesticide

. To the extent possible, data should conform to the quality standards in Appendix A of the Office of Pesticide Program’s ( OPP ) Standard Operating Proce dure : Inclusion of Impaired Water Body and Other Water Quality Data in OPP’s Registration Review Risk Assessment an d Management Process to ensure they can be used quantitatively or qualitatively in pesticide risk assessments. This information can be locat ed at:

http://www.epa.gov/oppsrrd1/registration_review/water_quality_sop.htm

H

. Trade Irritants

Through the Registration Review process, the Agency intends to solicit in formation on trade irritants and, to the extent feasible, take steps toward facilitating irritant resolution. Growers and other stakeholders are asked to comment on any trade irritant issues resulting from lack of Maximum Residue Limits (MRL’s) or dispari ties between United
States tolerances and MRL’s in key export markets, providing as much specificity as possible regarding the nature of the concern.

Pseudomonas aureofaciens strain Tx

- 1 active ingredient is not applied to food or feed in

the United State s. In addition, there are no known MRL’s (FAO and WHO 2010). Therefore, the Agency does not anticipate that current uses of th is active ingredient will pose concerns as trade irritants.

I

. Additional Information

Stakeholders are specifically asked to provide information and data that will assist the Agency in refining the risk assessments, including any species

- specific ecological effects determinations. The Agency is interested in obtaining the following information regarding the use of Pseudomonas a ureofaciens strain Tx

- 1 :

1

. Confirmation on the following label information:

a

. Sites of application

b

. Formulations

c

. Application methods and equipment

d

. Maximum application rates- 16

- e

. Frequency of application, application intervals and maximum

n umber of applications

f

. Geographic limitations on use

2

. Use or potential use distribution

3

. Use history

4

. Usage/use information for nonagricultural uses (e.g., materials preservation)

5

. Typical application interval

6

. State or local use restric tions

7

. Monitoring data

J

. Next Steps

After the 60

- day comment period closes, the Agency will review and respond to any comments received in a timely manner

, then issue a Final Work P lan for Case 60 09 : Pseudomonas aureofaciens

.- 17

-



### B IBLIOGRAPHY : Ps eudomonas aureofaciens strain Tx

- 1

MRID Studies

44337100 EcoSoil Systems Inc. (1997) Submission of Product Chemistry and Toxicity

Data in Support of the Application for Registration of Ping Fungicide.

Transmittal of 6 Studies.

44337101 Harrison, E. (1997) Ping Fungicide: Product Identity and Composition.

Unpublished study prepared by EcoSoil Systems, Inc. 81 p.

44337102 Harrison, E. (1997) Ping Fungicide: Analysis and Certification of Product

Ingredients. Unpublished study prepared by Encore Te chnologies. 12 p.

44337103 Harrison, E. (1997) Ping Fungicide: Physical and Chemical Properties.

Unpublished study prepared by EcoSoil Systems, Inc. 4 p.

44337104 Douds, D. (1997) An Acute Oral Toxicity Study in Rats with Pseudomonas

aureofaciens Strai n Tx

- 1: Final Report: Lab Project Number: 3446.1. Unpublished study prepared by Springborn Laboratories, Inc. 23 p.

44337105 Douds, D. (1997) A Primary Skin Irritation Study in Rabbits with Pseudomonas

aureofaciens Strain Tx

- 1: Final Report: Lab Project Number: 3446.2. Unpublished study prepared by Springborn Laboratories, Inc. 29 p.

44337106 Harrison, E. (1997) Justification for Using Data on P. fluorescens to Satisfy

Data Requirements for P. aureofaciens. Unpublished study prepared by EcoSoil Systems, Inc. 169 p.

44422400 Eco Soil Systems (1997) Submission of Product Chemistry and Environmental

Fate Data in Support of the Registration of the Biopesticide based on the Active Ingredient Pseudomonas aureofaciens strain Tx

- 1. Transmittal of 1 Study.

44 422401 Debus, T.; Odelson, D. (1997) Pseudomonas aureofaciens strain Tx

- 1:

Supplemental Product Identification Data to Support Registration: Lab Project Number: SC 3867. Unpublished study prepared by Eco Soil Systems, Inc. 17 p.

44437500 Eco Soil Systems , Inc. (1997) Submission of Product Chemistry Data in Support

of the Application for Registration of Biopesticide Pseudomonas aureofaciens

strain Tx

- 1. Transmittal of 1 Study.

44437501 Ourisson, P. (1997) Growth of Pseudomonas aureofaciens Strain Tx

- 1 an d Other

Microorganisms Using the BioJect Automated Microbial Fermentation and Injection System (Bioject): Lab Project Number: 245

- 502. Unpublished study prepared by Quality Associates, Inc. 87 p.

44461700 Eco Soil Systems, Inc. (1998) Submission of Toxic ity, Product Chemistry and

Hazard Evaluation Data in Support of the Application for Registration of Spot

-- 18

- Less Biofungicide. Transmittal of 3 Studies.

44461701 Odelson, D.; Debus, T. (1998) Spot

- Less Biofungicide: Response to EPA Letter

of December 17, 19 97: Manufacturing Process and Discussion of Formation of Impurities. Unpublished study prepared by Eco Soil Systems, Inc. 12 p. {OPPTS 885.1200 and 885.1300}

44461702 Odelson, D.; Debus, T. (1998) Spot

- Less Biofungicide: Response to EPA Letter

of Decembe r 17, 1997: Information to Satisfy Acute Toxicity Data Requirements. Unpublished study prepared by Eco Soil Systems, Inc. 6 p.

44461703 Odelson, D.; Debus, T. (1998) Spot

- Less Biofungicide: Response to EPA Letter

of December 17, 1997: Literature Cross Re ferences Supporting the Citation of Data Conducted on Pseudomonas fluorescens. Unpublished study prepared by Eco Soil Systems, Inc. 103 p.

44708700 Eco Soil Systems (1998) Submission of Efficacy Data in Support of the

Application for Registration of Spot

- Less Biofungicide. Transmittal of 1 Study.

44708701 Odelson, D. (1998) Addition of Non

- Target Bacteria and Pseudomonas

aureofaciens Strain Tx

- 1 into the BioJect Automated Microbial Fermentation and Injection System (BioJect): Lab Project Number: ESSI

- 10 9801. Unpublished study prepared by Eco Soil Systems. 31 p.

44713300 Eco Soil Systems, Inc. (1998) Submission of Efficacy Data in Support of the

Application for Registration of Spot

- Less Biofungicide. Transmittal of 1 Study.

44713301 Dwyer, P.; Vargas, J.; Bresnahan, J. et al. (1998) Compilation of Efficacy Data

for the Microbial Pesticide Pseudomonas aureofaciens Strain Tx

- 1: (Spot

- Less). Unpublished study prepared by Eco Soil Systems. 23 p.

45040800 Eco Soil Systems, Inc. (2000) Submission of Toxici ty Data in Support of the

Registration of Spot

- Less Biofungicide. Transmittal of 5 Studies.

45040801 Drottar, K.; Krueger, H. (2000) Pseudomonas aureofaciens Strain TX

- 1

Biopesticide: A Five Concentration Toxicity and Pathogenicity Test with the

Rainbow Trout (Oncorhynchus mykiss) Final Report: Lab Project Number:

479A

- 103. Unpublished study prepared by Wildlife International, Ltd. 30 p. {OPPTS 885.4200}

45040802 Drottar, K.; Krueger, H. (2000) Pseudomonas aureofaciens Strain TX

- 1

Biopesticide: A 21

- Day Life

- Cycle Toxicity and Pathogenicity Test with the Cladoceran (Daphnia magna): Final Report: Lab Project Number: 479A

- 101A. Unpublished study prepared by Wildlife International, Ltd. 35 p. {OPPTS 885.4250}

45040803 Drottar, K.; Krueger, H. (2000) Pseud omonas aureofaciens Strain TX

- 1

Biopesticide: A Five Concentration Toxicity and Pathogenicity Test with the Bluegill (Lepomis macrochirus): Final Report: Lab Project Number: 479A

- 102. Unpublished study prepared by Wildlife International, Ltd. 30 p. {OPPTS 885.4200}- 19

-

45040804 Gallagher, S.; Casey, C.; Beavers, J. (2000) Pseudomonas aureofaciens Strain

TX

- 1 Biopesticide: An Avian Oral Pathogenicity and Toxicity Study in the Northern Bobwhite: Lab Project Number: 479

- 100. Unpublished study prepared by Wildlif

e International, Ltd. 32 p. {OPPTS 885.4050}

45040805 Gallagher, S.; Casey, C.; Beavers, J. (2000) Pseudomonas aureofaciens Strain

TX

- 1 Biopesticide: An Avian Oral Pathogenicity and Toxicity Study in the Mallard: Lab Project Number: 479

- 102. Unpublished study prepared by Wildlife International, Ltd. 32 p.

45048500 Eco Soil Systems, Inc. (2000) Submission of Product Chemistry Data in Support

of the Registration of Spot

- Less Biofungicide. Transmittal of 1 Study.

45048501 Odelson, D. (2000) Analysis of B ioject Grown Pseudomonas aureofaciens

Strain Tx

- 1 (Spot

- Less Biofungicide) for Antibiotics. Unpublished study prepared by U.S. Dept. of Agriculture. 16 p. {OPPTS 885.1400}

45077600 Eco

- Soil Systems, Inc. (2000) Submission of Toxicity and Efficacy Data in

Support of the Registration of Spot

- Less Biofungicide. Transmittal of 2 Studies.

45077601 Swick, C. (2000) Review of Antibiotic Production by Pseudomonads.

Unpublished study prepared by Eco Soil Systems, Inc. 265 p.

45077602 Odelson, D. (2000) Evaluat ion of Dietary Effect(s) of Pseudomonas

aureofaciens Strain TX

- 1 (Spotless) on Adult Honey Bees (Apis mellifera L.): Lab Project Number: CAR 184

- 99. Unpublished study prepared by California Agricultural Research, Inc. 101 p. {OPPTS 885.4340}

45225200 Eco Soil Systems, Inc. (2000) Submission of Toxicity Data in Support of the

Registration of Spot

- Less Biofungicide. Transmittal of 2 Studies.

45225201 Bryan, R.; Porch, J.; Krueger, H. (2000) Pseudomonas Aureofaciens Strain TX

1: A Dietary Pathogenicity and Toxicity Study with the Ladybird Beetle (Hippodamia convergens): Final Report: Lab Project Number: 479

- 103B. Unpublished study prepared by Wildlife International, Ltd. 20 p.

45225202 Bryan, R.; Porch, J.; Krueger, H. (2000) Pseudomonas aureofaciens Stra in TX

1: A Dietary Pathogenicity and Toxicity Study with Parasitic Hymenoptera (Nasonia vitripennis): Final Report: Lab Project Number: 479

- 104B. Unpublished study prepared by Wildlife International, Ltd. 21 p.

45265200 Eco Soil Systems, Inc. (2000) Subm ission of Toxicity Data in Support of the

Registration of Spot

- Less Biofungicide. Transmittal of 1 Study.

45265201 Bryan, R.; Porch, J.; Krueger, H. (2000) Pseudomonas aureofaciens strain TX

- 1:

A Dietary Pathogenicity and Toxicity Study with Green Lacewi ng Larvae (Chrysoperla carnea): Lab Project Number: 479

- 105A. Unpublished study prepared by Wildlife International, Ltd. 17 p. {OPPTS 885.4340}- 20

- EPA Reviews

U .S. EPA 1998. Memorandum from Gail Tomimatsu, Ph.D. to Michael Mendelsohn

. Eco Soil System's Request to Bridge Ecotoxicity Studies from Pseudomonas

fluorescens (Dagger G. Biofungicide, EPA Reg. No. 55638

- 5) to a New Microbial Active Ingredient, Pseudomonas aureofaciens Strain Tx

- 1; DP Bar code: D244137; Case No. 061928. June 29, 1998

.

U .S. EPA 1999 a

. Registration Eligibility Decision: Pseudomonas aureofaciens Strain

Tx

- 1

. Environmental Protection Agency

, Office of Pesticide Programs. February 11 , 1999

.

U .S. EPA 1999 b

. Memorandum from Chris Wozniak, Ph.D to Michael Mendelsohn.

Assessm ent of Pseudomonas aureofaciens Strain Tx

- 1, Spot

- Less, a Biological Fungicide, as Applied Through Boom

- type Sprayers

. Environmental Protection Agency

, Office of Pesticide Programs. June 29, 1999.

U .S. EPA 2000

. Memorandum from Robyn Rose to Michael Mend elsohn. Review of study designed to evaluate the dietary effects of adult honey bees feeding on

Pseudomonas aureofaciens strain TX

- 1 (Spotless TM ) for EPA Reg. No. 7 0688

- 1 U.S. Environmental Protection Agency

, Office of Pesticide Programs. June 12, 2000

.

U .S. EPA 200 1. Memorandum from Robyn Rose to Michael Mendelsohn. Review of ecological studies including Dietary Pathogenicity/Toxicity for lady beetles, green lacewing larvae, and parasitic Hymenoptera for Pseudomonas aureofaciens strain TX

- 1. EPA Reg. No. 70688

- 1; ( MRID Nos 452252

- 01, 452652

- 01, and 452252

- 02 ). U.S. Environmental Protection Agency

, Office of Pesticide Programs. August 9, 2001

U .S. EPA 2002. Memorandum from Robyn Rose to Susanne Cerrelli. Review of Eco Soil Systems, Inc. response to BPP D c omments on green lacewing studies addressed in the April 9, 2001 review of Pseudomonas aureofaciens strain TX

- I. EPA Reg. No.

70688

- 1 ( MR I D No 452652

- 01 )

. U.S. Environmental Protection Agency

, Office of Pesticide Programs. February 4, 2002.

U .S. EPA 2003

. Memorandum from Joel Gagliardi, Ph.D. to Susanne Cerrelli. Review of

F ive

- batch analysis including a produc tion study, and background literature, to support unconditional registration of Spot

- less B iof ungicid e. November 21, 2003.

U .S. EPA 2007 a ., Russel l S. Jones, Ph.D. and Gail S. Tomimatsu, Ph.D. Data Evaluation Report of Pseudomonas aureofaciens Strain TX

- 1 Biopesticide: A Five

Concentration Toxicity and P athogenicity Test with the Rainbow Trout ( Oncorhynchus mykiss ). ( MRID # 45 0 408

- 01) U.S. Environm ental Protection Agency, Office of Pesticide Programs. November 6, 2007

.- 21

-

U .S. EPA 2007 b ., Russell S. Jones, Ph.D. and Gail S. Tomimatsu, Ph.D. Data Evaluation Report of Pseudomonas aureofaciens Strain TX

- 1 Biopesticide : A 21

- Day

Life

- Cycle Toxicity and Pathogenicity Test with the Cladoceran ( Daphnia magna ) ( MRID

# 45 0 408

- 02. ) U.S. Environmental Protection Agency, Office of Pesticide Programs.

November 6 , 2007

.

U .S. EPA 2008a., Russell S. Jones, Ph.D. and Gail S. Tomimatsu, Ph.D. Data Evaluation Repo rt of Pseudomonas aureofaciens Strain TX

- 1 Biopesticide: A Five

Concentration Toxicity and P athogenicity Test with the Bluegill ( Lepomis macrochirus ) ( MRID # 45 0 408

- 03) U.S. Environmental Protection Agency, Office of Pesticide Programs. January 10, 2008.

U .S. EPA 2008b. Russell S. Jones, Ph.D. and Gail S. Tomimatsu, Ph.D. Data Evaluation Report of Pseudomonas aureofaciens Strain TX

- 1 : An Avian Oral Pathogenicity and Toxicity Stu dy in Northern Bobwhite [MRID # 450408

- 04]

. U.S. Environmental Protection Age ncy, Office of Pesticide Programs. January 10, 2008.

U .S. EPA 2008 c

. , Russell S. Jones, Ph.D. and Gail S. Tomimatsu, Ph.D. Data Evaluation Report of Pseudomonas aureofaciens Strain TX

- 1 : An Avian Oral Pathogenicity and Toxicity Stu dy in Mallard ( MRID # 450408

- 05 ). U.S. Environmental Protection Agency, Office of Pesticide Programs. January 16, 2008

.

Other Information

Palleroni, N.J., 2005. Genus I. Pseud omonas , pp. 351

- 358; 362

- 363 in: Brenner, D. J., Krieg, N. R. and Staley, J.T. Bergey’s Manual o f Systematic Bacteriology, Volume 2, 2 nd
Ed. Springer, New York.

Johnson and Palleroni, 1989

. “ Deoxyribonucleic Acid Similarities among Pseudomonas Specie s .” International Journal of Systematic Bacteriology

, Vol. 39. No. 3 ( July 1989 )

, 230

- 235

.