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## T

ABLE 

## O F 

## C ONTENTS



### I. PRELIMINARY WORK PLAN

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

A. Overview

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

B. Statutory and Regulatory Authority

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

C. Background and Regulatory Information

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

D. Anticipated Data Needs and Risk Assessments

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

E. Risk to Threatened and Endangered Species

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

F. Endocrine Effects

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

G. Incidents

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

H. Summary of Anticipated Data Needs

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

I. Timeline

................................ ................................ ................................ ................................ ................................ . 17

J. Guidance for Commenters

................................ ................................ ................................ ................................ ... 17

K. Next Steps

................................ ................................ ................................ ................................ .............................. 19



### II. FACT SUMMARY FOR CASE 6010

................................ ................................ ............................ 19

III. BIBLIOGRAPHY

................................ ................................ ................................ ........................... 19

A. Studies Supporting Registration Review of Harpin Protein

................................ ................................ ............. 19

B. Studies Supporting Registration Review of Harpinαβ Protein

................................ ................................ ......... 23

C. Environmental Protection Agency Risk Assessment Memoranda

................................ ................................ ... 24

D. Other References

................................ ................................ ................................ ................................ .................. 26Harpin Protein & Harpinαβ Protein (Harpin Proteins) EPA

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## ACRONYMS AND ABBREVIATIONS

BPPD EPA’s Biopesticides and Pollution Prevention Division

C

. F

. R

. Code of Federal Regulations

DNA Deoxyribonucleic a cid

EDSP Endocrine Disruptor Screening Program

EP End

- use pesticide product

EPA U.S. Environmental Protection Agency (the Agency)

FFDCA Federal Food, Drug, and Cosmetic Act

FIFRA Federal Insecticide, Fungicide, and Rodenticide Act

FQPA Food Quality Protection Act

FR Federal Register

FWP Final Work Plan
gal Gallon

HR H ypersensitive response

IDS Incident Data System

kD K ilodalton
kg K ilogram

lb Pound

mg M illigram

MRID No. Master Record Identification Number : EPA's system of recording and tracking studies submitted to the Office of Pesticide Programs.

MRL Maximum R esidue Limit

OPP EPA’s Office of Pesticide Programs

oz Ounce

PC Code Pesti cide chemical c ode: a six

- digit number assigned by OPP to identify pesticide chemicals.

PWP Preliminary Work Plan

TGAI Technical grade of the active ingredient

TMDL Total Maximum Daily Load

U.S. United StatesHarpin Protein & Harpinαβ Protein (Harpin Proteins) EPA

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### I. PRELIMINARY WORK PLAN

A. Overview

The docket for the harpin proteins (case 6010) is now open, initiating the first public comment period for this R egistration R eview case (docket number EPA

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- 0641) , which includes the following biochemical pesticides: (1) harpin protein (PC Code 006477) and (2) harpinαβ protein (PC Code 006506). 1 

### These substances elicit a complex natural defense mechanism in plants and are

r egistered for use on a wide array of plants (e.g., fruiting vegetables, pome fruit , tobacco, and turf) in agricultural and residen t ial settings t o manag e plant disease, enhanc e plant growth, and suppress insect

, mite and nematode pests. This Preliminary Work Plan (PWP) document explains what the Environmental Protection Agency’s (EPA) Office of Pesticide Programs knows about t he harpin proteins, highlighting anticipated data and assessment needs, identifying types of information that would be especially useful to EPA in conducting the review, and providing an anticipated timeline for completing review of the harpin proteins.

The R egistration R eview process was designed to include a public participation component to solicit input from interested stakeholders. EPA intends, by sharing this information in the docket, to inform the public of what it knows about the harpin proteins and what types of new data or other information would be helpful for EPA to receive as it moves toward a decision on the harpin proteins.

EPA encourages all interested stakeholders to review the PWP and to provide comments and additional

information that w ill help EPA ’s decision

- making process for these biochemical pesticides. In addition to general areas on which persons may wish to comment, there are some areas identified in the PWP about

which EPA specifically seeks comments and information. Interested stakeholders could include the following: environmental nonprofit or interest groups; pesticide manufacturers; agricultural labor or commodity groups; commercial, institutional, residential, and othe r users of pesticides; or the public at large.

The PWP begins by discussing the statutory and regulatory authority for Registration Review. Next, it provides background on the harpin proteins , which includes a description of their mode of action, current ly registered pesticide products, application rates and methods, use sites, and tolerance exemption information. Then, it lists the anticipated data needs and risk assessments and a projected Registration Review timeline for the harpin proteins . Finally, t he PWP describes guidance for

commenters , explains the next steps EPA will be taking for this Registration Review, summarizes background information for this Registration Review, and lists supporting studies, risk assessment memoranda, and other documents.

Further information about this case, including background and supporting documents, is

available at http://www.regulations.gov (u nder “ SEARCH for: Rules, Comments, Adjudications or Supporting Documents, ” enter the docket identification number, w hich is indicated above)

.

1 EPA classified the harpin proteins as biochemical pesticides because they are derived from naturally occurring microorganisms, do not contain any persistent microbes, have a nontoxic mode of action, and do not alter the deoxyribonucleic acid (DNA) of treated plants ( 40 C.F.R. § 158.2000(a)(1) ). Although biochemical pesticides are typically managed by the Biochemical Pesticides Branch of the Biopesticides and Pollution Prevention Division (BPPD), the Microbial Pesticides Branch of BPPD has assumed responsibility of pesticides products containing harpin proteins because these active ingredients are derived f rom and produced commercially in bacteria.Harpin Protein & Harpinαβ Protein (Harpin Proteins) EPA

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### B

. Statutory and Regulatory Authority

The Food Quality Protection Act (FQPA) of 1996 mandated a Registration Review program

. All pesticides distributed or sold in the United States (U.S.) must generally be registered by EPA , based on scientific data showing that they will not cause unreasonable risks to human health or 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 statutory standard of no unreasonable adverse effects to human health or

the environment . Chan ges in science, public policy, and pesticide use practices will occur over time. Through the Registration Review program, EPA periodically reevaluates pesticides to make sure that

, as change occurs, products in the marketplace can continue to be used safel y. Informatio n on this program is provided on EPA’s website

. 2



### EPA is implementing the Registration Review program pursuant to Federal Insecticide, Fungicide, and Rodenticide Act (FIFRA) section 3(g) and will review each registered pesticide every 15

yea rs to determine whether it continues to meet the FIFRA standard for registration. The regulations governing Registration Review begin at 40 C.F.R. § 155.40. EPA will consider benefits information and data as required by FIFRA. The public phase of Registrat ion Review begins when the initial docket is opened for each case. The docket is EPA ’s opportunity to state what it knows about the pesticide and what additional risk analyses and data or information it believes are needed to make a Registration

Review dec ision. After reviewing and responding to comments and data received in the docket during

this initial comment period, EPA wil l develop a F inal W ork P lan (FWP) and anticipated schedule for the Registration Review of the harpin proteins

.

C

. Background and Regulatory Information

i. Summary of the Active Ingredients 3



### The h arpin protein s act by eliciting a complex natural defense mechanism i n plants, analogous to

a broad

- spectrum immune response in animals. This protective response makes the plant resistant to a wide range of fungal, bacterial, and viral diseases during growth and post

- harvest. Unli ke the action of most pesticides (e.g., conventional chemistrie s), the harpin proteins do not interact directly with disease pests. Consequently, disease pests are not expected to develop resistance to these active ingredients. In addition to protecting against various diseases, the harpin proteins enhance plant growth and improve yield and quality of many crops, including food crops, turf, and ornamentals.

While harpin protein is derived only from Erwinia amylovora , a bacterium that causes fire blight in apples and pears, harpinαβ protein is a combination of four harpin protein fragments derived from Erwinia amylovora (harpinN and harpinW)

, Ralstonia solanacearum (popA) , and Pseudomonas syringae (harpinZ)

. Commercial pr oduction of these biochemical pesticides begins when the DNA fragments of the harpin proteins are transferred from the appropriate bacterial sources, mentioned above,

to Escherichia coli K

- 12

— a nonpathogenic, debilitated bacterium that has no potential to colonize the mammalian gut and is unlikely to survive in the environment because of nutritional deficiencies. A t the end of the fermentation process, Escherichia coli K

- 12 cells are kill ed and lysed to release the harpin proteins and other cell constituents for formulation into pesticide products.

2 See http://www.epa.gov/oppsrrd1/registration_review/

. 3 Information in this section extracted from U.S. EPA (2005a) and/or U.S. EPA (2002a).Harpin Protein & Harpinαβ Protein (Harpin Proteins) EPA

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### ii. Harpin Protein and Harpinαβ Pesticide Products

In 2000, EPA registered the first harpin protein

- containing pesticide product for use on vegetabl es, field crops, small grains, trees, vines, ornamentals, and turf as a plant disease manager, insect suppressor, and plant growth enhancer

. After EPA approved the initial harpin protein pesticide product, the registrant of that product

, Eden Bioscience Corporation

, submitted 12 additional applications for pesticide products with the harpin protein active ingredients . Of these (plus the one registered in 2000)

, 1 was withdrawn by the applicant and thus neve r registered; 8 were registered b ut
have since been cancelled 4 

### for nonpayment of maintenance fees ; and 4 remain registered. T he currently registered harpin protein products are end

- use pesticide products , and the registrant of all these pesticide products is now Plant Health Care, Inc. (see Table 1).

4 See t he Federal Register of August 6, 2008 ( 73 FR 45758 ) and July 28, 2010 ( 75 FR 44240 ).Harpin Protein & Harpinαβ Protein (Harpin Proteins) EPA

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7 TABLE 1. Summary of Currently Registered Harpin Protein and Harpinαβ Protein Pesticide Products

EPA Registration

Number Product Name (% Active Ingredient

,

Formulation Type ) Date Registered Application Rate s Application Methods Use Sites

Harpin Protein (PC Code 006477)

71771

- 1 Messenger (3 % , Wettable Dry

Granule/Powder ) April 19, 2000 0.35

– 5 oz product/100

lb seed Seed Treatment Equipment

Vegetables , Field Crops, Tobacco,

Berries and Small Fruit, Trees, Vines, Turf, Ornamentals, and/or Other Crops 2.25

– 13.35 oz product/acre crop Chemigation (Sprinkler & Drip) and

Conventional Ground or Aerial Application Equipment 5

– 9 oz product/100 gal

water

4.5

– 6 oz product/100 gal water Preplant Dip

(1) 0.12 oz product/1 gal water (2) 2 oz product/25 gal

water Pump Sprayer, Mister, and Watering Can

71771

- 10 Mighty Plant with Messenger Gold ( 0.1 % , Wettable Dry

Granule/Powder ) July 20, 2007 13.25 oz product/25 gal

water Pump Sprayer and Watering Can

Harpinαβ Protein (PC Code 006506)

71771

- 3 ProAct ( 1 % , Wettable Dry

Granule/Powder ) February 11,

2005 0.1

– 5 oz product/100 lb

seed Seed Treatment Equipment

Vegetables , Field Crops, Tobacco,

Berries and Small Fruit, Trees, Vines, Turf, Ornamentals, and/or Other Crops 0.5

– 13 oz product/acre

crop Chemigation (Sprinkler & Drip) and

Conventional Ground or Aerial Application Equipment

1

– 9 oz product/100 gal

water Preplant Dip

71771

- 7 Harp

- N

- Tek (0.0125% , Wettable Dry Granule/Powder ) November 21,

2006 5

– 30 lb product/acre

crop Chemigation (Sprinkler & Drip) and

Conventional Ground or Aerial Application Equipment

1

– 9 oz product/100 gal

water Preplant Dip



### iii. Tolerance Actions

Given that many of the uses associated with the registered harpin protein pesticide products involve treating food or feed crops directly for the purposes of plant disease management,

insect /mite/nematode suppression, and plant growth enhancement

— meaning that pesticide residues

coul d be found in or on such food or feed

— Eden Bioscience Corporation petitioned EPA to establish

tolerance exemptions for the harpin proteins . In light of available toxicity and exposure data, EPA concluded that there was a reasonable certainty that no harm w ould result to the U.S. population, including infants and children, from aggregate exposure to residues of the harpin proteins . Therefore,

EPA established a tolerance exemption for residues of these active ingredients

. Since this toleranceHarpin Protein & Harpinαβ Protein (Harpin Proteins) EPA

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### exemption took effect after FQPA amended FIFRA in 1996, it did not require reassessment. The current

tolerance exemption expression for th e harpin protein active ingredients is as follows:

40 C.F.R. § 180. 1204 ( Harpin Proteins )

An exemption from the requirement of a tolerance is established for residues of individual harpin proteins that meet specified physiochemical and toxicological criteria when used as biochemical pesticides on all food commodities t o enhance plant growth, quality and yield, to im prove overall plant health, and to aid in pest management. The physiochemical and toxicological criteria identifying harpin proteins are as follows:

(a) Consists of a protein less than 100 kD in size, that is acidic (p H <7.0), glycine rich (>10%), and

contains no more than one cystine residue. (b) The source(s) of genetic material encoding the protein are bacterial plant pathogens not known to be mammalian pathogens. (c) Elicits the hypersensitive response (HR), which is characterized as rapid, localized cell death in plant tissue after infiltration of harpin into the intercellular spaces of plant leaves. (d) Possesses a common secondary structure consisting of α and β units that form an HR domain. (e) Is heat stable (retains HR activity when heated to 65°C for 20 minutes). (f) Is readily degraded by a proteinase representative of environmental conditions (no protein fragments >3.5 kD after 15 minutes degradation with Subtilisin A). (g) Exhibits a rat acute oral toxicity (LD 50 

### ) of greater than 5,000 mg product/kg body weight.

Federal R egister Citations

– Notice s of Filing ( 64 FR 49010, September 9, 1999 ; 69 FR 4151 , January 28, 2004 ) and Final Rules ( 65 FR 25660 , May 3, 2000 ; 69 FR 24992, May 5, 2004 )

D

. Anticipated Data Needs and Risk Ass essment s

Provided below are tables of the current data requirements for biochemical pesticides as set forth in 40 C.F.R. part 158, subpart U . They are arrayed by scientific discipline (i.e., product chemistry

, toxicology , and nontarget organisms and environmental fate) and descri ptions are included regarding how the data requirements have been fulfilled or where data gaps (anticipated data needs) have been

identified.

Presently

, both active ingredients have several data gaps in all three disciplines. Data gaps have

been identified for studies that were either not submitted (indicated with a “No

– Data Gap” designation

in Tables 2

– 5) or not performed using the appropriate test substance (indicated with a “No

– Data Gap” designation, along w ith a footnote explanation

, in Tables 2

– 5)

. For a succinct summary of data gaps, please see Table 6.Harpin Protein & Harpinαβ Protein (Harpin Proteins) EPA

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### i. Product Chemistry ( 40 C.F.R. § 158.2 030 )

TABLE 2

. Summary of Product Chemistry Data for the Harpin Proteins

Harmonized

G uideline

Number Data Requirement Active Ingredient Has Data Requirement

Been Addressed?

How? Source MRID /Accession

Number or

Other Reference

880.1100 Product Identity and Composition Harpin Protein Yes

– Study U.S. EPA (1999) U.S. EPA (2000 a ) U.S. EPA (2002 a ) U.S. EPA (2002b) 445424

- 01

448702

- 01

451 912

- 01

Harpinαβ Protein Y es

– Study U.S. EPA (2004 a ) U.S. EPA (2005 a ) 460470

- 01

880.1200 Description of Starting Materials,

Production and

Formulation Process Harpin Protein Yes

– Study U.S. EPA (1999) U.S. EPA (2000 a ) U.S. EPA (2002a) U.S. EPA (2002b) 445424

- 01

446546

- 01

446599

- 01

448702

- 01

454904

- 01

Harpinαβ Protein Yes

– Study U.S. EPA (2004 a ) U.S. EPA (2005 a ) U.S. EPA (200 7 ) 460470

- 01

472317

- 01

8 80.14 00 Discussion of

Formation of

Impurities Harpin Protein Yes

– Study U.S. EPA (1999) U.S. EPA (2000 a ) U.S. EPA (2002a) U.S. EPA (200 2b ) 445424

- 01

448702

- 01

454904

- 01

Harpinαβ Protein Yes

– Study U.S. EPA (2004 a ) U.S. EPA (2005 a ) 460470

- 01

830.1700 Preliminary Analysis Harpin Protein Yes

– Study U.S. EPA (1999) U.S. EPA (2000 a ) 445424

- 01

448702

- 01

Harpinαβ Protein No

– Data Gap 1 Not applicable

830.1750 Certified Limits Not applicable to the TGAI test substance.

830.1800 Enforcement Analytical Method Not applicable to the TGAI test substance.

1 For this test, 40 C.F.R. § 158.2030 requires that data be generated using the TGAI and EP. T he preliminary analysis studies on file for harpinαβ protein were done using only an EP (i.e., harpinαβ protein and inert ingredients). These studies are as follows: MRID Nos. 460470

- 01, 460470

- 02, 468799

- 01, 468800

- 01,

468801

- 01, and 468802

- 01

.Harpin Protein & Harpinαβ Protein (Harpin Proteins) EPA

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10 TABLE 3

. Summary of Physical and Chemical Characteristics for the Harpin Proteins

Harmonized

Guideline

Number Data Requirement Active Ingredient Has Data Requirement

Been Addressed?

How? Source MRID /Accession

Number or

Other Reference

830.6302 Color Harpin Protein Yes

– Study None 445424

- 01

445424

- 02

Harpinαβ Protein No

– Data Gap 1 Not applicable

830.6303 Physical State Harpin Protein Yes

– Study U.S. EPA (1999) 445424

- 01

Harpinαβ Protein No

– Data Gap 2

Not applicable 830.6304 Odor Harpin Protein No

– Data Gap

Harpinαβ Protein No

– Data Gap 1

830.6313 Stability to Normal

and Elevated Temperatures,

Metals and Metal

Ions Harpin Protein

No

– Data Gap
Harpinαβ Protein

830.6315 Flammability

Not applicable to the TGAI test substance. 830.6317 Storage Stability

830.6319 Miscibility

830.6320 Corrosion

Characteristics

830.7000 pH Harpin Protein Yes

– Study None 445424

- 01

Harpinαβ Protein No

– Data Gap 2 Not applicable

830.7050 UV/Visible Light Absorption Harpin Protein Yes

– Study U.S. EPA (2002a) 448702

- 05

Harpinαβ Protein No

– Data Gap Not applicable

830.7100 Viscosity Not applicable to the TGAI test substance.

830.7200 Melting Point/Melting Range Harpin Protein Not required 3

Harpinαβ Protein

No

– Data Gap

Not applicable 830.7220 Boiling Point/Boiling Range Harpin Protein
Harpinαβ Protein

830.7300 Density/Relative Density/Bulk Density Harpin Protein No

– Data Gap 4

Harpinαβ Protein No

– Data Gap 2

830.7520 Particle Size, Fiber Length, and

Diameter

Distribution Harpin Protein

No

– Data Gap Harpinαβ Protein

830.7550

830.7560

830.7570 Partition Coefficient (n

- Octanol/Water) Harpin Protein

Harpinαβ Protein

830.7840 Water Solubility Harpin Protein Harpinαβ ProteinHarpin Protein & Harpinαβ Protein (Harpin Proteins) EPA

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11 TABLE 3

. Summary of Physical and Chemical Characteristics for the Harpin Proteins

Harmonized

Guideline

Number Data Requirement Active Ingredient Has Data Requirement

Been Addressed?

How? Source MRID /Accession

Number or

Other Reference

830.7950 Vapor Pressure Harpin Protein No

– Data Gap Not applicable Harpinαβ Protein

1 For this test, 40 C.F.R. § 158.2030 requires that data be generated using the TGAI. The color and odor studies on file for harpinαβ protein were done using only an EP (i.e., harpinαβ protein and inert ingredients). These studies are as follows: MRID Nos. 460470

- 01, 468799

- 01, 468799

- 02, 468800

- 01, 468800

- 02,

468801

- 01, 468801

- 02, 468802

- 01, and 468802

- 02.

2 For this test, 40 C.F.R. § 158.2030 requires that data be generated using the TGAI and EP. The physical state , pH, and bulk density studies on file for harpinαβ protein were done using only an EP (i.e., harpinαβ protein and inert ingredients). These stu dies are as follows: MRID Nos. 460470

- 01, 460470

- 07,

468799

- 01, 468799

- 02, 468800

- 01, 468800

- 02, 468801

- 01, 468801

- 02, 468802

- 01, and 468802

- 02

. 3 Refer to test note #13 of 40 C.F.R. § 158.2030(e).

4 For this test, 40 C.F.R. § 158.2030 requires that data be generated using the TGAI a nd EP. The bulk density studies on file for harpin protein were done using only an EP (i.e., harpin protein and inert ingredients). These studies are as follows: MRID Nos. 448702

- 01, 448702

- 03, and 456157

- 01.



### ii. Toxicology ( 40 C.F.R. § 158. 2050 )

TABLE 4

. Summary of Toxicology Data for the Harpin Proteins

Harmonized

Guideline

Number Data Requirement Active Ingredient Has Data Requirement

Been Addressed?

How? Source MRID /Accession

Number or

Other Reference

870.1100 Acute Oral Toxicity

Harpin Protein Yes

– Study U.S. EPA (1999) U.S. EPA (2002a) 445424

- 03

445424

- 04

446914

- 03

446914

- 04

447449

- 01

447449

- 02 Harpinαβ Protein No

– Data Gap 1

Not applicable 870.1200 Acute Dermal Toxicity Harpin Protein No

– Data Gap 2

Harpinαβ Protein No

– Data Gap 1

870.1300 Acute Inhalation Toxicity Harpin Protein No

– Data Gap 2

Harpinαβ Protein No

– Data Gap 1

870.2400 Primary Eye Irritation Harpin Protein No

– Data Gap 2

Harpinαβ Protein No

– Data Gap 1

870.2500 Primary Dermal

Irritation Harpin Protein No

– Data Gap 2

Harpinαβ Protein No

– Data Gap 1

870.2600 Dermal Sensitization Harpin Protein Yes

– Waiver U.S. EPA (2002a) None Harpinαβ Protein U.S. EPA (2005a)

None Hypersensitivity

Incidents Harpin Protein Yes

– Study U.S. EPA (1999) U.S. EPA (2002a) 445424

- 08

448702

- 12 Harpinαβ Protein

870.3100 90

- Day Oral Harpin Protein Yes

– Waiver U.S. EPA (2002a) None Harpinαβ Protein U.S. EPA (2005a)

870.3250 90

- Day Dermal Harpin Protein

No

– Data Gap Not applicable Harpinαβ Protein

870.3465 90

- Day Inhalation Harpin Protein Harpinαβ ProteinHarpin Protein & Harpinαβ Protein (Harpin Proteins) EPA

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12 TABLE 4

. Summary of Toxicology Data for the Harpin Proteins

Harmonized

Guideline

Number Data Requirement Active Ingredient Has Data Requirement

Been Addressed?

How? Source MRID /Accession

Number or

Other Reference

870.3700 Prenatal Developmental Harpin Protein

Yes

– Waiver U.S. EPA (2002a)

None Harpinαβ Protein U.S. EPA (2005a)

870.5100 Bacterial Reverse

Mutation Test Harpin Protein U.S. EPA (2002a)

Harpinαβ Protein U.S. EPA (2005a)

87 0.5385

870.5375 In vitro Mammalian Cell Assay Harpin Protein U.S. EPA (2002a)

Harpinαβ Protein U.S. EPA (2005a)

1 For this test, 40 C.F.R. § 158.2050 requires that data be generated using the TGAI and EP. The acute oral toxicity , acute dermal toxicity, acute inhalation toxicity, primary eye irritation, and primary dermal irritation stud ies on file for harpinαβ protein w ere done using only an EP (i.e., harpinαβ protein and inert ingredients). These studies are as follows: MRID Nos. 460470

- 08, 46 2040

- 01, 462040

- 02, 462040

- 04, and 462827

- 01

. 2 For this test, 40 C.F.R. § 158.2050 requires that data be generated using the TGAI and EP. The acute dermal toxicity, acute inhalation toxicity, primary eye irritation, and primary dermal irritation studies on file for harpin protein was done using only an EP (i.e., harpin protein and inert ingredients). These studies are

as follows: MRID Nos. 445424

- 05, 445424

- 06, 445424

- 07, 447449

- 03, 447449

- 04, 447449

- 05, 448702

- 08, 448702

- 09, 448702

- 10, 448702

- 11, and 456 064

- 01.



### iii. Nontarget Organisms and Environment Fate ( 40 C.F.R. § 158.2 060 )

TABLE 5

. Summary of Nontarget Organisms Data for the Harpin Proteins

Harmonized

Guideline

Number Data Requirement Active Ingredient Has Data Requirement Been

Addressed? How? Source MRID /Accession

Number or Other

Reference

850.2100 Avian Acute Oral Toxicity Harpin Protein Yes

– Study U.S. EPA (1998) U.S. EPA (2002a) 445424

- 09

446914

- 02 Harpinαβ Protein Yes

– Study 1 U.S. EPA (2005a) U.S. EPA (2005b)

850.2200 Avian Dietary Toxicity Harpin Protein Yes

– Study U.S. EPA (1998) U.S. EPA (2002a) 445424

- 10

446914

- 01 Harpinαβ Protein Yes

– Study 1 U.S. EPA (2005a) U.S. EPA (2005b)

850.1075 Fish Acute Toxicity,

Freshwater Harpin Protein

No

– Data Gap 2 Not applicable Harpinαβ Protein

850.1010 Aquatic Invertebrate Acute Toxicity,

Freshwater Harpin Protein

Yes

– Study 1 U.S. EPA (1998) U.S. EPA (2002a) 446546

- 02 Harpinαβ Protein U.S. EPA (2004b)

850.4100 Terrestrial Plant Toxicity, Seedling Emergence Harpin Protein
No

– Data Gap 2

Not applicable Harpinαβ Protein

850.4150 Terrestrial Plant Toxicity, Vegetative Vigor Harpin Protein

No

– Data Gap

Harpinαβ ProteinHarpin Protein & Harpinαβ Protein (Harpin Proteins) EPA

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13 TABLE 5

. Summary of Nontarget Organisms Data for the Harpin Proteins

Harmonized

Guideline

Number Data Requirement Active Ingredient Has Data Requirement Been

Addressed? How? Source MRID /Accession

Number or Other

Reference

880.4350 Nontarget Insect Testing Harpin Protein

Yes

– Waiver U.S. EPA (2000b)

None

Harpinαβ Protein U.S. EPA (2005a) U.S. EPA (2004b)

1 Because of the close structural, chemical, and biological similarity of harpinαβ protein to harpin protein, EPA ’s safety findings for harpin protein are also applicable to harpinαβ protein.

2 For this test, 40 C.F.R. § 158.2060 requires that data be generated using the T GAI and, in some instances, the EP too

. T he only fish acute toxicity (freshwater ) and terrestrial plant (seedling emergence) studies on file for harpin protein and harpinαβ protein were done using an EP (i.e., harpin protein a nd inert ingredients)

. These studies are as follows: MRID Nos. 448702

- 20 and 448702

- 22.



### E

. Risk to Threatened and Endangered Species

EPA has not prepared a risk assessment that supports a complete endangered species

determination for the harpin protein s

. During Registration Review, EPA plans on conducting an ecological risk assessment t o determine whether use of the harpin protein s will have “no effect” or “may

affect” federally listed threatened or endangered species (listed species) and/ or thei r desig nated critical

habitat. When an assessment concludes that a pesticide’s use “may affect” a listed species and/ or its designated critical habitat, EPA will consult with the U.S. Fish and Wildlife Service and/or National Marine Fisheries Service (the Services), as appropriate.

F

. Endocrine Effects

As required by the Administrator under the Federal Food, Drug, and Cosmetic Act ( FFDCA) section 408(p), EP A 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 in order to determine whether certain substances (including pesticide active and other ingredients) m ay 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 t he 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 e strogenic 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. The harpin protein s are not among the group of 5 8 pesticide active ingredients on the initial list to be screened under

the EDSP.

EPA , as part of this Preliminary Work Plan , believes tha t the harpin protein s , the active ingredient s involved in this Registratio n Review case, likely are substance s that w ould not produce any effect in humans similar to an effect produced by a naturally occurring estrogenic substance. As such,

pu rsuant to Section 408(p)(4), EPA will determine in the future whether it can exempt the harpin protein s from the requirements of the Section 408(p) EDSP. In the event EPA does determine to exempt these substances from the EDSP, an order will be issued.Harpin Protein & Harpinαβ Protein (Harpin Proteins) EPA

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14 

### 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 ba ttery, please visit our website at http://www.epa.gov/endo/

.

G

. Incidents

From January 1, 1992 through September 17, 2012, t he Office of Pesticide Programs’ Incident Data System (IDS) indicates that there have been 3 reports of adverse effects to humans following exposure to a pesticide product containing harpin protei n or harpinαβ protein and 3 reports of adverse

effects to human s following exposure to both a pesticide product containing harpin protein and pesticide products containing other active ingredients. Of the 3 incident reports involving exposure to a pesticide product containing harpin protein or harpinαβ protein, two of the cases are minor (H

- D) 5 

### and the other is

moderate (H

- C)

. 6 

### Of the 3 incident reports involving exposure to both a pesticide product containing harpin protein and pesticide products containing other active ingredients, two of the cases are minor (H

D), 5 

### while the other case describes 3 fatalities (H

- A) 7 

### and 2 major human incidents (H

- B). 8 

### For the case with the fatalities and major incidents, 40 other pesticide products , besides the harpin protein pesticide product, are included in the report (e.g., Guthion 2L containing the organophosphate active ingredient, azinophos

- methyl , and Lorsban 15G containing the organophosphate active ingredient, chlorpyrifos )

. EPA also checked for incidents in its Ecological Incident Information System (version 2.1. 1. ) and , from January 1966 through August 25, 2011, found no reports of adverse field effects to nontarget plants and animals as a result of exposure t o the harpin proteins. EPA will consider any incident data or comments submitted in response to this Preliminary Work Plan.

5 H

- D is considered to be a minor human incident. In 40 C.F.R. § 159.184(c)(5)(i)(D), the H

- D classification is described as follows: “If the person alleged or exhibited some symptoms, but they were minimally traumatic. The symptoms resolved rapidly and usua lly involve skin, eye or respiratory irritation.”

6 H

- C is considered to be a moderate human incident. In 40 C.F.R. § 159.184(c)(5)(i)(C), the H

- C classification is described as follows: “If the person alleged or exhibited symptoms more pronounced, more prolonged or of a more systemic nature than minor symptoms. Usually some form of treatment of the person would have been indicated. Symptoms were not life threatening and the person has returne d to his/her

pre

- exposure state of health with no additional r esidual disability.

7 H

- A indicates that a person died (40 C.F.R. § 159.184(c)(5)(i)(A)),

8 H

- B is considered to be a major human incident. In 40 C.F.R. § 159.184(c)(5)(i)(B), the H

- B classification is described as follows: “If the person alleged or exhibi ted symptoms which may have been life

- threatening, or resulted in adverse reproductive effects or in residual disability.”Harpin Protein & Harpinαβ Protein (Harpin Proteins) EPA

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### H

. Summary of Anticipated Data Needs

Table 6 summarizes the anticipated data needs for the harpin protein s.

Table 6. Anticipated Data Needs for the Registration Review of the Harpin Proteins

Harmonized Guideline

Number Study Title Test

Material Estimated

Timeframe

(Months from Receipt of DCI)

Harpin Protein

830.6304 Odor

TGAI 8 months 830.6313 Stability to Normal and Elevated Temperatures,

Metals and Metal Ions

830.7220 Boiling Point/Boiling Range

830.7300 Density/Relative Density/Bulk Density

830.7520 Particle Size, Fiber Length, and Diameter Distribution

830.7550

830.7560

830.7570 Partition Coefficient (n

- Octanol/Water)

830.7840 Water Solubility

830.7950 Vapor Pressure

870.1200 Acuter Dermal Toxicity

870.1300 Acute Inhalation Toxicity

870.2400 Primary Eye Irritation

870.2500 Primary Dermal Irritation

870.3250 90

- day Dermal 24 months 870.3465 90

- day Inhalation

850.1075 Fish Acute Toxicity, Freshwater

12 months 850.4100 Terrestrial Plant Toxicity, Seedling Emergence

850.4150 Terrestrial Plant Toxicity, Vegetative Vigor
Harpinαβ Protein

830.1700 Preliminary Analysis

TGAI 8 months 830.6302 Color

830.6303 Physical State

830.6304 Odor

830.6313 Stability to Normal and Elevated Temperatures,

Metals and Metal Ions

830.7000 pH

830.7050 UV/Visible Light Absorption

830.7200 Melting Point/Melting Range

830.7300 Density/Relative Density/Bulk Density

830.7520 Particle Size, Fiber Length, and Diameter DistributionHarpin Protein & Harpinαβ Protein (Harpin Proteins) EPA

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16 Table 6. Anticipated Data Needs for the Registration Review of the Harpin Proteins

Harmonized Guideline

Number Study Title Test

Material Estimated

Timeframe

(Months from Receipt of DCI)

830.7550

830.7560

830.7570 Partition Coefficient (n

- Octanol/Water)

TGAI 8 months 830.7840 Water Solubility

830.7950 Vapor Pressure

870.1100 Acute Oral Toxicity

870.1200 Acuter Dermal Toxicity

870.1300 Acute Inhalation Toxicity

870.2400 Primary Eye Irritation

870.2500 Primary Dermal Irritation

870.3250 90

- day Dermal 24 months 870.3465 90

- day Inhalation

850.1075 Fish Acute Toxicity, Freshwater

12 months 850.4100 Terrestrial Plant Toxicity, Seedling Emergence

850.4150 Terrestrial Plant Toxicity, Vegetative Vigor

TGAI = technical grade of the active ingredientHarpin Protein & Harpinαβ Protein (Harpin Proteins) EPA

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### I

. Timeline

The projected timeline for case 6010

— harpin proteins

— Registration Review follows.

Activities Estimated Month/Year

Opening the Docket Open Docket and 60

- Day Public Comment Period for Harpin Proteins September 201 2

Close Public Comment Period November 201 2

Case Development

Issue Final Work Plan J uly 201 3

Issue Data Call

- in July 201 4

Data Submission July 201 6

Open 30

- Day Public Comment Period for Draft Risk Assessments January 2018

Close Public Comment Period February 201 8

Registration Review Decision

Open 60

- Day Public Comment Period for Proposed Registration Review

Decision June 201 8

Close Public Comment Period August 201 8

Final Decision September 201 8

*Estimated Total (years) 6

* This schedule is subject to revision should unforeseen issues arise during the registration review process. In the event an issue arises, such as the failure to acquire an EDSP exemption, an amended Final Work Plan will be issued at that time. That Final Work Plan will identify a ny newly anticipated data needs and set forth a new timeline.



### J

. Guidance for Commenters

The public is invited to comment on EPA ’s Preliminary Work P lan and rationale. The areas below highlight topics of special interest to EPA where comments, data submissions, or reference to sources of additional information could be of particular use. EPA will consider all comments

, as well as any additional information or data provi ded in a timely manner

, prior to issuing a Final Work P lan for

this case.

i. Environmental Justice

EPA seeks to achieve environmental justice, the fair treatment 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 policie s. To help address potential environmental justice issues, EPA 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 the harpin proteins , 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.Harpin Protein & Harpinαβ Protein (Harpin Proteins) EPA

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### ii. Water Quality

The harpin proteins are not identified as a cause of impairment for any water bodies listed as impaired under section 303(d) of the Clean Water Act. 2 

### In addition, no Total Maximum Daily Loads (TMDL s ) have been developed fo r the harpin proteins

. 3 

### More information on i mpaired water bodies and

TMDL s c an be found at EPA ’s website. 9 

### EPA invites submission of water quality data for these pesticides. To the extent possible, data should conform to the quality standards in Appendix A of the Office of Pesticide Program’s (OPP) Standard Operating Procedure: Inclusion of Impaired Water Body and Other Water Quality Data in OPP’s Registration Review Risk Assessment and Management

Process 4 

### to ensure they can be used quantitatively or qualitatively in pesticide risk assessments.

iii. Tra de Irritants

Through the Registration Review process, EPA intends to solicit information on trade irritants and, to the extent feasible, take steps toward facilitating irritant resolution. EPA will work to harmonize tolerance and international Maximum Residue Limits (MRLs) and may modify tolerance levels to do so, when possible. Growers and other stakeholders are asked to comment on any trade irritant issues resulting from lack of MRL s or dispar ities between U.S. tolerances and MRLs in key export markets

, providing as much specificity as possible regarding the nature of the concern.

The harpin proteins have an exemption from the requirem ent of a tolerance . Moreover, there are

n o known MRLs for the harpin protein s . Therefore, EPA does not anticipate that current uses of the harpin protein s will pose concerns as trade irritants.

iv. Additional Information

Stakeholders are also specifically asked to provide information and data that will assist EPA in refining the risk assessments

. EPA is interested in obtaining the following information regarding the harpin protein s :

 Any data, information, and/or scientific rationale that could be used to fill data gaps

 Confirmation on the following label information:

- Sites of application

- Formulations

- Application methods and equipment

- Maximum application rates

- Frequency of application, application intervals, and maximum number of applications

- Geographic limit ations on use

 Use or potential use distribution (e.g., acreage and geographical distribution of relevant use sites)

 Median and 90 th 

### percentile reported use rates (lbs active ingredient/1,000 square feet) from usage data

– national, state, and county

2 Based on information provided at http://www.epa.gov/owow/tmdl/

. 3 B ased on information provided at http://iaspub.epa.gov/tmdl_waters10/text_search.tmdl_search_form

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

. 4 This document can be found at http://www.epa.gov/oppsrrd1/registration_review/water_quality_sop.htm

.Harpin Protein & Harpinαβ Protein (Harpin Proteins) EPA

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###  Application timing (date of first application and application intervals)

– national, state, and

county

 Usage/use information for nonagricultural use s

 Typical application interval (days)

 State or local use restrictions

 Monitoring data

K

. Next S teps

After the 60

- day comment period closes, EPA will review and respond to any comments received in a timely manner and then issue a F inal W ork P lan for case 6010 : Harpin Proteins

.

II. FACT SUMMARY FOR CASE 6010

TABLE 7

. Background Information for the Harpin Proteins Registration Review

Active Ingredient (PC Code) First Registered Currently Registered

Pesticide Products 1 Publicly Available EPA

Science and Regulatory

Documents Additional Risk Assessment Needed? (Yes or No )

Product Chemistry Human

Health Ecological

Harpin Protein ( 006477 ) 2000 2 end

- use product s ( Messenger & Mighty Plant with Messenger

Gold ) Biopesticides Registration

Action Document

BPPD Fact Sheet Yes Yes Yes

Harpinαβ Protein (006506) 2005 2 end

- use products (ProAct & Harp

- N

- Tek) Biopesticides Registration

Action Document

BPPD Fact Sheet Yes Yes Yes

1 See Table 1 for additional information (e.g., use sites) on the currently registered products. Labels for these products can be found in the Pesticide Product Label System

.



### II I

. BIBLIOGRAPHY

A. Studies Supporting Registration Review of Harpin Protein

MRID No. Study Description

445424

- 01 Wei Z, Kelleher T. 1998

. Harpin Messenger Product Analysis Data and

Information . Unpublished study prepared by Eden Bioscience Corp oration . 138

pages.

445424

- 02 Hohstadt K, Kelleher T. 1998. Characterization of Test Substances Used in Toxicology Studies

– Harpin Messenger. Unpublished study prepared by Eden Bioscience Corporation. 7 pages.

445424

- 03 Hilaski R. 1998. An Acute Oral Toxicity Study of H arpin Protein Technical in Rats. Lab Project Number: 789

- 002. Unpublished study prepared by MPI Research. 40 pages.Harpin Protein & Harpinαβ Protein (Harpin Proteins) EPA

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### MRID No. Study Description

445424

- 04 Hilaski R. 1998. An Acute Oral Toxicity Study of Concentrated Harpin Protein

Technical in Rat s. Lab Project Number: 789

- 003. Unpublished study prepared by MPI Research. 45 pages.

445424

- 05 Hilaski R. 1998. An Acute Inhalation Toxicity Study of Harpin End Product in Rats. Lab Project Number: 789

- 006. Unpublished study prepared by MPI

Research. 64 pages.

445424

- 06 Hilaski R. 1998. An Eye Irritation Study of Harpin End Product in Rabbits. Lab Project Number: 789

- 005. Unpublished study prepared by MPI Research. 21

pages.

445424

- 07 Hilaski R. 1998. A Dermal Irritation Study of Harpin End Prod uct in Rabbits. Lab Project Number: 789

- 004. Unpublished study prepared by MPI Research. 18

pages.

445424

- 08 Kelleher T. 1998. Hypersensitivity Incidents

– Harpin Messenger. Unpublished study prepared by Eden Bioscience Corporation. 4 pages.

445424

- 0 9 Gallagher S, Grimes J, Beavers J. 1998. An Acute Oral Toxicity Study with the Northern Bobwhite: Harpin Protein Technical. Lab Project Number: 430

- 102 430/072897/QLD.NC/SUB430. Unpublished study prepared by Wildlife International Limited. 41 pages.

44 5424

- 10 Gallagher S, Grimes J, Beavers J. 1998. A Dietary LC 50 

### with the Northern Bobwhite: Harpin Protein Technical. Lab Project Number: 430

- 101 430/072997/QLCSDT.SC/SUB430. Unpublished study prepared by Wildlife International Limited. 56 pages.

446546

- 01 Wei Z, Schading B. 1998. Summary of Production Process Used to Produce Messenger. Unpublished study prepared by Eden Bioscience Corporation. 29

pages.

446546

- 02 Ward G, Bussard J. 1998. Acute Toxicity of Harpin Protein Technical to Daphnia

magna Under Static

- R enewal Test Conditions. Unpublis hed study prepared by ABC Laboratories, Inc. 33 pages

.

446599

- 01 Wei Z, Schading B. 1998. Messenger: Supplemental Information. Lab Project Number: 038E110. Unpublished study prepared by Eden Bioscienc e Corporation. 45 pages.Harpin Protein & Harpinαβ Protein (Harpin Proteins) EPA

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21 

### MRID No. Study Description

446914

- 01 Gallagher S, Grimes J, Beavers J. 1998. Harpin Protein Technical: A Dietary LC 50 

### Study with the Northern Bobwhite

– Amended Report. Lab Project

Number: 430

- 101. Unpublished study prepared by Wildlife International Limited. 58 pages.

446914

- 02 Gallagher S, Grimes J, Beavers J. 1998. Harpin Protein Technical: An Acute Oral Toxicity Study with the Northern Bobwhite

– Amended Report. Lab Project

Number: 430

- 102. Unpublished study prepared by Wildlife International Limited. 43 pages.

446914

- 03 Hilaski R. 1998. An Acute Oral Toxicity Study of Harpin Protein Technical in

Rats

– Amendment to the Final Report. Lab Project Number: 789

- 002. Unpublis hed study prepared by MPI Research. 45 pages.

446914

- 04 Hilaski R. 1998. An Acute Oral Toxicity Study of Concentrated Harpin Protein

Technical in Rats

– Amendment to the Final Report. Lab Project Number: 789

- 003. Unpublished study prepared by MPI Resea rch. 50 pages.

447449

- 01 Hilaski R. 1999. An Acute Oral Toxicity Study of Harpin Protein Technical in Rats (Amendment to MRID No. 446914

- 03)

– Revised Final Report. Lab Project

Number: 789

- 002. Unpublished study prepared by MPI Research. 53 pages.

447449

- 02 Hilaski R. 1999. An Acute Oral Toxicity Study of Concentrated Harpin Protein Technical in Rats (Amendment to MRID No. 446914

- 04)

– Revised Final Report. Lab Project Number: 789

- 003. Unpublished study prepared by MPI Research. 58

pages.

44744 9

- 03 Hilaski R. 1999. An Acute Inhalation Toxicity Study of Harpin End Product in Rats (Amendment to MRID No. 445424

- 05)

– Revised Final Report. Lab Project

Number: 789

- 006. Unpublished study prepared by MPI Research. 73 pages.

447449

- 04 Hilaski R. 19 99. An Eye Irritation Study of Harpin End Product in Rabbits (Amendment to MRID No. 445424

- 06)

– Revised Final Report. Lab Project

Number: 789

- 005. Unpublished study prepared by MPI Research. 30 pages.

447449

- 05 Hilaski R. 1999. A Dermal Irritation Stu dy of Harpin End Product in Rabbits (Amendment to MRID No. 445424

- 07)

– Revised Final Report. Lab Project

Number: 789

- 004. Unpublished study prepared by MPI Research. 27 pages.

448702

- 01 Wei Z, Niggemeyer J. 1999. Messenger Product Analysis. Unpublishe d study prepared by Eden Bioscience Corporation. 69 pages.Harpin Protein & Harpinαβ Protein (Harpin Proteins) EPA

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### MRID No. Study Description

448702

- 03 Jacobs R, Nixon W. 1999. Determination of the Bulk Density of Messenger Biopesticide. Lab Project Number: 430C

- 104. Unpublished study prepared by Wildlife International Limited. 21 pages.

448702

- 05 Jacobs R, Nixon W. 1999. Determination of the Ultraviolet

- Visible Absorption Spectrum of Harpin EA 

### . Lab Project Number: 430C

- 106. Unpublished study prepared by Wildlife International Limited. 38 pages.

448702

- 08 Findlay J. 1999. Acute Dermal Toxicity Study of Messenger in Rabbits (Limit Test)

– Final Report. Lab Project Number: 1062 SN5. Unpublished study prepared by ITT Research Institute. 26 pages.

448702

- 09 Sullivan D. 1999. Acute Nose

- On ly Inhalation Toxicity Study (Limit Test) of Messenger in Rats. Lab Project Number: 1062 SN6. Unpublished study prepared by ITT Research Institute. 32 pages.

448702

- 10 Findlay J. 1999. Acute Eye Irritation Study of Messenger in Rabbits

– Final Report. Lab Project Number: 1062 SN4. Unpublished study prepared by ITT Research Institute. 22 pages.

448702

- 11 Findlay J. 1999. Acute Dermal Irritation Study of Messenger in Rabbits

– Final Report. Lab Project Number: 1062 SN3. Unpublished study prepared by ITT Research Institute. 24 pages.

448702

- 12 Wei Z. 1999. Hypersensitivity Incidents (Messenger). Unpublished study prepared by Eden Bioscience Corporation. 4 pages.

448702

- 20 Palmer S, Kendall T, Nixon W et al . Messenger (Harpin 3% A.I.): A 96

- Hour

Static

- Renewal Acute Toxicity Test with the Rainbow Trout ( Onchoryhnchus mykiss ) (Final Report). Lab Project Number: 430A

- 102 430/032499/RBT 96H3/SUB430. Unpublished study prepared by Wil dlife International Limited. 53

pages.

448702

- 22 Porch J, Krueger H. 1999. Messenger (Harpin EA 

### 35 A.I.): A Toxicity Test to Determine the Effects of the Test Substance on Seedling Emergence of Ten Species of Plants (Final Report). Lab Project Number: 430

- 107

430/031299/SEEDEM

- 10/SUB430. Unpublished study prepared by Wildlife International Limited. 85 pages.

451912

- 01 Fan H. 2000. Supplement to Volume 2: Messenger Product Analysis (MRID No.

448702

- 01). Unpublished study prepared by Eden Bioscience Corporation. 10

pages.Harpin Protein & Harpinαβ Protein (Harpin Proteins) EPA

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### MRID No. Study Description

454904

- 01 Wei Z, Fan H. 2001. Messenger Manufacturing Process and Discussion of Formation of Unintentional Ingredients. Unpublished study prepared by Eden Bioscience Corporation. 18 pages.

456064

- 01 Kuhn J. 2001. Acute Eye Irritation Study in Rabbits: Messenger 3% + New Inert (Final Report). Lab Project Number: 6666

- 01. Unpublished study prepared by Stillmeadow, Inc. 17 pages.

456157

- 01 Lezotte F, Nixon W. 2001. Determination of the Colo r, Physical State, pH, and Bulk Density of Messenger Nursery and Ornamental (1% Harpin). Lab Project

Number: 430C

- 109. Unpublished study prepared by Wildlife International Limited. 18 pages.

B. Studies Supporting Registration Review of Harpinαβ Protein

M RID No. Study Description

460470

- 01 Fan H. 2003. EBC

- 351 Product Analysis. Unpublished study prepared by Eden Bioscience Corporation. 87 pages.

460470

- 02 Buttke J, Song X. 2003. Quantification of Active Ingredient in Five Batches of 1% Harpinαβ EBC

- 351. Project Number: EBC2/03/003. Unpublished study prepared by Eden Bioscience Corporation. 37 pages.

460470

- 07 Page

- Lester S. 2003 Bulk Density of EBC

- 351 (1% Harpinαβ). Project Number: EBC2/03/010. Unpublished study prepared by Eden Bioscience Co rporation. 20

pages.

460470

- 08 Kuhn J. 2003. Acute Oral Toxicity Study (UDP) in Rats

– EBC

- 351 (1% Harpinαβ) (Amended Final Report). Project Number: 7435/03. Unpublished study prepared by Stillmeadow, Inc. 12 pages.

462040

- 01 Kuhn J. 2003. Acute Derm al Toxicity Study in Rabbits (1% Harpinαβ)

– Amended Final Report. Project Number: 7436/03. Unpublished study prepared by Stillmeadow, Inc. 15 pages.

462040

- 02 Sullivan D, Rajendran N. 2003. Acute Nose

- Only Inhalation Toxicity Study of

EBC

- 351 (1% Har pinαβ) in Rats (Limit Test). Project Number: 2061/001/SN/1 2061/001. Unpublished study prepared by IIT Research Institute. 36 pages.

462040

- 04 Kuhn J. 2003. Acute Eye Irritation Study in Rabbits

– EBC

- 351 (1% Harpinαβ) (Final Report). Project Number: 7 438/03. Unpublished study prepared by Stillmeadow, Inc. 18 pages.Harpin Protein & Harpinαβ Protein (Harpin Proteins) EPA

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### MRID No. Study Description

462827

- 01 Kuhn J. 2003. Acute Dermal Irritation Study in Rabbits

– EBC

- 351 (1% Harpinαβ) (Final Report). Project Number: 7439/03 EBC02/03/003. Unpublished study prepared by Stillmeadow, Inc. and Eden Bioscience Corporation, 13 pages.

468799

- 01 Fan H, Betz F. 2006. EBC

- 282 Product Analysis. Unpublished study prepared by

Eden Bios cience Corporation. 16 pages.

468799

- 02 Kaminsky M. 2006. Product Chemisty

– End

- Use Product, EBC

- 282 (Final Report). Project Number: 9879/06. Unpublished study prepared by Stillmeadow, Inc. 12 pages.

468799

- 01 Fan H, Betz F. 2006. EBC

- 282 Product An alysis. Unpublished study prepared by Eden Bioscience Corporation. 16 pages.

468800

- 01 Fan H, Betz F. 2006. EBC

- 354 Product Analysis. Unpublished study prepared by Eden Bioscience Corporation. 11 pages.

468800

- 02 Kaminsky M. 2006. Product Chemisty

– End

- Use Product, EBC

- 584 (Final Report). Project Number: 9880/06. Unpublished study prepared by Stillmeadow, Inc. 12 pages.

468801

- 01 Fan H, Betz F. 2006. EBC

- 280 Product Analysis. Unpublished study prepared by Eden Bioscience Corporation. 16 pages.

468801

- 02 Kaminsky M. 2006. Product Chemisty

– End

- Use Product, EBC

- 280 (Final Report). Project Number: 9877/06. Unpublished study prepared by Stillmeadow, Inc. 12 pages.

468802

- 01 Fan H, Betz F. 2006. EBC

- 281 Product Analysis. Unpublished study prepa red by Eden Bioscience Corporation. 16 pages.

468802

- 02 Kaminsky M. 2006. Product Chemisty

– End

- Use Product, EBC

- 281 (Final Report). Project Number: 9878/06. Unpublished study prepared by Stillmeadow, Inc. 12 pages.

472317

- 01 Wei Z, Thenell S. 2007 . ProAct Manufacturing Process. Unpublished study prepared by Plant Health Care, Inc. 71 pages.

C. Environmental Protection Agency Risk Assessment Memoranda

U.S. EPA. 200 7 . ProAct ® 

### Containing 1% Harpin Alpha Beta Protein. Memorandum from J.V. Gagliardi,

P h.D. and J.L. Kough, Ph.D. to J. Kausch dated December 4, 2007.Harpin Protein & Harpinαβ Protein (Harpin Proteins) EPA

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### U.S. EPA. 2005 b . Environmental Risk Assessment for EBC

- 351 (Harpinαβ Protein), Eden Bioscience Corporation . Memorandum from H. Hill and Z. Vaituzis, Ph.D. to D. Horne, Ph.D. dated January 7, 2005.

U.S. EPA. 2004 a . Review of Product Characterization, Susceptibility to Proteases, an Analytical Detection Method, and Toxicology Data for EBC

- 351 (Harpinαβ) in Support of an Experimental Use Permit for this Novel Hybrid Harpin Protein from Eden Biosciences, Bothell, Washington. Memorandum from C.A. Wozniak, Ph.D. and J.L. Kough, Ph.D . to D. Horne, Ph.D. dated March 31, 2004

.

U.S. EPA. 2004b. Environmental Risk Assessment for Experimental Testing of EBC

- 351 (Harpinαβ Protein). Memorandum from H. Hill and Z. Vaituzis, Ph.D. to D. Horne, Ph.D. and D. Szuhay dated March 30, 2004.

U.S. EPA. 2002b. Review of Amended Manufacturing Process, Product Analysis, Formation of Unintentional Ingredients, Request to Waive Acute Mammalian Toxicity Studies a nd an Acute Eye Irritation Study for Messenger ® 

### (Harpin) from Eden Biosciences, Bothell, Washington. Memorandum from C.A. Wozniak, Ph.D. and J.L. Kough, Ph.D. to D. Horne, Ph.D. dated June 12, 2002.

U.S. EPA. 2000 a . Review of Product Characterization and Toxicology Data for Harpin MessengerTM in Support of a Section Three Registration for MessengerTM (Harpin) from Eden Biosciences, Bothell, Washington. Memorandum from C.A. Wozniak, Ph.D. and J.L. Kough, Ph.D. dated June 7, 2000.

U.S. EPA. 2000b. Ecological Risk Assessment. Memorandum from G.S. Tomimatsu, Ph.D. and Z. Vaituzis, Ph.D. to D. Horne, Ph.D. and P.O. Hutton, Ph.D. dated March 28, 2000.

U.S. EPA. 1999

. Review of Submission for Harpin MessengerTM Product Analysis Data and Information, Characterization of Test Substances Used in Toxicology Studies, an Acute Oral Toxicity Study of Harpin Protein Technical in Rats, an Acute Oral Toxicity Study of

Concentra ted Harpin Protein Technical in Rats, an Acute Inhalation Toxicity Study of Harpin End Product in Rats, an Eye Irritation Study of Harpin End Product in Rats, a Dermal Irritation Study of Harpin End Product in Rabbits, Hypersensitive Incidents, Messenger TM: Supplemental Information and Summary of Production Process Used to Produce MessengerTM in Support for an Experimental Use Permit for MessengerTM (Harpin) from Eden Biosciences, Bothell, Washington. Memorandum from C.A. Wozniak , Ph.D. and J.L. Kough, Ph .D. to D. Horne, Ph.D. dated August 23, 1999

.

U.S. EPA. 1998. Review of Submitted Materials for Ecotoxicology Risk Assessment for Experimental Testing of a New Biological Active Ingredient, Harpin (Messenger ® 

### , Eden Biosciences, Inc.).

Memorandum from G .S. Tomimatsu, Ph.D. and Z. Vaituzis, Ph.D. to D. Horne, Ph.D. and P.O. Hutton dated October 29, 1998.Harpin Protein & Harpinαβ Protein (Harpin Proteins) EPA

- HQ

- OPP

- 201 2

- 0641 www.regulations.gov

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### D. Other References

U.S. EPA. 2005 a . Harpinαβ Protein (PC Code 006506) Biopesticides Registration Action Document (Issued January 31, 2005). Available from http://www.epa.gov/opp00001/chem_search/reg_actions/registration/decision_PC

- 006506_31

- Jan

- 05.pdf

.

U .S. EPA. 2002 a . Harpin Protein (PC Code 006477) Biopesticides Registration Action Document (Issued March 31, 2002). Available from http://ww w.epa.gov/opp00001/chem_search/reg_actions/registration/decision_PC

- 006477_31

- Mar

- 02.pdf

.