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TABLE OF CONTENTS

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 ..................................................................................................................................................26
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ACRONYMS AND ABBREVIATIONS

BPPD EPA’s Biopesticides and Pollution Prevention Division

C.F.R. Code of Federal Regulations

DNA Deoxyribonucleic acid

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 Hypersensitive response

IDS Incident Data System 

kD Kilodalton

kg Kilogram

lb Pound

mg Milligram

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

of Pesticide Programs.

MRL Maximum Residue Limit

OPP EPA’s Office of Pesticide Programs

oz Ounce

PC Code Pesticide chemical code: 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 States
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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 Registration Review case (docket number EPA-HQ-OPP-2012-0641), 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 

registered for use on a wide array of plants (e.g., fruiting vegetables, pome fruit, tobacco, and turf) in 

agricultural and residential settings to manage plant disease, enhance 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 the 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 Registration Review 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 will 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 other 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, currently 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, the 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 (under “SEARCH for: Rules, Comments, Adjudications or 

Supporting Documents,” enter the docket identification number, which 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 from and produced commercially in bacteria.
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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. Changes 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 safely. Information 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 

years 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 Registration 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 decision. After reviewing and responding to comments and data received in the docket during 

this initial comment period, EPA will develop a Final Work Plan (FWP) and anticipated schedule for the 

Registration Review of the harpin proteins.

C. Background and Regulatory Information

i. Summary of the Active Ingredients3

The harpin proteins act by eliciting a complex natural defense mechanism in 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. Unlike the action of 

most pesticides (e.g., conventional chemistries), 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 production 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. At the 

end of the fermentation process, Escherichia coli K-12 cells are killed 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).
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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 

vegetables, 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 never registered; 8 were registered but 

have since been cancelled4 for nonpayment of maintenance fees; and 4 remain registered. The 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 the Federal Register of August 6, 2008 (73 FR 45758) and July 28, 2010 (75 FR 44240). 
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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 Rates 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 

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

tolerance exemption expression for the 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 to enhance plant growth, quality and yield, to improve 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 (pH<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 (LD50) of greater than 5,000 mg product/kg body weight.

Federal Register Citations – Notices 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 Assessments

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 descriptions 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 with a footnote explanation, in Tables 2–5). For a succinct summary of data gaps, 

please see Table 6. 
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i. Product Chemistry (40 C.F.R. § 158.2030)

TABLE 2. Summary of Product Chemistry 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

880.1100 Product Identity and 

Composition Harpin Protein Yes – Study

U.S. EPA (1999)

U.S. EPA (2000a)

U.S. EPA (2002a)

U.S. EPA (2002b)

445424-01

448702-01

451912-01

Harpinαβ Protein Yes – Study U.S. EPA (2004a)

U.S. EPA (2005a) 460470-01

880.1200 Description of 

Starting Materials, 

Production and 

Formulation Process

Harpin Protein Yes – Study 

U.S. EPA (1999)

U.S. EPA (2000a)

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 (2004a)

U.S. EPA (2005a)

U.S. EPA (2007)

460470-01

472317-01

880.1400 Discussion of 

Formation of 

Impurities Harpin Protein Yes – Study

U.S. EPA (1999)

U.S. EPA (2000a)

U.S. EPA (2002a)

U.S. EPA (2002b)

445424-01

448702-01

454904-01

Harpinαβ Protein Yes – Study U.S. EPA (2004a)

U.S. EPA (2005a) 460470-01

830.1700 Preliminary Analysis

Harpin Protein Yes – Study U.S. EPA (1999)

U.S. EPA (2000a)

445424-01

448702-01

Harpinαβ Protein No – Data Gap1 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. The 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.
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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 Gap1 Not applicable

830.6303 Physical State

Harpin Protein Yes – Study U.S. EPA (1999) 445424-01

Harpinαβ Protein No – Data Gap2

Not applicable

830.6304 Odor Harpin Protein No – Data Gap

Harpinαβ Protein No – Data Gap1

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 Gap2 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 required3

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 Gap4

Harpinαβ Protein No – Data Gap2

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 Protein
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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 studies 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 and 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 Gap1

Not applicable

870.1200 Acute Dermal Toxicity Harpin Protein No – Data Gap2

Harpinαβ Protein No – Data Gap1

870.1300 Acute Inhalation 

Toxicity

Harpin Protein No – Data Gap2

Harpinαβ Protein No – Data Gap1

870.2400 Primary Eye Irritation Harpin Protein No – Data Gap2

Harpinαβ Protein No – Data Gap1

870.2500 Primary Dermal 

Irritation

Harpin Protein No – Data Gap2

Harpinαβ Protein No – Data Gap1

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

Harpinαβ Protein 448702-12

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 Protein
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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)

870.5385

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 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. 460470-08, 462040-01, 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 456064-

01.

iii. Nontarget Organisms and Environment Fate (40 C.F.R. § 158.2060)

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 – Study1 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 – Study1 U.S. EPA (2005a)

U.S. EPA (2005b)

850.1075 Fish Acute Toxicity, 

Freshwater Harpin Protein

No – Data Gap2 Not applicable

Harpinαβ Protein

850.1010 Aquatic Invertebrate 

Acute Toxicity, 

Freshwater

Harpin Protein

Yes – Study1

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 Gap2

Not applicable

Harpinαβ Protein

850.4150 Terrestrial Plant 

Toxicity, Vegetative 

Vigor

Harpin Protein

No – Data Gap

Harpinαβ Protein
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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 TGAI and, in some instances, the EP too. The 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 and 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 proteins. During Registration Review, EPA plans on conducting an 

ecological risk assessment to determine whether use of the harpin proteins will have “no effect” or “may 

affect” federally listed threatened or endangered species (listed species) and/or their designated 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), EPA has developed the Endocrine Disruptor Screening Program (EDSP) and has begun 

to implement the screening program that is to be used to test all pesticides 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 produced by a “naturally occurring estrogen, or other such endocrine effects 

as the Administrator may designate.” 

FFDCA section 408(p)(4) authorizes the Administrator, by order, to exempt from the 

requirements of the 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 9 inert ingredients. The harpin 

proteins are not among the group of 58 pesticide active ingredients on the initial list to be screened under 

the EDSP.

EPA, as part of this Preliminary Work Plan, believes that the harpin proteins, the active 

ingredients involved in this Registration Review case, likely are substances that would not produce any 

effect in humans similar to an effect produced by a naturally occurring estrogenic substance. As such, 

pursuant to Section 408(p)(4), EPA will determine in the future whether it can exempt the harpin 

proteins 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. 
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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 at 

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

G. Incidents

From January 1, 1992 through September 17, 2012, the 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 protein or harpinαβ protein and 3 reports of adverse 

effects to humans 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 (HD),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 to 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 usually 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 returned to his/her 

pre-exposure state of health with no additional residual 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 exhibited symptoms which may have been life-threatening, or resulted in adverse reproductive effects or in residual 

disability.”
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H. Summary of Anticipated Data Needs

Table 6 summarizes the anticipated data needs for the harpin proteins.

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

850.4100 Terrestrial Plant Toxicity, Seedling Emergence 12 months

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 Distribution
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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

850.4100 Terrestrial Plant Toxicity, Seedling Emergence 12 months

850.4150 Terrestrial Plant Toxicity, Vegetative Vigor

TGAI = technical grade of the active ingredient
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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 2012

 Close Public Comment Period November 2012

 Case Development

 Issue Final Work Plan July 2013

 Issue Data Call-in July 2014

 Data Submission July 2016

 Open 30-Day Public Comment Period for Draft Risk Assessments January 2018

 Close Public Comment Period February 2018

 Registration Review Decision

 Open 60-Day Public Comment Period for Proposed Registration Review 

 Decision 

June 2018

 Close Public Comment Period August 2018

 Final Decision September 2018

*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 any 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 Plan 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 provided in a timely manner, prior to issuing a Final Work Plan 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 policies. 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.
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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 

(TMDLs) have been developed for the harpin proteins.

3 More information on impaired water bodies and 

TMDLs can 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 

Process4 to ensure they can be used quantitatively or qualitatively in pesticide risk assessments.

iii. Trade 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 MRLs or disparities 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 requirement of a tolerance. Moreover, there are 

no known MRLs for the harpin proteins. Therefore, EPA does not anticipate that current uses of the 

harpin proteins 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 proteins: 

 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 limitations on use

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

sites)

 Median and 90th 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 Based 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.
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 Application timing (date of first application and application intervals) – national, state, and 

county

 Usage/use information for nonagricultural uses

 Typical application interval (days)

 State or local use restrictions

 Monitoring data

K. Next Steps

After the 60-day comment period closes, EPA will review and respond to any comments 

received in a timely manner and then issue a Final Work Plan 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 Products1

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 products

(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.

III. 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 Corporation. 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 Harpin Protein Technical in 

Rats. Lab Project Number: 789-002. Unpublished study prepared by MPI 

Research. 40 pages.
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MRID No. Study Description

445424-04 Hilaski R. 1998. An Acute Oral Toxicity Study of Concentrated Harpin Protein 

Technical in Rats. 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 Product 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.

445424-09 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.

445424-10 Gallagher S, Grimes J, Beavers J. 1998. A Dietary LC50 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-Renewal Test Conditions. Unpublished 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 Bioscience Corporation. 

45 pages.
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MRID No. Study Description

446914-01 Gallagher S, Grimes J, Beavers J. 1998. Harpin Protein Technical: A Dietary LC50

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. 

Unpublished 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 Research. 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.

447449-03 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. 1999. 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 Study 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. Unpublished study 

prepared by Eden Bioscience Corporation. 69 pages.
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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 HarpinEA. 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-Only 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 Wildlife International Limited. 53 

pages.

448702-22 Porch J, Krueger H. 1999. Messenger (HarpinEA 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.
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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 Color, 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

MRID 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 Corporation. 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 Dermal 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% Harpinαβ) 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: 7438/03. Unpublished study prepared by 

Stillmeadow, Inc. 18 pages.
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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 Bioscience 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 Analysis. 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 prepared 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. 2007. ProAct® Containing 1% Harpin Alpha Beta Protein. Memorandum from J.V. Gagliardi, 

Ph.D. and J.L. Kough, Ph.D. to J. Kausch dated December 4, 2007.
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U.S. EPA. 2005b. 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. 2004a. 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 and 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. 2000a. 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 

Concentrated 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, MessengerTM: 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-2012-0641

www.regulations.gov

26

D. Other References

U.S. EPA. 2005a. 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. 2002a. Harpin Protein (PC Code 006477) Biopesticides Registration Action Document 

(Issued March 31, 2002). Available from 

http://www.epa.gov/opp00001/chem_search/reg_actions/registration/decision_PC-006477_31-

Mar-02.pdf.