BIOPESTICIDES REGISTRATION ACTION DOCUMENT



## Pasteuria nishizawae

– Pn1

Pesticide Chemical (PC) Code: 016455

U.S. Environmental Protection Agency Office of Pesticide Programs Biopesticides and Poll ution Prevention Division

February 28 , 2012Pasteuria nishizawae

– Pn1 Page 2 of 34 Biopesticides Registration Action Document

TABLE OF CONTENTS

I. EXECUTIVE SUMMARY

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

II. ACTIVE INGREDIENT OV ERVIEW

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

III. REGULATORY BACKGROUN D

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

A. Applications for Pesticide Product Registration

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

B

. Food Tolerance Exemption

................................ ................................ ................................ .............. 7

IV. RISK ASSESSMENT

................................ ................................ ................................ ................................ .... 7

A. Product Analysis Assessment (40 CFR § 158.2120)

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

B. Human Health Assessment (40 CFR § 158.2140)

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

C. Environmental Assessment (40 CFR § 158.2150)

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

V. ENVIRONMENTAL JUSTIC E

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

VI. RISK MANAGEMENT DECI SION

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

VII. ACTIONS REQU IRED OF THE REGISTRA NT

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

VIII. GLOSSARY OF ACRONYMS AND ABBREVIATIONS

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

IX. BIBLIOGRAPHY

................................ ................................ ................................ ................................ ....... 20

A. Studies Submitted to Support the Pasteuria nishizawae

– Pn1 Pesticide

Product Registrations

................................ ................................ ................................ ..................... 20

B. Environmental Protection Agency Risk Assessme nt Memoranda

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

C. Other References

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

APPENDIX A. MICROBI A L PESTICIDES DATA RE QUIREMENTS

................................ ............................ 26

APPENDIX B. PESTICI DE PRODUCTS

................................ ................................ ................................ .............. 34Pasteuria nishizawae

– Pn1 Page 3 of 34 Biopesticides Registration Action Document

BIOPESTICIDES REGISTRATION ACTION DOCUMENT TEAM

Office of Pesticide Programs ( OPP) Biopesticides and Pollution Prevention Division

Microbial Pesticides Branch

Science Reviews

Joel V. Gagliardi , Ph.D. Product Analysis, Human Health

John L. Kough, Ph.D. Product Analysis, Human Health Shannon Borges Environmental Effects Gail Tomimatsu, Ph.D. Environmental Effects

Regulations

Sheryl K. Reilly, Ph.D. Chief, Microbial Pesticides Branch

Jeannine Kausch Regulatory Action LeaderPasteuria nishizawae

– Pn1 Page 4 of 34 Biopesticides Registration Action Document

I. EXECUTIVE SUMMARY

In July and August 2010

, Pasteuria Biosc ience, Inc. submitted application s for a manufacturing

use p esticide product, Soyacyst Tech (EPA File Symbol 85004

- A), and two end

- use pesticide product s
, Soyacyst Tech+ (EPA File Symbol 85004

- T ) and Soyacyst LF (EPA File Symbol

85004

- O), to the United Sta tes Environmental Protection Agen cy (EPA) under section 3 of the Federal Insecticide, Fungicide, and Rodenticide Act (FIFRA)

. Concurrently with these applications, Pasteuria Bioscience, Inc. filed a petition for a tolerance exemption for residues of

Pasteu ria nishizawae

– Pn1 , the new active ingredient contained in all three proposed pesticide produc ts

. Background

Pasteuria , a genus of bacteria, includes several species that have shown potential in controlling plant

- parasitic nematodes that attack and cause signific ant damage to many valuable agricultural

crops (see, e.g., the Federal Register of December 28, 1994 ( 59 Federal Register ( FR ) 66740 ) and June 30, 2010 ( 75 FR 37734 ) for F inal R ules that established tolerance exemptions for residues of the nematicides, Pasteuria penetrans (40 Code of Federal Regulations ( CFR ) 180.1135) and Pasteuria usgae (40 CFR 180.12 90), respectively) . These gram

- positive, mycelial, endospore

forming bacteria are obligate parasites (i.e., organisms that depend on particular hosts to complete their own life cycle) of nematodes and water fleas. 1 Pasteuria species are ubiquitous in

most environments and are found in nematodes in at least 80 countries on 5 continents, as well as on islands in the Atlantic, Pacific, and Indian Oceans. Higher population densities often occur in areas where there is an ample supply of nematode hosts (e.g., wh ere crops susceptible to nematodes are cultivated) ( Centitas and Dickson 2004; Noel 2008; Tain et al . 2007 ). Pasteuria
nishizawae

– Pn1 was specifically isolated from an Illinois soybean field in the mid

- 2000s

.

Although endospores of Pasteuria nishizawae have been observed to attach to the cuticle of three

nematodes of the genus Heterodera and one nematode of the genus Globodera , it is known only to infect and complete its life cycle within the female soybean cyst nematode ( Heterodera glycines ) . In the fo llowing manner, Pasteuria nishizawae

– Pn1 exerts a pesticidal effect on the soybean cyst nematode through parasitism that ultimately results in the death of infected females:

(1) Endospores attach to the cuticle of a juvenile soybean cyst nematode female

. (2) O nce a soybean cyst nematode female invades soybean roots, Pasteuria
nishizawae

– Pn1 produces a germ tube that penetrates the body of the nematode. (3) Primary and secondary microcolonies of Pasteuria nishizawae

– Pn1 develop and proliferate within the body of the nematode, causing its death

.

In light of the demonstrated nematicidal capabilities and host specificity of Pasteuria nishizawae

– Pn1, Pasteuria Bioscience, Inc. proposed to register several pesticide products that could be applied to soybean or it s seed to control the soybean cyst nematode.

EPA s cientists reviewed product analysis, toxicology, and nontarget organism data and information (40 CFR §§ 158.2120, 158.2140, and 158.2150, respectively) submitted to support

the registration of the t hree pr oposed Pasteuria nishizawae

– Pn1 pesticide products. Overall,

1 Pasteuria ramosa is the only Pasteuria species that is known to parasitize water fleas.Pasteuria nishizawae

– Pn1 Page 5 of 34 Biopesticides Registration Action Document

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

, and allow for registration under FIFRA section 3(c)(5)

.

T he p roduct analysis data requirements for Pasteuria nishizawae

– Pn1 , including product chemistry and composition, analysis of samples, and physical and chemical characteri stics, were fulfilled by acceptable guideline studies. Product Analysis

Adequat e mamm alian toxicology data and information were submitted to support the Pasteuria
nishizawae

– Pn1 pesticide products. A cute oral, pulmonary, and injection toxicity/pathogenicity

studies showed t hat, at a single high dose, Pasteuria nishizawae

– Pn1 is n ot toxic and/o r

p athogenic via these route s of exposure

. Moreover, the applicant reported that no hypersensitivity

incidents occurred during rese arch, development, or testing of Pasteuria nishizawae

– Pn1 . In

l ight of the results of the acute toxicity/path ogenicity data and the absence of hypersensitivity incidents, testing at higher tiers (i.e., Tiers II and III) was not required. Toxicology

EPA concludes that there is a reasonable certainty that no harm will result to the U.S. population,

includ ing infants and children, from aggregate exposure to residues of Pasteuria nishizawae

– Pn1 . No dietary risks are expected from use of Pasteuria nishizawae

– Pn1 as an active ingredient in pesticide products. Significant exposure to Pasteuria nishizawae

– Pn1 through

food and drinking water is not anticipated due to : (1) the proposed application methods of the

end

- use pesticide products (soil directed, soil incorporated , and/or seed directed ; no aquatic applications ) ; ( 2 ) the filtering effect of many partic ulate soil types; and ( 3 ) the conditions (e.g., filtration and pH adjustments) water is subjected to in wastewater treatment systems and drinking

water facilities

. Should Pasteuria nishizawae

– Pn1 be present on food or in drinking water, its specificity f or the s oybean cyst nematode and supporting acute oral toxicity and pathogenicity

d ata indicate that human exposure to this bacterium is not likely to be a concern. Tolerance Exemption

Despite the low toxicological profile of Pasteuria nishizawae

– Pn1

, baseline personal protective equipment (PPE) is required for handlers that may be exposed to the active ingredient, due to their occupation, for prolonged periods or numerous times . Handlers working with Pasteuria
nishizawae

– Pn1 in agricultural settings are directed to wear a long

- sleeved shirt, long pants, socks, shoes, and a dust/mist filtering respirator meeting National Institute for Occupational Safety and Health (NIOSH) standards of at least N

- 95, R

- 95, or P

- 95. EPA may require

additional PPE (e.g. , protective eyewear) , other than the standard described above, on a product

specific basis. Occupational Exposure

Data and other i nformation (e.g., scientific literature) submitted by the applicant to support requests to waive nontarget organism testing for Pasteuria nishizawae

– Pn1 are sufficient to

fulfill the relevant microbial pesticide data requirements and for risk assessment purposes. Further testing of nontarget organisms at higher tier levels (i.e., Tiers II, III, and IV) is not required. EPA pe rformed an environment al risk assessment based on data and other information provided by the ap plicant

, and determined that adverse effects to nontarget organisms are not anticipated from the proposed pesticidal uses of Pasteuria nishizawae

– Pn1 . Moreover , EPA Nontarget OrganismsPasteuria nishizawae

– Pn1 Page 6 of 34 Biopesticides Registration Action Document

made a “No Effect” determination for direct and indirect effects to listed species and their

d esignated critical habitats resulting from these same proposed pesticidal uses.

On October 1, 2009, EPA announced a new policy to provide a more meaningful opportunity for the public to participate i n major registration decisions before they occur. According to this policy, EPA intends to provide a public comment period prior to making a registration decision for, at minimum, the fol lowing types of appli cations: new active ingredients; first food uses; first outdoor uses; first residential use s; or any other registration actions for which EPA believes there may be significant public interest

. Public Participation

Consistent with the policy of making re gistration actions more transparent, the proposed

pe sticide products containing Pasteuria nishizawae

– Pn1

, a new active ingredient, and allowing for this active ingredient’s first outdoor and food uses were subject to a 30

- day comment period. During this comment period, no comments were received on EPA’s preliminary decision to register the Pasteuria nishizawae

– Pn1 pesticide products, Soyacyst Tech , Soyacyst Tech+, and Soyacyst LF. Therefore, EPA maintained that, based upon the risk assessment and inform ation submitted in support of registration of such pesticide products, it was appropriate to issue the Soyacyst Tech, Soyacyst Tech+, and Soyacyst LF registrations under FIFRA section 3(c)(5). The

basis for this decision can be found in the risk assessment for Pasteuria nishizawae

– Pn1, which is characterized throughout this Biopesticides Registration Action Document (BRAD).

II. ACTIVE INGREDIENT OVERVIEW

Biological Name: Pasteuria nishizawae

– Pn1

Culture Deposit: American Type Culture Collect ion in Manassas

, Virginia

under Accession N umber SD

- 5833

OPP Chemical Code: 016455

Type of Pesticide: Microbial Pesticide

– Nematicide

See Appendix B for specific information (e.g., use sites, applicati on rate s , methods of application, formulation type s , and target pests) regarding

the registered pesticide products containing this active ingredient.

III. REGULATORY BACKGROUND

A. Applications for Pesticide Product Registration

In July and August 2 010

, MacIntosh and Associates , Inc. (address: 1203 Hartford Avenue; Saint Paul, Minnesota 55116

- 1622 ) , on behalf of Pasteuria Bioscience , Inc. (address: 12085 Research Drive, Suite 185; Alachua, Florida 32615 ), submitted application s to register a manufact uring

-Pasteuria nishizawae

– Pn1 Page 7 of 34 Biopesticides Registration Action Document

use pesticide product, Soyacyst Tech (EPA File Symbol 85004

- A )

, and two end

- use pesticide product s
, Soyacyst Tech+ (EPA File Symbol 85004

- T ) and Soyacyst LF (EPA File Symbol

85004

- O ), under FIFRA section 3. On November 24 , 2010 ( 75 FR 71697 ) and February 2, 2011 ( 76 FR 5805 )

, EPA announced receipt of these application s to register pesticide produc t s containing a new active ingredient and opened a 3 0

- day public comment period pursuant to the provisions of FIFRA section 3(c)(4). No comme nts were received following these publication s

.

B . Food Tolerance Exemption

Concurrent with its registration a pplication s and under Federal Food, Drug, and Cosmetic Act (FFDCA) section 408(d), Pasteuria Bioscience , Inc. submitted a petition to establish an exemption from the requirement of a tolerance for Pasteuria nishizawae

– Pn1 (Pesticide Petition (PP) 0F7749 )

. In the Federal Register of February 4, 2011 ( 76 FR 6465 ), EPA announced that

Pasteuria Bioscience , Inc. proposed to establish an exemption from the requirement of a tolerance f or residues of the microbial pesticide

, Pasteuria nishizawae

– Pn1

, in or on all raw agricultural crops and opened a 30

- day comment period. Two comments were received following this publication and are addressed in the preamble to the F inal R ule

.

On Feb ruary 15, 2012 , EPA established an exemption from the requirement of a tolerance for

residues of Pasteuria nishizawae

– Pn1 in or on all food commodities when applied as a nematicide and used in accordance with good agricultural practices (40 CFR § 180.131 1; 77 FR

8736 ).

IV. RISK ASSESSMENT

In the Federal Register of October 26, 2007, EPA issued a Final Rule on the data requirements to support registration of microbial pestici des and updated the definition for microbial pesticides ( 72 FR 61002 )

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

evaluated for this BRAD were consid ered in light of these requirements.

The classifications that are found for each data submission are assigned by EPA science

reviewers and are an indication of the usefulness of the information contained in the documents for risk assessment. A rating of “ acceptable ” indicates the study is scientifically sound and is

useful for risk assessment. A “ supplemental ” rating indicates the data provide some information that can be useful for risk assessment. The studies may have certain aspects determined not to be scientifically acceptable (“ supplemental : upgradeable ”). If a study is rated as “supplemental: upgradeable ,” EPA always provides an indication of what is lacking or what can be provided to change the rating to “ acceptable .” If there is simply a “ supplemen tal ” rating, the reviewer will

often state that t he study is not required by 40 C FR P art 158. Both “ acceptable” and “supplemental ” studies may be used in the risk assessment process as appropriate. An “unacceptable ” rating indicates that new data must be s ubmitted.

For the acu te toxicity data requirements, Toxicity C ategories are assigned based on the hazard(s) identified from studies and/or other infor mation submitted to EPA in support of a pesticide registration. The active ingredient or particular prod uct is classified into Toxicity Category I, II, III, or IV, where Toxicity Category I indicates the highest toxicity and Toxicity Category IV indicates the lowest toxicity (see 40 CFR § 156.62 )

.Pasteuria nishizawae

– Pn1 Page 8 of 34 Biopesticides Registration Action Document

A. Product Analysis Assessment ( 40 CFR § 158.2120 )

A ll product analysis data requirements for Pasteuria nishizawae

– Pn1 have been fulfilled. Refer to Tables 1, 2, 3, and 4 in Appendix A fo r a summary of the data requirements, including both generic and product

- specific information.

B. Human Health Assessment ( 40 CFR § 158.2140 )

1. Toxicity

All toxicology data requirements for Pasteuria nishizawae

– Pn1 have been fulfilled

. Acceptable Tier I mammalian toxicology data and information support registration of the Pasteuria
nishizawa e

– Pn1 pesticide products

. Furthermore, Tier II and Tier III studies were not required

for Pasteuria nishizawae

– Pn1

, based on the lack of acute toxicity/pathogenicity in the Tier I

studies.

For a summary of the generic toxicology data requirements desc ribed in sections IV(B)(1)(a) and IV(B)(1)(b), as well as additional product

- specific data submitted to support the individual registrations, refer to Tables 5 and 6 in Appendix A

.

a. Acute Toxicity/Pathogenicity

– Tier I

Acute Oral Toxicity/Pathogenicity

– Rat ( Harmonized Guideline 885.3050; Master Record Identification Number (MRID No.) 481517

- 09 ) : Groups of rats were treated with live test

substance at 1.6 x 10 9 spores/rat

, were treated with killed (autoclaved) test subs tance, or were in 1 of 2 untreated control groups. No adverse clinical signs were observed in any test animals. At
necropsy, no treatment

- related gross observations were noted. Organ weights primarily did not

differ , though in male rats treated with live t est substance

, brain weights on day s 7, 14, and 21 and spleen weight on day 21 were slightly but significantly decreased. In female rats from the same group, the only statistical ly significant difference was heavier stomach/small intestine weight on day 7. No evidence of pathogenicity or toxicity was found from oral administration of
Pasteuria nishizawae

– Pn1 to rats. Although clearance and infectivity were not measured, EPA believes these endpoints are not a concern given Pasteuria nishizawae

– Pn1’s well

- established host specificity for the soybean cyst nematode . This study was rated supplemental

.

Acute Pulmonary Toxicity/Pathogenicity

– Rat ( Harmonized Gui deline 885.3150; MRID No.

481517

- 10 ) : Groups of rats were treated with live test substance at 1.6 x 10 8 spores/rat , were treated with killed (autoclaved) test substance, or were untreated controls. No adverse clinical signs were observed in any test animals. At necropsy, no treatment

- related gross observations were noted. Organ weights primarily did no t differ , though in male rats treated with live test

substance , brain weights on days 7 , 14, and 21 and liver weight on day 21 were slightly but significantly decreased. In female rats from the same group, there were no statistically significant

difference s in organ weights. No evidence of pathogenicity or toxicity was found from pulmonary administration of Pasteuria nishizawae

– Pn1 to rats. Although clearance and infectivity were not measured, EPA believes these endpoints are not a concern given Pasteuria
nishizawae

– Pn1’s well

- established host specificity for the soybean cyst nematode . This study was rated supplemental

.Pasteuria nishizawae

– Pn1 Page 9 of 34 Biopesticides Registration Action Document

Acute Injection Toxicity/Pathogenicity (Intra venous )

– Rat (Harmonized Guideline 885.3200;

MRID No . 481517

- 1 1 ) : Groups of rats were tre ated with live test substance at 1.0 x 10 9
spores/rat , were treated with killed (autoclaved) test substance, or were untreated controls. No adverse clinical signs were observed in any test animals. At necropsy, no treatment

- related gross

observations were noted. Organ weights primarily did not differ , though in males rats treated with live test substance, spleen weights on days 14 and 21 and liver weight on day 21 were slightly but significantly decreased. In female rats from the same group, there were no

s tatistically significant differences in organ weights. No evidence of pathogenicity or toxicity was

found from intravenous administration of Pasteuria nishizawae

– Pn1 to rats. Although clearance and infectivity were not measured, EPA believes these endpoi nts are not a concern given
Pasteuria nishizawae

– Pn1’s well

- established host specificity for the soybean cyst nematode

. This study was rated supplemental

.

Hypersensitivity Incidents ( Harmonized Guideline 885.3400; MRID No. 481517

- 12 ) : The applicant rep orted that n o hypersensitivity incidents, including immediate

- type or delayed

- type
reactions of humans and domestic animals

, occurred during research, development, or testing of
Pasteuria nishizawae

– Pn1 . Any future hypersensitivity incidents must be repo rted to EPA (refer to test note #3 of 40 CFR § 158.2140(d))

.

Cell Culture ( Harmonized Guideline 885.3500) : This study is not required because Pasteuria
nishizawae

– Pn1 is not a virus (refer to test note #4 of 40 CFR § 158.2140(d)).

b. Acute Toxicology and Subchronic Toxicity/Pathogenicity

– Tier II; Reproductive Fertility Effects, Carcinogenicity, Immunotoxicity, and Infectivity/Pathogenicity Analysis

– Tier III

Tier II and Tier III studies were not required for Pasteuria nishizawae

– Pn1

, b ased on the lack of acute toxicity/pathogenicity in the Tier I studies.

c. Endocrine Disruptors

As required under FFDCA section 408(p), EPA has developed the Endocrine Disruptor Screening Program (EDSP) to d etermine 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 desig nate.” The EDSP employs a two

- tiered approach to making the statutorily required determinations. Tier 1 consists of a battery of 11 screening assays to identify the potential of a chemical substance to interact with the estrogen, androgen, or thyroid (E, A , or T) hormonal systems. Chemicals that go through Tier 1 screening and are found to have the potential to interact with E, A, or T hormonal systems will proceed to the next stage of the EDSP where EPA will determine which, if any, of the Tier 2 tests are necessary based on the available data. Tier 2 testing is designed to identify any adverse endocrine

- related effects caused by the substance, and establish a quantitative relationship between the dose and the E, A, or T effect.

Between October 2009 and Fe bruary 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. This list of chemicals was selected based on the potential for human exposure through pathways su ch asPasteuria nishizawae

– Pn1 Page 10 of 34 Biopesticides Registration Action Document

food and water, residential activity, and certain post

- application agricultural scenarios. This list should not be construed as a list of known or likely endocrine disruptors.

Pasteuria nishizawae

– Pn1 is not among the group of 58 pesticide active ingredients on the initial list to be screened under the EDSP. Under FFDCA section 408(p), EPA must screen all pesticide chemicals. Accordingly, EPA anticipates issuing future EDSP orders/data call

- ins for all pesticide active ingredients.

For further in formation on the status of the EDSP, the policies and procedures, the list of 67 chemicals, the test guidelines and the Tier 1 screening battery, please visit our website: http://www.epa.gov/endo/

.

2

. Federal Foo d, Drug, and Cosmetic Act (FFDCA ) Considerations

Section 408(c)(2)(A)(i) of FFDCA allows EPA to establish an exemption from the requirement for a tolerance (the legal limit for a pesticide chemical residue in or on a food) only if EPA

determines that the exemption is “safe.” Section 408(c)(2)(A)(ii) of FFDCA defines “safe” to mean that “there is a reasonable certainty that no harm will result from aggregate exposure to the pesticide chemical residue, including all anticipated dietary exposures and all othe r exposures for which there is reliable information.” This includes exposure through drinking water and in

resi dential settings

, but does not include occupational exposure. Pursuant to section 408(c)(2)(B) of FFDCA, in establishing or maintaining in effect an exemption from the requirement of a tolerance, EPA must take into account the factors set forth in section 408(b)(2)(C) of FFDCA, which require EPA to give special consideration to exposure of infants and children to the pesticide chemical residue in e stablishing a tolerance exemption

, and to “ensure that there is a reasonable certainty that no harm will result to infants and children from aggregate exposure to the pesticide chemical residue....” Additionally, section 408(b)(2)(D) of FFDCA requires that EPA consider “available information concerning the cumulative effects of [a particular pesticide's]

. . . residues and other substances that have a common mechanism of toxicity.”

EPA performs a number of analyses to determine the risks from aggregate expo sure to pesticide residues. First, EPA determines the toxicity of pesticides. Second, EPA examines exposure to the pesticide through food, drinking water, and through other exposures that occur as a result of pesticide use in residential settings.

Consiste nt with section 408(b)(2)(D) of FFDCA, EPA has reviewed the available scientific data and other relevant information, and considered its validity, completeness, and reliability and the relationship of this information to human risk. EPA has also considered available information concerning the variability of the sensitivities of major identifiable subgroups of consumers, including infants and children.

Based on the acute toxicity/pathogenicity data and information discussed previously and presented in Table s 5 and 6 in Appendix A , t he data required for a FFDCA risk assessment for
Pasteuria nishizawae

– Pn1 have been fulfilled

.Pasteuria nishizawae

– Pn1 Page 11 of 34 Biopesticides Registration Action Document

a. Aggregate Exposure

In examining aggregate exposure, section 408 of FFDCA directs EPA to consi der available information concerning exposures from the pesticide residue in food and all other nonoccupational exposures, including drinking water from ground water or surface water and exposure through pesticide use in gardens, lawns, or buildings (resi dential and other indoor uses).

Food Exposure and Risk Characterization : Dietary exposure to Pasteuria nishizawae

– Pn1

, a naturally occurring soil bacterium (Atibalentja et al

. 2004; Noel et al . 2005; Sayre et al . 1991), is anticipated to be negligible

. For o ptimal control of the target pest

, soybean cyst nematode ( Heterodera glycines ), Pasteuria nishizawae

– Pn1 is appli ed in a manner that facilitates spore

movement into or spore placement near th e root zone of potentially affected plants . This requires

that end users take certain actions , depending on the treatment type, that would inevitably

minimize the amount of Pasteuria nishizawae

– Pn1 residues on above

- ground food commodities. That is, although Pasteuria nishizawae

– Pn1 can be applied to soil , pl ants, or seeds, some s e eds are incorporated into the soil immediately after treatment ( at

- planting, hopper box, planter box, or slurry box seed treatment s )

, and pesticide applications made to plants or the

soil are always followed by irrigation to incorpor ate Pasteuria nishizawae

– Pn1 into the soil

. In

instances where food commodities develop underground or where treated seed is diverted for food or feed purposes or to process into oil , exposure to Pasteuria nishizawae

– Pn1 residues is a more likely scena rio. Regardless of the situation, s hould Pasteuria nishizawae

– Pn1 be present on food, its specificity for the soybean cyst nematode and available data indicate no toxicity

, pathogenicity , and/or infectivity is likely to occur with any dietary exposure th at results from pesticide application s made in accordance with good agricultural practice s (see section IV(B)(1)(a) and Table s 5 and 6 in Appendix A ).

Drinking Water Exposure and Risk Characterization : Exposure to residues of Pasteuria
nishizawae

– Pn1 in consumed drinking water is possible but not likely. The proposed use patterns for Pasteuria nishizawae

– Pn1 are soil directed

, soil incorporated

, and/or seed directed, thereby limiting contact with surface water by drift and runoff

. Furthermore, ground water is not expected to have significant exposure to Pasteuria nishizawae

– Pn1 since , like other microorganism s , this microbial pesticide would likely be filtered out by the particulate nature of many soil types (Aislabie et al . 2001; DeFelice et al . 1993; Pang et al . 2008)

. I f Pasteuria
nishizawae

– Pn1 were to be transferred to surface or ground water s (e.g., through spray drift or runoff) that are intended for eventual human consumption and directed to wastewater treatment systems or drinking water facilities

, it may not survive some of the cond itions water is subjected to in such systems or facilities , including chlor ination, pH adjustments, and filtration (Center s for Disease Control and Prevention 2009; U.S. EPA 2004)

. In the remote likelihood that Pasteuria
nishizawae

– Pn1 is present in drinking water (e.g., water not subject to treatment systems or facilities), its specificity for the soybean cyst nematode and available data indicate no toxicity

, pathogenicity , and/or i nfectivity is likely to occur with any drinking water exposure that results from pesticide application s made in accordance with good agricultural practice s (see section IV(B)(1)(a) and Table s 5 and 6 in Appendix A )

.

Non

- occu pational, Residential Risk Characterization : Given Pasteuria nishizawae ’ s natural

occurrence in soil (Atibalentja et al . 2004; Noel et al . 2005; Sayre et al . 1991)

, non

- occupational
exposure to the bacterium is likely already occurring

. Additional exposure to Pasteuria
nishizawae

– Pn1 due to pesticidal applications is not expected because all proposed pesticidePasteuria nishizawae

– Pn1 Page 12 of 34 Biopesticides Registration Action Document

end

- use products are labeled for use in distinct agricultural settings. Even if non

- occupational
exposures were to occur (e.g., eventual expansion of use sites) , such exposures would not exceed

EPA’s level of concern in light of Pasteuria nishizawae

– Pn1’s specificity for the soybean cyst

nematode and test results that indicated Pasteuria nishizawae

– Pn1 is not toxic (acute dermal toxicity and acut e pulmonary toxicity/pathogenicity ), is essentially non

- irritating (primary dermal irritation), and is not pathogenic (acute pulmonary toxicity/pathogenicity) (see section IV(B)(1)(a) and Table s 5 and 6 in Appendix A ).

b. Cumulative Effects from Substances with a Common Mechanism of Toxicity

Section 408(b)(2)(D)(v) of FFDCA requires that, when considering whether to establish, modify, or revoke a tolerance exemption, EPA consider “available information concerning the cumul ative effects of [a particular pesticide’s]

. . . residues and other substances that have a common mechanism of toxicity.”

No mechanism of toxicity in mammals has been identified for Pasteuria nishizawae

– Pn1, and
Pasteuria nishizawae

– Pn1 does not appea r to produce a toxic metabolite against the target pest

. For the purposes of the tolerance action , EPA has assumed that Pasteuria nishizawae

– Pn1 does

not have a common mechanism of toxi city with other substances. Therefore

, section 408(b)(2)(D)(v) of FFD CA does not apply. For information regarding EPA ’s efforts to determine which chemicals have a common mechanism of toxicity and to evaluate the cumulative effects o f such chemicals, see EPA ’s website at http://www.epa.gov/pesticides/cumulative

.

c. Determination of Safety for the U.S. Population, Infants and Children

FFDCA section 408(b)(2)(C) provides that, i n considering the establishment of a tolerance or tolerance exemption for a pesticide che mical residue, EPA shall assess the available information about consumption patterns among infants and children, special susceptibility of infants and children to pesticide chemical residues, and the cumulative effects on infants and children of the

residu es and other substances with a common mechanism of toxicity. In addition, FFDCA section 408(b)(2)(C) provides that EPA shall apply an additional tenfold (10X) margin of exposure ( safety ) for infants and children in the case of threshold effects to account for prenatal and postnatal toxicity and the completeness of the database on toxicity and exposure unless EPA determines that a different margin of exposure ( safety ) will be safe for infants and children. This additional margin of exposure ( safety ) is commo nly referred to as the Food Quality Protection Act Safety Factor. In applying this provision, EPA either retains the default value of 10X or uses a different additional safety factor when reliable data available to EPA support the choice of a

different fac tor.

Based on the acute toxicity an d pathogenicity data discussed in section IV(B)(1)(a) and Tables 5

and 6 in Appendix A

, as well as Pasteuria nishizawae

– Pn1’s host specificit y for the soybean cyst nematode , EPA concludes that there are no threshold effects of concern to infants, children,

or adults when Pasteuria nishizawae

– Pn1 is used as labeled in accordance with good agricultural practices. As a result, EPA concludes that no additional margin of exposure (safety)

is necessary to protect infants and children and that not adding any additional margin of exposure (safety) will be safe for infants and children.

Moreover, based on the same data and EPA analysis as presented directly above, the Agency isPasteuria nishizawae

– Pn1 Page 13 of 34 Biopesticides Registration Action Document

able to conclude t hat there is a reasonable certainty that no harm will result to the U.S. population, including infants and children, from aggregate exposure to the residues of Pasteuria
nishizawae

– Pn1 when it is used as labeled and in accordance w ith good agricultural p ractices as

a nematicide . Such exposure includes all anticipated dietary exposures and all other exposures for which there is reliable information. EPA has arrived at this conclusion because, considered collectively, the data and information available on P asteuria nishizawae

– Pn1 do not

demons trate toxic, pathogenic , and/or infective potential to mammals, including infants and

children.

3. Occupational Exposure and Risk Characterization

Handler exposure to Pasteuria nishizawae

– Pn1 is not expected to pose any undue risk. Regardless, appropriate personal protective equipment and precautionary statements are required on pesticide product labels to mitigate any potential risks to pesticide handlers due to prolonged or numerous exposures. Handlers applying Pasteuria nishizawae

– Pn1 end

- use pesticide products in agricultural settings must wear a long

- sleeved shirt, long pants, socks, shoes, and a dust/mist filtering respirator meeting NIOSH standards of at least N

- 95, R

- 95, or P

- 95.

Additional PPE (e.g., pr otective eyewear) , other than the standard described above, may be required on a product

- specific basis.

4. Human Health Risk Characterization

EPA considered human exposure to Pasteuria nishizawae

– Pn1 in light of the standard for registration in FIFRA and the relevant safety factors in FFDCA. A determination has been made

that no unreason able adverse effects to the U.S. population in general, and to infants and children in particu lar, will result when Pasteuria nishizawae

– Pn1 pesticide products are us ed in

accordance with EPA

- approved labeling.

C. Environmental Assessment ( 40 CFR § 158.2150 )

Data and other i nformation (e.g., scientific literature) submitted by the applicant to support requests to waive nontarget organism testing for Pasteuria nishizawae

– Pn1 are sufficient to

fulfill the relevant microbial pesticide data requirements and for risk assessment purposes. Further testing of nontarget organisms at higher tier levels (i.e., Tiers I I, III, and IV) is not required . EPA performed an environmental risk assessment based on data and other information provided by the ap plicant

, and determin ed that adverse effects to nontarget organisms are not anticipated from the proposed pesticidal uses of Pasteuria nishizawae

– Pn1

. Moreover, EPA made a “No Effect” determination for direct and indirect effects to listed species and their designated critic al habitats resulting from these same proposed pesticidal uses.

For a summary of the generic data requirements described in sections IV(C)(1), refer to Table 7

in Appendix A

.Pasteuria nishizawae

– Pn1 Page 14 of 34 Biopesticides Registration Action Document

1. Ecological Exposure and Risk Charac terization

a. Terrestrial Animals and Plants

Birds ( Harmonized Guideline 885.4050)

, Wild Mammals (Harmonized Guideline 885.4150)

, Nontarget Plants (Harmonized Guideline 885.4300), Nontarget Insects ( Harmonized Guideline 885.4340)

, and Honey Bees (Ha rmonized Guideline 885.4380) (MRID No. 481517

- 17 ) : The proposed end

- use pesticide products are for use on soybeans. The liquid formulation end

- use pesticide product may be applied by soil

- directed spray or chemigation, including both soil

directed and foli age

- directed (e.g., ov erhead sprinkler) chemigation. Applications are to be

followed with a sufficient amount of water to move the Pasteuria nishizawae

– Pn1 spores into the root zone. Therefore, both formulations will primarily deposit Pasteuria nishizawa e

– Pn1 into the soil. While this type of application reduces the potential for exposure to many nontarget species, exposure to nontarget species consuming, contacting, or living in the soil will not necessarily be eliminated.

Data and other i nformation s ubmitted by Pasteuria Bioscience, Inc. to support its waiver requests provides sufficient information to conclude that adverse effects are not expected in birds,

non target insects, and honey bees as a result of exposure to Pasteuria nishizawae

– Pn1

. Paste uria spp. are widespread in their distribution throughout t he world . They are known only as parasites of plant

- parasitic nematodes, with the exception of Pasteuria ramosa that is known only to parasitize Daphnia spp. (Atibalentja et al

. 2004 ; Chen and Dick son 1998; Sayre et al

. 1991). Pasteuria nishizawae is considered to be an obligate parasite of Heterodera glycines and is known only to infect and complete its life cycle within females of Heterodera glycines ( Noel et
al . 2005 ; Sayre et al . 1991 ). Therefor e, Pasteuria nishizawae

– Pn1 is not expected to be toxic or pathogenic to birds, nontarget insects

, or honey bees if they are exposed as a result of the proposed pesticidal applications.

An acute oral toxicity/pathogenicity study with laboratory rats (MR ID No. 481517

- 09) and an acute oral toxicity study (MRID No. 481517

- 13) are available to evaluate the potential effects of

Pasteuria nishizawae

– Pn1 on wild mammals. In the acute oral toxicity/pathogenicity study,

l aboratory rats were dosed with approxima tely 1.6 x 10 9 spores/animal and were observed for 21 days. No signs of toxic or pathogenic effects were found as a result of the exposure. This study was determined to be supplemental because clearance and infectivity were not observed. The

acute oral tox icity study showed no evidence of toxicity in rats dosed with 5

, 000 milligrams per kilogram bodyweight , and this study was determined to be acceptable. These studies show that toxicity/pathogenicity of Pasteuria nishizawae

– Pn1 to laboratory rats is not e xpected. There is no reason that the test animals in these studies would not be adequate models with which to determine potential effects to wild mammals, so adverse effects to wild mammals are not expected as a result of the proposed pesticidal applicatio ns of Pasteuria nishizawae

– Pn1.

Non target plant testing is not required because Pasteuria nishizawae is not related to any known plant pathogen. Adverse effects on plants are not expected to result from the proposed pesticidal applications of Pasteuria nishizawae

– Pn1

.

Additionally, there are no reports of toxicity or pathogenicity of Pasteuria nishizawae to nontarget terrestrial animals or plants. To support its data waiver rationale, the applicantPasteuria nishizawae

– Pn1 Page 15 of 34 Biopesticides Registration Action Document

performed a search within the Agricola and PubMed dat abases, along with several biological journals (i.e., Journal of Bacteriology, Applied and Environmental Microbiology, FEMS Microbiology Ecology, Soil Biology and Bioch emistry) for the period of 1980

– 2010. The search was conducted using “ Pasteuria ” as the search word, and returned no reports of acute toxicity

data on birds , freshwater fish, freshwater invertebrates, nontarget insects, and honey bees . The Biopesticides and Pollution Prevention Division also conducted a search within the

Environmental Informa tion Database that is available to EPA. This database simultaneously searches the Agricola, Biosis Previews, CAB Abstracts, Energy Science and Technology, General Science Abstracts, and the National Technical Information Service literature search

databases . The search was performed with the term “ Pasteuria ” coupled with each of the search

terms of “ bird, ” “mammal,” “invertebrate,” “arthropod,” “insect,” “honey bee,” and “plant” for all of the years available (1926

– present) . The search was also performed wit h the search term

“ Pasteuria nishizawae .” No unanticipated reports of adverse effects of Pasteuria nishizawae or other Pasteuria species in nontarget organisms were found in any of these searches. 2 Any reports of effects to nontarget organisms related to P asteuria nishizawae that are found in the future may trigger the need for testing; however, given the specificity of Pasteuria spp. for their hosts as is currently understood, such reports are not expected.

Based on the information above and available dat a, adverse effects are not expected to occur to terrestrial animals or plants as a result of the proposed pesticidal applications of Pasteuria
nishizawae

– Pn1.

b. Aquatic Animals and Plants

Freshwater Fish ( Harmonized Guideline 885.4200) , Freshwater Invertebrates ( Harmonized Guideline 885.4240)

, Estuarine/Marine Fish and Inverte brates (Harmonized Guideline 885.4280) , and Nontarget Plants (Harmoniz ed Guideline 885.4300) (MRID No

. 481517

- 17 ) : Since the proposed applications are intended to deposit Paste uria nishizawae

– Pn1 in the soil, some runoff of the active ingredient to freshwater and marine/estua rine environments is expected. A pplications of the liquid formulation and seed treatment , however, are intended primarily to put Pasteuria nishizawae

– Pn 1 in the root zone of the treated plants, so these amounts are not likely to be significant. Pasteuria nishizawae and other Pasteuria spp. are naturally found in soil, so some natural presence of Pasteuria nishizawae in surface waters is expected. To date, no information is available on the size of naturally occurring populations of

Pasteuria nishizawae

.

Data waiver rationale was submitted to fulfill data requirements for and support effects

conclusions f or freshwater aquatic organisms . The rationale prov ides sufficient information to conclude that adverse effects are not expected in freshwater fish or freshwater invertebrates as a result of exposure to Pasteuria nishizawae

– Pn1 . The rationale was similar to that presented for terrestrial organisms in tha t Pasteuria spp. are found in soils worldwide

, and Pasteuria
nishizawae is known only to parasitize females of Heterodera glycines

. Pasteuria ramosa is a closely related species that parasitizes Daphnia magna and other Daphnia spp. (Ebert et al

. 1996); how ever, it is widely understood that all other known Pasteuria spp. are obligate parasites

of soil

- dwelling nematodes (Atibalentja et al

. 2004; Chen and Dickson 1998; Sayre et al . 1991).

2 Pasteuria ramosa is already recognized as paras itizing water fleas and is not considered an “unanticipated” report.Pasteuria nishizawae

– Pn1 Page 16 of 34 Biopesticides Registration Action Document

There are no available reports in the literature documenting pathoge nicity or toxicity to freshwater fish or invertebrates as a result of exposure to Pasteuria nishizawae

. A search

, si milar to the one described for terrestrial animals and plants, was performed for aquatic animals and plants. In addition to the s earch terms used for terrestrial animals and plants , the terms “fish,” “aquatic,” “freshwater,” “estuarine,” and “marine” were also searched. No reports of adverse

effects of Pasteuria nishizawae

– Pn1 in aquatic nontarget organisms were found in any of these

searche s. As noted above, reports of adverse effects that may be found in the future may trigger the need for testing; however, none are anticipated due to the specificity of Pasteuria spp. for

their nematode hosts.

B ased on the information submitted, adverse ef fects to freshwater and marine/estuarine fish, invertebrates, and plants are not expected as a result of exposure to proposed pesticidal applications of Pasteuria nishizawae

– Pn1.

2. Environmental Fate Data

As t he data and information provided are s ufficient to fulfill the Tier I nontarget organism data requirements and allow for nontarget organism risk asse ssment for Pasteuria nishizawae

– Pn1

, f urther testing at higher tier levels (i.e., Tiers II, III, and IV) is not required.

3

. Threatened and Endangered Species Assessment

Since EPA has determined that no effects are ant icipated for any non target species exposed to
Pasteuria nishizawae

– Pn1 as a result of the proposed applications, effects to threatened and endangered species and their design ated critical ha bitats are also not expected. Therefore, a “No Effect” determination is made for direct and indirect effects to listed species and their designated critical habitats resulting from the proposed pesticidal uses of Pasteuria nishizawae

– Pn1

, as labeled.

V. 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

— with respect to the development, implementation, and enfo rcement of environmental laws, regulations, and policies. Fair treatment means that no group of people, including racial, ethnic, or socioeconomic groups, should bear a disproportionate share of the negative environmental consequences resulting from indust rial, municipal, and commercial operations or the execution of federal, state, local, and tribal environmental programs and policies. Meaningful involvement means that (1) potentially affected community residents have an appropriate opportunity to particip ate in decisions about a proposed activity that will affect their environment and/or health; (2) the public’s contribution can influence the regulatory agency’s decision; (3) the concerns of all participants involved will

be considered in the decision

- maki ng process; and (4) the decision

- makers seek out and facilitate the involvement of those potentially affected. EPA has this goal for all communities and persons

across the United States.

To help address potential environmental justice issues, during the 30

- day public participation comment period, EPA sought information on any groups or segments of the population who, as aPasteuria nishizawae

– Pn1 Page 17 of 34 Biopesticides Registration Action Document

result their location, cultural practices, or other factors, may have atypical, unusually high
exposure to Pasteuria nishizawae

– Pn1

, compared to the general population. No public comments were received on this particular matter

.

For additional information regarding environmental justice issues, please visit EPA’s web site at http://www.epa.gov/compliance/environmentaljustice/index.html

.

V I

. RISK MANAGEMENT DECISION

Section 3(c)(5) of FIFRA permits for the registration of a pesticide provided that all the following determinations are made:

(1) Its compos ition is such as to warrant the proposed claims for it; (2) Its labeling and other material required to be submitted comply with the requirements of FIFRA; (3) It will perform its intended function without unreasonable adverse effects on

th e environment; AND (4) When used in accordance with widespread and commonly recognized practice, it will not generally cause unreasonable adverse effects on the

environment.

To satisfy criterion 1, the Pasteuria nishizawae

– Pn1 pesticide pr oducts have well

- known properties. EPA has no knowledge that would contradict the claims made on the Soyacyst Tech, Soyacyst Tech+ , and Soyacyst LF label s, and such pesticide products are not expected to cause

unreasonable adverse effects on the environmen t when used according to the ir respective label instructions. Criterion 2 is satisfied by the current product label s, as well as the data and

information presented in this document. It is believed that the Pasteuria nishizawae

– Pn1 pesticide products will not cause any unreasonable adverse effects on the environment, and Soyacyst Tech+ and Soyacyst LF ( end

- use pesticide product s ) , in particular, are likel y to p rovide protection against soybean cyst nematode as claimed, satisfying criterion 3. Criterion 4 i s satisfied in that the Pasteuria nishizawae

– Pn1 pesticide products are not expected to cause unreasonable adverse effects when used according to label instructions. Therefore, Soyacyst

Tech

, Soyacyst Tech+ , an d Soyacyst LF

, containing Pasteuria nishizaw ae

– Pn1 as a new active ingredient, are eligible for registration under FIFRA section 3(c)(5) for the labeled uses.

V I I

. ACTIONS REQUIRED OF THE REGISTRANT

A. Final Printed Labeling

Before releasing pesticide products containing Pasteuria nishizawa e

– Pn1 for shipment, the registrant is required to provide appropriate final printed labeling to EPA

.Pasteuria nishizawae

– Pn1 Page 18 of 34 Biopesticides Registration Action Document

B. Terms of Registration

No additional data are being required as terms of registration.

C. Reporting of Adverse Effects and Hypersensitivity In cidents

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

that certain specific data are required to be reported to EPA as a requirement for maintaining the federal registration for a pesticide product. A bri ef summary of these types of data are described

below.

Reports of all incidents of adverse effects to the environment must be submitted to EPA under the provisions stated in FIFRA section 6(a)(2). Additionally, all incidents of hypersensitivity (including both suspected and confirmed incidents) must be reported to EPA under the provisions of 40 CFR § 158.2140(d).Pasteuria nishizawae

– Pn1 Page 19 of 34 Biopesticides Registration Action Document

VIII. GLOSSARY OF ACRONYMS AND ABBREVIATIONS

BRAD Biopesticides Registration Action Document

CFR Code of Federal Regulations

cSt centist okes

EDSP Endocrine Disruptor Screening Program

EP end

- use pesticide product

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

FFDCA Federal Food, Drug, and Cosmetic Act

FIFRA Federal Insecticide, Fungicide, and Rodenticide Act

FR Federal R egister g/mL grams per milliliter lb/gal pounds per gallon

LD 50 median lethal dose. A statistically derived single dose that can be expected to cause death in 50% of the test animals when administered by the route indicated (oral, dermal, or inhalation ). It is expressed as a weight of substance per unit weight of animal (e.g., mg/kg). mg/kg milligram s per kilogram

MP manufacturing

- use pesticide product

MRID No. Master Record Identification Number

NIOSH National Institute for Occupational Safety and H ealth

OPP Office of Pesticide Programs

PC Code Pesticide Chemical Code

PP Pesticide Petition

PPE personal protective equipment

TGAI technical grade of the active ingredient

U.S. United States

w/w weight to weightPasteuria nishizawae

– Pn1 Page 20 of 34 Biopesticides Registration Action Document

I X. BIBLIOGR APHY

A. Studies Submitted to Support the Pasteuria nishizawae

– Pn1 Pesticide Product Registrations

MRID No. Study Information

48 1517

- 01 MacIntosh S, Smith K . 2010. Product Identity: Pasteuria nishiz awae

– Pn1. Project Number: PBI/2010/001 . Unpublished study prepared by MacIntosh & Associates, Inc. and Pasteuria Bioscience, Inc., 90 pages.

481517

- 02 Smith K. 2010. Manufacturing Process for Pasteuria nishizawae

– Pn1. Project

Number: P BI/2010/002. Unpublished study prepared by Pasteuria Bioscience, Inc., 36 pages.

481517

- 03 MacIntosh S. 2010. Discussion of Formation of Unintentional Ingredients:
Pasteuria nishizawae

– Pn1. Project Number: 14060/10. Unpublished study

pre pared by MacIntosh & Associates, Inc., 29 pages.

481517

- 04 Smith K. 2010. Analysis of Samples/Enforcement Methods for Pasteuria
nishizawae

– Pn1. Unpublished study prepared by Pasteuria Bioscience, Inc., 62

pages.

481517

- 05 Smith K. 2010. Cer tification of Limits for Pasteuria nishizawae

– Pn1. Project Number: PBI/2010/004. Unpublished study prepared by Pasteuria Bioscience, Inc., 8 pages.

481517

- 06 Kaminsky M. 2010. Pasteuria nishizawae

– Pn1 Product Chemistry: Final Report.

P roject Number: 13885/10. Unpublished study prepared by Stillmeadow, Inc., 16

pages.

481517

- 07 Smith K. 2010. Product Stability to Elevated Temperatures: Pasteuria nishizawae

– Pn1. Project Number: 115/54. PBI/2010/005. Unpublished study prepare d by Pasteuria Bioscience, Inc., 8 pages.

481517

- 08 Smith K. 2010. Storage Stability & Corrosion Characteristics for Pasteuria
nishizawae . Project Number: 13886/10, PBI/2010/002. Unpublished study prepared by Pasteuria Bioscience, Inc., 16 pages.

481517

- 09 Gingras B. 2010. Toxicity/Pathogenicity Testing of Pasteuria nishizawae

– Pn1 Following Acute Oral Challenge in Rats. Project Number: 2262/SN4. Unpublished study prepared by IIT Research Institute, 58 pages.

481517

- 10 Gingra s B. 2010. Toxicity/Pathogenicity Testing of Pasteuria nishizawae

– Pn1 Following Acute Intratracheal Challenge in Rats

– Final Report. Project Number: 2262/SN5. Unpublished study prepared by IIT Research Institute, 53 pages.Pasteuria nishizawae

– Pn1 Page 21 of 34 Biopesticides Registration Action Document

MRID No. Study Information

481517

- 11 Gingras B. 2010. Toxicity/Pathogenicity Testing of Pasteuria nishizawae

– Pn1 Following Acute Injection Challenge in Rats. Project Number: 2262/SN6. Unpublished study prepared by I IT Research Institute, 56 pages.

481517

- 12 MacIntosh S. 2010. Hypersensitivity Incidents: Pasteuria nishizawae

– Pn1. Project Number: PBI/2010/007. Unpublished study prepared by MacIntosh & Associates, Inc., 7 pages.

481517

- 13 Sullivan D. 201 0. Acute Oral Toxicity Study of Pasteuria nishizawae

– Pn1 in Rats. Project Number: 2262/SN11. Unpublished study prepared by IIT Research Institute, 28 pages.

481517

- 14 Sullivan D. 2010. Acute Dermal Toxicity Study of Pasteuria nishizawae

– Pn1 in Rabbits. Project Number: 2262/SN12. Unpublished study prepared by IIT Research Institute, 30 pages.

481517

- 15 Sullivan D. 2010. Acute Eye Irritation Study of Pasteuria nishizawae

– Pn1 in

Rabbits. P roject Number: 2262/SN13. Unpublished study prepared by IIT Research Institute, 25 pages.

481517

- 16 Sullivan D. 2010. Acute Dermal Irritation Study of Pasteuria nishizawae

– Pn1 in Rabbits. Project Number: 2262/SN14. Unpublished study prepared by IIT Research Institute, 24 pa ges.

481517

- 17 MacIntosh S, Smith K, Hewlett T. 2010. Request for Waiver from the Requirement to Conduct Guideline Studies (Avian Oral Toxicity, Freshwater Fish Toxicity/Pathogenicity, Freshwater Invertebrate Toxicity/Pathogenicity, Non

Ta rget Insect Testing, and Honey Bee Testing) for Pasteuria nishizawae

– Pn1. Project Number: PBI/2010/014. Unpublished study prepared by MacIntosh & Associates, Inc., 29 pages.

482103

- 01 MacIntosh S, Smith K. 2010. Product Identity and Manufact uring Process:
Pasteuria nishizawae

– Pn1 (Liquid Formulation; End Product). Project Number: PBI/2010/020. Unpublished study prepared by Pasteuria Bioscience, Inc. and MacIntosh & Associates, Inc., 17 pages.

482103

- 02 MacIntosh S. 2010. Di scussion of Formation of Unintentional Ingredients:
Pasteuria nishizawae

– Pn1 (Liquid Formulation; End Product). Unpublished study prepared by MacIntosh & Associates, Inc., 6 pages.

482103

- 03 Smith K. 2010. Analysis of Samples/Enforcement Meth ods for Pasteuria
nishizawae

– Pn1 (Liquid Formulation; End Product). Unpublished study prepared by Pasteuria Bioscience, Inc., 20 pages.Pasteuria nishizawae

– Pn1 Page 22 of 34 Biopesticides Registration Action Document

MRID No. Study Information

482103

- 04 Smith K. 2010. Certif ication of Limits for Pasteuria nishizawae

– Pn1 (Liquid Formulation; End Product). Project Number: PBI/2010/022. Unpublished study prepared by Pasteuria Bioscience, Inc., 8 pages.

482103

- 05 Kaminsky M. 2010. Pasteuria nishizawae

– Liquid Formu lation: Product Chemistry

– Final Report. Project Number: 14070/10. Unpublished study prepared by Stillmeadow, Inc., 8 pages.

482103

- 06 Smith K. 2010. Storage Stability: Interim Report Pasteuria nishizawae

– Pn1 (Liquid Formulation; End Pr oduct). Project Number: PBI/2010/023. Unpublished study prepared by Pasteuria Bioscience, Inc., 8 pages.

482103

- 07 Smith K. 2010. Corrosion Characteristics for Pasteuria nishizawae

– Pn1 (Liquid Formulation; End Product). Project Number: PBI/20 10/024. Unpublished study prepared by Pasteuria Bioscience, Inc., 11 pages.

482103

- 08 MacIntosh S. 2010. Hypersensitivity Incidents: Pasteuria nishizawae

– Pn1 (Liquid Formulation; End Product). Project Number: PBI/2010/024. Unpublished

st udy prepared by MacIntosh & Associates, Inc., 11 pages.

482103

- 09 Sullivan D. 2010. Acute Oral Toxicity Study of Pasteuria nishizawae

– Pn1 (End Product) in Rats. Project Number: 2262/SN15. Unpublished study prepared by IIT Research Institute, 28 pages.

482103

- 10 Sullivan D. 2010. Acute Dermal Toxicity Study of Pasteuria nishizawae

– Pn1 (End Product) in Rabbits. Project Number: 2262/SN20. Unpublished study prepared by IIT Research Institute, 28 pages.

482103

- 11 Sullivan D. 2010. A cute Eye Irritation Study of Pasteuria nishizawae

– Pn1 (End Product) in Rabbits. Project Number: 2262/SN21. Unpublished study prepared by IIT Research Institute, 22 pages.

482103

- 12 Sullivan D. 2010. Acute Dermal Irritation Study of Pasteuria nishizawae

– Pn1 (End Product) in Rabbits. Project Number: 2262/SN22. Unpublished study prepared by IIT Research Institute, 22 pages.

482744

- 02 Kaminsky M. 2010. Pasteuria nishizawae

– Pn1 and Pasteuria nishizawae

– Seed

Treatment: Product Chemistry

– Final Report. Project Number: 14451/10. Unpublished study prepared by Stillmeadow, Inc., 10 pages.

482744

- 05 MacIntosh S. 2010. Request for Waivers from the Requirement to Conduct Toxicology Guideline Study: Acute Inhalation Toxici ty for Pasteuria nishizawae

– Pn1 & Pasteuria nishizawae

– Seed Treatment. Project Number: PBI/2010/027. Unpublished study prepared by MacIntosh & Associates, Inc., 7 pages.Pasteuria nishizawae

– Pn1 Page 23 of 34 Biopesticides Registration Action Document

MRID No. Study Informatio n

482744

- 06 MacIntosh S. 2010. Request for Waivers from the Requirement to Conduct Toxicology Guideline Study: Acute Inhalation Toxicity for Pasteuria nishizawae

– Liquid Formulation. Project Number: PBI/2010/028. Unpublished study

prepare d by MacIntosh & Associates, Inc., 7 pages.

485915

- 01 MacIntosh S, Smith K. 2011. Product Identity: Pasteuria nishizawae

– Pn1. Unpublished study prepared by Pasteuria Bioscience, Inc., 6 pages.

485915

- 02 Smith K. 2011. Manufacturing Process fo r Pasteuria nishizawae

– Pn1: Supplement to MRID No. 481517

- 02. Unpublished study prepared by Pasteuria Bioscience, Inc., 7 pages.

485915

- 03 MacIntosh S. 2011. Discussion of Formation of Unintentional Ingredients

– Pasteuria nishizawae

– P n1: Supplement to MRID No. 481517

- 03. Unpublished study prepared by MacIntosh & Associates, Inc., 10 pages.

485915

- 04 Smith K. 2011. Analysis of Samples/Enforcement Methods for Pasteuria
nishizawae

– Pn1: Supplement to MRID No. 481517

- 04. Unpub lished study prepared by Pasteuria Bioscience, Inc., 14 pages.

485915

- 05 Smith K. 2011. Storage Stability: Pasteuria nishizawae

– Pn1

– Final Report. Project Number: PBI/2010/002. Unpublished study prepared by Pasteuria Bioscience, Inc., 1 3 pages.

485915

- 06 Kaminsky M. 2011. Pasteuria nishizawae

– Pn1: Corrosion Characteristics

– Final Report. Project Number: 13886/10. Unpublished study prepared by Stillmeadow, Inc., 10 pages.

486052

- 01 Smith K. 2011. Storage Stability Pasteur ia nishizawae

– Liquid Formulation: Supplement to MRID No. 482103

- 06

– Final Report. Project Number: PBI/2010/023. Unpublished study prepared by Pasteuria Bioscience, Inc., 8 pages.

486052

- 02 Kaminsky M . 2011. Corrosion Characteristics Pasteuri a nishizawae

– Liquid

Formulation: Final Report. Project Number: 13887/10. Unpublished study prepared by Stillmeadow

, Inc., 11 pages.

486461

- 01 Smith K. 2011. New Sources of Inert Ingredients Pasteuria nishizawae

– Pn1 (TGAI/MP) and Pasteu ria nishizawae

– Seed Treatment (EP). Unpublished study prepared by Pasteuria Bioscience, Inc., 51 pages.

486461

- 02 Smith K. 2011. New Sources of Inert Ingredients Pasteuria nishizawae

– Pn1 (Liquid Formulations; EP) and Pasteuria nishizawae

– Stabilized Seed Treatment (EP). Unpublished study prepared by Pasteuria Bioscience, Inc., 101 pages.Pasteuria nishizawae

– Pn1 Page 24 of 34 Biopesticides Registration Action Document

B. Environmental Protection Agency Risk Assessment Memoranda

U.S. EPA. 2011a. Pasteuria nishizawae strain Pn1 . Memorandum from J.V. Gagliardi , Ph .D. and J.L. Kough, Ph.D. to J. Kausch dated August 16 , 2011.

U.S. EPA. 2011b. Environmental Risk Assessment of Pasteuria nishizawae Pn1 (PC 016455) for a Section 3 Registration of the Technical Product (EPA File Symbol 85004

- A) and Two End Use Products ( EPA File Symbols 85004

- T and 85004

- O) for Control of the Soybean Cyst Nematode in Soybeans. Memorandum from S. Borges and G. Tomimatsu, Ph.D. to J. Kausch dated September 15, 2011.

U.S. EPA. 2011c. Pasteuria nishizawae strain Pn1 from Pasteuria Bioscience , Inc . Memorandum from J.V. Gagliardi, Ph.D. and J.L. Kough, Ph.D. to J. Kausch dated December 20, 2011.

C. Other References

Aislabie J, Smith JJ, Fraser R, McLeod M. 2001. Leaching of bacterial indicators of faecal contamination through four New Zea land soils. Australian Journal of Soil Research

39:1397

– 1406.

Atibalentja N

, Jakstys BP, Noel GR. 2004. Life cycle, ultrastructure, and host specificity of the

North American isolate of Pasteuria that parasitizes the s oybean cyst nematode,

Heterodera glycines

. Journal of Nematology 36: 171

– 180.

Centers for Disease Control and Prevention. 2009. Drinking Water

- Water Treatment. Available

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

.

Cetintas R, Dickson DW. 2004. Persistence and suppressiveness of Pasteuria penetrans to Meloidogyne arenaria Race 1. Journal of Nematology 36:540

– 549.

Chen ZX, Dickson DW . 1998. Review of Pasteuria penetrans : biology, ecology, an d biological control potential. Journal of Nematology 30: 313

– 340.

Ciancio A, Bonsignore R, Vov las N, Lamberti F . 1994. Host records and spore morphometrics of Pasteuria penetrans group parasites of nematodes. Journal of Invertebrate Pathology

63: 260

– 267.

DeFelice K, Wollenhaupt N, Buchholz D. 1993. Aquifers and Soil Filter Effect. Available from http://extension.missouri.edu/p/WQ24

.

Ebert D, Rainey P, Embley TM, Scholz D . 1996. Development, life cycle, ultrastructure, and phylogenetic position of Pasteuria ramosa Metchnikoff 1888: rediscovery of an obligate endoparasite of Daphnia magna Straus. Philosophical Transactions of the Royal Society of London series B . 351:1689

– 1701.

Noel GR . 2008. IPM of soybean cyst nematode in the USA. In : Integrated Management and Biocontrol of Vegetable and Grain Crops Nematodes. Eds. A. Cianio and K.G. Mukerji. Springer. Pages 119

– 126.Pasteuria nishizawae

– Pn1 Page 25 of 34 Biopesticides Registration Action Document

Noel GR, Atibalentja N, Domier LL . 2005. Emended des cription of Pasteuria nishizawae

. International Journal of Systematic and Evolutionary Microbiology 55:1681

– 1685.

Pang L, M cLeod M, Aislabie J, Šimůnek J, Close M, Hector R. 2008. Modeling transport of

microbes in ten undisturbed soils under effluent irr igation. Vadose Zone Journal 7:97

– 111.

Sayre RM, Wergin WP, Schmidt JM, Starr MP . 1991. Pasteuria nishizawae sp. nov., a mycelial and endospore

- forming bacterium parasitic on cyst nematodes of genera Heterodera and

Globodera

. Research in Microbiology 14 : 551

– 564.

Sayre RM, Starr MP . 1985. Pasteuria penetrans (ex Thorne, 1940) nom. rev., comb. n., sp. n ., a mycelial and endospore

- forming bacterium parasitic in plant

- parasitic nematodes. Proceedings of the Helminthological Society of Washington 52: 149

– 1 65.

Sturhan D. 1988. New host and geographical records of nematode

- parasitic bacteria of the Pasteuria penetrans group. Nematologica 34: 350

– 356.

Tain B, Yang J, Zhang K

- Q. 2007. Bacteria used in the biological control of plant

- parasitic nematodes: popu lations, mechanisms of action, and future prospects. FEMS Microbiology Ecology 61:197

– 213.

U.S. EPA. 2004. Primer for Municipal Wastewater Treatment Systems. EPA 832

- R

- 04

- 001.

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

.Pasteuria nishizawae

– Pn1 Page 26 of 34 Biopesticides Registration Action Document

A PPENDIX A. MICROBIAL PESTICIDES DATA REQUIREMENTS (40 CFR PART 158

– SUBPART V )

TABLE 1. Product Analysis Data Requirements for the Technical Grade of the Active Ingredient (TGAI)
, Pasteur ia nishizawae

– Pn1 , and the Manufacturing

- Use Pesticide Product ( MP )
, Soyacyst Tech (40 CFR § 158.2 120 )

Harmonized

G uideline

N umber Data Requirement Results MRID No. Pasteuria nishizawae

– Pn1 Soyacyst Tech

885.1100 Product Identity N/A Submitt ed data fulfill the requirement for product identity. Soyacyst Tech

contains 99.88 % by weight Pasteuria
nishizawae

– Pn1 (contains at least 1 x

10 8 spores per gram). 481517

- 01

485915

- 01

486461

- 01

885.1200 Manufacturing

Process Submitted data fulfill th e requiremen t for manufacturing process

. 481517

- 02

485915

- 02

N/A Deposition of a Sample in a Nationally Recognized Culture

Collection Pasteuria nishizawae

– Pn1 is on deposit with the American Type Culture Collection in Manassas, Virginia under Accession Number

SD

- 5833

. N/A 481517

- 01

485915

- 01

885.1300 Discussion of

Formation of

Unintentional

Ingredients Submitted data fulfill the requiremen t for discussion of formation of unintentional ingredients

. 481517

- 03

485 915

- 03

885.1400 Analysis of S amples Submit ted data fulfill the requirement for analysis of samples. 481517

- 04

485915

- 04

8 85 .1 50 0 Certification of

L imits N/A Limits listed o n the confidential

statement of formula are adequate/a cceptable. 481517

- 05Pasteuria nishizawae

– Pn1 Page 27 of 34 Biopesticides Registration Action Document

TABLE 2

. Product Analysis Data Requirements for the End

- Use Pesticide Product ( E P s ), Soyacyst Tech+ and Soyacyst LF (40 CFR § 158.2120)

Harmonized

Guideline

Number Data Requirement Results

MRID No. Soyacyst Tech+ * Soyacyst LF **

885.1100 Product Identity Submitted data fulfil l the requirement for product identity. Soyacyst Tech+

contains 99.88 % by weight
Pasteuria nishizawae

– Pn1 (contains at least 1 x 10 8
spores per gram). Submitted data fulfill the requirement for product identity. Soyacyst LF contains

33.29 % by weight Pa steuria
nishizawae

– Pn1 ( contains at

least 1.3 x 10 7 spores per gram)

. 481517

- 01*

485915

- 01*

486461

- 01*

482103

- 01**

486461

- 02**

885.1200 Manufacturing Process Submitted data fulfill the requirement for manufacturing

process. 481517

- 02*

485915

- 02*

482103

- 01**

N/A Deposition of a Sample in a Nationally Recognized

Culture Collection N/A N/A

885.1300 Discussion of Formation of Unintentional Ingredients Submitted data fulfill the requirement for discussion of formation of unintentional ingredients. 481517

- 0 3*

485915

- 03*

482103

- 02**

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

. 481517

- 04*

485915

- 04*

482103

- 03**

885.1500 Certification of Limits Limits listed on the confidential statement of formula are adequate/ acceptable. 481517

- 05*

482103

- 04**Pasteuria nishizawae

– Pn1 Page 28 of 34 Biopesticides Registration Action Document

TABLE 3

. Physical and Chemical Characteristics for the Technical Grade of the Active Ingredient (TGAI), Pasteuria nishizawae

– Pn1 , and the Manufacturing

- Use Pesticide Product (MP), Soyacyst Tech ( 40 CFR § 158.2120)

H armonized

G uideline N umber Data Requirement Results MRID No. Pasteuria nishizawae

– Pn1 Soyacyst Tech

830.6302 1 Color Light brown/caramel brown

481517

- 06 830.6303 1 Physical State Liquid

830.6304 1 Odor Faint odor/similar to dog food

830.6313 1 Sta bility to Normal and

E levated Temperatures, Metals,

and Metal I ons Stable up to 14 days at 54°C without loss of viability 481517

- 07

830.6317 Storage Stability St able for one year when stored at 4°C 481517

- 08

485915

- 05

830.6319 Miscibility N/A Not require d because Soyacyst Tech is not an

e mulsifiable liquid form of a microbial pesticide (refer to test note #2 of 40 CFR § 158.2120(d)). N/A

830.6320 Corrosion

Characteristics N/A Not corrosive to packaging

materials in a one

- year study. 481517

- 08

485915

- 06

830.7000 1 pH 4.69

– 5.21 (1% w/w) 481517

- 06

830.7100 Viscosity N/A 1.570 cSt (20°C) 1.046 cSt (40°C) 482744

- 02

830.7300 1 Density/Relative Density/Bulk Density

(Specific Gravity) 1.04 g/ mL (or 1.04 g/cm 3 ) 481517

- 06

1 According to 40 CFR § 158.2120, these data are only required for the technical grade of the active ingredient. Since Pasteuria

Bioscience

, Inc. included this informati on with its application for Soyacyst Tech , it is summarized appropriately in this table.Pasteuria nishizawae

– Pn1 Page 29 of 34 Biopesticides Registration Action Document

TABLE 4. Physical and Chemical Characteristics for the End

- Use Pesticide Product s (EP s ), Soyacyst Tech+ and Soyacyst LF (40 CFR § 158.2120)

Harmonized

Guideline Number Data Requirement Results MRID No. Soyacyst Tech+ * Soyacyst LF **

830.6302 1 Color Light bro wn/caramel

brown N/A 481517

- 06

830.6303 1 Physical State Liquid N/A 481517

- 06

830.6304 1 Odor Faint odor/similar to dog

food N/A 481517

- 06

830.6313 1 Stability to Normal and Elevated Temperatures, Metals, and Metal Ions Stable up to 14 days at

54°C without loss of viability N/A 481517

- 07

830.6317 Storage Stability St able for one year when stored at 4°C 481517

- 08*

485915

- 05*

482103

- 06**

486052

- 01**

830.6319 Miscibility Not required because the end

- use pesticide products, Soyacyst Tech+ and Soyacyst LF , are not emulsifiable liquid forms of microbial pesticides (refer to test note #2 of 40 CFR § 158.2120(d)). N/A

830.6320 Corrosion Characteristics Not corrosive to packaging materials in a one

- year study. 481517

- 08*

485915

- 06*

482103

- 07**

486052

- 02**

830.700 0 pH 4.69

– 5.21 (1% w/w) 1 3.5

– 4.5 2 481517

- 06 *

830.7100 Viscosity 1.570 cSt (20°C) 1.046 cSt (40°C) 318.2 cSt (20°C) 255.15 cSt (40°C) 482744

- 02*

482103

- 05**

830.7300 Density/Relative Density/Bulk Density

(Specific Gravity) 1.04 g/mL (or 1.04 g/ cm 3 ) 1 8.5 l b/gal 2 481517

- 06 *

1 According to 40 CFR § 158.2120, these data are only required for the technical grade of the active ingredient. Since Pasteuri a

Bioscience, Inc. included this information with its applications for Soyacyst Tech+, it is summarized app ropriately in this table.

2 Obtained from the confidential statement of formula for the end

- use pesticide product.Pasteuria nishizawae

– Pn1 Page 30 of 34 Biopesticides Registration Action Document

T ABLE 5

. Toxicology Data Requirements for the Technical Grade of the Active Ingredient (TGAI)
, Pasteuria nishizawae

– Pn1 , and

the Manufactu ring

- Use Pesticide Product (MP)
, Soyacyst Tech (40 CFR § 158.2140)

Harmonized

Guideline

N umber Data Requirement Results MRID N o. Pasteuria nishizawae

– Pn1 Soyacyst Tech

Tier I

885.3050 Acute Oral Toxicity/Pathogenicity Not toxic and/or pathogenic t o rats
when administered by oral gavage in a single dose of 1.6 x 10 9 spores per animal. Although clearance and infectivity were not measured, EPA believes these endpoints are not a concern given Pasteuria nishizawae

– Pn1’s well

- established host specifici ty for the soybean cyst nematode

. Classification: Supplemental N/A 481517

- 09

885.3150 Acute Pulmonary Toxicity/Pathogenicity Not toxic and/or pathogenic to rats when administered by intratracheal instillation in a single dose of 1.6 x 10 8
spores per anim al. Although clearance and infectivity were not measured, EPA believes these endpoints are not a concern given Pasteuria nishizawae

– Pn1’s well

- established host specificity for the soybean cyst nematode.

Classification: Supplemental N/A 481517

- 10

885.320 0 Acute Injection Toxicity/Pathogenicity (Intra peritoneal ) N ot toxic and/or pathogenic to rats
when administered intravenously in a single dose of 1.0 x 10 9 spores per animal

. Although clearance and infectivity were not measured, EPA believes these endpoin ts are not a concern given Pasteuria nishizawae

– Pn1’s well

- established host specificity for the soybean cyst nematode.

Classification: Supplemental N/A 481517

- 11

885.3400 Hypersensitivity

Incidents The applicant reported that n o hypersensitivity inciden ts, including

immediate

- type or delayed

- type reactions of humans and domestic animals

, occurred durin g research, development, or testing of the TGAI or MP

. Any future hypersensitivity incidents must be reported to EPA (refer to test note #3 of 40 CFR § 158 .2140(d))

. 481517

- 12

885.3500 Cell Culture Not required because Pasteuria
nishizawae

– Pn1 is not a virus (refer to test note #4 of 40 CFR § 158.2140(d)). N/A N/A

870.1100 Acute Oral Toxicity N/A Oral LD 50 combined (male and female rats) > 5,000 mg/kg

C lassification: Acceptable

TOXICITY CATEGORY I V 481517

- 13

870.1200 Acute Dermal Toxicity N/A Dermal LD 50 combined (male and female rabbits) > 2,000 mg/kg

Classification: Acceptable

TOXICITY CATEGORY IV 481517

- 14Pasteuria nishizawae

– Pn1 Page 31 of 34 Biopesticides Registration Action Document

T ABLE 5

. Toxicology Data Requirements for the Technical Grade of the Active Ingredient (TGAI)
, Pasteuria nishizawae

– Pn1 , and

the Manufactu ring

- Use Pesticide Product (MP)
, Soyacyst Tech (40 CFR § 158.2140)

Harmonized

Guideline

N umber Data Requirement Results MRID N o. Pasteuria nishizawae

– Pn1 Soyacyst Tech

870.1300 Acute Inhalation Toxicity N/A Waiv ed based on the results of

MRID No

. 481517

- 10 and

because this formulation contains inert ingredients that are not expected to be of toxicological

conce rn (refer to test note #5 of 40 CFR § 158.2140(d))

. Classification: Acceptable

TOXICITY CATEGORY III 482 744

- 05

870.2400 Acute Eye Irritation N/A Soyacyst Tech was essentially

non

- irritating to the eyes of

rabbits. Classification: Acceptable

TOXICITY CATEGORY I V 481517

- 15

870.2500 Primary Dermal

Irritation N/A Soyacyst Tech was essentially

non

- irritating to the skin of

rabbits. Classification: Acceptable

TOXICITY CATEGORY I V 481517

- 16

Tiers II and III Not required for Pasteuria nishizawae

– Pn1 based on the lack of acute toxicity/pathogenicity in the Tier I studies.

TABLE 6

. Toxicology Data Requirement s for the End

- Use Pesticide Product s (EPs)
, Soyacyst Tech+ and Soyacyst LF (40 CFR § 158.2140)

Harmonized

Guideline

Number Data Requirement Results

MRID No. Soyacyst Tech+ * Soyacyst LF **

885.3050 Acute Oral Toxicity/Pathogenicity N/A N/A

885.3150 A cute Pulmonary Toxicity/Pathogenicity N/A N/A

885.3200 Acute Injection Toxicity/Pathogenicity N/A N/A

885.3400 Hypersensitivity

Incidents The applicant reported than n o hypersensitivity incidents, including

immediate

- type or delayed

- type reactions of hum ans and domestic animals

, occurred during research, development, or testing of the EP s

. Any future hypersensitivity incidents must be reported to EPA (refer to test note #3 of 40 CFR § 158.2140(d)). 481517

- 12*

482103

- 08**

885.3500 Cell Culture N/A N/A

87 0.1100 Acute Oral Toxicity Oral LD 50 combined (male and female rats) > 5,000 mg/kg Classification: Acceptable

TOXICITY CATEGORY I V 481517

- 13*

482103

- 09**

870.1200 Acute Dermal Toxicity Dermal LD 50 combined (male and female rabbits) > 2,000 mg/kg

Classific ation: Acceptable

TOXICITY CATEGORY IV 481517

- 14*

482103

- 10**Pasteuria nishizawae

– Pn1 Page 32 of 34 Biopesticides Registration Action Document

TABLE 6

. Toxicology Data Requirement s for the End

- Use Pesticide Product s (EPs)
, Soyacyst Tech+ and Soyacyst LF (40 CFR § 158.2140)

Harmonized

Guideline

Number Data Requirement Results

MRID No. Soyacyst Tech+ * Soyacyst LF **

870.1300 Acute Inhalation Toxicity Waived based on the results of MRID No

. 481517

- 10 and because th ese formulation s contain inert ingredients that are not expected to be of toxicological conce r n (refer to test note #5 of 40 CFR § 158.2140(d)). Classification: Acceptable

TOXICITY CATEGORY III 482744

- 05*

482744

- 06**

870.2400 Acute Eye Irritation The EP test substances were essentially non

- irritating to the eyes of rabbits. Classification: Accepta ble

TOXICITY CATEGORY I V 481517

- 15*

482103

- 11**

870.2500 Primary Dermal

Irritation The EP test substances were essentially non

- irritating to the skin of rabbits. Classification: Acceptable

TOXICITY CATEGORY I V 481517

- 16*

482103

- 12**

TABLE 7

. Nontarget Organism Toxicity and Environmental Fate Data Requirements for the Technical Grade of the Active Ingredient (TGAI), Pasteuria ni s hizawae

– Pn1 (40 CFR § 158.2150)

Harmonized Guideline

Number Data Requirement Results MRID No.

Tier I

885.4050 Avian Oral T oxicity Data and other information provide sufficient

information to determine that toxicity/pathogenicity to avian wildlife is not expected as a result of the proposed pesticidal

uses. Classification: Acceptable 481517

- 17

885.4100 Avian Inhalation

Toxici ty/Pathogenicity Not required as the nature of the microbial pesticide does not indicate potential pathogenicity to birds or relatedness to any known bird pathogens (refer to test note #3 of 40 CFR § 158.2150(e)). N/A

885.4150 Wild Mammal Toxicity/Pathoge nicity Tests required by 40 CFR § 158.2140 are adequate and appropriate for assessment of

hazards to wild mammals. Studies submitted with laboratory rats indicate no adverse effects due to oral exposure. Classification: Acceptable for wild mammal

risk asse ssment N/A

885.4200 Freshwater Fish Toxicity/Pathogenicity Data and other information provide sufficient

information to determine that toxicity/pathogenicity to freshwater fish and invertebrates is not expected as a result of the proposed pesticidal uses. Classification: Acceptable 481517

- 17

885.4240 Freshwater Invertebrate Toxicity/Pathogenicity

885.4280 Estuarine/Marine Fish and Invertebrate Testing Not required as the microbial pesticide will not be applied directly to water and is not expected to

e nter marine/estuarine environments in amounts that would result in significant concentrations (refer to test note #6 of 40 CFR § 158.2150(e)). N/A

885.4300 Nontarget Plant Testing Not required as the microbial pesticide is not taxonomically related to any known plant pathogens (refer to test note #7 of 40 CFR § 158.2150(e)). N/APasteuria nishizawae

– Pn1 Page 33 of 34 Biopesticides Registration Action Document

TABLE 7

. Nontarget Organism Toxicity and Environmental Fate Data Requirements for the Technical Grade of the Active Ingredient (TGAI), Pasteuria ni s hizawae

– Pn1 (40 CFR § 158.2150)

Harmonized Guideline

Number Data Requirement Results MRID No.

885.4340 Nontarget Insect Testing Data and other information provide sufficient

information to determine that toxicity/pathogenicity to honey bees and nontarget insects is not exp ected as a result of the proposed pesticidal uses. Classification: Acceptable 481517

- 17

885.4380 Honey Bee Testing

Tiers II, III, and IV Not required for Pasteuria nishizawae

– Pn1 based on the acceptability of the data and other information provided for Tier I

.Pasteuria nishizawae

– Pn1 Page 34 of 34 Biopesticides Registration Action Document

APPENDIX B. PESTICIDE PRODUCTS

EPA Registration

Number Registration

Name Percentage

Active Ingredient Formulation Type Use Site(s) Method(s) of Application Application Rate (s) Target Pest

85004

- 6 Soyacys t
Tech 99.88% Technical N/A N/A N/A N/A

85004

- 7 Soyacyst

Tech+ 99.88% End Use

– Liquid Soybean Seed Treatment (Preplant or Commercial) 2

– 5 fluid ounces of Soyacyst Tech+ per 100 pounds of soybean seed Soybean cyst

nematode ( Heterodera glycines )

85004

- 9 Soyacyst LF 33.29% End Use

– Liquid Soybean Seed Treatment (Preplant or Commercial) 2

– 5 fluid ounces of Soyacyst LF per 100 pounds of soybean seed Soybean cyst

nematode ( Heterodera glycines ) Ground application equipment or chemigation 1.3

– 64 fluid o unces of Soyacyst LF per 1,000 square feet of soybean