BIOPESTICIDES REGISTRATION ACTION DOCUMENT



## Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph 3

Pesticide Chemical (PC) Code: 016 457

U.S. Environmental Protection Agency Office of Pesticide Program s Biopesticides and Pollution Prevention Division

May 23, 2013Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 Page 2 of 36

Bio pesticides Registration Action Document

TABLE OF CONTENTS

I. EXECUTIVE SUMMARY

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

II. ACTIVE INGREDIENT OV ERVIEW

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

III. REGULATORY BACKGROUN D

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

IV. RISK ASSESSMENT

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

A. Product Analysis Assessment (40 C.F.R. § 158.2120)

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

B. Human Health Assessment (40 C.F.R. § 158.2140)

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

C. Environmental Assessment (40 C.F.R. § 158.2150)

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

V

. ENVIRONMENTAL JUSTIC E

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

VI. RISK MANAGEMENT DECI SION

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

VII. ACTIONS REQUIRED OF THE REGISTRANT

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

A. Final Printed Labeling

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

B. Terms of Registration

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

C. Reporting of Adverse Effects and Hypersensitivity Incidents

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

VIII. GLOSSARY OF ACRONYMS AND ABBREVIATIONS

................................ ................................ .................... 1 7

IX. BI BLIOGRAPHY

................................ ................................ ................................ ................................ ...................... 18

A. S tudies Submitted or Cited to Support the Paste uria spp. ( Hoplolaimus galeatus nematode)

– Ph3

Pesticide P roduct Registrations

................................ ................................ ................................ ......................... 18

B. Environmental Protection Agency Risk Assessment Memoranda

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

C. Other References

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

APPENDIX A. MICROBI AL PESTICIDES DATA R EQUIREMENTS

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

APPENDIX B. PESTICI DE PRODUCTS

................................ ................................ ................................ ............................ 36Pasteuria spp. ( Hoplolaimus galeatus nematode)

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Bio pesticides Registration Action Document

BIOPESTICIDES REGISTRATIO N ACTION DOCUMENT TEAM

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

Microbial Pesticides Branch

Science Review

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

John L. Kough, Ph.D. Pr oduct Analysis, Human Health Shannon Borges Environmental Effects

Regulat ory Review

Kimberly Nesci Chief, Microbial Pesticides Branch

Jeannine Kausch Regulatory Action LeaderPasteuria spp. ( Hoplolaimus galeatus nematode)

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Bio pesticides Registration Action Document

I. EXECUTIVE SUMMARY

Background

In March 201 2

, Pasteuria B ioscience, Inc. submitted application s for a manufacturing

- use p esticide product, Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 ( EPA File Symbol : 85004

- RN )

, and an end

- use pesticide product

, NewPro ( EPA File Symbol : 85004

- RR )

, to the U .S. Environmen tal Protection Agen cy (EPA) under section 3 of the Federal Insecticide, Fungicide, and Rodenticide Act (FIFRA)

. These pesticide products contain a new active ingredient, Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3.

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

parasitic nematodes that attack and cause significant damage to many valuable agricultural crops . (See the Federal Register (FR) publications of December 28, 1994 ( 59 FR 66740 ), June 30, 2010 ( 75 FR

37734 ), February 15, 2012 ( 77 FR 8736 ) , and July 9, 2012 ( 77 FR 40271 ) for Final Rules that established tolerance exemptions for residues of the nematicides, Pasteuria penetrans (40 Code of Federal Regulations (C

. F

. R

. ) § 180.1135), Pasteuria usgae (40 C

. F

. R

. § 180.1290), Pasteuria
nishizawae

– Pn1 (40 C

. F

. R

. § 180.1311), and Pasteuria spp. ( Rotylenchulus reniformis nematode)

– Pr3 (40 C.F.R. § 180.1316 )

, respectively .) These gram

- pos itive, 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

nema todes in at least 80 countries on 5 continents, as well as on islands in the Atlantic, Pacific, and

Indian Oceans (Chen and Dickson 1998; Ciancio et al . 1994; Sturhan 1988)

. H igher population densities

often occur in areas where there is an ample supply of nematode hosts (e.g., where crops susceptible to nematodes are cultivated) ( Centitas and Dickson 2004; Noel 2008; Tain et al . 2007 ). Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph 3 was specifically isolated from turfgrass soil samples collected

at th e DeBary Golf Course in central Florida

.

Endospores of Pasteuria spp. ( Hoplolaimus galeatus nematode)

– P h 3 attach to the lance nematode ( Hoplolaimus galeatus ) at all life stages, except the egg stage . After an endospore attaches to the cuticle

of a nema tode host , a germ tube penetrates the cuticle , and growth and sporogenesis begin in the pseudocoelom of the nematode. The nematode is eventually filled with cells, mycelial hyphae, and sporangia, which lead s to its death. In light of the demonstrated nemat icidal capabilities and host specificity of Pasteuria spp. ( Hoplolaimus galeatus nematode)

– P h 3 , Pasteuria Bioscience , Inc. proposed to register a manu facturing

- use pesticide product and a n end

- use pesticide product that is

intended for use as a soil or f oliar spray on t urfgrass to control the lance nematode

. The end

- use pesticide product also contains Pasteuria usgae

– B l 1, another active ingredient that is present in three currently registered pesticide products 2 and is intended to control the sting nema tode ( Belonolaimus longicaudatus ).

The EPA ’s s cientists reviewed product analysis, toxicology, and nontarget organism data and information ( 40 C

. F

. R

. §§ 158.2120, 158.2140, and 158.2150, respectively) submitted to support the

1 Pasteuria ramosa is the only Pasteuria species that is known to parasitize water fleas; it is not an active ingredient in any pesticides.

2 The currently registered pesticide products containing Pasteuria usgae

– Bl1 as an active ingredi ent are Pasteuria usgae

– BL1 (EPA Reg. No. 85004

- 1), Econem TM (EPA Reg. No. 85004

- 2), and Pasteuria usgae

– Liquid Formulation (EPA Reg.

No. 85004

- 3).Pasteuria spp. ( Hoplolaimus galeatus nematode)

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Bio pesticides Registration Action Document

registration of the two prop osed Pasteuria spp. ( Hoplolaimus galeatus nematode)

– P h 3 pesticide products for nonfood /nonfeed use on turfgrass , specifically St. Augustine Grass and /or Bermuda Grass not to be fed to or grazed by livestock. Overall, such data and information are adequat e for risk
assessment purposes , fulfill the current microbial pesticide data requirements

, and support registration of the proposed products under FIFRA section 3(c)(5)

.

Product Analysis The applicant fulfilled m ost of the product analysis data requiremen ts for Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3, including product chemistry and composition, analysis of samples, and physical and chemical characteristics, by acceptable guideline studies. As a term of registration, the EPA is requiring a one

- year storage stability study

.

Toxicology The applicant submitted adequate mamm alian toxicology data and information to support the Pasteuria spp. ( Hoplolaimus galeatus nematode)

– P h 3 pesticide products. An a cute oral toxicity/pathogenicity study showed t hat, at a single high dose, Pasteuria spp. ( Hoplolaimus galeatus nematode)

– P h 3 is not

toxic and/or p athogenic via this route of exposure

. Based on the applicant’s scientific rationale, which

included an independent survey of published scien tific litera ture that showed no adverse effects from human exposure to any Pasteuria species and citation to previous toxicological studies done on other registered Pasteuria species 3 that resulted in no reported incidences of toxicity and/or pathogenicity, the

EPA wa ived the requirement for acute pulmonary and acute injection toxicity/pathogenicity testing.
Moreover, the applicant reported that no hypersensitivity incidents occurred during rese arch, development, or testing of Pasteuria spp. ( Hoplolaimus galeatus nemat ode )

– P h 3. In l ight of the results

of the acute toxicity/pathogenicity data and the absence of hypersensitivity incidents, the EPA is not requiring testing at higher tiers (i.e., Tiers II and III)

.

Occupational Exposure Despite the low toxicological prof ile of Pasteuria spp. ( Hoplolaimus galeatus nematode)

– P h 3

, the EPA is requiring baseline personal protective equipment (PPE) for handlers who, due to their occupation, may be exposed to the active ingredient for prolonged periods and/ or numerous times

. H andlers working with Pasteuria spp. ( Hoplolaimus galeatus nematode)

– P h 3 in agricultural and/or

commercial settings are directed to wear a long

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

- 95, R

- 95, or P

- 95. For future products, the EPA may require additional

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

- specific basis.

Nontarget Organisms

The EPA found that s cientific rationale submitted by the applicant are sufficient to fulfill the nontarget organism 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. The EPA performed an environment al risk

assessment

, based on information provided by the ap plicant

, and determined that adverse effects to
nontarg et organisms are not expected from the proposed pesticidal uses of Pas teuria spp. ( Hoplolaimus galeatus nematode)

– P h 3 . Moreover, the EPA made a “No Effect” determination for direct and indirect effects to listed species and their d esignated critical habitats resulting from these same proposed

3 Pasteuria Bioscience, Inc. previously conducted a full battery of toxicological testing on Pasteuria usgae

– Bl1

, Pasteuria
nishizawae

– Pn1, and Pasteuria spp. ( Rotylenchulus reniformis nematode)

– Pr3.Pasteuria spp. ( Hoplolaimus galeatus nematode)

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Bio pesticides Registration Action Document

pesticidal uses.

Public P articipation

On Octob er 1, 2009, the EPA announced a policy to provide a more meaningful opportunity for the public to participate i n major registration decisions before they occur. According to this policy, the EPA provides a public comment period prior t o making a registration decision for the following types of appli cations: new active ingredients; first food uses; first outdoor uses; first residential use s; or any other registration actions for which the EPA believes there may be significant public inte rest

.

Consistent with the policy of making registration actions more transparent, the proposed pe sticide products containing Pasteuria spp. ( Hoplolaimus galeatus nematode)

– P h 3

, a new active ingredient, and allowing for this active ingredient’s first o utdoor uses were subject to a 15

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

the Pasteuria spp. ( Hoplolaimus galeatus nematode)

– P h 3 pesticide products, Pasteuria spp. ( Hoplolaimu s galeatus nematode)

– Ph3 and NewPro

. Therefore, the EPA maintained that, based upon the risk assessment and information submitted in support of such pesticide products, it was appropriate

to issue the Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 and NewPro registrations under FIFRA section 3(c)(5)

. The basis for this decision can be found in the risk assessment for Pasteuria spp. ( Hoplolaimus galeatus nematode)

– P h 3 , which is ch aracterized throughout this Biopesticides Registration Action Documen t (BRAD)

.

II. ACTIVE INGREDIENT OVERVIEW

Biological Name: Pasteuria spp. ( Hoplolaimus galeatus nematode)

– P h 3

Culture Deposit: American Type Culture Collection in Manassas

, Virginia

under Accession N umber SD

- 583 2

OPP Chemical Code: 01645 7

Type of Pesticide: Microbial Pesticide

– Nematicide

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

the r egistered pesticide products containing this active ingredient.Pasteuria spp. ( Hoplolaimus galeatus nematode)

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Bio pesticides Registration Action Document

III. REGULATORY BACKGROUND

In March 2012

, 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 manufacturing

- use pesticide product,

Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 (EPA File Symbol: 85004

- RN)

, and an end

- use pesticide produc t, NewPro (EPA File Symbol: 85004

- RR)

, under FIFRA section 3. On September 28, 2012 ( 77 FR 59611 )

, the EPA announced receipt of these application s to register pesticide product 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 th is publication.

IV. RISK ASSESSMENT

In the Federal Register s of October 26, 2007 ( 72 FR 60988 ) and August 30, 2012 ( 77 FR 52610 )

, the

EPA issued Final Rule s on the data requirements to support registration of microbial pesticides and updated the definition for microbial pesticides

. The data and information evaluated for this BRAD were considered in light of these requirements.

The classifications that are found for each data submiss ion are assigned by the EPA ’s 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

assessmen t. 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 “supple mental: upgradeable ,” the EPA always provides an indication of what is lacking or what can be provided to change the rating to “ acceptable .” If there is simply a “ supplemental ” rating, the reviewer will often state that 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 submitted.

For the acu te toxicity data requirements, Toxicity C ategories are assigne d based on the hazard(s) identified from studies and/or other infor mation submitted to the EPA in support of a pesticide registration. The active ingredient or particular product is classified into Toxicity Category I, II, III, or IV, where Toxicity Categ ory I indicates the highest toxicity and Toxicity Category IV indicates the lowest toxicity (see section II of U.S. EPA (2012b) )

.

A. Product Analysis Assessment ( 40 C

. F

. R

. § 158.2120 )

With the exception of a one

- year storage stability study, which the EPA will require as a term of registration, the applicant fulfilled all product analysis data requiremen ts for Pasteuria spp. ( Hoplolaimus galeatus nematode )

– P h 3 . Refer to Tables 1, 2, 3, and 4 in Appendix A for a summary of

the data requirements, including both generic and product

- specific information.Pasteuria spp. ( Hoplolaimus galeatus nematode)

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Bio pesticides Registration Action Document

B. Human Health Assessment ( 40 C

. F

. R

. § 158.2140 )

1. Toxicity

The applicant fulfilled a ll toxicology data requirement s for Pasteuria spp. ( Hoplolaimus galeatus

nematode )

– P h3

. Acceptable Tier I mammalian toxicology data and information support registration of

the Pasteuria spp. ( Hoplolaimus galeatus nematode )

– P h 3 pesticide products

. Furthermore, the EPA is not requiri ng Tier II and Tier III studies for Pasteuria spp. ( Hoplolaimus galeatus nematode )

– P h 3 based on the lack of acute toxicity/pathogenicity in the Tier I studies.

For a summary of the generic toxicology data requirements described in sections IV(B)(1)(a) a nd 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/Pathogenici ty

– Rat ( Harmonized Guideline 885.3050; Master Record Identification Number (MRID No.) 487876

- 12 ) : Groups of fasted, 2

- month

- old, Sprague

- Dawley, albino rats (12 per sex) were given a single oral dose of 1 milliliter (mL) of Pasteuria spp. ( Hoplolaimus ga leatus nematode)

– Ph3 (3 x 10 8 spores per mL). The animals were observed for a period of up to 21 days, with interim scheduled sacrifices on Days 3, 7, 14, and 21

. In total, 4 tre atment groups were compared: 1 group was dosed wit h the active test substanc e, 1 group was dosed with the autoclaved test substance,

and 2 groups were untreated controls. Enumeration and clearance could not be determined because there is no agar plate method for culturing Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3. There was no mortality and were no signs of pharmacologic and/or toxicologic effects observed in any animal during the study. The gross necropsy conducted at termination of the study revealed no external or

internal observable abnormalities. Differences noted i n statistical comparison of relative organ weights between treatment groups were small, though significant; such comparison, however, did not indicate
any treatment

- related effects. Based on the results of this study, Pasteuria spp. ( Hoplolaimus galeatus

n ematode)

– Ph3 does not appear to be toxic and/or pathogenic in rats when dosed by oral gavage at 3 x 10 8 spores per animal. The EPA rated this study as supplemental

.

Acute Pulmonary Toxicity/Pathogenicity ( Harmonized Gui deline 885.3150; MRID No. 487876

- 13 ) and Acute Injection Toxicity/Pathogenicity (Harmonized Guideline 885.3200; MRID No

. 487876

- 13 ) : Upon

consideration of the applicant’s scientific rationale, which included an independent survey of public

scientific literature (covering food safety and m ammalian health effects) that showed no adverse effects from human exposure to any Pasteuria species and citation to previous toxicological studies done on other registered Pasteuria species that resulted in no reported incidence of toxicity and/or pathoge nicity,

the EPA waived acute pulmonary and acute injection toxicity/pathogenicity testing for Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3.

The toxicological studies conducted with other registered Pasteuria species are briefly described below:

Acute Pulmonary Toxicity/Pathogenicity Testing

Pasteuria usgae

– Bl1

, MRID No. 474267

- 10 : In an acute pulmonary toxicity and pathogenicity assessment, there were no test substance

- related significant adverse effects seen in rats receiving a singlePasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 Page 9 of 36

Bio pesticides Registration Action Document

dose of approximately 1

– 3 x 10 8 spores of Pasteuria usgae

– Bl1 . One dosed female exhibited pale lungs. Additionally, one untreated control female lost weight by D ay 21

, and another untreated control female lost weight by D ay 14 but gained weight by D ay 21. One m icrobial pest control agent ( MPCA )

- treat ed male did not gain weight by D ay 7 but gained weight thereafter. All other animals gained weight throughout the study. Based on these results, Pasteuria usgae

– Bl1 d id not appear to be toxic and/or pathogenic in r ats when dosed at approximately 1

– 3 x 10 8 spores per animal. Since microbial enumeration was not performed because the test material would not grow on agar media, the infectivity was uncertain; h owever, because the spores are highly specific to the sting n ematode, infectivity is unlikely to be a concern. The EPA rated this study as acceptable and classified Pasteuria usgae

– Bl1 as Toxicity C ategory IV.

Pasteuria spp. (Rotylenchulus reniformis nematode)

– Pr3, MRID No. 481460

- 10 : Groups of fasted, 45

- to 4 8

- day

- old CD ® rats (3 per sex per group) were exposed by the intratracheal route to Pasteuria spp. ( Rotylenchulus reniformis nematode)

– Pr3 at a dose of approximately 1.5 x 10 8 spores per animal. The animals were then observed for up to 21 days. Three mal es and three females were treated with

autoclaved Pasteuria spp. ( Rotylenchulus reniformis nematode)

– Pr3 as controls, while three males and three females were not treated and served as naïve controls. Based on the results of this study, Pasteuria spp. ( R otylenchulus reniformis nematode)

– Pr3 does not appear to be toxic and/or pathogenic in rats when dosed at approximately 1.5 x 10 8 spores per animal. Pasteuria spp. ( Rotylenchulus reniformis nematode)

– Pr3 showed no clinical signs or major active ingredi ent

- related tissue abnormalities and no evidence of toxicity or pathogenicity to rats following a single intratracheal instillation of 1.5 x 10 8

spores per rat. The EPA rated this study as supplemental

.

Pasteuria nishizawae

– Pn1, MRID No. 481517

- 10 : Grou ps of rats were treated with live test substance

at 1.6 x 10 8 spores per 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

g ross observations were noted. Organ weights primarily did not 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 sa me group, there were no statistically significant differences in organ weights. No evidence of pathogenicity or toxicity was found from pulmonary

administration of Pasteuria nishizawae

– Pn1 to rats. The EPA rated this study as supplemental

.

Acute Injecti on Toxicity/Pathogenicity Testing

Pasteuria usgae

– Bl1, MRID No. 474267

- 11 : There were no treatment

- related significant adverse effects seen in rats receiving a single intravenous dose of 10 8 Pasteuria usgae

– Bl1 spores. One treated female lost weight b y D ay 7 but gained weight prior to sacrifice on D ay 14. All other animals gained weight throughout the study. All animals survived and appeared normal during the study. No abnormalities were observed in any animal at necropsy or in harvested organs. No sig nificant variations in organ weight were found between different groups or sexes. The acute intravenous median lethal dose ( LD 50 ) o f Pasteuria usgae

– Bl1 was greater than 1 x 10 8 spores per animal in male and female rats. Pasteuria usgae

– Bl1 d id not app ear to be toxic and/or pathogenic in rats when dosed at 10 8 spores per animal. Since microbial enumeration was not performed because the test material would not grow on agar media, the infectivity was uncertain; h owever, because the spores are highly speci fic to the sting nematode, infectivity is unlikely to be a concern. The EPA rated this study as acceptable.Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 Page 10 of 36

Bio pesticides Registration Action Document

Pasteuria spp. (Rotylenchulus reniformis nematode)

– Pr3, MRID No . 481460

- 1 1 : Groups of fasted, 45

- to 48

- day

- old CD ® rats (3 per sex per group) w ere exposed by the intravenous route to Pasteuria spp. ( Rotylenchulus reniformis nematode)

– Pr3 at a dose of approximately 1 x 10 7 spores per animal. The animals were then observed for up to 21 days. Three males and three females were treated with

autocla ved Pasteuria spp. ( Rotylenchulus reniformis nematode)

– Pr3 as controls, while three males and three females were not treated and served as naïve controls. Based on the results of this study, Pasteuria spp. ( Rotylenchulus reniformis nematode)

– Pr3 does n ot appear to be toxic and/or pathogenic in rats when dosed at approximately 1 x 10 7 spores per animal. Pasteuria spp. ( Rotylenchulus reniformis nematode)

– Pr3 showed no active ingredient

- related clinical signs or major tissue abnormalities and no

evidence of toxicity or pathogenicity to rats following a single intravenous injection of 1 x 10 7 spores per

rat. The EPA rated this study as supplemental

.

Pasteuria nishizawae

– Pn1, MRID No . 481517

- 1 1 : Groups of rats were treated with live test substance

at 1.0 x 10 9 spores per 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 d id not differ , though in male rats treated with live test substance, spleen weights on D ays 14 and 21 and liver weight on D ay 21 were slightly but significantly decreased. In female rats from the same group, there were no statistically significant

differen ces in organ weights. No evidence of pathogenicity or toxicity was found from intravenous

administration of Pasteuria nishizawae

– Pn1 to rats. The EPA rated this study as supplemental

.

The EPA believes all these data and information, when taken together, clearly indicate that Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 would not be toxic, infective, and/or pathogenic through the inhalation or injection routes of exposure and that further testing is not necessary.

Hypersensitivity Incidents ( Harm onized Guideline 885.3400; MRID No. 487876

- 11 ) : The applicant reported 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 spp

. ( Hoplolaimus galeatus nematode)

– Ph3 . Any future hypersensitivity incidents must be reported to the EPA (refer to test note #3 of 40 C

. F

. R

. § 158.2140(d))

.

Cell Culture ( Harmonized Guideline 885.3500) : This study is not required because Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 is not a virus (refer to test note #4 of 40 C

. F

. R

. § 158.2140(d)).

b. Acute Toxicology and Subchronic Toxicity/Pathogenicity

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

– Tier III

The EPA is not requiring Tier II and Tier III studies for Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 based on the lack of acute toxicity/pathogenicity in the Tier I studies.

c. Endocrine Disrupto rs

As required by the Administrator under the Federal Food, Drug, and Cosmetic Act ( FFDCA ) section 408(p), the EPA has developed the Endocrine Disru ptor Screening Program (EDSP) and has begun to implement the sc reening 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 orPasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 Page 11 of 36

Bio pesticides Registration Action Document

wildlife similar to an effect produced by a “naturally occurring e strogen, 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 sub stance 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, the EPA issued test orders/data ca ll

- ins for the first group of 67 chemicals, which contains 58 pesticide active ingredients and 9 inert ingredients. Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 is not among the group of 58 pesticide active ingredients on the

initial list to be scr eened under the EDSP.

The EPA believes tha t Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 , the active ingredient described in this document, likely is a substance that would not produce any effect in humans similar to an effect produced by a natur ally occurring estrogenic substance. As such, pu rsuant to Section 408(p)(4),

the EPA will determine in the future whether it can exempt Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 from the requirements of the Section 408(p) EDSP. In the event the EPA does determine to exempt this substance from the EDSP, an order will be issued.

For further information on the status of the EDSP, the policies and procedures, the list of 67 chemicals, the test guidelines and the Tier 1 screening battery, please vis it our websit e at http://www.epa.gov/endo/

.

2 . Occupational Exposure and Risk Characterization

Handler exposure to Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 is not expected to pose any undue risk. Reg ardless, the EPA is requiring appropriate personal protective equipment and

precau tionary statements on pesticide product labels to mitigate any potential risks to pesticide handlers due to prolonged and/ or numerous exposures. Handlers applying Pasteuria s pp. ( Hoplolaimus galeatus nematode)

– Ph3 end

- use pesticide products in agricultural and/or commercial settings must wear a long

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

NIOSH standards of at least N

- 95, R

- 95, or P

- 95. For future products, the EPA may require a dditional

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

- specific basis.

3 . Human Health Risk Characterization

The EPA considered human exposure t o Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 in light of the standard for registration in FIFRA. 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 from incidental exposures when Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 pesticide products are applied to turfgrass in accordance with the EPA’s approved labeling. This is a nonfood use. In addition

, this is a nonfeed use as turfgrass t reated with Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 is not to be fed to or grazed by livestock.Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 Page 12 of 36

Bio pesticides Registration Action Document

C. Environmental Assessment ( 40 C

. F

. R

. § 158.2150 )

The EPA found that s cientific rationale submitted by the applicant are sufficient to fulfill the nontarget organism data requirements and for risk assessment purposes. Further testing of nontarge t organisms at higher tier levels (i.e., Tiers II, III, and IV) is not required. The EPA performed an environment al risk

assessment

, based on information provided by the ap plicant, and determined that adverse effects to nontarget organisms are not expected from the proposed pesticidal uses of Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3. Moreover, the EPA made a “No Effect” determination for direct and indirect effects to listed species and their designated critical habitats resulting from these sam e proposed pesticidal uses.

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

.

1. Ecological Exposure and Risk Characterization

a. Terrestrial Animals an d Plants

Birds ( Harmonized Guideline s 885.4050 and 885.4100 )

, Wild Mammals (Harmonized Guideline 885.4150) , Nontarget Plants (Harmonized Guideline 885.4300), Nontarget Insects ( Harmonized Guideline 885.4340)

, and Honey Bees (Harmonized Guideline 885.4380) (MRID No. 487876

- 14 ) : The proposed end

- use pesticide product is for use on turf , and it is applied by directed spray to soil (preplant)

or turf

. Applications are to be followed with a sufficient amount of water to move the Pasteuria spp. ( Hoplolaimus gale atus nematode)

– Ph3 spores into the root zone. 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.

Sci entific rationale 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 spp. ( Hoplolaimus galeatus nematode)

– Ph3

. Pasteuria species are widesprea d in their distribution throughout t he world ( Ciancio et
al . 1998; Ciancio et al . 1994; Giblin

- Davis et al . 1990; Sayre and Starr 1985; Sturhan 1988) . They are known only as parasites of plant

- parasitic nematodes, with the exception of Pasteuria ramosa tha t is known only to parasitize Daphnia s pecies (Atibalentja et al

. 2004 ; Chen and Dickson 1998; Sayre et al

. 1991). Therefore, Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 is not expected to be toxic or pathogenic to birds, nontarget insects

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

An acute oral toxicity/pathogenicity study with laboratory rats (MRID No. 487876

- 12 ) is available to
evaluate the potential effects of Pasteuria spp. ( Hoplolaimus galeatus ne matode)

– Ph3 on wild

mammals. T he acute oral toxicity/pathogenicity study is summarized in section IV(B)(1)(a) of this

BRAD . No signs of toxic or pathogenic effects were observed in that study. The study was determined to be supplemental because clearance and infectivity were not observed , since Pasteuria species cannot be

cultured outside of their hosts

. There is no reason that the test animals in th is study 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 applications of Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3

.Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 Page 13 of 36

Bio pesticides Registration Action Document

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

of Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3

.

Additionally, there are no reports of toxicity or pathogenicity of Pasteuria spp. ( Hoplolaimus g aleatus nematode)

– Ph3 to nontarget terrestrial animals or plants. To support its data waiver rationale, the applicant performed a search within the Agricola and PubMed databases, along with several biological journals (i.e., Journal of Bacteriology, Appl ied and Environmental Microbiology, FEMS Microbiology Ecology, Soil Biology and Bioch emistry) for the period of 1980

– 201 2 . The search was conducted using

“ Pasteuria ” as the search word, and returned no reports of acute toxicity data on birds , freshwater fi sh, freshwater invertebrates, nontarget insects, and honey bees

. The EPA also conducted a search within the Thomson Reuters Web of Knowledge Database, which simultaneously searches the Web of Science, Current Contacts Connect, Medline, and Journal Citation Reports databases. The search was performed

with the term “ Pasteuria ” coupled with each of the search terms of “ bird, ” “mammal,” “invertebrate,” “arthrop od,” “insect,” “honey bee,” “plant pathogen, ” and “phytopathogen” for all of the years available

. No u nanticipated reports of adverse effects of Pasteuria species in nontarget organisms were found in any of these searches. 4 Any reports of effects to nontarget organisms related to Pasteuria species that are

found in the future may trigger the need for testi ng; however, given the specificity of these bacteria for their hosts as is currently understood, such reports are not expected.

Based on the rationale above and available information , adverse effects are not expected to occur to terrestrial animals or pla nts as a result of the proposed pesticidal applications of Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3

. Additionally, adverse effects to terrestrial nontarget organisms

were not identified for Pasteuria usgae

– B l 1

, 5 and because of the specificity of Pasteuria species for their hosts, no interactions between these bacteria are expected. Therefore, additional risk is not anticipated for terrestrial nontarget species resulting from the proposed pesticidal uses of the end

- use pesticide product contain ing both Pasteuria species ( Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 and Pasteuria usgae

– Bl1 )

.

b. Aquatic Animals and Plants

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

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

. 487876

- 14 ) : A pplications of the end

- use pesticide product are intended primarily to put Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 in the root zone of the treated plants, so runoff into aquatic areas is not likely to be significant. Pasteuria sp ecies are naturally found in soil, so some natural presence of Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 in surface waters is expected. To date, no information is available on the size of naturally occurring populations of Pasteuria species

.

Scientific rationale was submitted to fulfill data requirements for and support effects conclusions f or freshwater aquatic orga nisms . The rationale provides sufficient information to conclude that adverse effects are not expected in freshwater fish or freshwater invertebrates as a result of exposure to Pasteuria

4 Pasteuria ramosa is already recognized as parasitizing water fleas and is not considered an “unanticipated” report.

5 Memorandum from S. Borges throug h Z. Vaituzis to J. Kausch, Subject: Environmental risk assessment of Pasteuria

usgae ...for control of sting nematodes in turf and strawberries, dated April 8, 2009 (U.S. EPA 2009).Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 Page 14 of 36

Bio pesticides Registration Action Document

spp. ( Hoplolaimus galeatus nematode)

– Ph3

. The rationale was similar to that presented for terrestrial organisms in that Pasteuria species are found in soils worldwide. Pasteuria ramosa is a closely related species that parasitizes Daphnia magna and other Daphnia species (Ebert et al . 1996); however, it is widely understoo d that all other known Pasteuria sp ecies are obligate parasites of soil

- dwelling nematodes (Atibalentja et al

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

There are no available reports in the literature documenting pathogenicity or toxicity to fre shwater fish or invertebrates as a result of exposure to Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3

. 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 earc h terms used for terrestrial animals and plants , the terms “fish,” “aquatic,” “freshwater,” “estuarine,” and “marine” were also searched. No unanticipated reports of

adverse effects of Pasteuria species in aquatic nontarget organisms were found in any of t hese searches. 6
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 species for their nematode

hosts.

B ased on the rationale above a nd information submitted, adverse effects to freshwater and marine/estuarine fish, invertebrates, and plants are not expected as a result of exposure to proposed pesticidal applications of Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3

. As noted abov e, due

to the specificity of Pasteuria sp ecies for their hosts, no interactions between these bacteria are expected. Therefore, additional risk to aquatic nontarget species is not anticipated for the proposed uses of the end

use pesticide product c ontainin g both Pasteuria species ( Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 and Pasteuria usgae

– Bl1 )

.

2. Environmental Fate Data

As t he rationale provided are sufficient to fulfill the Tier I nontarget organism data requirements and

allow for nont arget organism risk asse ssment for Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3

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

.

3

. Threatened and Endangered Species Assessment

Since the EPA has de termined that no effects are ant icipated for any non target species exposed to

Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 as a result of the proposed pesticidal applications, effects to federally listed threatened and endangered species and their designated 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 spp. ( Hoplo laimus galeatus nematode)

– Ph3 , as labeled.

6 Pasteuria ramosa is already recognized as parasitizing water fleas and is not considered an “unanticipated” report.Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 Page 15 of 36

Bio pesticides Registration Action Document

V. ENVIRONMENTAL JUSTICE

The EPA seeks to achieve environmental justice

— the fair treatment and meaningful involvement of all people regardless of race, color, national origin, or income

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

- making process; and (4)

the decision

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

To help address potent ial environmental justice issues, during the 15

- day public participation comment period, the EPA sought information on any groups or segments of the population who, as a result of their location, cultural practices, or other factors, may have atypical, unu sually high exposure to Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 , compared to the general population. No public comments were received on this particular matter.

For additional information regarding environmental justice issues, please visit t he 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 p esticide provided that all the following

determinations are made:

(1) Its composition 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 wi ll perform its intended function without unreasonable adverse effects on the environment; AND (4) When used in accordance with widespread and commonly recognized practice, it will not generally cause unreasonable adverse effects on the

environment.

To satisfy criterion 1, the Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 pesticide products

have well

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

Pasteuria spp. ( Hoplolaimus galeatus nemato de)

– Ph3 and NewPro label s, and such pesticide products

are not expected to cause unreasonable adverse effects on the environment when used according to the ir respective label instructions. Criterion 2 is satisfied by the current product label s, as well a s the data and

information presented in this document. It is believed that the Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 pesticide products will not cause any unreasonable adverse effects on the environment,

and NewPro ( end

- use pesticide product ) , in particular, is likel y to p rovide protection against lance andPasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 Page 16 of 36

Bio pesticides Registration Action Document

sting nematode s as claimed, satisfying criterion 3. Criterion 4 is satisfied in that the Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 pesticide products are not expected to cause u nreasonable adverse effects when used according to label instructions. Therefore, Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 an d NewPro

, containing Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 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 spp. ( Hoplolaimus galeatus nematode)

– P h 3 for shipment, the re gistrant is required to provide appropriate final printed labeling to the EPA

.

B. Terms of Registration

The following item will be a term of the Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 registration :

(1) Storage Stability (Harmonized Guideline 830.6317) : Within one year of registration, Pasteuria Bioscience, Inc. must provide the results of a one

- year storage stability study.

The following items will be terms of the NewPro registration:

(1) Analysis of Samples (Harmonized Guideline 885.1400) : Within 18 months of registration, Pasteuria Bioscience, Inc. must submit four (4) additional batch analyses to allow the EPA to confirm that the current NewPro production facilities are following the approved manufacturing process and meeting the specific ations for acceptable com mercial product.

(2) Storage Stability (Harmonized Guideline 830.6317) : Within one year of registration, Pasteuria Bioscience, Inc. must provide the results of a one

- year storage stability study.

(3) Corrosion Characteristics ( Harmonized Guideline 830.6320) : Within one year of registration, Pasteuria Bioscience, Inc. must provide the results of a one

- year corrosion characteristics study.

C. Reporting of Adverse Effects and Hypersensitivity Incidents

Notwithstanding the inform ation stated in the previous sections, it should be clearly understood that

certain specific data are required to be reported to the EPA as a requirement for maintaining the federal registration for a pesticide product. A brief summary of these types of da ta are described below.

Reports of all incidents of adverse effects to the environment must be submitted to the EPA under the provisions stated in FIFRA section 6(a)(2). Additionally, all incidents of hypersensitivity (including both suspected and confirm ed incidents) must be reported to the EPA under the provisions of 40 C

. F

. R

. § 158.2140(d).Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 Page 17 of 36

Bio pesticides Registration Action Document

VIII. GLOSSARY OF ACRONYMS AND ABBREVIATIONS

BRAD Biopesticides Registration Action Document

C

. F

. R

. Code of Federal Regulations

cSt centistokes

EDSP Endocrine Di sruptor Screening Program

EP end

- use pesticide product

EPA United States Environmental Protection Agency

FFDCA Federal Food, Drug, and Cosmetic Act

FIFRA Federal Insecticide, Fungicide, and Rodenticide Act

FR Federal Register g/cm 3 grams per cubic centimet er 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

mL milliliter

MP manufacturing

- use pesticide product

MPCA microbial pest control agent

MRID No. Master Record Identification Number

NIOSH National Institute for Occupat ional Safety and Health

OPP Office of Pesticide Programs

PC Code Pesticide Chemical Code

PPE personal protective equipment

TGAI technical grade of the active ingredient

U.S. United StatesPasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 Page 18 of 36

Bio pesticides Registration Action Document

I X. BIBLIOGRAPHY

A. Studies Submitted or Cited to Support the Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph 3 Pesticide Product Registrations

MRID No. Study Information

474267

- 09 Crutchfield V. 2008. Acute Oral Toxicity/Pathogenicity Study in Rats (MPCA) : Pasteuria

usgae Spores

– Liquid Suspension

– Final Report. Project Number: 11463/07. Unpublished study prepared by Stillmeadow, Inc., 15 pages.

474267

- 10 Crutchfield V. 2008. Acute Pulmonary Toxicity/Pathogenicity Study in Rats (MPCA): Pasteuria usgae Spores

– Liquid Suspension

– Final Report. Project Number: 11465/07. Unpublished study prepared by Stillmeadow, Inc., 15 pages.

474267

- 11 Crutchfield V. 2008. Acute Injection Toxicity/Pathogenicity Study in Rats (MPCA): Pasteuria usgae Spores

– Liquid Suspension

– Final Report. Project Number: 11466/07. Unpublished study prepared by Stillmeadow, Inc., 12 pages.

474267

- 12 Crutchfield V. 2008. Acute Dermal Toxicity/Pathogenicity Study in Rats (MPCA): Pasteuria usgae Spores

– Liquid Suspension

– Final Report. Project Number: 1146 4/07. Unpublished study prepared by Stillmeadow, Inc., 12 pages.

476504

- 05 Harder J. 2009. Acute Eye Irritation Study of Pasteuria usgae Liquid Formulation (EP) in Rabbits. Project Number: 2229/001/SNI, 2229/001, 1. Unpublished study prepared by IIT

Res earch Institute, 27 pages.

476504

- 06 Harder J. 2009. Acute Dermal Irritation Study of Pasteuria usgae Liquid Formulation (EP) in Rabbits. Project Number: 2229/001, 2, 2229/001/SN2. Unpublished study prepared by IIT Research Institute, 25 pages.

481460

- 10 Gingras B. 2010. Toxicity/Pathogenicity Testing of Pasteuria reniformis PR3 Following Acute Intratracheal Challenge in Rats (Final Report). Project Number: 2262/SN2, 2262. Unpublished study prepared by IIT Research Institute, 55 pages.

481460

- 11 Gi ngras B. 2010. Toxicity/Pathogenicity Testing of Pasteuria reniformis PR3 Following Acute Injection Challenge in Rats. Project Number: 2262/SN3, 2262. Unpublished study prepared by IIT Research Institute, 57 pages.Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 Page 19 of 36

Bio pesticides Registration Action Document

MRID No. Study Information

481460

- 13 Sullivan D. 2010. Acute Oral Toxicity Study of Pasteuria reniformis PR3 in Rats. Project Number: 2262, 2262/SN7. Unpublished study prepared by IIT Research Institute, 28

pages.

481460

- 14 Sullivan D. 2010. Acute Dermal Toxicity Study of Pasteuria reniformis PR3 in Rabbits. Project Number: 2262, 2262/SN8. Unpublished study prepared by IIT Research Institute, 30 pages.

481460

- 15 Sullivan D. 2010. Acute Eye Irritation Study of Pasteuria reniformis PR3 in Rabbits. Project Number: 2262, 2262/SN9. Unpublished study prepared by IIT Rese arch Institute, 25 pages.

481460

- 16 Sullivan D. 2010. Acute Dermal Irritation Study of Pasteuria reniformis PR3 in Rabbits. Project Number: 2262, 2262/SN10. Unpublished study prepared by IIT Research Institute, 25 pages.

481517

- 10 Gingras B. 2010. To xicity/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.

481517

- 11 Gingras B. 2010. Toxicity/Pathog enicity Testing of Pasteuria nishizawae Pn1 Following Acute Injection Challenge in Rats. Project Number: 2262/SN6. Unpublished study prepared by IIT Research Institute, 56 pages.

481517

- 13 Sullivan D. 2010. Acute Oral Toxicity Study of Pasteuria nishiza wae 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 prep ared by IIT Research Institute, 30

pages.

481517

- 15 Sullivan D. 2010. Acute Eye Irritation Study of Pasteuria nishizawae Pn1 in Rabbits. Project Number: 2262/SN13. Unpublished study prepared by IIT Research Institute, 25

pages.Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 Page 20 of 36

Bio pesticides Registration Action Document

MRID No. Study Information

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

pages.

482061

- 09 Sullivan D. 2010. Acute Oral Toxicity Study of Pasteuria reniformis PR3 (End Product) in Rats. Project Number: 2262/SN15. Unpublished study prepared by IIT Research Institute, 28 pages.

482061

- 10 Sullivan D. 2010. Acute Dermal Toxicity Study of Pasteuria reniformis PR3 (End Product) in Rabbits. Project Number: 2262/SN16, 2262, 16. Unpublished study prepared by IIT Research Institute, 28 pages.

482061

- 11 Sullivan D. 2010. Acute Eye Irritation Study of Pasteuria reniformis PR3 (End Product) in Rabbits. Project Number: 2262, 17, 2262/SN17. Unpublished study prepared by IIT

Resea rch Institute, 22 pages.

482061

- 12 Sullivan D. 2010. Acute Dermal Irritation Study of Pasteuria reniformis PR3 (End Product) in Rabbits. Project Number: 2262, 18, 2262/SN17. Unpublished study prepared by IIT Research Institute, 22 pages.

482103

- 09 Su llivan D. 2010. Acute Oral Toxicity Study of Pasteuria nishizawae PN1 (End Product) in Rats. Project Number: 2262/SN15 , 2262, 15 . 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, 20. Unpublished study prepared by IIT Research Institute, 28 pages.

482103

- 11 Sullivan D. 2010. Acute Eye Irritation Study of Pasteuria nishizawae PN1 (End Product)

in Rabbits . Project Number: 2262/SN21, 2262, 21. 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, 2262, 22. U npublished study prepared by IIT Research Institute, 22 pages.Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 Page 21 of 36

Bio pesticides Registration Action Document

MRID No. Study Information

487875

- 01 MacIntosh S, Smith K. 2012. Product Identity and Manufacturing Process for NewPro [A Mixed Isolate End Product Containing: Pasteuria spp. ( Hoplolaimus nematode)

– PH3

and Candidatu s Pasteuria usgae

– BL1]. Unpublished study prepared by MacIntosh & Associates, Inc. and Pasteuria Bioscience, Inc., 16 pages.

487875

- 02 Smith K. 2012. Discussion of Formation of Unintentional Ingredients: NewPro (Liquid Formulation; End Product). Unpub lished study prepared by Pasteuria Bioscience, Inc., 6

pages.

487875

- 03 Smith K. 2012. Analysis of Samples/Enforcement Methods for NewPro [A Mixed Isolate End Product Containing: Pasteuria spp. ( Hoplolaimus nematode)

– PH3 and Candidatus Pasteuria usgae

– BL1]. Unpublished study prepared by Pasteuria Bioscience, Inc., 14

pages.

487875

- 05 Kaminsky M. 2012. Mixed Pasteuria spp. (PH3 + BL1) Liquid Formulation (EP) Product Chemistry: Final Report. Project Number: 15964/12. Unpublished study prepared by

St illmeadow, Inc., 9 pages.

487875

- 06 Smith K. 2012. Storage Stability for NewPro [A Mixed Isolate End Product Containing:

Pasteuria spp. ( Hoplolaimus nematode)

– PH3 and Candidatus Pasteuria usgae

– BL1]. Project Number: PBI/171/31. Unpublished study pre pared by Pasteuria Bioscience, Inc., 11 pages.

487875

- 07 Smith K. 2012. Corrosion Characteristics for NewPro [A Mixed Isolate End Product Containing: Pasteuria spp. ( Hoplolaimus nematode)

– PH3 and Candidatus Pasteuria

usgae

– BL1]. Project Number: 1596 3/12. Unpublished study prepared by Stillmeadow, Inc., 11 pages.

487875

- 08 Smith K. 2012. Hypersensitive Incidents: NewPro, a Mixed Isolate End Product (PH3 + BL1). Project Number: 15963/12. Unpublished study prepared by Pasteuria Bioscience, Inc., 6 pa ges.

487875

- 10 MacIntosh S, Smith K. 2012. Satisfaction of Data Requirements with Public Literature and/or Scientific Rationale: Freshwater Fish Toxicity/Pathogenicity and Freshwater Invertebrate Toxicity/Pathogenicity for NewPro. Unpublished study prep ared by MacIntosh & Associates, Inc. and Pasteuria Bioscience, Inc., 26 pages.Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 Page 22 of 36

Bio pesticides Registration Action Document

MRID No. Study Information

487876

- 01 Schmidt L, Smith K, MacIntosh S. 2012. Product Identity for Pasteuria spp. ( Hoplolaimus nematode)

– PH3. Unpublished study prepared by MacIntosh & Associates, Inc. and Pasteuria Bioscience, Inc., 107 pages.

487876

- 02 Smith K. 2012. Manufacturing Process for Pasteuria spp. ( Hoplolaimus nematode)

– PH3. Unpublished study prepared by Pasteuria Bioscience, Inc., 19 pages.

487876

- 03 MacIntosh S. 2012. Discussion of Formation of Unintentional Ingredients: Pasteuria spp. ( Hoplolaimus nematode)

– PH3. Unpublished study prepared by MacIntosh & Associates, Inc., 7 pages.

487876

- 04 Smith K. 2012. Analysis of Samples/Enforcement Methods for Pasteuria spp. ( Hoplolaimus ne matode)

– PH3. Project Number: PBI/09/2008. Unpublished study prepared by MacIntosh & Associates, Inc., 82 pages.

487876

- 06 Kaminsky M. 2012. Pasteuria spp. ( Hoplolaimus nematode): Product Chemistry (Group B) Final Report. Project Number: 15962/12. Unpu blished study prepared by Stillmeadow, Inc., 21 pages.

487876

- 07 Kaminsky M. 2012. Pasteuria spp. ( Hoplolaimus nematode): Product Chemistry (Group B) Final Report. Project Number: 15686/11. Unpublished study prepared by Stillmeadow, Inc., 15 pages.

48 7876

- 08 Smith K. 2012. Product Stability to Elevated Temperatures: Pasteuria spp. ( Hoplolaimus nematode)

– PH3. Project Number: PBI/STUDY/171/33. Unpublished study prepared by Pasteuria Bioscience, Inc., 8 pages.

487876

- 09 Smith K. 2012. Storage Stabil ity for Pasteuria spp. ( Hoplolaimus nematode)

– PH3. Project Number: PBI/151/96. Unpublished study prepared by Pasteuria Bioscience, Inc., 11 pages.

487876

- 10 Smith K. 2011. Corrosion Characteristics for Pasteuria spp. ( Hoplolaimus nematode)

– PH3. Proj ect Number: 15685/11. Unpublished study prepared by Stillmeadow, Inc., 11

pages.Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 Page 23 of 36

Bio pesticides Registration Action Document

MRID No. Study Information

487876

- 11 Smith K. 2012. Hypersensitivity Incidents: Pasteuria spp. ( Hoplolaimus nematode)

– PH3. Unpublished study prepared by Pasteuria Bioscience, Inc., 6 pages.

48787 6

- 12 Monds K. 2012. Pasteuria spp. ( Hoplolaimus nematode): Acute Oral Toxicity/Pathogenicity Study in Rats

– Final Report. Project Number: 15695/11. Unpublished study prepared by Stillmeadow, Inc., 31 pages.

487876

- 13 MacIntosh S. 2012. Satisfaction of Data Requirements with Public Literature and/or Scientific Rationale: Acute Pulmonary Toxicity/Pathogenicity, Acute Injection Toxicity/Pathogenicity, Acute Oral Toxicity, Acute Dermal Toxicity, Acute Inhalation Toxicity, Acute Eye Irritation Toxicity, and Acute Dermal Irritation Toxicity for Pasteuria spp. ( Hoplolaimus nematode)

– PH3. Unpublished study prepared by MacIntosh & Associates, Inc., 31 pages.

487876

- 14 MacIntosh S, Smith K, Hewlett T. 2012. Satisfaction of Data Requirements with Public

Lite rature and/or Scientific Rationale: Avian Oral Toxicity, Freshwater Fish Toxicity/Pathogenicity, Freshwater Invertebrate Toxicity/Pathogenicity, Nontarget Insect Testing, and Honey Bee Testing for Pasteuria spp. ( Hoplolaimus nematode)

– PH3. Unpublished st udy prepared by Pasteuria Bioscience, Inc. and MacIntosh & Associates, Inc., 29 pages.

488957

- 01 MacIntosh S, Smith K. 2012. Satisfaction of Data Requirements with Public Literature and/or Scientific Rationale: Acute Oral Toxicity, Acute Dermal Toxici ty, Acute Inhalation Toxicity, Acute Eye Irritation Toxicity, and Acute Dermal Irritation Toxicity for NewPro. Unpublished study prepared by MacIntosh & Associates, Inc. and Pasteuria Bioscience, Inc., 29 pages.

488957

- 02 Smith K. 2012. Certification o f Limits for NewPro. Unpublished study prepared by Pasteuria Bioscience, Inc., 9 pages.

490450

- 01 Smith K. 2013. Final Report: Storage Stability

– Pasteuria spp. ( Hoplolaimus galeatus nematode)

– PH3. Project Number: PBI/157/96. Unpublished study prepar ed by Pasteuria Bioscience, Inc., 9 pages.

490450

- 02 Kaminsky M. 2012. Pasteuria spp. ( Hoplolaimus nematode) Corrosion Characteristics. Project Number: 15685/11. Unpublished study prepared by Stillmeadow, Inc., 12 pages.Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 Page 24 of 36

Bio pesticides Registration Action Document

MRID No. Study Information

490450

- 03 Smith K. 2013. Rev ised Certification of Limits for Pasteuria spp. ( Hoplolaimus nematode)

– PH3: Replaces MRID No. 487876

- 05. Unpublished study prepared by Pasteuria Bioscience, Inc., 8 pages.

B. Environmental Protection Agency Risk Assessment Memoranda

U.S. EPA. 201 3 a. Pasteuria spp. ( Hoplolaimus galeatus nematode)

– P h3

– MUP and NewPro Containing the Ph3 MUP and Pasteuria usgae

– BL1 MUP, EPA Reg. No. 85004

- 1

. Memorandum from J.V. Gagliardi , Ph.D. through J.L. Kough, Ph.D. to J. Kausch dated January 22, 2013

.

U.S. EPA . 201 3b . Environmental Risk Assessment of Pasteuria spp. ( Hoplolaimus galeatus nematode)

– P h 3 (PC 01645 7 ) for a Section 3 Registration of the Technical Product (EPA File Symbol

85004

- RN ) and End Use Pr oduct (EPA File Symbol 85004

- RR ) for Control of the La nce Nematode in Turf . Memorandum from S. Borges to J. Kausch dated March 4, 2013

.

U.S. EPA. 2012 a

. Pasteuria spp. ( Rotylenchulus reniformis nematode)

– Pr3. Memorandum from J.V. Gagliardi, Ph.D. through J.L. Kough, Ph.D. to J. Kausch dated March 2, 2012.

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

U.S. EPA. 2010. Review of Additional Product Chemistry and Human Health Data for Tolerance

Petition fo r Pasteuria usgae

– Liquid Formulation (EPA Reg. No. 85004

- G)

– Pasteuria

Biosciences. Memorandum from S. Schüssel through J.L. Kough, Ph.D. to J. Kausch dated February 25, 2010.

U.S. EPA. 2009 a . Environmental Risk Assessment of Pasteuria usgae (PC 006545) for Section 3 Registrations of the Technical Product ( Pasteuria usgae Spores

– Liquid Suspension, EPA File Symbol 85004

- R) and End Use Products ( Pasteuria usgae Spores

– Clay Granule, EPA File Symbol 85004

- E; Pasteuria usgae

– Liquid Formulation , E PA File Symbol 85004

- G ) for Control of Sting Nematodes in Turf and Strawberries. Memorandum from S. Borges through Z. Vaituzis, Ph.D. to J. Kausch dated April 8, 2009.

U.S. EPA. 2009b. Review of Product Chemistry and Human Health Studies for Pasteuria usg ae

– Liquid Suspension to Support the Registration of Technical Grade Active Ingredient (EPA Reg. No. 85004

- R). Memorandum from S. Schüssel through J.L. Kough, Ph.D. to J. Kausch dated April 9, 2009.

C. Other References

Atibalentja N, Jakstys B P, Noel GR. 2004. Life cycle, ultrastructure, and host specificity of the

North American isolate of Pasteuria that parasitizes the soybean cyst nematode, Heterodera glycines

. Journal of Nematology 36:171

– 180.Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 Page 25 of 36

Bio pesticides Registration Action Document

Cetintas R, Dickson DW. 2004. Persistence an d 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, and biological control potential. Journal of Nematology 30:313

– 340.

C iancio A, Farfan V, Torres EC, Grasso G. 1998. Observations on a Pasteuria isolate parasitic on Hoplolaimus galeatus in Peru. Journal of Nematology 30:206

– 210.

Ciancio A, Bonsignore R, Vovlas N, Lamberti F. 1994. Host records and spore morphometrics of

P asteuria penetrans group parasites of nematodes. Journal of Invertebrate Pathology

63:260

– 267.

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

– 1701.

Giblin

- Davis RM, McDaniel LL, Bilz FG. 1990. Isolates of the Pasteuria penetrans group from phytoparasitic nema todes in bermudagrass turf. Supplement to the Journal of Nematology

22:750

– 762.

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.

S pringer. Pages 119

– 126.

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

– 165.

Sturha n 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: populations, me chanisms of action, and future prospects. FEMS Microbiology Ecology 61:197

– 213.

U.S. EPA. 2012b. Chapter 7 of the Label Review Manual (Precautionary Statements) (Revised July 2012) . Available from http://www.epa.gov/oppfead1/labeling/lrm/chap

- 07.pdf

.Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 Page 26 of 36 Biopesticides Registration Acti on Document

A PPENDIX A. MICROBIAL PESTICIDES DATA REQUIREMENTS (40 C

. F

. R

. PART 158

– SUBPART V )

TABLE 1. Product Analysis Data Requirements for the Technical Grade of the Active Ingredient (TGAI) / Manufacturing

- Use Pesticide Product (MP)
, Pasteuria spp. ( Hoplolaimus galeatus nematode)

– P h 3 (40 C

. F

. R

. § 158.2 120 )

Harmonized

G uideline

N umber Data Requirement Results MRID No. TGAI MP

885.1100 Product Identity N/A Submitt ed data fulfill the r equirement for product identity. Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 contains 99.88 % by weight Pasteuri a spp. ( Hoplolaimus galeatus nematode)

– P h 3 (contains

at least 1 x 10 8 spores per mL ). 487876

- 01

885.1200 Manufacturing Process Sub mitted data fulfill the requiremen t for manufacturing process

. 487876

- 02

885.1250 Deposition of a Sample in a Nationally Recognized Culture

Collection Pasteuria spp. ( Hoplolaimus galeatus nematode)

– P h 3

is on deposit with the American Type Culture Collec tion in Manassas, Virginia under Accession Number SD

583 2

. N/A 487876

- 01

885.1300 Discussion of

Formation of

Unintentional

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

. 487876

- 03

885.1400 Ana lysis of Samples Submitted data fulfill the requirement for analysis of samples. 487876

- 04

885.1500 Certification of Limits N/A Limits listed on the confidential statement of formula are adequate/acceptable. 490450

- 03Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 Page 27 of 36 Biopesticides Registration Acti on Document

TABLE 2

. Product Analysis Data Requirements for the End

- Use Pesticide Product ( E P), NewPro (40 C

. F

. R

. § 158.2120)

Harmonized

Guideline

Number Data Requirement Results MRID No.

885.1100 Product Identity Submitted data fulfill the requirement for product identity. NewPro contains 1 6.6650% by weight Pasteuria usgae

– B l 1 and 16.6450% by weight Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 (contains at least 3 .3 x 10 7

spores (both isolates combined) per mL )

. 487875

- 01

885.1200 Manufacturing Process Submitted data fulfill the requirement for manufacturing process.

885.1250 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 requiremen t for discussion of formatio n of unintentional ingredients. 487875

- 02

885.1400 Analysis of Samples Submitted data fulfill the requirement for analysis of samples for purposes of FIFRA section 3(c)(5) registration. As a term of the NewPro registration, the EPA is requiring confirmato ry batch analyses be submitted within 18 months of registration

. 487875

- 03

885.1500 Certification of Limits Limits listed on the confidential statement of formula are adequate/acceptable. 488957

- 02Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 Page 28 of 36 Biopesticides Registration Acti on Document

TABLE 3

. Physical and Chemical Characteristics for the Technical Grade of the Active Ingredient (TGAI) / Manufacturing

- Use Pesticide Product (MP), Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 ( 40 C

. F

. R

. § 158.2120)

Harmonized G uideline

N umber Data Requirement Results MRID No.

TGAI MP

830.6302 Colo r Light brown, brown, or red

- brown N/A 487876

- 06

830.6303 Physical State Liquid N/A

830.6304 Odor Agar broth, yeasty N/A

830.6313 Stability to Normal and E levated Temperatures, Metals,

and Metal I ons After 14 days at 54°C, no loss of potency. N/A 4878 76

- 08

830.6317 Storage Stability Study is in progress. As a term of the Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 registration, the EPA is requiring the results of a one

- year storage stability study be submitted within one year of registration. 487876

- 09

490450

- 01

830.6319 Miscibility N/A Not required because Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 is not an emulsifiable liquid form of a microbial pesticide (refer to test note # 3

of 40 C

. F

. R

. § 158.2120(d)). N/A

830.6320 Corrosion Characteristics N/A Stable at least one year at 20°C in packaging. 487876

- 10

490450

- 02

830.7000 pH 4.28

– 5.01 (1% w/w) N/A 487876

- 06

830.7100 Viscosity N/A 1. 39

– 1.51 cSt (20°C)

0.93

– 1.02 cSt (40°C) 487876

- 07

830.7300 Density/Relative Density/Bulk

Densi ty (Specific Gravity) 1.04 g/cm 3 N/A 487876

- 06Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 Page 29 of 36 Biopesticides Registration Acti on Document

TABLE 4. Physical and Chemical Characteristics for the End

- Use Pesticide Product (EP), NewPro (40 C

. F

. R

. § 158.2120)

Harmonized Guideline

Number Data Requirement Results MRID No.

830.6302 Col or

N/A N/A 830.6303 Physical State

830.6304 Odor

830.6313 Stability to Normal and Elevated Temperatures, Metals, and Metal

Ions

830.6317 Storage Stability Study is in progress. As a t erm of the NewPro registration, the EPA is requiring the

result s of a one

- year storage stability study be submitted within one year of registration. 487875

- 06

830.6319 Miscibility Not required becaus e NewPro is not an emulsifiable liquid form of a microbial pesticide (refer to test note #3 of 40 C

. F

. R

. § 158.2120(d))

. N/A

830.6320 Corrosion Characteristics Study is in progress. As a term of the NewPro registration, the EPA is requiring the

results of a one

- year corrosion characteristics study be submitted within one year of registration. 487875

- 07

830.7000 pH 3.5

– 4 .5 1 N/A

830.7100 Viscosity 106.03 cSt (20°C)

62.52 cSt (40°C) 487875

- 05

830.7300 Density/Relative Density/Bulk Density (Specific Gravity) 8.5 lb/gal 1 N/A

1 Obtained from the confidential statement of formula for the end

- use pesticide product.Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 Page 30 of 36 Biopesticides Registration Acti on Document

T ABLE 5

. Toxicology Data Requirements for the for the Technical Grade of the Active Ingredient (TGAI) / Manufacturing

- Use Pesticide Product (MP), Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 (40 C

. F

. R

. § 158.2140)

Harmonized

Guideline

N umber Data Requirement Results MRID N o. TGAI MP

Tier I

885.3050 Acute Oral Toxicity/Pathogenicity Not toxic and/or pathogenic to rats when administered by oral gavage in a single dose of 3 x 10 8 spores per animal.

Classification: Supplemental N/A 487876

- 12

885.3150 Acute Pulmonary Toxicity/Pathogenicity Waived based on the applicant’s scientific rationale, which

included an independent survey of published scientific literature (covering food safety and mammalian health effects) that showed no adverse eff ects from human
exposure to any Pasteuria species and cited to previous acute pulmonary toxicity/pathogenicity studies done on other registered Pasteuria species (MRID Nos. 474267

- 10,

481460

- 10, and 481517

- 10) that resulted in no reported

incidence of toxi city and/or pathogenicity.

Classification: Acceptable N/A 487876

- 13

885.3200 Acute Injection Toxicity/Pathogenicity Waived based on the applicant’s scientific rationale, which

included an independent survey of published scientific literature (covering fo od safety and mammalian health effects) that showed no adverse effects from human
exposure to any Pasteuria species and cited to previous acute injection toxicity/pathogenicity studies done on other registered Pasteuria species (MRID Nos. 474267

- 11,

481460

- 11, and 481517

- 11) that resulted in no reported incidence of toxicity and/or pathogenicity.

Classification: Acceptable N/A 487876

- 13

885.3400 Hypersensitivity

Incidents The applicant reported that n o hypersensitivity incidents, including immediate

- type or delayed

- type reactions of humans

and domestic animals

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

. Any future hypersensitivity incidents must be reported to the EPA (refer to test note #3 of 40 C

. F

. R

. § 158.2140(d))

. 487876

- 11

88 5.3500 Cell Culture Not required because Pasteuria spp. ( Hoplolaimus galeatus nematode)

– P h 3 is not a virus (refer to test note #4 of 40

C

. F

. R

. § 158.2140(d)). N/A N/APasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 Page 3 1 of 36 Biopesticides Registration Acti on Document

T ABLE 5

. Toxicology Data Requirements for the for the Technical Grade of the Active Ingredient (TGAI) / Manufacturing

- Use Pesticide Product (MP), Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 (40 C

. F

. R

. § 158.2140)

Harmonized

Guideline

N umber Data Requirement Results MRID N o. TGAI MP

870.1100 Acute Oral Toxicity N/A Waived based on the applicant’s scientific rationale, which

included an independent survey of published literature (covering food safety and mammalian health effects) that showed no adverse effects from human exposure to any

Pasteuria species

, the results of MRID No. 487876

- 12,

and because this formulation c ontains inert ingredients that are not expected to be of toxicological concern (refer to test note #5 of 40 C.F.R. § 158.2140(d)). Classification: Acceptable

TOXICITY CATEGORY IV 1 487876

- 13

870.1200 Acute Dermal Toxicity N/A Waived based on the applicant’ s scientific rationale, which

included an independent survey of published literature (covering food safety and mammalian health effects) that showed no adverse effects from human exposure to any

Pasteuria species

, and because this formulation contains

iner t ingredients that are not expected to be of toxicological concern (refer to test note #5 of 40 C.F.R. § 158.2140(d)). Classification: Acceptable

TOXICITY CATEGORY I II 1 487876

- 13

870.1300 Acute Inhalation Toxicity N/A Waived based on the applicant’s scien tific rationale, which

included an independent survey of published literature (covering food safety and mammalian health effects) that showed no adverse effects from human exposure to any

Pasteuria species and cited to previous acute pulmonary toxicity/pat hogenicity studies done on other registered

Pasteuria species (MRID Nos. 474267

- 10, 481460

- 10,

and 481517

- 10) that resulted in no reported incidence of toxicity, and because this formulation contains inert ingredients that are not expected to be of toxicol ogical concern (refer to test note #5 of 40 C.F.R. § 158.2140(d)). Classification: Acceptable

TOXICITY CATEGORY III 487876

- 13Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 Page 32 of 36 Biopesticides Registration Acti on Document

T ABLE 5

. Toxicology Data Requirements for the for the Technical Grade of the Active Ingredient (TGAI) / Manufacturing

- Use Pesticide Product (MP), Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 (40 C

. F

. R

. § 158.2140)

Harmonized

Guideline

N umber Data Requirement Results MRID N o. TGAI MP

870.2400 Acute Eye Irritation N/A Waived based on the applicant’s scientific rationale, which

included an independent survey of p ublished literature (covering food safety and mammalian health effects) that showed no adverse effects from human exposure to any

Pasteuria species, and because this formulation contains inert ingredients that are not expected to be of toxicological concer n (refer to test note #5 of 40 C.F.R. § 158.2140(d)). Classification: Acceptable

TOXICITY CATEGORY III 1 487876

- 13

870.2500 Primary Dermal Irritation N/A Waived based on the applicant’s scientific rationale, which

included an independent survey of publishe d literature (covering food safety and mammalian health effects) that showed no adverse effects from human exposure to any

Pasteuria species, and because this formulation contains inert ingredients that are not expected to be of toxicological concern (refe r to test note #5 of 40 C.F.R. § 158.2140(d)). Classification: Acceptable

TOXICITY CATEGORY IV 1 487876

- 13

Tiers II and III Not required for Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph 3 based on the lack of acute toxicity/pathogenicity in the Tier I studies.

1 The manufacturing

- use pesticide product, Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3, was assigned this T oxicity C ategory because of the similarity between this product (inert

ingredient types and amounts only) and other previou sly registered Pasteuria species products that are supported by product

- specific data (i.e., MRID Nos. 481460

- 13 (oral toxicity ) , 481517

- 13 (oral toxicity )

, 481460

- 14 (dermal toxicity ), 481517

- 14 (dermal toxicity ), 481460

- 15 (eye irritation), 481517

- 15 (eye irritation), 481460

- 16 (dermal irritation), and 481517

- 16 (dermal irritation))

. Regarding the active ingredient component, it is not expected to contribute any toxicity to the manufacturing

- use pesticide product because of the lack of adverse effec ts of Pasteuri a species on humans in the published literature and a multitude of studies done with other registered Pasteuria species that resulted in no reported incidence of toxicity.Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 Page 33 of 36 Biopesticides Registration Acti on Document

TABLE 6

. Toxicology Data Requirements for the End

- Us e Pesticide Product (EP )
, NewPro (40 C

. F

. R

. § 158.2140)

Harmonized

Guideline

Number Data Requirement Results MRID No.

885.3050 Acute Oral Toxicity/Pathogenicity

N/A N/A 885.3150 Acute Pulmonary Toxicity/Pathogenicity

885.3200 Acute Injection Toxicity /Pathogenicity

885.3400 Hypersensitivity

Incidents The applicant reported than n o hypersensitivity incidents, including immediate

- type or delayed

- type reactions of humans and

domestic animals

, occurred during research, development, or testing of the EP. Any future hypersensitivity incidents must be reported to the EPA (refer to test note #3 of 40 C

. F

. R

. § 158.2140(d)). 487875

- 08

885.3500 Cell Culture N/A N/A

870.1100 Acute Oral Toxicity Waived based on the applicant’s scientific rationale, which includ ed an independent survey of published literature (covering food safety and mammalian health effects) that showed no adverse effects from human exposure to any Pasteuria species, the

results of MRID No s . 487876

- 12 and 474267

- 09 , and because this formulation contains inert ingredients that are not expected to be of toxicological concern (refer to test note #5 of 40 C.F.R. § 158.2140(d)). Classification: Acceptable

TOXICITY CATEGORY IV 1 488957

- 01

870.1200 Acute Dermal Toxicity Waived based on the applicant’s scientific rationale, which included an independent survey of published literature (covering food safety and mammalian health effects) that showed no adverse effects from human exposure to any Pasteuria species and

cited to the results of MRID No. 474267

- 1 2

, and because this formulation contains inert ingredients that are not expected to be of toxicological concern (refer to test note #5 of 40 C.F.R. § 158.2140(d)). Classification: Acceptable

TOXICITY CATEGORY III 1 488957

- 01

870.1300 Acute Inhalation

Toxic ity Waived based on the applicant’s scientific rationale, which included an independent survey of published literature (covering food safety and mammalian health effects) that showed no adverse effects from human exposure to any Pasteuria species and

cited to previous acute pulmonary toxicity/pathogenicity studies done on other registered Pasteuria species (MRID Nos.

474267

- 10, 481460

- 10, and 481517

- 10) that resulted in no reported incidence of toxicity, and because this formulation contains inert ingredien ts that are not expected to be of toxicological concern (refer to test note #5 of 40 C.F.R. § 158.2140(d)). Classification: Acceptable

TOXICITY CATEGORY III 488957

- 01

870.2400 Acute Eye Irritation Waived based on the applicant’s scientific rationale, whic h included an independent survey of published literature (covering food safety and mammalian health effects) that showed no adverse effects from human exposure to any Pasteuria species, and because this formulation contains inert ingredients that are not e xpected to be of toxicological concern (refer to test note #5 of 40 C.F.R. § 158.2140(d)). Classification: Acceptable

TOXICITY CATEGORY IV 1 488957

- 01Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 Page 34 of 36 Biopesticides Registration Acti on Document

TABLE 6

. Toxicology Data Requirements for the End

- Us e Pesticide Product (EP )
, NewPro (40 C

. F

. R

. § 158.2140)

Harmonized

Guideline

Number Data Requirement Results MRID No.

870.2500 Primary Dermal

Irritation Waived based on the applicant’s scientific rationale, which included a n independent survey of published literature (covering food safety and mammalian health effects) that showed no adverse effects from human exposure to any Pasteuria species, and because this formulation contains inert ingredients that are not expected to b e of toxicological concern (refer to test note #5 of 40 C.F.R. § 158.2140(d)). Classification: Acceptable

TOXICITY CATEGORY IV 1 488957

- 01

1 The end

- use pesticide product, NewPro, was assigned this Toxicity Category because of the similarity between this p roduct (inert ingredient types and amounts only) and other previously

registered Pasteuria species products that are supported by product

- specific data (i.e., MRID Nos. 482061

- 09 (oral toxicity), 482103

- 09 (oral toxicity), 482061

- 10 (dermal toxicity), 4 82103

- 10 (dermal toxicity), 476504

- 05 (eye irritation), 482061

- 11 (eye irritation), 482103

- 11 (eye irritation), 476504

- 06 (dermal irritation), 482061

- 12 (d ermal irritati on), and 482103

- 12 (dermal irritation)). Regarding the two active ingredient comp onents, they are not expected to contribute any toxicity to the end

- use pesticide product because of the lack of adverse effects of Pasteuria species on humans in the published literature and a multitude of studies done with other registered Pasteuria s pecies that resulted in no reported incidence of toxicity.Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 Page 35 of 36 Biopesticides Registration Acti on Document

TABLE 7

. Nontarget Organism Toxicity and Environmental Fate Data Requirements for the Technical Grade of the Active Ingredient (TGAI), Pasteuria spp. ( Hoplolaimus gale atus nematode)

– Ph3 (40 C

. F

. R

. § 158.2150) 1

Harmonized Guideline

Number Data Requirement Results MRID No.

Tier I

885.4050 Avian Oral Toxicity The rationale provide s sufficient information to determine that toxicity/pathogenicity to avian

wildlife is no t expected. Classification: Acceptable 487876

- 14

885.4100 Avian Inhalation Toxicity/Pathogenicity Not required as the nature of the microbial pesticide does not indicate potential pathogenicity to birds or relatedness to any known bird pathogens (refer to test note #3 of 40 C.F.R. § 158.2150(e)). N/A

885.4150 Wild Mammal Toxicity/Pathogenicity Tests required by 40 C.F.R. § 158.2140 are adequate and appropriate for assessment of hazards to wild mammals. Studies submitted with laboratory rats indicate no ad verse effects due to oral exposure. Classification: Acceptable for wild mammal risk assessment N/A

885.4200 Freshwater Fish Toxicity/Pathogenicity The rationale provide s sufficient information to determine that toxicity/pathogenicity to

freshwater fish an d invertebrates is not expected. Classification: Acceptable 487876

- 14

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 t o water and is not expected to enter marine/estuarine environments in amounts that would result in significant concentrations (refer to test note #6 of 40 C.F.R. § 158.2150(e)). Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 is an obligate parasite o f plant

- parasitic nematodes inhabiting terrestrial soil. 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 C.F.R. § 158.2150(e)) and adverse effects to plants are not expected. N/A

885.4340 Nontarget Insect Testing The rationale provide s sufficient information to determine that toxicity/pathogenicity to honey

bees and nontarget insects is not expected. Classification: Acceptable 487876

- 14

88 5.4380 Honey Bee Testing

Tiers II, III, and IV Not required for Pasteuria spp. ( Hoplolaimus galeatus nematode)

– P h 3 based on the acceptability of the rationale provided for Tier I

. 1 Additional rationale was submitted for the end

- use pesticide produc t, NewPro

, in MRID No. 487875

- 10. Data were not specifically required on the end

- use pesticide product, and the rationale

was determined to be supplemental.Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 Page 36 of 36 Biopesticides Registration Acti on Document

APPENDIX B. PESTICIDE PRODUCTS

EPA Registration

Number Registration Name Percentage

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

85004

- 10 Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 99.88 0 % Technical N/A

85004

- 11 NewPro 16.6650% 1

16.6450% 2 End Use

– Liquid Turfgrass Ground application equipment or chemigation 1.3

– 64 fluid ounces of NewPro per 1,000 square feet of turfgrass Lance Nematode ( Hoplolaimus galeatus )

Sting Nematode ( Bel o nolaimus longicaudatus )

1 Pasteuria usgae

– B 1 1

2 Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3