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 Progr ams Biopesticides and Pollution Prevention Division

DRAFT

– March 2 6 , 2013Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 Page 2 of 36 Biopesticides Registration Action Document

TABLE OF CONTENTS

I. EXECUTIVE SUMMARY

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

II. ACTIV E INGREDIENT OVERVIE W

................................ ................................ ................................ ........ 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

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

VII. ACTIONS REQUIRED OF THE REGISTRANT

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

VIII. GLOSSARY OF ACRONYMS AND ABBREVIATIONS

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

IX. BIBLIOGRAPHY

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

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

– P h 3 Pesticide Product Registrations

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

B. Environmental Protection Agency Risk Assessme nt Memoranda

................................ ............. 25

C. Other References

................................ ................................ ................................ ............................ 25

APPENDIX A. MICROBI AL PESTICIDES DATA R EQUIREMENTS

................................ ............................ 27

APPENDIX B. PESTICI DE P RODUCTS

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

– Ph3 Page 3 of 36 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

Regulations

Kimberly Nesci Chief, Microbial Pesticides Branch

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

– Ph3 Page 4 of 36 Biopesticides Registration Action Document

I. EXECUTIVE SUMM ARY

Background

In March 201 2

, Pasteuria Bioscience, 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. Environmental Protection Agen cy (EPA or the Agency ) under section 3 of the Federal Insecticide, Fungicide, and Rodenticide Act (FIFRA)

. These pesticide products contain a new active ingredient, Pasteuria spp. ( Hoplolaimus ga leatus 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 (F R) 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)

– P r3 (40 C.F.R. § 180.1316 )

, 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 a re 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 (Chen and Dickson 1998; Ciancio et al . 1994; Sturhan 1988)

. H igher population densitie s 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 specifica lly isolated from turfgrass soil samples collected at the 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 sta ge . After an endospore

attaches to the cuticle of a nematode 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, whic h lead s to its death. In light of the demonstrated nematicidal 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 p esticide product that is intended for use as a soil or foliar 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 pesti cide products 2 and is intended to control the sting nematode ( Belonolaimus longicaudatus ).

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

. F

. R

. §§ 158.2120, 158.2140, and 158.2150, respectively) sub mitted to support

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

– P h 3

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 ingredient 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)

– Ph3 Page 5 of 36 Biopesticides Registration Action Document

pesticide products for nonfood use on turfgrass

. Overall, such data and information are adequate for risk assessment purposes , fulfill the current m icrobial pesticide data requirements

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

.

Product Analysis Most of the product analysis data requirements for Pasteuria spp. ( Hoplolaimus galeatus nematode)

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

- year storage stability study

.

Toxicology Adequate mamm alian tox icology data and information were submitted 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 literature that showed no adverse effects from human exposure to any Pasteuria sp ecies and

citation to previous toxicological studies done on other registered Pasteuria species 3 that resulted in no reported incidences of toxicity and/or pathogenicity, EPA waived the requirement for acute pulmonary and acute injection toxicity/pathogeni city testing. Moreover, the applicant reported that no hypersensitivity incidents occurred during rese arch, development, or testing of Pasteuria spp. ( Hoplolaimus galeatus nematode )

– P h 3. In l ight of the results of the acute toxicity/pathogenicity data an d the absence of hypersensitivity incidents, testing at higher tiers (i.e., Tiers II and III) was not required.

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

– P h 3

, baseline personal protecti ve equipment (PPE) is required for handlers that may be exposed to the active ingredient, due to their occupation, for prolonged periods and/ or numerous times . Handlers working with Pasteuria spp. ( Hoplolaimus galeatus nematode)

– P h 3 in agricultural setti ngs are directed to wear a long

- sleeved shirt, long pants, socks, shoes, waterproof gloves, 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. For future pro ducts, EPA may require additional PPE (e.g., protective eyewear) , other than the standard described above, on a product

- specific basis.

Nontarget Organisms

Scientific rationale submitted by the applicant is sufficient to fulfill the nontarget organism d ata 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 performed an environment al risk

assessment based on information provided by the ap plicant

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

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

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)

– Ph3 Page 6 of 36 Biopesticides Registration Action Document

Public Participation

On Octob er 1, 2009, EPA announced a policy to provide a more meaningful opportunity for the public to participate i n major registrat ion decisions before they occur. According to this policy, EPA provides a public comment period prior to 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 EPA believes there may be significant public

interest

.

Consistent with the policy of making registration actions more transparent, the proposed

pe sticide products containing Pasteuria spp. ( Hoplolaimus g aleatus nematode)

– P h 3

, a new active ingredient, will be subject to a comment period. In addition to containing a new active ingredient, the registration of these pesticide products would result in the first outdoor uses for

Pasteuria spp. ( Hoplolaimus ga leatus nematode)

– P h 3 . The docket identification number, associated with these registration actions and accessed through either http://www.regulations.gov

or http://www.epa.gov/pesticides/regulating/registration

- status.html , is EPA

- HQ

- OPP

- 201 2

- 0609 . The following documents are available for comment in EPA

- HQ

- OPP

- 201 2

- 0 609 : (1)

draft Pasteuria spp. ( Hoplolaimus galeatus nematode)

– P h 3 Biopesticides Registration Action Document (BRAD); (2) environmental risk assessment for Pasteuria spp. ( Hoplolaimus galeatus nematode)

– P h 3 ; (3) draft label for the manufacturing

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

– Ph3 (EPA File Symbol : 85004

- RN ); and (4) draft label for

the end

- use pesticide product, NewPro (EPA File Sym bol : 85004

- RR). While a final decision on registration is contingent upon review and consideration of public comments, EPA presently believes that, based upon the risk assessment and information submitted in support of Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 and NewPro

, it is appropriate to issue these registrations. The basis for this preliminary decision can be found in the risk assessm ent for

Pasteuria spp. ( Hoplolaimus galeatus nematode)

– P h 3 , which is characterized throughout this

BRAD.

II. ACTIVE INGREDIENT OVERVIEW

Biological Name: Pasteuria spp. ( Hoplolaimus galeatus nematode)

– P h 3

Culture Deposit: American Type Cultu re 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 proposed pesticide products containing this active ingredient.Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 Page 7 of 36 Biopesticides Registration Action Document

III. REGULATORY BACKGROUND

In March 2012

, MacIntosh and Associates , Inc. (address: 1203 Ha rtford 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. ( Hoplolai mus galeatus nematode)

– Ph3 (EPA File Symbol:

85004

- RN)

, and an end

- use pesticide product, NewPro (EPA File Symbol: 85004

- RR)

, under

FIFRA section 3. On September 28, 2012 ( 77 F R 59611 )

, 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 )

, 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 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 ” i ndicates 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

scientifical ly 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 “ 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.

Fo r 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 product is classi fied 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 C

. F

. R

. § 156.62 )

.

A. Product Analysis Assessment ( 40 C

. F

. R

. § 158.2120 )

With the exception of a one

- year storage stability study, which EPA will require as a term of registration, all product analysis data requirements for Pasteuria spp. ( Hoplolaimus galeatus

nematode )

– P h 3 have been fulfilled . 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)

– Ph3 Page 8 of 36 Biopesticides Registration Action Document

B. Human Health Assessment ( 40 C

. F

. R

. § 158.2140 )

1. Toxicity

All toxicology data requirements for Pasteuria spp. ( Hoplolaimus galeatus nematode )

– P h3

have been fulfilled

. Acce ptable Tier I mammalian toxicology data and information support registration of the Pasteuria spp. ( Hoplolaimus galeatus nematode )

– P h 3 pesticide products

. Furthermore, Tier II and Tier III studies were not required for Pasteuria spp. ( Hoplolaimus galeatu s 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) 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.) 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 galeatus nematode)

– Ph3 (3 x 10 8 spores per mL). The animals were obser ved 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 substance, 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 toxicologi c 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 in statistical comparison of relative organ weights between
treatment g roups 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 nematode)

– Ph3 does not appear to be toxic and/or pathogenic in rats w hen dosed by oral gavage at 3 x 10 8 spores per animal. This study was rated 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 an the applicant’s scientific rationale, which included an independent survey of public scientific literature (covering food safety and mammalian health effects) that showed no adverse effects from human exposur e 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 pathogenicity, EPA waived acute pulmonary and acute injection toxicity/pathogenic ity testing for Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3.Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 Page 9 of 36 Biopesticides Registration Action Document

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

below:

Acute Pulmonary Toxicity/Pathogenicity Testing

Pasteuria usgae

– Bl1

, M RID 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 single dose of approximately 1

– 3 x 10 8 spores of Pasteuria usgae

– Bl1 . One dosed fema le 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 microbial 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 rats when dosed at approximately 1

– 3 x 10 8 spores per animal. Since micro bial 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 nematode, infectivity is unlikely to be a

concern. This study was rated a cceptable, and Pasteuria usgae

– Bl1 was classified as Toxicity

C ategory IV.

Pasteuria spp. (Rotylenchulus reniformis nematode)

– Pr3, MRID No. 481460

- 10 : Groups of fasted, 45

- to 48

- 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 males and three

females were treated with autoclaved Pasteuria spp. ( Rotylenc hulus 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 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 ingredient

- related tissue abnormalities and no evidence of toxicity or pathogenic ity to rats following a single intratracheal instillation of 1.5 x 10 8 spores per rat. This study was rated supplemental

.

Pasteuria nishizawae

– Pn1, MRID No. 481517

- 10 : Groups of rats were treated with live test

substance at 1.6 x 10 8 spores per rat , wer e 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 m ale 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 differences in organ weights. N o evidence of pathogenicity or toxicity was found from pulmonary administration of Pasteuria nishizawae

– Pn1 to rats. This study was rated supplemental

.

Acute Injection 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

– Bl1Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 Page 10 of 36 Biopesticides Registration Action Document

spores. One treated female lost weight by 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 significant variations in organ weight were found between different groups or sexes.

The ac ute 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 appear to be toxic and/or pathogenic in rats when dosed at 10 8 spores per animal. Since mic robial 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 nematode, infectivity is unlikely to be a concern. This study was rat ed acceptable.

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) were 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 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. ( Rotylenchulus reniformis nematode)

– Pr3 does not appear to be toxic and/or pathogenic in rats when dosed at approximately 1 x 10 7 spores

per a nimal. 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. This study was rated 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 did not differ , though

in male rats treated with live test substance, spleen weights on D ays 14 and 21 a nd liver weight

on D ay 21 were slightly but significantly decreased. In female rats from the same group, there were no statistically significant differences in organ weights. No evidence of pathogenicity or toxicity was found from intravenous administratio n of Pasteuria nishizawae

– Pn1 to rats. This study was rated supplemental

.

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 ( Harmonized Guideline 885.3400; MRID No. 487876

- 11 ) : The applicant reported that n o hypersensitivity incidents, includi ng 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 EPA (refer to t est 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)).Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 Page 11 of 36 Biopesticides Registration Action Document

b. Acute T oxicology 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 spp. ( Hopl olaimus galeatus nematode)

– Ph3

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

c. Endocrine Disruptors

As required by the Administrator under the Federal Food, Drug, and Cosmetic Act ( FFDCA ) section 408(p), EPA has developed the Endocrine Disru ptor Screening Program (EDSP) and has begun to implement the screening program that is to be used to test all pesticides in order to

determine whether certain substances (including pesticide active and other ingredients) may have an effect in humans or wildlife similar to an effect produced by a “naturally occurring estrogen, or other such endocrine effects as the Administrator may designate.”

FFDCA section 408(p)(4) authorizes the Administr ator, by order, to exempt from the requirements of the Estrogenic Substances Screening Program a biologic substance or other substance if a determination is made that the substance is not anticipated to produce any effect in humans similar to an effect pro duced by a naturally occurring estrogenic substance.

Between October 2009 and February 2010, EPA issued test orders/data call

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

– Ph3 is not among the group of 58 pesticide active ingredients on the initial list to be screened under the EDSP.

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

– Ph3 , the active ingredient

described in t his document, likely is a substance that would not produce any effect in humans similar to an effect produced by a naturally occurring estrogenic substance. As such, pu rsuant to Section 408(p)(4), EPA will determine in the future whether it can exempt Past euria spp. ( Hoplolaimus galeatus nematode)

– Ph3 from the requirements of the Section 408(p) EDSP. In the event 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 pol icies and procedures, the list of 67 chemicals, the test guidelines and the Tier 1 screening battery, please visit our websit e at http://www.epa.gov/endo/

.

2 . Occupational Exposure and Risk Characterization

Hand ler exposure to Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 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 potenti al risks to pesticide handlers due to prolonged and/ or numerous exposures. Handlers applying Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 end

- use pesticide products in agricultural 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,

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

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

– Ph3 Page 12 of 36 Biopesticides Registration Action Document

3 . Human Health Risk Characterization

EPA considered human exposure to 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 t he 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 EPA

- approved labeling. This is a nonfood use. In addition this is a nonfeed use as turfgrass treated with

Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 is not to be fed to or grazed by livestock.

C. Environmental Assessment ( 40 C

. F

. R

. § 158.2150 )

Scientific rationale submitted by the applicant is 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. EPA performed an environment al risk

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

– Ph3. Moreover, EPA made a “No Effect” determination for direct and indirect effects to listed species and their designated cri tical 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

.

1. Ecological Exposure and Risk Character ization

a. Terrestrial Animals and 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 Hon ey 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

mo ve the Pasteuria spp. ( Hoplolaimus galeatus 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 w ill not necessarily be

eliminated.

Scientific 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 widespread 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,

wit h the exception of Pasteuria ramosa that 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 pathoge nic to birds, nontarget insects

, or honey bees if they are exposed as a result of the proposed pesticidal applications.Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 Page 13 of 36 Biopesticides Registration Action Document

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 nematode)

– 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 determin ed 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
potent ial 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

.

Non target plant testing is not required because Pasteuria species are not related to any known plant pathogen. 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 pathoge nicity of Pasteuria spp. ( Hoplolaimus galeatus 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 journa ls (i.e., Journal of Bacteriology, Applied 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 acut e toxicity data on birds , freshwater fish, freshwater invertebrates, nontarget insects, and honey bees

. EPA also conducted a search within the Thomson Reuters Web of Knowledge Database, which simultaneously searches the Web of Science, Current Contacts Con nect, 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” f or all of the years

available . No unanticipated 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 fu ture may trigger the need for testing; 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 plants 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 th e end

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

– Ph3 and Pasteuria usgae

– Bl1 )

.

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

. 5 Memorandum from S. Borges through Z. Vaituzis to J. Kausch, Subject: Environmental risk asse ssment 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 Biopesticides Registration Action Document

b. Aquatic Animals and Plants

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

armonized Guideline 885.4240)

, Estuarine/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 P asteuria 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 con clusions

f

or freshwater aquatic organisms . 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 spp. ( Hoplolaimus galeatus nemato de)

– 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 . 19 96); however, it is widely understood 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 document ing pathogenicity or toxicity to freshwater 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 earch 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 nontar get organisms were found in any of these 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 ho sts.

B ased on the rationale above and 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 gal eatus nematode)

– Ph3

. As noted above, 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 ontaining 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 nontarget organism risk asse ssment for Pasteuria spp. ( Hoplolaimus galeatus

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 3 6 Biopesticides Registration Action Document

nematode)

– Ph3 , f urther testing at higher tier levels (i.e., Tiers II, III, and IV) is not required.

3

. Threatened and Endangered Species Asses sment

Since EPA has determined 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 endang ered 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 o f Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 , 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

— wi th 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 ne gative environmental 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 communit y residents have 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 partic ipants involved will

be considered in the decision

- making 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 addr ess potential environmental justice issues, EPA seeks information on any groups or segments of the population who, as a result their location, cultural practices, or other factors, may have atypical, unusually high exposure to Pasteuria spp. ( Hoplolaimus g aleatus nematode)

– Ph3 , compared to the general population. Please comment if you are aware of any subpopulations that may have atypical, unusually high exposure compared to the general population.

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

.Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 Page 16 of 36 Biopesticides Registration Action Document

V I

. RISK MANAGEMENT DECISION

Section 3(c)(5) of FIFRA per mits for the registration of a pesticide 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

r equirements of FIFRA; (3) It will 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 unreasonab le adverse effects on the

environment.

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

– Ph3 pesticide products have well

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

made on the Pasteuria spp. ( Hoplolaimus galeatus nematode)

– 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 curre nt product label s, as well as 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 NewPr o ( end

- use pesticide product) , in particular, is likel y to p rovide protection against lance and 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 unreasonable 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 i ngredient, 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 registrant is required to provide appropriate final printed labeling to

EPA

.

B. Terms of Registration

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

– Ph3 registration :

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

- year storage stability study.Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 Page 17 of 36 Biopesticides Registration Action Document

The following items will be terms of the NewPro registration:

(1) Analysis of Samples (Harmoni zed Guideline 885.1400) : Within 18 months of registration, Pasteuria Bioscience, Inc. must submit four (4) additional batch analyses to allow EPA to confirm that the current NewPro production facilities are following the approved manufacturing process and meeting the specifications 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) Corrosi on 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

Notwit hstanding 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 brief summary of t hese 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 C

. F

. R

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

– Ph3 Page 18 of 36 Biopesticides 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 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 Fe deral Register g/cm 3 grams per cubic centimeter 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 Occupational 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 19 of 36 Biopesticides 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 S tudy 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 (MPC A): 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 usg ae 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 S uspension

– Final Report. Project Number: 11464/07. 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 Research 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 Gingras 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 pa ges.Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 Page 20 of 36 Biopesticides 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, 22 62/SN9. Unpublished study prepared by IIT Research 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 Institu te, 25 pages.

481517

- 10 Gingras 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.

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 IIT Research Institute, 56 pages.

481517

- 13 Sullivan D. 2010. 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.

Pro ject 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. Project Number: 2262/SN13. Unpublished study prepared by IIT Research Institute, 25

pages.Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 Page 21 of 36 Biopesticides 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 T oxicity 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 Research 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 Sullivan 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 Sul livan 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 Pas teuria 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 R abbits. Project Number: 2262/SN22, 2262, 22. Unpublished study prepared by IIT Research Institute, 22 pages.Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 Page 22 of 36 Biopesticides 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 s pp. ( Hoplolaimus nematode)

– PH3

and Candidatus 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: N ewPro (Liquid Formulation; End Product). Unpublished 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 Numb er: 15964/12. Unpublished study prepared by Stillmeadow, 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].

Pro ject Number: PBI/171/31. Unpublished study prepared 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 Candidatu s Pasteuria

usgae

– BL1]. Project Number: 15963/12. 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 stud y prepared by Pasteuria Bioscience, Inc., 6 pages.

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/Pa thogenicity for NewPro. Unpublished study prepared by MacIntosh & Associates, Inc. and Pasteuria Bioscience, Inc., 26 pages.Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 Page 23 of 36 Biopesticides 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 s tudy 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/Enforcem ent Methods for Pasteuria spp. ( Hoplolaimus nematode)

– 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. Unpublished 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.

487876

- 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 pag es.

487876

- 09 Smith K. 2012. Storage Stability 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 Pas teuria spp. ( Hoplolaimus nematode)

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

pages.Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 Page 24 of 36 Biopesticides Registration Action Document

MRID No. Study Information

487876

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

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

487876

- 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 Inhalatio n 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. Sati sfaction of Data Requirements with Public Literature 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 study 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 Ratio nale: Acute Oral Toxicity, Acute Dermal Toxicity, 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 of 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. Projec t Number: PBI/157/96. Unpublished study prepared 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, In c., 12 pages.Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 Page 25 of 36 Biopesticides Registration Action Document

MRID No. Study Information

490450

- 03 Smith K. 2013. Revised 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 Agenc y 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 (E PA File Symbol 85004

- RR ) for Control

of the Lance Nematode in Turf . Memorandum from S. Borges to J. Kausch dated March 4, 2013

.

U.S. EPA. 2012. Pasteuria spp. ( Rotylenchulus reniformis nematode)

– Pr3. Memorandum from J.V. Gagliardi, Ph.D. through J.L. Ko ugh, 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 H uman Health Data for Tolerance

Petition for 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 . Environm ental 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 850 04

- E; Pasteuria usgae

– Liquid Formulation , EPA 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 Chemist ry and Human Health Studies for Pasteuria usgae

– 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 BP, 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 26 of 36 Biopesticides Registration Action Document

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, and biological control po tential. Journal of Nematology 30:313

– 340.

Ciancio 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. 199 4. Host records and spore morphometrics of Pasteuria 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: rediscovery 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 Pasteu ria penetrans group from phytoparasitic nematodes 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 N ematodes. Eds. A. Cianio and K.G. Mukerji. Springer. 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

Globo dera

. 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 Helminthologi cal Society of Washington 52:149

– 165.

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 o f plant

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

– 213.Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 Page 27 of 36 Biopesticides Registration Action Document

A PPENDIX A. MICROBIAL PESTICIDES DATA REQUIREMENTS (40 C

. F

. R

. PART 158

– SUBPART V )

TABLE 1. Product Analysis Data Requ irements 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 M P

885.1100 Product Identity N/A Submitt ed data fulfill the requirement 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 Submitted 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 nem atode)

– P h 3 is on deposit with the American Type Culture Collection 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 Analysis 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 adeq uate/acceptable. 490450

- 03Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 Page 28 of 36 Biopesticides Registration Action 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 Sub mitted data fulfill the requirement for product identity. NewPro contains 16.6650% by weight Pasteuria usgae

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

– Ph3 (contains

at least 1 x 10 8 spores per mL of each isolate )

. 4878 75

- 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 S ubmitted data fulfill the requiremen t for discussion of formation 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, EPA is requiring confirmatory 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

- 02

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 Color 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 Me tal I ons After 14 days at 54°C, no loss of potency. N/A 487876

- 08

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

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

- year storage stabi lity 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 #2 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 Density

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

- 06Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 Page 29 of 36 Biopesticides Registration Action 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 Color

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

re gistration, EPA is requiring the results 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 t est note #2

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, EPA is requiring the results of a one

- year corrosion characteristics study be submitted within one year of registra tion. 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 en d

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

– Ph3 Page 30 of 36 Biopesticides Registration Action 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 anim al.

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 mammali an 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) t hat resulted in no reported incidence of toxicity 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 food 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 P asteuria 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 hypersen sitivity 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 EPA (refer to test note #3 of 40 C

. F

. R

. § 158.2140(d))

. 487876

- 11

885.3500 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 31 of 36 Biopesticides Registration Action 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 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 487876

- 13

870.1200 Acute Dermal

To xicity 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

, a nd 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 I II 1 487876

- 13

870.1300 Acute Inhalation 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 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 487876

- 13Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 Page 32 of 36 Biopesticides Registration Action 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 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 n ot 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 487876

- 13

870.2500 Primary Dermal

Irritation N/A Waived based on the applicant’s scientific rationale, which i ncluded 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 expe cted 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

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

– Ph 3, based on the lack o f 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 previously 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 to xicity ), 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 pe sticide product because of the lack of adverse effects 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 Action Document

TABLE 6

. Toxicology Data Requirements for the End

- Use 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 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 bas ed 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 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

870.120 0 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 Pasteur ia species and cited to the results of MRID No.

474267

- 12

, 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 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 effect s 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 toxicit y, 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 488957

- 01

870.2400 Acute Eye Irritation Wa ived 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 th is 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

- 01Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3 Page 34 of 36 Biopesticides Registration Action Document

TABLE 6

. Toxicology Data Requirements for the End

- Use 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 o n 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 formulati on 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

1 The end

- use pesticide product, NewPro, was assigned this To xicity Category because of the similarity between this product (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 toxicit y), 482103

- 09 (oral toxicity), 482061

- 10 (dermal toxicity), 482103

- 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 components, 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 a nd 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 35 of 36 Biopesticides Registration Action Document

TABLE 7

. Nontarget Organism Toxicity and Environmental Fate Data Requirements for the Te chnical Grade of the Active Ingredient (TGAI), Pasteuria spp. ( Hoplolaimus galeatus 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 sufficie nt information to determine that toxicity/pathogenicity to avian wildlife is not expected. Classification: Acceptable 487876

- 14

885.4100 Avian Inhalation Toxicity/Pathogenicity Not required as the nature of the microbial pesticide does not indicate potent ial 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 adverse effects due to oral exposure. Classification: Acceptable for wild mammal

risk assessment N/A

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

freshwater fish and 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 to 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 of 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

885.4380 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 fo r Tier I

. 1 Additional rationale was submitted for the end

- use pesticide product, 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 Action Document

APPEN DIX B. PESTICIDE PRODUCTS

EPA File Symbol Registration

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

85004

- RN Pasteuria spp. ( Hoplolaimus

galeatus nematode)

– Ph3 99.88% Technica l N/A

85004

- RR NewPro 16.6650% 1

16.6450% 2 End Use

– Liquid Turfgrass (not

to be fed to or

grazed by livestock) Ground application equipment or chemigation 1.3

– 64 fluid ounces of NewPro per 1,000 square feet of

turfgrass Lance Nematode ( Hoplolaimus galeatu s )

Sting Nematode ( Bel o nolaimus longicaudatus )

1 Pasteuria usgae

– B 1 1

2 Pasteuria spp. ( Hoplolaimus galeatus nematode)

– Ph3