BIOPESTICIDES REGISTRATION ACTION DOCUMENT

Pasteuria spp. (Hoplolaimus galeatus nematode) – Ph3

Pesticide Chemical (PC) Code: 016457

U.S. Environmental Protection Agency

Office of Pesticide Programs

Biopesticides and Pollution Prevention Division

DRAFT – March 26, 2013
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Biopesticides Registration Action Document

TABLE OF CONTENTS

I. EXECUTIVE SUMMARY...........................................................................................................................4

II. ACTIVE INGREDIENT OVERVIEW........................................................................................................6

III. REGULATORY BACKGROUND ..............................................................................................................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 JUSTICE.................................................................................................................15

VI. RISK MANAGEMENT DECISION..........................................................................................................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) – Ph3 Pesticide Product Registrations.................................19

B. Environmental Protection Agency Risk Assessment Memoranda .............................................25

C. Other References............................................................................................................................25

APPENDIX A. MICROBIAL PESTICIDES DATA REQUIREMENTS............................................................27

APPENDIX B. PESTICIDE PRODUCTS..............................................................................................................36
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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 Leader
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Biopesticides Registration Action Document

I. EXECUTIVE SUMMARY

Background

In March 2012, Pasteuria Bioscience, Inc. submitted applications for a manufacturing-use 

pesticide 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 Agency (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 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 grampositive, mycelial, endospore-forming bacteria are obligate parasites (i.e., organisms that depend 

on particular hosts to complete their own life cycle) of nematodes and water fleas.1 Pasteuria 

species are ubiquitous in most environments and are found in nematodes in at least 80 countries 

on 5 continents, as well as on islands in the Atlantic, Pacific, and Indian Oceans (Chen and 

Dickson 1998; Ciancio et al. 1994; Sturhan 1988). Higher 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) – Ph3 was specifically isolated from turfgrass soil 

samples collected at the DeBary Golf Course in central Florida. 

Endospores of Pasteuria spp. (Hoplolaimus galeatus nematode) – Ph3 attach to the lance 

nematode (Hoplolaimus galeatus) at all life stages, except the egg stage. 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, which leads to its death. In light of the demonstrated 

nematicidal capabilities and host specificity of Pasteuria spp. (Hoplolaimus galeatus nematode) 

– Ph3, Pasteuria Bioscience, Inc. proposed to register a manufacturing-use pesticide product and 

an end-use pesticide product that is intended for use as a soil or foliar spray on turfgrass to 

control the lance nematode. The end-use pesticide product also contains Pasteuria usgae – Bl1, 

another active ingredient that is present in three currently registered pesticide products2 and is 

intended to control the sting nematode (Belonolaimus longicaudatus). 

EPA scientists 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 registration of the two proposed Pasteuria spp. (Hoplolaimus galeatus nematode) – Ph3 

 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), EconemTM (EPA Reg. No. 85004-2), and Pasteuria usgae – Liquid 

Formulation (EPA Reg. No. 85004-3).
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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 microbial 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 product 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 mammalian toxicology data and information were submitted to support the Pasteuria

spp. (Hoplolaimus galeatus nematode) – Ph3 pesticide products. An acute oral

toxicity/pathogenicity study showed that, at a single high dose, Pasteuria spp. (Hoplolaimus 

galeatus nematode) – Ph3 is not toxic and/or pathogenic via this route of exposure. Based on the 

applicant’s scientific rationale, which included an independent survey of published scientific 

literature that showed no adverse effects from human exposure to any Pasteuria species and 

citation to previous toxicological studies done on other registered Pasteuria species3 that resulted 

in no reported incidences of toxicity and/or pathogenicity, EPA waived the requirement for acute 

pulmonary and acute injection toxicity/pathogenicity testing. Moreover, the applicant reported 

that no hypersensitivity incidents occurred during research, development, or testing of Pasteuria 

spp. (Hoplolaimus galeatus nematode) – Ph3. In light of the results of the acute 

toxicity/pathogenicity data and 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) – Ph3, 

baseline personal protective 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) – Ph3 in agricultural settings 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 products, 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 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 environmental risk 

assessment based on information provided by the applicant, and determined that adverse effects 

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 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.
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Public Participation

On October 1, 2009, EPA announced a policy to provide a more meaningful opportunity for the 

public to participate in major registration 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 applications: new active ingredients; first food uses; first outdoor uses; first residential 

uses; 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 

pesticide products containing Pasteuria spp. (Hoplolaimus galeatus nematode) – Ph3, 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 galeatus nematode) – Ph3. 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-2012-

0609. The following documents are available for comment in EPA-HQ-OPP-2012-0609: (1) 

draft Pasteuria spp. (Hoplolaimus galeatus nematode) – Ph3 Biopesticides Registration Action 

Document (BRAD); (2) environmental risk assessment for Pasteuria spp. (Hoplolaimus galeatus

nematode) – Ph3; (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 Symbol: 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 assessment for

Pasteuria spp. (Hoplolaimus galeatus nematode) – Ph3, which is characterized throughout this 

BRAD. 

II. ACTIVE INGREDIENT OVERVIEW

Biological Name: Pasteuria spp. (Hoplolaimus galeatus nematode) – Ph3

Culture Deposit: American Type Culture Collection in Manassas, Virginia 

under Accession Number SD-5832

OPP Chemical Code: 016457

Type of Pesticide: Microbial Pesticide – Nematicide

See Appendix B for specific information (e.g.,

use sites, application rates, methods of application, 

formulation types, and target pests) regarding

the proposed pesticide products containing this 

active ingredient.
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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 applications to register a manufacturing-use

pesticide product, Pasteuria spp. (Hoplolaimus 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 FR 59611), EPA announced receipt of these

applications to register pesticide products containing a new active ingredient and opened a 30-

day public comment period pursuant to the provisions of FIFRA section 3(c)(4). No comments 

were received following this publication. 

IV. RISK ASSESSMENT

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

EPA issued Final Rules 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” indicates the study is scientifically sound and is 

useful for risk assessment. A “supplemental” rating indicates the data provide some information 

that can be useful for risk assessment. The studies may have certain aspects determined not to be 

scientifically acceptable (“supplemental: upgradeable”). If a study is rated as “supplemental: 

upgradeable,” EPA always provides an indication of what is lacking or what can be provided to 

change the rating to “acceptable.” If there is simply a “supplemental” rating, the reviewer will 

often state that the study is not required by 40 CFR Part 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 acute toxicity data requirements, Toxicity Categories are assigned based on the 

hazard(s) identified from studies and/or other information submitted to 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 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) – Ph3 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.
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B. Human Health Assessment (40 C.F.R. § 158.2140)

1. Toxicity

All toxicology data requirements for Pasteuria spp. (Hoplolaimus galeatus nematode) – Ph3

have been fulfilled. Acceptable Tier I mammalian toxicology data and information support 

registration of the Pasteuria spp. (Hoplolaimus galeatus nematode) – Ph3 pesticide products. 

Furthermore, Tier II and Tier III studies were not required for Pasteuria spp. (Hoplolaimus 

galeatus nematode) – Ph3, 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 108 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 

treatment groups were compared: 1 group was dosed with 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 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 in 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 nematode) – Ph3 does not appear to be toxic and/or pathogenic in rats when dosed by 

oral gavage at 3 x 108 spores per animal. This study was rated supplemental.

Acute Pulmonary Toxicity/Pathogenicity (Harmonized Guideline 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 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 pathogenicity, EPA waived acute pulmonary and acute 

injection toxicity/pathogenicity testing for Pasteuria spp. (Hoplolaimus galeatus nematode) –

Ph3. 
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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 single dose of approximately 1–3 x 108 spores of Pasteuria usgae – Bl1. One dosed female 

exhibited pale lungs. Additionally, one untreated control female lost weight by Day 21, and 

another untreated control female lost weight by Day 14 but gained weight by Day 21. One 

microbial pest control agent (MPCA)-treated male did not gain weight by Day 7 but gained 

weight thereafter. All other animals gained weight throughout the study. Based on these results, 

Pasteuria usgae – Bl1 did not appear to be toxic and/or pathogenic in rats when dosed at 

approximately 1–3 x 108 spores per animal. Since microbial enumeration was not performed 

because the test material would not grow on agar media, the infectivity was uncertain; however, 

because the spores are highly specific to the sting nematode, infectivity is unlikely to be a 

concern. This study was rated acceptable, and Pasteuria usgae – Bl1 was classified as Toxicity

Category 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 108

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 1.5 x 108

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 

pathogenicity to rats following a single intratracheal instillation of 1.5 x 108 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 108 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, 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. No 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 108 Pasteuria usgae – Bl1 
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Biopesticides Registration Action Document

spores. One treated female lost weight by Day 7 but gained weight prior to sacrifice on Day 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 acute intravenous median lethal dose (LD50) of Pasteuria usgae – Bl1 was greater than 1 x 

108 spores per animal in male and female rats. Pasteuria usgae – Bl1 did not appear to be toxic 

and/or pathogenic in rats when dosed at 108 spores per animal. Since microbial enumeration was 

not performed because the test material would not grow on agar media, the infectivity was 

uncertain; however, because the spores are highly specific to the sting nematode, infectivity is 

unlikely to be a concern. This study was rated acceptable.

Pasteuria spp. (Rotylenchulus reniformis nematode) – Pr3, MRID No. 481460-11: 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 107

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 107 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 107 spores per rat. This study 

was rated supplemental.

Pasteuria nishizawae – Pn1, MRID No. 481517-11: Groups of rats were treated with live test 

substance at 1.0 x 109 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 Days 14 and 21 and liver weight 

on Day 21 were slightly but significantly decreased. In female rats from the same group, there 

were no statistically significant differences in organ weights. No evidence of pathogenicity or 

toxicity was found from intravenous administration 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 no 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 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)).
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b. Acute Toxicology and Subchronic Toxicity/Pathogenicity – Tier II;

 Reproductive Fertility Effects, Carcinogenicity, Immunotoxicity, and 

 Infectivity/Pathogenicity Analysis – Tier III

Tier II and Tier III studies were not required for Pasteuria spp. (Hoplolaimus 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 Disruptor Screening Program (EDSP) and has 

begun to implement the screening program that is to be used to test all pesticides in order to 

determine whether certain substances (including pesticide active and other ingredients) may have 

an effect in humans or wildlife similar to an effect produced by a “naturally occurring estrogen, 

or other such endocrine effects as the Administrator may designate.” 

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

requirements of the Estrogenic Substances Screening Program a biologic substance or other 

substance if a determination is made that the substance is not anticipated to produce any effect in 

humans similar to an effect produced by a naturally occurring estrogenic substance. 

Between October 2009 and February 2010, EPA issued test orders/data call-ins for the first group 

of 67 chemicals, which contains 58 pesticide active ingredients and 9 inert ingredients. 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 that 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 naturally occurring estrogenic substance. As such, pursuant to 

Section 408(p)(4), 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 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 visit our website 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. Regardless, appropriate personal protective equipment and precautionary 

statements are required on pesticide product labels to mitigate any potential risks to pesticide 

handlers due to prolonged 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, 

additional PPE (e.g., protective eyewear), other than the standard described above, may be 

required on a product-specific basis.
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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 

unreasonable adverse effects to the U.S. population in general, and to infants and children in 

particular, 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 environmental risk 

assessment based on information provided by the applicant, and determined that adverse effects 

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 critical 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 Characterization 

a. Terrestrial Animals and Plants

Birds (Harmonized Guidelines 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 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 will not necessarily be 

eliminated.

Scientific rationale provides sufficient information to conclude that adverse effects are not 

expected in birds, nontarget insects, and honey bees as a result of exposure to Pasteuria spp. 

(Hoplolaimus galeatus nematode) – Ph3. Pasteuria species are widespread in their distribution 

throughout the 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 that is known only to parasitize Daphnia species 

(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 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. The 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 this 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.

Nontarget 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 pathogenicity 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 journals (i.e., Journal of Bacteriology, Applied and Environmental 

Microbiology, FEMS Microbiology Ecology, Soil Biology and Biochemistry) for the period of 

1980–2012. The search was conducted using “Pasteuria” as the search word, and returned no 

reports of acute toxicity data on birds, freshwater fish, freshwater invertebrates, nontarget insects, 

and honey bees. 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,” 

“arthropod,” “insect,” “honey bee,” “plant pathogen,” and “phytopathogen” for 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 future 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 – Bl1,

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 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 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

Biopesticides Registration Action Document

b. Aquatic Animals and Plants

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

Guideline 885.4240), Estuarine/Marine Fish and Invertebrates (Harmonized Guideline 

885.4280), and Nontarget Plants (Harmonized Guideline 885.4300) (MRID No. 487876-14):

Applications 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 species 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 

for 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 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 understood that all other known Pasteuria

species 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 

freshwater fish or invertebrates as a result of exposure to Pasteuria spp. (Hoplolaimus galeatus

nematode) – Ph3. A search, similar to the one described for terrestrial animals and plants, was 

performed for aquatic animals and plants. In addition to the search 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 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 hosts.

Based 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 galeatus nematode) – Ph3. As 

noted above, due to the specificity of Pasteuria species 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 containing both Pasteuria

species (Pasteuria spp. (Hoplolaimus galeatus nematode) – Ph3 and Pasteuria usgae – Bl1).

2. Environmental Fate Data 

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

and allow for nontarget organism risk assessment 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 36

Biopesticides Registration Action Document

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

3. Threatened and Endangered Species Assessment

Since EPA has determined that no effects are anticipated for any nontarget 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 habitats 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. (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—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 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 community 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 participants 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 address 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 galeatus 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 justice 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

VI. RISK MANAGEMENT DECISION

Section 3(c)(5) of FIFRA permits for the registration of a pesticide provided that all the 

following determinations are made:

(1) Its 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 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 unreasonable 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 labels, and

such pesticide products are not expected to cause unreasonable adverse effects on the 

environment when used according to their respective label instructions. Criterion 2 is satisfied by 

the current product labels, 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 NewPro (end-use pesticide 

product), in particular, is likely to provide protection against lance and sting nematodes 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 and 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. 

VII. ACTIONS REQUIRED OF THE REGISTRANT

A. Final Printed Labeling

Before releasing pesticide products containing Pasteuria spp. (Hoplolaimus galeatus nematode) 

– Ph3 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 Stability (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 (Harmonized 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 commercial 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 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 these types of data are described 

below.

Reports of all incidents of adverse effects to the environment must be submitted to EPA under 

the provisions stated in FIFRA section 6(a)(2). Additionally, all incidents of hypersensitivity 

(including both suspected and confirmed incidents) must be reported to EPA under the 

provisions of 40 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 Federal Register

g/cm3 grams per cubic centimeter

lb/gal pounds per gallon

LD50 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 milligrams 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 States
Pasteuria spp. (Hoplolaimus galeatus nematode) – Ph3 Page 19 of 36

Biopesticides Registration Action Document

IX. BIBLIOGRAPHY

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

 nematode) – Ph3 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: 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 pages.
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, 2262/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 Institute, 

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. 

Project Number: 2262/SN12. Unpublished study prepared by IIT Research Institute, 30 

pages.

481517-15

Sullivan D. 2010. Acute Eye Irritation Study of Pasteuria nishizawae Pn1 in Rabbits. 

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 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 

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

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. 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 spp. (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: NewPro (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 Number: 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]. 

Project 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 Candidatus 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 study 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/Pathogenicity 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 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 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 pages.

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 Pasteuria 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 by 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 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 

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 Rationale: 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. Project 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, Inc., 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 Agency Risk Assessment Memoranda

U.S. EPA. 2013a. Pasteuria spp. (Hoplolaimus galeatus nematode) – Ph3 – 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. 2013b. Environmental Risk Assessment of Pasteuria spp. (Hoplolaimus galeatus 

nematode) – Ph3 (PC 016457) for a Section 3 Registration of the Technical Product (EPA 

File Symbol 85004-RN) and End Use Product (EPA 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. 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 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. 2009a. 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, 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 Chemistry 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 potential. 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. 1994. 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 Pasteuria 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 Nematodes. 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 

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.

Sturhan D. 1988. New host and geographical records of nematode-parasitic bacteria of the 

Pasteuria penetrans group. Nematologica 34:350–356.

Tain B, Yang J, Zhang K-Q. 2007. Bacteria used in the biological control of plant-parasitic 

nematodes: 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

APPENDIX 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) – Ph3 (40 C.F.R. § 158.2120)

Harmonized 

Guideline 

Number 

Data Requirement

Results

MRID No.

TGAI MP

885.1100 Product Identity N/A

Submitted data fulfill the requirement

for product identity. Pasteuria spp. 

(Hoplolaimus galeatus nematode) –

Ph3 contains 99.88% by weight 

Pasteuria spp. (Hoplolaimus galeatus

nematode) – Ph3 (contains at least 1 x 

108 spores per mL).

487876-01

885.1200 Manufacturing 

Process Submitted data fulfill the requirement for manufacturing process. 487876-02

885.1250

Deposition of a 

Sample in a 

Nationally 

Recognized Culture 

Collection

Pasteuria spp. (Hoplolaimus 

galeatus nematode) – Ph3 is on 

deposit with the American Type 

Culture Collection in Manassas, 

Virginia under Accession Number 

SD-5832.

N/A 487876-01

885.1300

Discussion of 

Formation of 

Unintentional 

Ingredients

Submitted data fulfill the requirement 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 

adequate/acceptable.

490450-03
Pasteuria 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 (EP), 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 16.6650% by weight Pasteuria usgae – Bl1 and 16.6450% by 

weight Pasteuria spp. (Hoplolaimus galeatus nematode) – Ph3 (contains 

at least 1 x 108 spores per mL of each isolate). 

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 requirement 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 

Guideline Number Data Requirement

Results

MRID No.

TGAI MP

830.6302 Color Light brown, brown, or redbrown 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 

Elevated

Temperatures, Metals,

and Metal Ions

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 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 #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/cm3 N/A 487876-06
Pasteuria 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 term of the NewPro 

registration, 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 because NewPro 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

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 registration.

487875-07

830.7000 pH 3.5–4.51 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/gal1 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 Action Document

TABLE 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 

Number

Data Requirement

Results

MRID No.

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 108 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 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/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 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 no hypersensitivity incidents, including immediatetype or delayed-type reactions of humans and domestic animals, occurred during 

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) –

Ph3 is not a virus (refer to test note #4 

of 40 C.F.R. § 158.2140(d)).

N/A N/A
Pasteuria spp. (Hoplolaimus galeatus nematode) – Ph3 Page 31 of 36

Biopesticides Registration Action Document

TABLE 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 

Number

Data Requirement

Results

MRID No.

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 IV1

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 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 III1

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 

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

487876-13
Pasteuria spp. (Hoplolaimus galeatus nematode) – Ph3 Page 32 of 36

Biopesticides Registration Action Document

TABLE 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 

Number

Data Requirement

Results

MRID No.

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 not expected to be of 

toxicological concern (refer to test note 

#5 of 40 C.F.R. § 158.2140(d)).

Classification: Acceptable

TOXICITY CATEGORY III1

487876-13

870.2500 Primary Dermal 

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 not expected to be of 

toxicological concern (refer to test note 

#5 of 40 C.F.R. § 158.2140(d)).

Classification: Acceptable

TOXICITY CATEGORY IV1

487876-13

Tiers II and III

Not required for Pasteuria spp. (Hoplolaimus galeatus nematode) – Ph3, 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 Toxicity 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. 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 effects of Pasteuria 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

885.3150 N/A N/A Acute Pulmonary

Toxicity/Pathogenicity

885.3200 Acute Injection 

Toxicity/Pathogenicity

885.3400 Hypersensitivity 

Incidents

The applicant reported than no 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 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 Nos. 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 IV1

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. 

474267-12, 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 III1

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 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 

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

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 not expected to be of toxicological concern (refer 

to test note #5 of 40 C.F.R. § 158.2140(d)).

Classification: Acceptable

TOXICITY CATEGORY IV1

488957-01
Pasteuria 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 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 not expected to be of toxicological concern (refer 

to test note #5 of 40 C.F.R. § 158.2140(d)).

Classification: Acceptable

TOXICITY CATEGORY IV1

488957-01

1 The end-use pesticide product, NewPro, was assigned this Toxicity 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 toxicity), 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 (dermal irritation), 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 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 35 of 36

Biopesticides Registration Action Document

TABLE 7. Nontarget Organism Toxicity and Environmental Fate Data Requirements for the Technical 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 provides sufficient 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 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 adverse effects due to 

oral exposure.

Classification: Acceptable for wild mammal 

risk assessment

N/A

885.4200 Freshwater Fish 

Toxicity/Pathogenicity

The rationale provides sufficient 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 provides 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) – Ph3, based on the acceptability of the rationale provided for 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

APPENDIX 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% Technical 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 galeatus)

Sting Nematode

(Belonolaimus 

longicaudatus)

1 Pasteuria usgae – B11 2 Pasteuria spp. (Hoplolaimus galeatus nematode) – Ph3