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

May 23, 2013
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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........................................................................................................................15

VII. ACTIONS REQUIRED OF THE REGISTRANT..................................................................................................16

A. Final Printed Labeling .......................................................................................................................................16

B. Terms of Registration.........................................................................................................................................16

C. Reporting of Adverse Effects and Hypersensitivity Incidents........................................................................16

VIII. GLOSSARY OF ACRONYMS AND ABBREVIATIONS ....................................................................................17

IX. BIBLIOGRAPHY......................................................................................................................................................18

A. Studies Submitted or Cited to Support the Pasteuria spp. (Hoplolaimus galeatus nematode) – Ph3 

 Pesticide Product Registrations.........................................................................................................................18

B. Environmental Protection Agency Risk Assessment Memoranda .................................................................24

C. Other References................................................................................................................................................24

APPENDIX A. MICROBIAL PESTICIDES DATA REQUIREMENTS..........................................................................26

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 Review

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

John L. Kough, Ph.D. Product Analysis, Human Health

Shannon Borges Environmental Effects

Regulatory Review

Kimberly Nesci Chief, Microbial Pesticides Branch

Jeannine Kausch Regulatory Action Leader
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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) 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 plantparasitic nematodes that attack and cause significant damage to many valuable agricultural crops. (See 

the Federal Register (FR) publications of December 28, 1994 (59 FR 66740), June 30, 2010 (75 FR 

37734), February 15, 2012 (77 FR 8736), and July 9, 2012 (77 FR 40271) for Final Rules that 

established tolerance exemptions for residues of the nematicides, Pasteuria penetrans (40 Code of 

Federal Regulations (C.F.R.) § 180.1135), Pasteuria usgae (40 C.F.R. § 180.1290), Pasteuria 

nishizawae – Pn1 (40 C.F.R. § 180.1311), and Pasteuria spp. (Rotylenchulus reniformis nematode) –

Pr3 (40 C.F.R. § 180.1316), respectively.) These gram-positive, mycelial, endospore-forming bacteria 

are obligate parasites (i.e., organisms that depend on particular hosts to complete their own life cycle) of 

nematodes and water fleas.1 Pasteuria species are ubiquitous in most environments and are found in 

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

Indian Oceans (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). 

The EPA’s 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

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

products for nonfood/nonfeed use on turfgrass, specifically St. Augustine Grass and/or Bermuda Grass 

not to be fed to or grazed by livestock. Overall, such data and information are 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

The applicant fulfilled 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, by acceptable guideline studies. As a term of registration, the EPA 

is requiring a one-year storage stability study.

Toxicology

The applicant submitted adequate mammalian toxicology data and information 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, the 

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, the EPA is not 

requiring testing at higher tiers (i.e., Tiers II and III).

Occupational Exposure

Despite the low toxicological profile of Pasteuria spp. (Hoplolaimus galeatus nematode) – Ph3, the 

EPA is requiring baseline personal protective equipment (PPE) for handlers who, due to their 

occupation, may be exposed to the active ingredient for prolonged periods and/or numerous times. 

Handlers working with Pasteuria spp. (Hoplolaimus galeatus nematode) – Ph3 in agricultural and/or 

commercial settings are directed to wear a long-sleeved shirt, long pants, socks, shoes, waterproof 

gloves, and a dust/mist filtering respirator meeting National Institute for Occupational Safety and Health 

(NIOSH) standards of at least N-95, R-95, or P-95. For future products, the EPA may require additional 

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

Nontarget Organisms

The EPA found that scientific rationale submitted by the applicant are sufficient to fulfill the nontarget 

organism data requirements and for risk assessment purposes. Further testing of nontarget organisms at 

higher tier levels (i.e., Tiers II, III, and IV) is not required. The EPA performed an 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, the EPA made a “No Effect” determination for direct and indirect 

effects to listed species and their designated critical habitats resulting from these same proposed 

 3 Pasteuria Bioscience, Inc. previously conducted a full battery of toxicological testing on Pasteuria usgae – Bl1, Pasteuria 

nishizawae – Pn1, and Pasteuria spp. (Rotylenchulus reniformis nematode) – Pr3.
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pesticidal uses. 

Public Participation

On October 1, 2009, the 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, the 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 the 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,

and allowing for this active ingredient’s first outdoor uses were subject to a 15-day comment period. 

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

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

(Hoplolaimus galeatus nematode) – Ph3 and NewPro. Therefore, the EPA maintained that, based upon 

the risk assessment and information submitted in support of such pesticide products, it was appropriate 

to issue the Pasteuria spp. (Hoplolaimus galeatus nematode) – Ph3 and NewPro registrations under 

FIFRA section 3(c)(5). The basis for this decision can be found in the risk assessment for Pasteuria spp. 

(Hoplolaimus galeatus nematode) – Ph3, which is characterized throughout this Biopesticides 

Registration Action Document (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 registered 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), the 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), the 

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 the EPA’s science reviewers 

and are an indication of the usefulness of the information contained in the documents for risk 

assessment. A rating of “acceptable” indicates the study is scientifically sound and is useful for risk 

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,” the EPA always 

provides an indication of what is lacking or what can be provided to change the rating to “acceptable.” If 

there is simply a “supplemental” rating, the reviewer will often state that 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 the EPA in support of a pesticide 

registration. The active ingredient or particular product is classified into Toxicity Category I, II, III, or 

IV, where Toxicity Category I indicates the highest toxicity and Toxicity Category IV indicates the 

lowest toxicity (see section II of U.S. EPA (2012b)). 

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

With the exception of a one-year storage stability study, which the EPA will require as a term of 

registration, the applicant fulfilled all product analysis data requirements for Pasteuria spp. 

(Hoplolaimus galeatus nematode) – Ph3. 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

The applicant fulfilled all toxicology data requirements for Pasteuria spp. (Hoplolaimus galeatus

nematode) – Ph3. Acceptable Tier I mammalian toxicology data and information support registration of

the Pasteuria spp. (Hoplolaimus galeatus nematode) – Ph3 pesticide products. Furthermore, the EPA is 

not requiring Tier II and Tier III studies for Pasteuria spp. (Hoplolaimus galeatus nematode) – Ph3

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

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. The EPA rated this study as 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 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, 

the EPA waived acute pulmonary and acute injection toxicity/pathogenicity testing for Pasteuria spp. 

(Hoplolaimus galeatus nematode) – Ph3. 

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

Acute Pulmonary Toxicity/Pathogenicity Testing

Pasteuria usgae – Bl1, MRID No. 474267-10: In an acute pulmonary toxicity and pathogenicity 

assessment, there were no test substance-related significant adverse effects seen in rats receiving a single 
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Biopesticides Registration Action Document

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. The EPA rated this study as acceptable and classified Pasteuria usgae – Bl1 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. The EPA rated this study as 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. The EPA rated this study as 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 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. The EPA rated this study as acceptable.
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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. The EPA rated this study as 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. The EPA rated this study as supplemental.

The EPA believes all these data and information, when taken together, clearly indicate that Pasteuria 

spp. (Hoplolaimus galeatus nematode) – Ph3 would not be toxic, infective, and/or pathogenic through 

the inhalation or injection routes of exposure and that further testing is not necessary.

Hypersensitivity Incidents (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 the 

EPA (refer to test note #3 of 40 C.F.R. § 158.2140(d)).

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

(Hoplolaimus galeatus nematode) – Ph3 is not a virus (refer to test note #4 of 40 C.F.R. § 158.2140(d)).

b. Acute Toxicology and Subchronic Toxicity/Pathogenicity – Tier II;

 Reproductive Fertility Effects, Carcinogenicity, Immunotoxicity, and 

 Infectivity/Pathogenicity Analysis – Tier III

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

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

c. Endocrine Disruptors

As required by the Administrator under the Federal Food, Drug, and Cosmetic Act (FFDCA) section 

408(p), the 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 
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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, the 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. 

The 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), 

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

nematode) – Ph3 from the requirements of the Section 408(p) EDSP. In the event the EPA does 

determine to exempt this substance from the EDSP, an order will be issued. 

For further information on the status of the EDSP, the policies and procedures, the list of 67 chemicals, 

the test guidelines and the Tier 1 screening battery, please 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, the EPA is requiring appropriate personal protective equipment and 

precautionary statements 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 and/or commercial settings must wear a longsleeved shirt, long pants, socks, shoes, waterproof gloves, and a dust/mist filtering respirator meeting 

NIOSH standards of at least N-95, R-95, or P-95. For future products, the EPA may require additional 

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

3. Human Health Risk Characterization

The 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 the EPA’s 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.
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C. Environmental Assessment (40 C.F.R. § 158.2150)

The EPA found that scientific rationale submitted by the applicant are sufficient to fulfill the nontarget 

organism data requirements and for risk assessment purposes. Further testing of nontarget organisms at 

higher tier levels (i.e., Tiers II, III, and IV) is not required. The EPA performed an 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, the 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.

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

Biopesticides Registration Action Document

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. The EPA also conducted a search within the

Thomson Reuters Web of Knowledge Database, which simultaneously searches the Web of Science, 

Current Contacts Connect, Medline, and Journal Citation Reports databases. The search was performed 

with the term “Pasteuria” coupled with each of the search terms of “bird,” “mammal,” “invertebrate,” 

“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).

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 

 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

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 enduse 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 nematode) –

Ph3, the EPA is not requiring further testing at higher tier levels (i.e., Tiers II, III, and IV). 

3. Threatened and Endangered Species Assessment

Since the 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.

 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

V. ENVIRONMENTAL JUSTICE

The EPA seeks to achieve environmental justice—the fair treatment and meaningful involvement of all 

people regardless of race, color, national origin, or income—with respect to the development, 

implementation, and enforcement of environmental laws, regulations, and policies. Fair treatment means 

that no group of people, including racial, ethnic, or socioeconomic groups, should bear a 

disproportionate share of the negative 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. The EPA has 

this goal for all communities and persons across the United States. 

To help address potential environmental justice issues, during the 15-day public participation comment 

period, the EPA sought information on any groups or segments of the population who, as a result of their 

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

spp. (Hoplolaimus galeatus nematode) – Ph3, compared to the general population. No public comments 

were received on this particular matter.

For additional information regarding environmental justice issues, please visit the EPA’s web site at 

http://www.epa.gov/compliance/environmentaljustice/index.html.

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. The 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 
Pasteuria spp. (Hoplolaimus galeatus nematode) – Ph3 Page 16 of 36

Biopesticides Registration Action Document

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 the EPA.

B. Terms of Registration

The following item will be a term of the Pasteuria spp. (Hoplolaimus galeatus nematode) – Ph3

registration:

(1) Storage Stability (Harmonized Guideline 830.6317): Within one year of registration, Pasteuria 

Bioscience, Inc. must provide the results of a one-year storage stability study.

The following items will be terms of the NewPro registration:

(1) Analysis of Samples (Harmonized Guideline 885.1400): Within 18 months of registration, Pasteuria 

Bioscience, Inc. must submit four (4) additional batch analyses to allow the EPA to confirm that the 

current NewPro production facilities are following the approved manufacturing process and meeting 

the 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 the 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 the 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 the EPA under the provisions of 40 C.F.R. § 

158.2140(d).
Pasteuria spp. (Hoplolaimus galeatus nematode) – Ph3 Page 17 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

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 18 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 19 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 20 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 21 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 22 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 23 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 24 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. 2012a. 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 25 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.

U.S. EPA. 2012b. Chapter 7 of the Label Review Manual (Precautionary Statements) (Revised July 

2012). Available from http://www.epa.gov/oppfead1/labeling/lrm/chap-07.pdf.
Pasteuria spp. (Hoplolaimus galeatus nematode) – Ph3 Page 26 of 36

Biopesticides Registration 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 SD5832.

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 27 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 3.3 x 107

spores (both isolates combined) per mL). 487875-01

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

885.1250

Deposition of a Sample 

in a Nationally 

Recognized Culture 

Collection

N/A N/A

885.1300

Discussion of 

Formation of 

Unintentional 

Ingredients

Submitted data fulfill the 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, the 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
Pasteuria spp. (Hoplolaimus galeatus nematode) – Ph3 Page 28 of 36

Biopesticides Registration Action Document

TABLE 3. Physical and Chemical Characteristics for the Technical Grade of the Active Ingredient (TGAI)/Manufacturing-Use Pesticide Product (MP), Pasteuria spp. 

(Hoplolaimus galeatus nematode) – Ph3 (40 C.F.R. § 158.2120)

Harmonized Guideline 

Number 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 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, the EPA is requiring the results of a one-year 

storage stability study be submitted within one year of registration.

487876-09

490450-01

830.6319 Miscibility N/A

Not required because Pasteuria spp. 

(Hoplolaimus galeatus nematode) – Ph3 

is not an emulsifiable liquid form of a 

microbial pesticide (refer to test note #3

of 40 C.F.R. § 158.2120(d)).

N/A

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

packaging.

487876-10

490450-02

830.7000 pH 4.28–5.01 (1% w/w) N/A 487876-06

830.7100 Viscosity N/A 1.39–1.51 cSt (20°C)

0.93–1.02 cSt (40°C) 487876-07

830.7300 Density/Relative Density/Bulk 

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, the 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 #3 of 40 C.F.R. § 158.2120(d)). N/A

830.6320 Corrosion Characteristics

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

results of a one-year corrosion characteristics study be submitted within one year 

of registration.

487875-07

830.7000 pH 3.5–4.51 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 immediate-type 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 the 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 the EPA (refer to test note #3 of 40 C.F.R. § 158.2140(d)).

487875-08

885.3500 Cell Culture N/A N/A

870.1100 Acute Oral Toxicity

Waived based on the applicant’s scientific rationale, which 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 

Registration 

Number

Registration Name

Percentage 

Active 

Ingredient

Formulation 

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

85004-10

Pasteuria spp. 

(Hoplolaimus galeatus

nematode) – Ph3

99.880% Technical N/A

85004-11 NewPro 16.6650%1

16.6450%2

End Use –

Liquid Turfgrass Ground application equipment 

or chemigation

1.3–64 fluid ounces of NewPro 

per 1,000 square feet of 

turfgrass

Lance Nematode

(Hoplolaimus galeatus)

Sting Nematode

(Belonolaimus longicaudatus)

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