BIOPESTICIDES REGISTRATION ACTION DOCUMENT

Pasteuria nishizawae – Pn1 

Pesticide Chemical (PC) Code: 016455

U.S. Environmental Protection Agency

Office of Pesticide Programs

Biopesticides and Pollution Prevention Division

February 28, 2012
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TABLE OF CONTENTS

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

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

III. REGULATORY BACKGROUND ..............................................................................................................6

A. Applications for Pesticide Product Registration ............................................................................6

B. Food Tolerance Exemption ..............................................................................................................7

IV. RISK ASSESSMENT....................................................................................................................................7

A. Product Analysis Assessment (40 CFR § 158.2120) .......................................................................8

B. Human Health Assessment (40 CFR § 158.2140)...........................................................................8

C. Environmental Assessment (40 CFR § 158.2150).........................................................................13

V. ENVIRONMENTAL JUSTICE.................................................................................................................16

VI. RISK MANAGEMENT DECISION..........................................................................................................17

VII. ACTIONS REQUIRED OF THE REGISTRANT....................................................................................17

VIII. GLOSSARY OF ACRONYMS AND ABBREVIATIONS ......................................................................19

IX. BIBLIOGRAPHY .......................................................................................................................................20

A. Studies Submitted to Support the Pasteuria nishizawae – Pn1 Pesticide 

 Product Registrations.....................................................................................................................20

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

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

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

APPENDIX B. PESTICIDE PRODUCTS..............................................................................................................34
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BIOPESTICIDES REGISTRATION ACTION DOCUMENT TEAM

Office of Pesticide Programs (OPP)

Biopesticides and Pollution Prevention Division

Microbial Pesticides Branch

Science Reviews

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

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

Shannon Borges Environmental Effects

Gail Tomimatsu, Ph.D. Environmental Effects

Regulations

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

Jeannine Kausch Regulatory Action Leader
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I. EXECUTIVE SUMMARY

In July and August 2010, Pasteuria Bioscience, Inc. submitted applications for a manufacturinguse pesticide product, Soyacyst Tech (EPA File Symbol 85004-A), and two end-use pesticide 

products, Soyacyst Tech+ (EPA File Symbol 85004-T) and Soyacyst LF (EPA File Symbol 

85004-O), to the United States Environmental Protection Agency (EPA) under section 3 of the 

Federal Insecticide, Fungicide, and Rodenticide Act (FIFRA). Concurrently with these 

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

Pasteuria nishizawae – Pn1, the new active ingredient contained in all three proposed pesticide 

products. 

Background

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, e.g., the Federal Register of December 28, 1994 (59 Federal Register (FR) 66740) and 

June 30, 2010 (75 FR 37734) for Final Rules that established tolerance exemptions for residues 

of the nematicides, Pasteuria penetrans (40 Code of Federal Regulations (CFR) 180.1135) and 

Pasteuria usgae (40 CFR 180.1290), respectively). These gram-positive, mycelial, endosporeforming bacteria are obligate parasites (i.e., organisms that depend on particular hosts to 

complete their own life cycle) of nematodes and water fleas.1 Pasteuria species are ubiquitous in 

most environments and are found in nematodes in at least 80 countries on 5 continents, as well as 

on islands in the Atlantic, Pacific, and Indian Oceans. Higher population densities often occur in 

areas where there is an ample supply of nematode hosts (e.g., where crops susceptible to 

nematodes are cultivated) (Centitas and Dickson 2004; Noel 2008; Tain et al. 2007). Pasteuria 

nishizawae – Pn1 was specifically isolated from an Illinois soybean field in the mid-2000s. 

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

nematodes of the genus Heterodera and one nematode of the genus Globodera, it is known only 

to infect and complete its life cycle within the female soybean cyst nematode (Heterodera 

glycines). In the following manner, Pasteuria nishizawae – Pn1 exerts a pesticidal effect on the 

soybean cyst nematode through parasitism that ultimately results in the death of infected females: 

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

(2) Once a soybean cyst nematode female invades soybean roots, Pasteuria 

nishizawae – Pn1 produces a germ tube that penetrates the body of the nematode.

(3) Primary and secondary microcolonies of Pasteuria nishizawae – Pn1 develop and 

proliferate within the body of the nematode, causing its death.

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

– Pn1, Pasteuria Bioscience, Inc. proposed to register several pesticide products that could be 

applied to soybean or its seed to control the soybean cyst nematode.

EPA scientists reviewed product analysis, toxicology, and nontarget organism data and 

information (40 CFR §§ 158.2120, 158.2140, and 158.2150, respectively) submitted to support 

the registration of the three proposed Pasteuria nishizawae – Pn1 pesticide products. Overall, 

 1 Pasteuria ramosa is the only Pasteuria species that is known to parasitize water fleas. 
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such data and information are adequate for risk assessment purposes, fulfill the current microbial 

pesticide data requirements, and allow for registration under FIFRA section 3(c)(5).

The product analysis data requirements for Pasteuria nishizawae – Pn1, including product 

chemistry and composition, analysis of samples, and physical and chemical characteristics, were 

fulfilled by acceptable guideline studies.

Product Analysis

Adequate mammalian toxicology data and information were submitted to support the Pasteuria 

nishizawae – Pn1 pesticide products. Acute oral, pulmonary, and injection toxicity/pathogenicity 

studies showed that, at a single high dose, Pasteuria nishizawae – Pn1 is not toxic and/or 

pathogenic via these routes of exposure. Moreover, the applicant reported that no hypersensitivity 

incidents occurred during research, development, or testing of Pasteuria nishizawae – Pn1. 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.

Toxicology

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

including infants and children, from aggregate exposure to residues of Pasteuria nishizawae –

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

ingredient in pesticide products. Significant exposure to Pasteuria nishizawae – Pn1 through 

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

end-use pesticide products (soil directed, soil incorporated, and/or seed directed; no aquatic 

applications); (2) the filtering effect of many particulate soil types; and (3) the conditions (e.g., 

filtration and pH adjustments) water is subjected to in wastewater treatment systems and drinking 

water facilities. Should Pasteuria nishizawae – Pn1 be present on food or in drinking water, its

specificity for the soybean cyst nematode and supporting acute oral toxicity and pathogenicity 

data indicate that human exposure to this bacterium is not likely to be a concern.

Tolerance Exemption

Despite the low toxicological profile of Pasteuria nishizawae – Pn1, baseline personal protective 

equipment (PPE) is required for handlers that may be exposed to the active ingredient, due to 

their occupation, for prolonged periods or numerous times. Handlers working with Pasteuria 

nishizawae – Pn1 in agricultural settings are directed to wear a long-sleeved shirt, long pants, 

socks, shoes, and a dust/mist filtering respirator meeting National Institute for Occupational 

Safety and Health (NIOSH) standards of at least N-95, R-95, or P-95. EPA may require 

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

Occupational Exposure

Data and other information (e.g., scientific literature) submitted by the applicant to support 

requests to waive nontarget organism testing for Pasteuria nishizawae – Pn1 are sufficient to 

fulfill the relevant microbial pesticide data requirements and for risk assessment purposes. 

Further testing of nontarget organisms at higher tier levels (i.e., Tiers II, III, and IV) is not 

required. EPA performed an environmental risk assessment based on data and other information 

provided by the applicant, and determined that adverse effects to nontarget organisms are not 

anticipated from the proposed pesticidal uses of Pasteuria nishizawae – Pn1. Moreover, EPA

Nontarget Organisms
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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. 

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

the public to participate in major registration decisions before they occur. According to this 

policy, EPA intends to provide a public comment period prior to making a registration decision 

for, at minimum, the 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.

Public Participation

 

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

pesticide products containing Pasteuria nishizawae – Pn1, a new active ingredient, and allowing 

for this active ingredient’s first outdoor and food uses were subject to a 30-day comment period. 

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

register the Pasteuria nishizawae – Pn1 pesticide products, Soyacyst Tech, Soyacyst Tech+, and 

Soyacyst LF. Therefore, EPA maintained that, based upon the risk assessment and information 

submitted in support of registration of such pesticide products, it was appropriate to issue the 

Soyacyst Tech, Soyacyst Tech+, and Soyacyst LF registrations under FIFRA section 3(c)(5). The 

basis for this decision can be found in the risk assessment for Pasteuria nishizawae – Pn1, which 

is characterized throughout this Biopesticides Registration Action Document (BRAD). 

II. ACTIVE INGREDIENT OVERVIEW

Biological Name: Pasteuria nishizawae – Pn1

Culture Deposit: American Type Culture Collection in Manassas, Virginia 

under Accession Number SD-5833

OPP Chemical Code: 016455

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.

III. REGULATORY BACKGROUND

A. Applications for Pesticide Product Registration

In July and August 2010, 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-
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use pesticide product, Soyacyst Tech (EPA File Symbol 85004-A), and two end-use pesticide 

products, Soyacyst Tech+ (EPA File Symbol 85004-T) and Soyacyst LF (EPA File Symbol 

85004-O), under FIFRA section 3. On November 24, 2010 (75 FR 71697) and February 2, 2011

(76 FR 5805), EPA announced receipt of these 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 these publications. 

B. Food Tolerance Exemption

Concurrent with its registration applications and under Federal Food, Drug, and Cosmetic Act 

(FFDCA) section 408(d), Pasteuria Bioscience, Inc. submitted a petition to establish an 

exemption from the requirement of a tolerance for Pasteuria nishizawae – Pn1 (Pesticide Petition

(PP) 0F7749). In the Federal Register of February 4, 2011 (76 FR 6465), EPA announced that 

Pasteuria Bioscience, Inc. proposed to establish an exemption from the requirement of a 

tolerance for residues of the microbial pesticide, Pasteuria nishizawae – Pn1, in or on all raw 

agricultural crops and opened a 30-day comment period. Two comments were received following 

this publication and are addressed in the preamble to the Final Rule. 

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

residues of Pasteuria nishizawae – Pn1 in or on all food commodities when applied as a 

nematicide and used in accordance with good agricultural practices (40 CFR § 180.1311; 77 FR 

8736).

IV. RISK ASSESSMENT

In the Federal Register of October 26, 2007, EPA issued a Final Rule on the data requirements to 

support registration of microbial pesticides and updated the definition for microbial pesticides 

(72 FR 61002). The rule became effective on December 26, 2007. The data and information 

evaluated for this BRAD were 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 CFR § 156.62). 
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A. Product Analysis Assessment (40 CFR § 158.2120)

All product analysis data requirements for Pasteuria nishizawae – Pn1 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. 

B. Human Health Assessment (40 CFR § 158.2140)

1. Toxicity

All toxicology data requirements for Pasteuria nishizawae – Pn1 have been fulfilled. Acceptable 

Tier I mammalian toxicology data and information support registration of the Pasteuria 

nishizawae – Pn1 pesticide products. Furthermore, Tier II and Tier III studies were not required 

for Pasteuria nishizawae – Pn1, based on the lack of acute toxicity/pathogenicity in the Tier I 

studies.

For a summary of the generic toxicology data requirements 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.) 481517-09): Groups of rats were treated with live test 

substance at 1.6 x 109 spores/rat, were treated with killed (autoclaved) test substance, or were in 

1 of 2 untreated control groups. No adverse clinical signs were observed in any test animals. At 

necropsy, no treatment-related gross observations were noted. Organ weights primarily did not 

differ, though in male rats treated with live test substance, brain weights on days 7, 14, and 21

and spleen weight on day 21 were slightly but significantly decreased. In female rats from the 

same group, the only statistically significant difference was heavier stomach/small intestine 

weight on day 7. No evidence of pathogenicity or toxicity was found from oral administration of 

Pasteuria nishizawae – Pn1 to rats. Although clearance and infectivity were not measured, EPA 

believes these endpoints are not a concern given Pasteuria nishizawae – Pn1’s well-established 

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

Acute Pulmonary Toxicity/Pathogenicity – Rat (Harmonized Guideline 885.3150; MRID No. 

481517-10): Groups of rats were treated with live test substance at 1.6 x 108 spores/rat, were 

treated with killed (autoclaved) test substance, or were untreated controls. No adverse clinical 

signs were observed in any test animals. At necropsy, no treatment-related gross observations 

were noted. Organ weights primarily did not differ, though in 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. Although clearance and 

infectivity were not measured, EPA believes these endpoints are not a concern given Pasteuria

nishizawae – Pn1’s well-established host specificity for the soybean cyst nematode. This study 

was rated supplemental.
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Acute Injection Toxicity/Pathogenicity (Intravenous) – Rat (Harmonized Guideline 885.3200; 

MRID No. 481517-11): Groups of rats were treated with live test substance at 1.0 x 109

spores/rat, were treated with killed (autoclaved) test substance, or were untreated controls. No 

adverse clinical signs were observed in any test animals. At necropsy, no treatment-related gross 

observations were noted. Organ weights primarily did not differ, though in males rats treated 

with live test substance, spleen weights on days 14 and 21 and liver weight on day 21 were 

slightly but significantly decreased. In female rats from the same group, there were no 

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

found from intravenous administration of Pasteuria nishizawae – Pn1 to rats. Although clearance 

and infectivity were not measured, EPA believes these endpoints are not a concern given 

Pasteuria nishizawae – Pn1’s well-established host specificity for the soybean cyst nematode. 

This study was rated supplemental.

Hypersensitivity Incidents (Harmonized Guideline 885.3400; MRID No. 481517-12): The 

applicant 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 nishizawae – Pn1. Any future hypersensitivity incidents must be reported to EPA (refer 

to test note #3 of 40 CFR § 158.2140(d)).

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

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

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

 Reproductive Fertility Effects, Carcinogenicity, Immunotoxicity, and 

 Infectivity/Pathogenicity Analysis – Tier III

Tier II and Tier III studies were not required for Pasteuria nishizawae – Pn1, based on the lack of 

acute toxicity/pathogenicity in the Tier I studies.

c. Endocrine Disruptors

As required under FFDCA section 408(p), EPA has developed the Endocrine Disruptor 

Screening Program (EDSP) to 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.” The EDSP employs a two-tiered approach to making the statutorily required 

determinations. Tier 1 consists of a battery of 11 screening assays to identify the potential of a 

chemical substance to interact with the estrogen, androgen, or thyroid (E, A, or T) hormonal 

systems. Chemicals that go through Tier 1 screening and are found to have the potential to 

interact with E, A, or T hormonal systems will proceed to the next stage of the EDSP where EPA 

will determine which, if any, of the Tier 2 tests are necessary based on the available data. Tier 2 

testing is designed to identify any adverse endocrine-related effects caused by the substance, and 

establish a quantitative relationship between the dose and the E, A, or T effect.

Between October 2009 and 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. This list 

of chemicals was selected based on the potential for human exposure through pathways such as 
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food and water, residential activity, and certain post-application agricultural scenarios. This list 

should not be construed as a list of known or likely endocrine disruptors.

Pasteuria nishizawae – Pn1 is not among the group of 58 pesticide active ingredients on the 

initial list to be screened under the EDSP. Under FFDCA section 408(p), EPA must screen all 

pesticide chemicals. Accordingly, EPA anticipates issuing future EDSP orders/data call-ins for 

all pesticide active ingredients. 

For further 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: 

http://www.epa.gov/endo/.

2. Federal Food, Drug, and Cosmetic Act (FFDCA) Considerations

Section 408(c)(2)(A)(i) of FFDCA allows EPA to establish an exemption from the requirement 

for a tolerance (the legal limit for a pesticide chemical residue in or on a food) only if EPA 

determines that the exemption is “safe.” Section 408(c)(2)(A)(ii) of FFDCA defines “safe” to 

mean that “there is a reasonable certainty that no harm will result from aggregate exposure to the 

pesticide chemical residue, including all anticipated dietary exposures and all other exposures for 

which there is reliable information.” This includes exposure through drinking water and in 

residential settings, but does not include occupational exposure. Pursuant to section 408(c)(2)(B) 

of FFDCA, in establishing or maintaining in effect an exemption from the requirement of a 

tolerance, EPA must take into account the factors set forth in section 408(b)(2)(C) of FFDCA, 

which require EPA to give special consideration to exposure of infants and children to the 

pesticide chemical residue in establishing a tolerance exemption, and to “ensure that there is a 

reasonable certainty that no harm will result to infants and children from aggregate exposure to 

the pesticide chemical residue....” Additionally, section 408(b)(2)(D) of FFDCA requires that

EPA consider “available information concerning the cumulative effects of [a particular 

pesticide's] . . . residues and other substances that have a common mechanism of toxicity.”

EPA performs a number of analyses to determine the risks from aggregate exposure to pesticide 

residues. First, EPA determines the toxicity of pesticides. Second, EPA examines exposure to the 

pesticide through food, drinking water, and through other exposures that occur as a result of 

pesticide use in residential settings.

Consistent with section 408(b)(2)(D) of FFDCA, EPA has reviewed the available scientific data 

and other relevant information, and considered its validity, completeness, and reliability and the 

relationship of this information to human risk. EPA has also considered available information 

concerning the variability of the sensitivities of major identifiable subgroups of consumers, 

including infants and children.

Based on the acute toxicity/pathogenicity data and information discussed previously and 

presented in Tables 5 and 6 in Appendix A, the data required for a FFDCA risk assessment for 

Pasteuria nishizawae – Pn1 have been fulfilled.
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a. Aggregate Exposure

In examining aggregate exposure, section 408 of FFDCA directs EPA to consider 

available information concerning exposures from the pesticide residue in food and all other 

nonoccupational exposures, including drinking water from ground water or surface water and 

exposure through pesticide use in gardens, lawns, or buildings (residential and other indoor uses).

Food Exposure and Risk Characterization: Dietary exposure to Pasteuria nishizawae – Pn1, a 

naturally occurring soil bacterium (Atibalentja et al. 2004; Noel et al. 2005; Sayre et al. 1991), is

anticipated to be negligible. For optimal control of the target pest, soybean cyst nematode 

(Heterodera glycines), Pasteuria nishizawae – Pn1 is applied in a manner that facilitates spore 

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

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

minimize the amount of Pasteuria nishizawae – Pn1 residues on above-ground food 

commodities. That is, although Pasteuria nishizawae – Pn1 can be applied to soil, plants, or 

seeds, some seeds are incorporated into the soil immediately after treatment (at-planting, hopper 

box, planter box, or slurry box seed treatments), and pesticide applications made to plants or the 

soil are always followed by irrigation to incorporate Pasteuria nishizawae – Pn1 into the soil. In 

instances where food commodities develop underground or where treated seed is diverted for 

food or feed purposes or to process into oil, exposure to Pasteuria nishizawae – Pn1 residues is a 

more likely scenario. Regardless of the situation, should Pasteuria nishizawae – Pn1 be present 

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

pathogenicity, and/or infectivity is likely to occur with any dietary exposure that results from 

pesticide applications made in accordance with good agricultural practices (see section 

IV(B)(1)(a) and Tables 5 and 6 in Appendix A).

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

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

patterns for Pasteuria nishizawae – Pn1 are soil directed, soil incorporated, and/or seed directed, 

thereby limiting contact with surface water by drift and runoff. Furthermore, ground water is not 

expected to have significant exposure to Pasteuria nishizawae – Pn1 since, like other

microorganisms, this microbial pesticide would likely be filtered out by the particulate nature of 

many soil types (Aislabie et al. 2001; DeFelice et al. 1993; Pang et al. 2008). If Pasteuria 

nishizawae – Pn1 were to be transferred to surface or ground waters (e.g., through spray drift or 

runoff) that are intended for eventual human consumption and directed to wastewater treatment 

systems or drinking water facilities, it may not survive some of the conditions water is subjected 

to in such systems or facilities, including chlorination, pH adjustments, and filtration (Centers for 

Disease Control and Prevention 2009; U.S. EPA 2004). In the remote likelihood that Pasteuria 

nishizawae – Pn1 is present in drinking water (e.g., water not subject to treatment systems or 

facilities), its specificity for the soybean cyst nematode and available data indicate no toxicity, 

pathogenicity, and/or infectivity is likely to occur with any drinking water exposure that results 

from pesticide applications made in accordance with good agricultural practices (see section 

IV(B)(1)(a) and Tables 5 and 6 in Appendix A).

Non-occupational, Residential Risk Characterization: Given Pasteuria nishizawae’s natural 

occurrence in soil (Atibalentja et al. 2004; Noel et al. 2005; Sayre et al. 1991), non-occupational 

exposure to the bacterium is likely already occurring. Additional exposure to Pasteuria 

nishizawae – Pn1 due to pesticidal applications is not expected because all proposed pesticide 
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end-use products are labeled for use in distinct agricultural settings. Even if non-occupational 

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

EPA’s level of concern in light of Pasteuria nishizawae – Pn1’s specificity for the soybean cyst 

nematode and test results that indicated Pasteuria nishizawae – Pn1 is not toxic (acute dermal 

toxicity and acute pulmonary toxicity/pathogenicity), is essentially non-irritating (primary dermal 

irritation), and is not pathogenic (acute pulmonary toxicity/pathogenicity) (see section 

IV(B)(1)(a) and Tables 5 and 6 in Appendix A).

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

Section 408(b)(2)(D)(v) of FFDCA requires that, when considering whether to establish, modify, 

or revoke a tolerance exemption, EPA consider “available information concerning the cumulative 

effects of [a particular pesticide’s] . . . residues and other substances that have a common 

mechanism of toxicity.”

No mechanism of toxicity in mammals has been identified for Pasteuria nishizawae – Pn1, and 

Pasteuria nishizawae – Pn1 does not appear to produce a toxic metabolite against the target pest.

For the purposes of the tolerance action, EPA has assumed that Pasteuria nishizawae – Pn1 does 

not have a common mechanism of toxicity with other substances. Therefore, section 

408(b)(2)(D)(v) of FFDCA does not apply. For information regarding EPA’s efforts to determine 

which chemicals have a common mechanism of toxicity and to evaluate the cumulative effects of 

such chemicals, see EPA’s website at http://www.epa.gov/pesticides/cumulative.

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

FFDCA section 408(b)(2)(C) provides that, in considering the establishment of a tolerance or 

tolerance exemption for a pesticide chemical residue, EPA shall assess the available information 

about consumption patterns among infants and children, special susceptibility of infants and 

children to pesticide chemical residues, and the cumulative effects on infants and children of the 

residues and other substances with a common mechanism of toxicity. In addition, FFDCA 

section 408(b)(2)(C) provides that EPA shall apply an additional tenfold (10X) margin of 

exposure (safety) for infants and children in the case of threshold effects to account for prenatal 

and postnatal toxicity and the completeness of the database on toxicity and exposure unless EPA 

determines that a different margin of exposure (safety) will be safe for infants and children. This 

additional margin of exposure (safety) is commonly referred to as the Food Quality Protection 

Act Safety Factor. In applying this provision, EPA either retains the default value of 10X or uses 

a different additional safety factor when reliable data available to EPA support the choice of a 

different factor.

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

and 6 in Appendix A, as well as Pasteuria nishizawae – Pn1’s host specificity for the soybean 

cyst nematode, EPA concludes that there are no threshold effects of concern to infants, children, 

or adults when Pasteuria nishizawae – Pn1 is used as labeled in accordance with good 

agricultural practices. As a result, EPA concludes that no additional margin of exposure (safety) 

is necessary to protect infants and children and that not adding any additional margin of exposure 

(safety) will be safe for infants and children.

Moreover, based on the same data and EPA analysis as presented directly above, the Agency is 
Pasteuria nishizawae – Pn1 Page 13 of 34

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able to conclude that there is a reasonable certainty that no harm will result to the U.S. 

population, including infants and children, from aggregate exposure to the residues of Pasteuria 

nishizawae – Pn1 when it is used as labeled and in accordance with good agricultural practices as 

a nematicide. Such exposure includes all anticipated dietary exposures and all other exposures 

for which there is reliable information. EPA has arrived at this conclusion because, considered 

collectively, the data and information available on Pasteuria nishizawae – Pn1 do not 

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

children. 

3. Occupational Exposure and Risk Characterization

Handler exposure to Pasteuria nishizawae – Pn1 is not expected to pose any undue risk. 

Regardless, appropriate personal protective equipment and precautionary statements are required 

on pesticide product labels to mitigate any potential risks to pesticide handlers due to prolonged 

or numerous exposures. Handlers applying Pasteuria nishizawae – Pn1 end-use pesticide

products in agricultural settings must wear a long-sleeved shirt, long pants, socks, shoes, and a 

dust/mist filtering respirator meeting NIOSH standards of at least N-95, R-95, or P-95. 

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

required on a product-specific basis.

4. Human Health Risk Characterization

EPA considered human exposure to Pasteuria nishizawae – Pn1 in light of the standard for 

registration in FIFRA and the relevant safety factors in FFDCA. A determination has been made 

that no unreasonable adverse effects to the U.S. population in general, and to infants and children 

in particular, will result when Pasteuria nishizawae – Pn1 pesticide products are used in 

accordance with EPA-approved labeling.

C. Environmental Assessment (40 CFR § 158.2150)

Data and other information (e.g., scientific literature) submitted by the applicant to support 

requests to waive nontarget organism testing for Pasteuria nishizawae – Pn1 are sufficient to

fulfill the relevant microbial pesticide data requirements and for risk assessment purposes. 

Further testing of nontarget organisms at higher tier levels (i.e., Tiers II, III, and IV) is not 

required. EPA performed an environmental risk assessment based on data and other information

provided by the applicant, and determined that adverse effects to nontarget organisms are not 

anticipated from the proposed pesticidal uses of Pasteuria nishizawae – Pn1. Moreover, EPA 

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

designated 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.
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1. Ecological Exposure and Risk Characterization 

a. Terrestrial Animals and Plants

Birds (Harmonized Guideline 885.4050), Wild Mammals (Harmonized Guideline 885.4150), 

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

885.4340), and Honey Bees (Harmonized Guideline 885.4380) (MRID No. 481517-17): The 

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

pesticide product may be applied by soil-directed spray or chemigation, including both soildirected and foliage-directed (e.g., overhead sprinkler) chemigation. Applications are to be 

followed with a sufficient amount of water to move the Pasteuria nishizawae – Pn1 spores into 

the root zone. Therefore, both formulations will primarily deposit Pasteuria nishizawae – Pn1 

into the soil. While this type of application reduces the potential for exposure to many nontarget 

species, exposure to nontarget species consuming, contacting, or living in the soil will not 

necessarily be eliminated.

Data and other information submitted by Pasteuria Bioscience, Inc. to support its waiver requests

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 nishizawae – Pn1.

Pasteuria spp. are widespread in their distribution throughout the world. They are known only as 

parasites of plant-parasitic nematodes, with the exception of Pasteuria ramosa that is known 

only to parasitize Daphnia spp. (Atibalentja et al. 2004; Chen and Dickson 1998; Sayre et al. 

1991). Pasteuria nishizawae is considered to be an obligate parasite of Heterodera glycines and 

is known only to infect and complete its life cycle within females of Heterodera glycines (Noel et 

al. 2005; Sayre et al. 1991). Therefore, Pasteuria nishizawae – Pn1 is not expected to be toxic or 

pathogenic to birds, nontarget insects, or honey bees if they are exposed as a result of the 

proposed pesticidal applications.

An acute oral toxicity/pathogenicity study with laboratory rats (MRID No. 481517-09) and an 

acute oral toxicity study (MRID No. 481517-13) are available to evaluate the potential effects of 

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

laboratory rats were dosed with approximately 1.6 x 109 spores/animal and were observed for 21 

days. No signs of toxic or pathogenic effects were found as a result of the exposure. This study 

was determined to be supplemental because clearance and infectivity were not observed. The 

acute oral toxicity study showed no evidence of toxicity in rats dosed with 5,000 milligrams per 

kilogram bodyweight, and this study was determined to be acceptable. These studies show that 

toxicity/pathogenicity of Pasteuria nishizawae – Pn1 to laboratory rats is not expected. There is 

no reason that the test animals in these studies would not be adequate models with which to 

determine potential effects to wild mammals, so adverse effects to wild mammals are not 

expected as a result of the proposed pesticidal applications of Pasteuria nishizawae – Pn1.

Nontarget plant testing is not required because Pasteuria nishizawae is not related to any known 

plant pathogen. Adverse effects on plants are not expected to result from the proposed pesticidal 

applications of Pasteuria nishizawae – Pn1.

Additionally, there are no reports of toxicity or pathogenicity of Pasteuria nishizawae to 

nontarget terrestrial animals or plants. To support its data waiver rationale, the applicant 
Pasteuria nishizawae – Pn1 Page 15 of 34

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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 1980–2010. The search 

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

data on birds, freshwater fish, freshwater invertebrates, nontarget insects, and honey bees. The 

Biopesticides and Pollution Prevention Division also conducted a search within the 

Environmental Information Database that is available to EPA. This database simultaneously 

searches the Agricola, Biosis Previews, CAB Abstracts, Energy Science and Technology, 

General Science Abstracts, and the National Technical Information Service literature search 

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

terms of “bird,” “mammal,” “invertebrate,” “arthropod,” “insect,” “honey bee,” and “plant” for 

all of the years available (1926–present). The search was also performed with the search term 

“Pasteuria nishizawae.” No unanticipated reports of adverse effects of Pasteuria nishizawae or 

other Pasteuria species in nontarget organisms were found in any of these searches.2 Any reports 

of effects to nontarget organisms related to Pasteuria nishizawae that are found in the future may

trigger the need for testing; however, given the specificity of Pasteuria spp. for their hosts as is 

currently understood, such reports are not expected.

Based on the information above and available data, adverse effects are not expected to occur to 

terrestrial animals or plants as a result of the proposed pesticidal applications of Pasteuria 

nishizawae – Pn1.

b. Aquatic Animals and Plants

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

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

885.4280), and Nontarget Plants (Harmonized Guideline 885.4300) (MRID No. 481517-17):

Since the proposed applications are intended to deposit Pasteuria nishizawae – Pn1 in the soil, 

some runoff of the active ingredient to freshwater and marine/estuarine environments is 

expected. Applications of the liquid formulation and seed treatment, however, are intended 

primarily to put Pasteuria nishizawae – Pn1 in the root zone of the treated plants, so these 

amounts are not likely to be significant. Pasteuria nishizawae and other Pasteuria spp. are 

naturally found in soil, so some natural presence of Pasteuria nishizawae in surface waters is 

expected. To date, no information is available on the size of naturally occurring populations of 

Pasteuria nishizawae. 

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

conclusions 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 nishizawae – Pn1. The rationale was similar to that presented for 

terrestrial organisms in that Pasteuria spp. are found in soils worldwide, and Pasteuria 

nishizawae is known only to parasitize females of Heterodera glycines. Pasteuria ramosa is a 

closely related species that parasitizes Daphnia magna and other Daphnia spp. (Ebert et al. 

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

of soil-dwelling nematodes (Atibalentja et al. 2004; Chen and Dickson 1998; Sayre et al. 1991). 

 2 Pasteuria ramosa is already recognized as parasitizing water fleas and is not considered an “unanticipated” report.
Pasteuria nishizawae – Pn1 Page 16 of 34

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There are no available reports in the literature documenting pathogenicity or toxicity to 

freshwater fish or invertebrates as a result of exposure to Pasteuria nishizawae. 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 reports of adverse 

effects of Pasteuria nishizawae – Pn1 in aquatic nontarget organisms were found in any of these 

searches. As noted above, reports of adverse effects that may be found in the future may trigger 

the need for testing; however, none are anticipated due to the specificity of Pasteuria spp. for 

their nematode hosts.

Based on the 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 nishizawae – Pn1. 

2. Environmental Fate Data 

As the data and information provided are sufficient to fulfill the Tier I nontarget organism data 

requirements and allow for nontarget organism risk assessment for Pasteuria nishizawae – Pn1, 

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 nishizawae – Pn1 as a result of the proposed applications, effects to 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 nishizawae – Pn1, as 

labeled.

V. ENVIRONMENTAL JUSTICE

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

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

implementation, and 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, during the 30-day public participation 

comment period, EPA sought information on any groups or segments of the population who, as a 
Pasteuria nishizawae – Pn1 Page 17 of 34

Biopesticides Registration Action Document

result their location, cultural practices, or other factors, may have atypical, unusually high 

exposure to Pasteuria nishizawae – Pn1, compared to the general population. No public 

comments were received on this particular matter.

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

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

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 nishizawae – Pn1 pesticide products have well-known 

properties. EPA has no knowledge that would contradict the claims made on the Soyacyst Tech, 

Soyacyst Tech+, and Soyacyst LF 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 nishizawae – Pn1

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

Soyacyst Tech+ and Soyacyst LF (end-use pesticide products), in particular, are likely to provide 

protection against soybean cyst nematode as claimed, satisfying criterion 3. Criterion 4 is 

satisfied in that the Pasteuria nishizawae – Pn1 pesticide products are not expected to cause 

unreasonable adverse effects when used according to label instructions. Therefore, Soyacyst 

Tech, Soyacyst Tech+, and Soyacyst LF, containing Pasteuria nishizawae – Pn1 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 nishizawae – Pn1 for shipment, the 

registrant is required to provide appropriate final printed labeling to EPA.
Pasteuria nishizawae – Pn1 Page 18 of 34

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B. Terms of Registration

No additional data are being required as terms of registration.

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 CFR § 158.2140(d).
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VIII. GLOSSARY OF ACRONYMS AND ABBREVIATIONS

BRAD Biopesticides Registration Action Document

CFR 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/mL grams per milliliter

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

MP manufacturing-use pesticide product

MRID No. Master Record Identification Number

NIOSH National Institute for Occupational Safety and Health

OPP Office of Pesticide Programs

PC Code Pesticide Chemical Code

PP Pesticide Petition

PPE personal protective equipment

TGAI technical grade of the active ingredient

U.S. United States

w/w weight to weight
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IX. BIBLIOGRAPHY

A. Studies Submitted to Support the Pasteuria nishizawae – Pn1 Pesticide Product

 Registrations

MRID No. Study Information

481517-01 MacIntosh S, Smith K. 2010. Product Identity: Pasteuria nishizawae – Pn1. 

Project Number: PBI/2010/001. Unpublished study prepared by MacIntosh & 

Associates, Inc. and Pasteuria Bioscience, Inc., 90 pages.

481517-02 Smith K. 2010. Manufacturing Process for Pasteuria nishizawae – Pn1. Project 

Number: PBI/2010/002. Unpublished study prepared by Pasteuria Bioscience, 

Inc., 36 pages.

481517-03 MacIntosh S. 2010. Discussion of Formation of Unintentional Ingredients: 

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

prepared by MacIntosh & Associates, Inc., 29 pages.

481517-04 Smith K. 2010. Analysis of Samples/Enforcement Methods for Pasteuria 

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

pages.

481517-05 Smith K. 2010. Certification of Limits for Pasteuria nishizawae – Pn1. Project 

Number: PBI/2010/004. Unpublished study prepared by Pasteuria Bioscience, 

Inc., 8 pages.

481517-06 Kaminsky M. 2010. Pasteuria nishizawae – Pn1 Product Chemistry: Final Report. 

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

pages.

481517-07 Smith K. 2010. Product Stability to Elevated Temperatures: Pasteuria nishizawae

– Pn1. Project Number: 115/54. PBI/2010/005. Unpublished study prepared by 

Pasteuria Bioscience, Inc., 8 pages.

481517-08 Smith K. 2010. Storage Stability & Corrosion Characteristics for Pasteuria 

nishizawae. Project Number: 13886/10, PBI/2010/002. Unpublished study 

prepared by Pasteuria Bioscience, Inc., 16 pages.

481517-09 Gingras B. 2010. Toxicity/Pathogenicity Testing of Pasteuria nishizawae – Pn1 

Following Acute Oral Challenge in Rats. Project Number: 2262/SN4. 

Unpublished study prepared by IIT Research Institute, 58 pages.

481517-10 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.
Pasteuria nishizawae – Pn1 Page 21 of 34

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MRID No. Study Information

481517-11 Gingras B. 2010. Toxicity/Pathogenicity Testing of Pasteuria nishizawae – Pn1 

Following Acute Injection Challenge in Rats. Project Number: 2262/SN6. 

Unpublished study prepared by IIT Research Institute, 56 pages.

481517-12 MacIntosh S. 2010. Hypersensitivity Incidents: Pasteuria nishizawae – Pn1. 

Project Number: PBI/2010/007. Unpublished study prepared by MacIntosh & 

Associates, Inc., 7 pages.

481517-13 Sullivan D. 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.

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.

481517-17 MacIntosh S, Smith K, Hewlett T. 2010. Request for Waiver from the 

Requirement to Conduct Guideline Studies (Avian Oral Toxicity, Freshwater Fish 

Toxicity/Pathogenicity, Freshwater Invertebrate Toxicity/Pathogenicity, NonTarget Insect Testing, and Honey Bee Testing) for Pasteuria nishizawae – Pn1. 

Project Number: PBI/2010/014. Unpublished study prepared by MacIntosh & 

Associates, Inc., 29 pages.

482103-01 MacIntosh S, Smith K. 2010. Product Identity and Manufacturing Process: 

Pasteuria nishizawae – Pn1 (Liquid Formulation; End Product). Project Number: 

PBI/2010/020. Unpublished study prepared by Pasteuria Bioscience, Inc. and 

MacIntosh & Associates, Inc., 17 pages.

482103-02 MacIntosh S. 2010. Discussion of Formation of Unintentional Ingredients: 

Pasteuria nishizawae – Pn1 (Liquid Formulation; End Product). Unpublished 

study prepared by MacIntosh & Associates, Inc., 6 pages.

482103-03 Smith K. 2010. Analysis of Samples/Enforcement Methods for Pasteuria 

nishizawae – Pn1 (Liquid Formulation; End Product). Unpublished study prepared 

by Pasteuria Bioscience, Inc., 20 pages.
Pasteuria nishizawae – Pn1 Page 22 of 34

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MRID No. Study Information

482103-04 Smith K. 2010. Certification of Limits for Pasteuria nishizawae – Pn1 (Liquid 

Formulation; End Product). Project Number: PBI/2010/022. Unpublished study 

prepared by Pasteuria Bioscience, Inc., 8 pages.

482103-05 Kaminsky M. 2010. Pasteuria nishizawae – Liquid Formulation: Product 

Chemistry – Final Report. Project Number: 14070/10. Unpublished study 

prepared by Stillmeadow, Inc., 8 pages.

482103-06 Smith K. 2010. Storage Stability: Interim Report Pasteuria nishizawae – Pn1 

(Liquid Formulation; End Product). Project Number: PBI/2010/023. Unpublished 

study prepared by Pasteuria Bioscience, Inc., 8 pages.

482103-07 Smith K. 2010. Corrosion Characteristics for Pasteuria nishizawae – Pn1 (Liquid 

Formulation; End Product). Project Number: PBI/2010/024. Unpublished study 

prepared by Pasteuria Bioscience, Inc., 11 pages.

482103-08 MacIntosh S. 2010. Hypersensitivity Incidents: Pasteuria nishizawae – Pn1 

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

study prepared by MacIntosh & Associates, Inc., 11 pages.

482103-09 Sullivan D. 2010. Acute Oral Toxicity Study of Pasteuria nishizawae – Pn1 (End 

Product) in Rats. Project Number: 2262/SN15. Unpublished study prepared by IIT 

Research Institute, 28 pages.

482103-10 Sullivan D. 2010. Acute Dermal Toxicity Study of Pasteuria nishizawae – Pn1 

(End Product) in Rabbits. Project Number: 2262/SN20. Unpublished study 

prepared by IIT Research Institute, 28 pages.

482103-11 Sullivan D. 2010. Acute Eye Irritation Study of Pasteuria nishizawae – Pn1 (End 

Product) in Rabbits. Project Number: 2262/SN21. Unpublished study prepared by 

IIT Research Institute, 22 pages.

482103-12 Sullivan D. 2010. Acute Dermal Irritation Study of Pasteuria nishizawae – Pn1 

(End Product) in Rabbits. Project Number: 2262/SN22. Unpublished study 

prepared by IIT Research Institute, 22 pages.

482744-02 Kaminsky M. 2010. Pasteuria nishizawae – Pn1 and Pasteuria nishizawae – Seed 

Treatment: Product Chemistry – Final Report. Project Number: 14451/10. 

Unpublished study prepared by Stillmeadow, Inc., 10 pages.

482744-05 MacIntosh S. 2010. Request for Waivers from the Requirement to Conduct 

Toxicology Guideline Study: Acute Inhalation Toxicity for Pasteuria nishizawae

– Pn1 & Pasteuria nishizawae – Seed Treatment. Project Number: PBI/2010/027. 

Unpublished study prepared by MacIntosh & Associates, Inc., 7 pages.
Pasteuria nishizawae – Pn1 Page 23 of 34

Biopesticides Registration Action Document

MRID No. Study Information

482744-06 MacIntosh S. 2010. Request for Waivers from the Requirement to Conduct 

Toxicology Guideline Study: Acute Inhalation Toxicity for Pasteuria nishizawae

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

prepared by MacIntosh & Associates, Inc., 7 pages.

485915-01 MacIntosh S, Smith K. 2011. Product Identity: Pasteuria nishizawae – Pn1. 

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

485915-02 Smith K. 2011. Manufacturing Process for Pasteuria nishizawae – Pn1: 

Supplement to MRID No. 481517-02. Unpublished study prepared by Pasteuria 

Bioscience, Inc., 7 pages.

485915-03 MacIntosh S. 2011. Discussion of Formation of Unintentional Ingredients –

Pasteuria nishizawae – Pn1: Supplement to MRID No. 481517-03. Unpublished 

study prepared by MacIntosh & Associates, Inc., 10 pages.

485915-04 Smith K. 2011. Analysis of Samples/Enforcement Methods for Pasteuria 

nishizawae – Pn1: Supplement to MRID No. 481517-04. Unpublished study 

prepared by Pasteuria Bioscience, Inc., 14 pages.

485915-05 Smith K. 2011. Storage Stability: Pasteuria nishizawae – Pn1 – Final Report. 

Project Number: PBI/2010/002. Unpublished study prepared by Pasteuria 

Bioscience, Inc., 13 pages.

485915-06 Kaminsky M. 2011. Pasteuria nishizawae – Pn1: Corrosion Characteristics –

Final Report. Project Number: 13886/10. Unpublished study prepared by 

Stillmeadow, Inc., 10 pages.

486052-01 Smith K. 2011. Storage Stability Pasteuria nishizawae – Liquid Formulation: 

Supplement to MRID No. 482103-06 – Final Report. Project Number: 

PBI/2010/023. Unpublished study prepared by Pasteuria Bioscience, Inc., 8 pages.

486052-02 Kaminsky M. 2011. Corrosion Characteristics Pasteuria nishizawae – Liquid 

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

prepared by Stillmeadow, Inc., 11 pages.

486461-01 Smith K. 2011. New Sources of Inert Ingredients Pasteuria nishizawae – Pn1 

(TGAI/MP) and Pasteuria nishizawae – Seed Treatment (EP). Unpublished study 

prepared by Pasteuria Bioscience, Inc., 51 pages.

486461-02 Smith K. 2011. New Sources of Inert Ingredients Pasteuria nishizawae – Pn1 

(Liquid Formulations; EP) and Pasteuria nishizawae – Stabilized Seed Treatment 

(EP). Unpublished study prepared by Pasteuria Bioscience, Inc., 101 pages.
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B. Environmental Protection Agency Risk Assessment Memoranda

U.S. EPA. 2011a. Pasteuria nishizawae strain Pn1. Memorandum from J.V. Gagliardi, Ph.D. and 

J.L. Kough, Ph.D. to J. Kausch dated August 16, 2011.

U.S. EPA. 2011b. Environmental Risk Assessment of Pasteuria nishizawae Pn1 (PC 016455) for 

a Section 3 Registration of the Technical Product (EPA File Symbol 85004-A) and Two 

End Use Products (EPA File Symbols 85004-T and 85004-O) for Control of the Soybean 

Cyst Nematode in Soybeans. Memorandum from S. Borges and G. Tomimatsu, Ph.D. to 

J. Kausch dated September 15, 2011.

U.S. EPA. 2011c. Pasteuria nishizawae strain Pn1 from Pasteuria Bioscience, Inc. Memorandum 

from J.V. Gagliardi, Ph.D. and J.L. Kough, Ph.D. to J. Kausch dated December 20, 2011.

C. Other References

Aislabie J, Smith JJ, Fraser R, McLeod M. 2001. Leaching of bacterial indicators of faecal 

contamination through four New Zealand soils. Australian Journal of Soil Research

39:1397–1406.

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

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

Heterodera glycines. Journal of Nematology 36:171–180.

Centers for Disease Control and Prevention. 2009. Drinking Water - Water Treatment. Available 

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

Cetintas R, Dickson DW. 2004. Persistence and suppressiveness of Pasteuria penetrans to

Meloidogyne arenaria Race 1. Journal of Nematology 36:540–549.

Chen ZX, Dickson DW. 1998. Review of Pasteuria penetrans: biology, ecology, and biological 

control potential. Journal of Nematology 30:313–340.

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.

DeFelice K, Wollenhaupt N, Buchholz D. 1993. Aquifers and Soil Filter Effect. Available from 

http://extension.missouri.edu/p/WQ24.

Ebert D, Rainey P, Embley TM, Scholz D. 1996. Development, life cycle, ultrastructure, and

phylogenetic position of Pasteuria ramosa Metchnikoff 1888: rediscovery of an obligate

endoparasite of Daphnia magna Straus. Philosophical Transactions of the Royal Society 

of London series B. 351:1689–1701.

Noel GR. 2008. IPM of soybean cyst nematode in the USA. In: Integrated Management and 

Biocontrol of Vegetable and Grain Crops Nematodes. Eds. A. Cianio and K.G. Mukerji. 

Springer. Pages 119–126. 
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Noel GR, Atibalentja N, Domier LL. 2005. Emended description of Pasteuria nishizawae. 

International Journal of Systematic and Evolutionary Microbiology 55:1681–1685.

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

microbes in ten undisturbed soils under effluent irrigation. Vadose Zone Journal 7:97–

111.

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

and endospore-forming bacterium parasitic on cyst nematodes of genera Heterodera and

Globodera. Research in Microbiology 14:551–564.

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

mycelial and endospore-forming bacterium parasitic in plant-parasitic nematodes. 

Proceedings of the Helminthological Society of Washington 52:149–165.

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. 2004. Primer for Municipal Wastewater Treatment Systems. EPA 832-R-04-001. 

Available from http://www.epa.gov/npdes/pubs/primer.pdf.
Pasteuria nishizawae – Pn1 Page 26 of 34

Biopesticides Registration Action Document

APPENDIX A. MICROBIAL PESTICIDES DATA REQUIREMENTS 

(40 CFR PART 158 – SUBPART V)

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

Pn1, and the Manufacturing-Use Pesticide Product (MP), Soyacyst Tech (40 CFR § 158.2120)

Harmonized 

Guideline 

Number 

Data Requirement

Results

MRID No.

Pasteuria nishizawae – Pn1 Soyacyst Tech

885.1100 Product Identity N/A

Submitted data fulfill the requirement

for product identity. Soyacyst Tech 

contains 99.88% by weight Pasteuria 

nishizawae – Pn1 (contains at least 1 x

108 spores per gram).

481517-01

485915-01

486461-01

885.1200 Manufacturing 

Process Submitted data fulfill the requirement for manufacturing process. 481517-02

485915-02

N/A

Deposition of a 

Sample in a 

Nationally 

Recognized Culture 

Collection

Pasteuria nishizawae – Pn1 is on 

deposit with the American Type 

Culture Collection in Manassas, 

Virginia under Accession Number 

SD-5833.

N/A 481517-01

485915-01

885.1300

Discussion of 

Formation of 

Unintentional 

Ingredients

Submitted data fulfill the requirement for discussion of formation of 

unintentional ingredients.

481517-03

485915-03

885.1400 Analysis of Samples Submitted data fulfill the requirement for analysis of samples. 481517-04

485915-04

885.1500 Certification of 

Limits N/A

Limits listed on the confidential 

statement of formula are 

adequate/acceptable.

481517-05
Pasteuria nishizawae – Pn1 Page 27 of 34

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TABLE 2. Product Analysis Data Requirements for the End-Use Pesticide Product (EPs), Soyacyst Tech+ and Soyacyst LF

(40 CFR § 158.2120)

Harmonized

Guideline 

Number

Data Requirement

Results

MRID No. Soyacyst Tech+* Soyacyst LF**

885.1100 Product Identity

Submitted data fulfill the 

requirement for product 

identity. Soyacyst Tech+ 

contains 99.88% by weight 

Pasteuria nishizawae – Pn1 

(contains at least 1 x 108

spores per gram). 

Submitted data fulfill the 

requirement for product 

identity. Soyacyst LF contains 

33.29% by weight Pasteuria 

nishizawae – Pn1 (contains at 

least 1.3 x 107 spores per gram).

481517-01*

485915-01*

486461-01*

482103-01**

486461-02**

885.1200 Manufacturing Process Submitted data fulfill the requirement for manufacturing 

process.

481517-02*

485915-02*

482103-01**

N/A

Deposition of a Sample in a 

Nationally Recognized 

Culture Collection

N/A N/A

885.1300 Discussion of Formation of 

Unintentional Ingredients

Submitted data fulfill the requirement for discussion of 

formation of unintentional ingredients.

481517-03*

485915-03*

482103-02**

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

481517-04*

485915-04*

482103-03**

885.1500 Certification of Limits Limits listed on the confidential statement of formula are 

adequate/acceptable.

481517-05*

482103-04**
Pasteuria nishizawae – Pn1 Page 28 of 34

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TABLE 3. Physical and Chemical Characteristics for the Technical Grade of the Active Ingredient (TGAI), Pasteuria nishizawae –

Pn1, and the Manufacturing-Use Pesticide Product (MP), Soyacyst Tech (40 CFR § 158.2120)

Harmonized 

Guideline Number Data Requirement

Results

MRID No.

Pasteuria nishizawae – Pn1 Soyacyst Tech

830.63021

Color Light brown/caramel brown

481517-06 830.63031 Physical State Liquid

830.63041 Odor Faint odor/similar to dog food

830.63131

Stability to Normal and 

Elevated

Temperatures, Metals,

and Metal Ions

Stable up to 14 days at 54°C without loss of viability 481517-07

830.6317 Storage Stability Stable for one year when stored at 4°C 481517-08

485915-05

830.6319 Miscibility N/A

Not required because

Soyacyst Tech is not an 

emulsifiable liquid form of a 

microbial pesticide (refer to 

test note #2 of 40 CFR § 

158.2120(d)).

N/A

830.6320 Corrosion 

Characteristics N/A Not corrosive to packaging materials in a one-year study.

481517-08

485915-06

830.70001 pH 4.69–5.21 (1% w/w) 481517-06

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

1.046 cSt (40°C) 482744-02

830.73001

Density/Relative 

Density/Bulk Density 

(Specific Gravity)

1.04 g/mL (or 1.04 g/cm3

) 481517-06

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

 Bioscience, Inc. included this information with its application for Soyacyst Tech, it is summarized appropriately in this table.
Pasteuria nishizawae – Pn1 Page 29 of 34

Biopesticides Registration Action Document

TABLE 4. Physical and Chemical Characteristics for the End-Use Pesticide Products (EPs), Soyacyst Tech+ and Soyacyst LF 

(40 CFR § 158.2120)

Harmonized

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

830.63021 Color Light brown/caramel 

brown N/A 481517-06

830.63031 Physical State Liquid N/A 481517-06

830.63041 Odor Faint odor/similar to dog 

food N/A 481517-06

830.63131

Stability to Normal and 

Elevated Temperatures, 

Metals, and Metal Ions

Stable up to 14 days at 

54°C without loss of 

viability

N/A 481517-07

830.6317 Storage Stability Stable for one year when stored at 4°C

481517-08*

485915-05*

482103-06**

486052-01**

830.6319 Miscibility

Not required because the end-use pesticide products, 

Soyacyst Tech+ and Soyacyst LF, are not emulsifiable 

liquid forms of microbial pesticides (refer to test note #2 

of 40 CFR § 158.2120(d)).

N/A

830.6320 Corrosion Characteristics Not corrosive to packaging materials in a one-year study.

481517-08*

485915-06*

482103-07**

486052-02**

830.7000 pH 4.69–5.21 (1% w/w)1 3.5–4.52 481517-06*

830.7100 Viscosity 1.570 cSt (20°C)

1.046 cSt (40°C)

318.2 cSt (20°C)

255.15 cSt (40°C)

482744-02*

482103-05**

830.7300

Density/Relative 

Density/Bulk Density 

(Specific Gravity)

1.04 g/mL (or 1.04 g/cm3

)

1 8.5 lb/gal2 481517-06*

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

 Bioscience, Inc. included this information with its applications for Soyacyst Tech+, it is summarized appropriately in this table. 2 Obtained from the confidential statement of formula for the end-use pesticide product.
Pasteuria nishizawae – Pn1 Page 30 of 34

Biopesticides Registration Action Document

TABLE 5. Toxicology Data Requirements for the Technical Grade of the Active Ingredient (TGAI), Pasteuria nishizawae – Pn1, and 

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

Harmonized

Guideline 

Number

Data Requirement

Results

MRID No.

Pasteuria nishizawae – Pn1 Soyacyst Tech

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 1.6 x 109 spores per 

animal. Although clearance and 

infectivity were not measured, EPA 

believes these endpoints are not a 

concern given Pasteuria nishizawae –

Pn1’s well-established host specificity 

for the soybean cyst nematode.

Classification: Supplemental

N/A 481517-09

885.3150 Acute Pulmonary 

Toxicity/Pathogenicity

Not toxic and/or pathogenic to rats 

when administered by intratracheal 

instillation in a single dose of 1.6 x 108

spores per animal. Although clearance 

and infectivity were not measured, EPA 

believes these endpoints are not a 

concern given Pasteuria nishizawae –

Pn1’s well-established host specificity 

for the soybean cyst nematode. 

Classification: Supplemental

N/A 481517-10

885.3200

Acute Injection 

Toxicity/Pathogenicity 

(Intraperitoneal)

Not toxic and/or pathogenic to rats

when administered intravenously in a 

single dose of 1.0 x 109 spores per 

animal. Although clearance and 

infectivity were not measured, EPA 

believes these endpoints are not a 

concern given Pasteuria nishizawae –

Pn1’s well-established host specificity 

for the soybean cyst nematode. 

Classification: Supplemental

N/A 481517-11

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 or MP. Any 

future hypersensitivity incidents must be reported to EPA (refer to test note 

#3 of 40 CFR § 158.2140(d)). 

481517-12

885.3500 Cell Culture

Not required because Pasteuria 

nishizawae – Pn1 is not a virus (refer to 

test note #4 of 40 CFR § 158.2140(d)).

N/A N/A

870.1100 Acute Oral Toxicity N/A

Oral LD50 combined (male and 

female rats) > 5,000 mg/kg

Classification: Acceptable

TOXICITY CATEGORY IV

481517-13

870.1200 Acute Dermal Toxicity N/A

Dermal LD50 combined (male and 

female rabbits) > 2,000 mg/kg

Classification: Acceptable

TOXICITY CATEGORY IV

481517-14
Pasteuria nishizawae – Pn1 Page 31 of 34

Biopesticides Registration Action Document

TABLE 5. Toxicology Data Requirements for the Technical Grade of the Active Ingredient (TGAI), Pasteuria nishizawae – Pn1, and 

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

Harmonized

Guideline 

Number

Data Requirement

Results

MRID No.

Pasteuria nishizawae – Pn1 Soyacyst Tech

870.1300 Acute Inhalation 

Toxicity N/A

Waived based on the results of 

MRID No. 481517-10 and 

because this formulation contains 

inert ingredients that are not 

expected to be of toxicological 

concern (refer to test note #5 of 

40 CFR § 158.2140(d)).

Classification: Acceptable

TOXICITY CATEGORY III

482744-05

870.2400 Acute Eye Irritation N/A

Soyacyst Tech was essentially 

non-irritating to the eyes of 

rabbits.

Classification: Acceptable

TOXICITY CATEGORY IV

481517-15

870.2500 Primary Dermal 

Irritation N/A

Soyacyst Tech was essentially 

non-irritating to the skin of 

rabbits.

Classification: Acceptable

TOXICITY CATEGORY IV

481517-16

Tiers II and III

Not required for Pasteuria nishizawae – Pn1 based on the lack of acute toxicity/pathogenicity in the Tier I studies.

TABLE 6. Toxicology Data Requirements for the End-Use Pesticide Products (EPs), Soyacyst Tech+ and Soyacyst LF

(40 CFR § 158.2140)

Harmonized

Guideline 

Number

Data Requirement

Results

MRID No. Soyacyst Tech+* Soyacyst LF**

885.3050 Acute Oral 

Toxicity/Pathogenicity N/A N/A

885.3150 Acute Pulmonary 

Toxicity/Pathogenicity N/A N/A

885.3200 Acute Injection 

Toxicity/Pathogenicity N/A N/A

885.3400 Hypersensitivity 

Incidents

The applicant reported than no hypersensitivity incidents, including 

immediate-type or delayed-type reactions of humans and domestic animals, 

occurred during research, development, or testing of the EPs. Any future 

hypersensitivity incidents must be reported to EPA (refer to test note #3 of 

40 CFR § 158.2140(d)).

481517-12*

482103-08**

885.3500 Cell Culture N/A N/A

870.1100 Acute Oral Toxicity

Oral LD50 combined (male and female rats) > 5,000 mg/kg

Classification: Acceptable

TOXICITY CATEGORY IV

481517-13*

482103-09**

870.1200 Acute Dermal 

Toxicity

Dermal LD50 combined (male and female rabbits) > 2,000 mg/kg

Classification: Acceptable

TOXICITY CATEGORY IV

481517-14*

482103-10**
Pasteuria nishizawae – Pn1 Page 32 of 34

Biopesticides Registration Action Document

TABLE 6. Toxicology Data Requirements for the End-Use Pesticide Products (EPs), Soyacyst Tech+ and Soyacyst LF

(40 CFR § 158.2140)

Harmonized

Guideline 

Number

Data Requirement

Results

MRID No. Soyacyst Tech+* Soyacyst LF**

870.1300 Acute Inhalation 

Toxicity

Waived based on the results of MRID No. 481517-10 and because these

formulations contain inert ingredients that are not expected to be of 

toxicological concern (refer to test note #5 of 40 CFR § 158.2140(d)).

Classification: Acceptable

TOXICITY CATEGORY III

482744-05*

482744-06**

870.2400 Acute Eye Irritation

The EP test substances were essentially non-irritating to the eyes of rabbits.

Classification: Acceptable

TOXICITY CATEGORY IV

481517-15*

482103-11**

870.2500 Primary Dermal 

Irritation

The EP test substances were essentially non-irritating to the skin of rabbits.

Classification: Acceptable

TOXICITY CATEGORY IV

481517-16*

482103-12**

TABLE 7. Nontarget Organism Toxicity and Environmental Fate Data Requirements for the Technical Grade of the Active 

Ingredient (TGAI), Pasteuria nishizawae – Pn1 (40 CFR § 158.2150)

Harmonized Guideline 

Number Data Requirement Results MRID No.

Tier I

885.4050 Avian Oral Toxicity

Data and other information provide sufficient 

information to determine that 

toxicity/pathogenicity to avian wildlife is not 

expected as a result of the proposed pesticidal

uses.

Classification: Acceptable

481517-17

885.4100 Avian Inhalation 

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 CFR § 

158.2150(e)).

N/A

885.4150 Wild Mammal 

Toxicity/Pathogenicity

Tests required by 40 CFR § 158.2140 are 

adequate and appropriate for assessment of 

hazards to wild mammals. Studies submitted with 

laboratory rats indicate no adverse effects due to 

oral exposure.

Classification: Acceptable for wild mammal 

risk assessment

N/A

885.4200 Freshwater Fish 

Toxicity/Pathogenicity

Data and other information provide sufficient 

information to determine that 

toxicity/pathogenicity to freshwater fish and 

invertebrates is not expected as a result of the 

proposed pesticidal uses.

Classification: Acceptable

481517-17

885.4240 Freshwater Invertebrate 

Toxicity/Pathogenicity

885.4280 Estuarine/Marine Fish and 

Invertebrate Testing

Not required as the microbial pesticide will not be 

applied directly to water and is not expected to 

enter marine/estuarine environments in amounts 

that would result in significant concentrations 

(refer to test note #6 of 40 CFR § 158.2150(e)).

N/A

885.4300 Nontarget Plant Testing

Not required as the microbial pesticide is not 

taxonomically related to any known plant 

pathogens (refer to test note #7 of 40 CFR § 

158.2150(e)).

N/A
Pasteuria nishizawae – Pn1 Page 33 of 34

Biopesticides Registration Action Document

TABLE 7. Nontarget Organism Toxicity and Environmental Fate Data Requirements for the Technical Grade of the Active 

Ingredient (TGAI), Pasteuria nishizawae – Pn1 (40 CFR § 158.2150)

Harmonized Guideline 

Number Data Requirement Results MRID No.

885.4340 Nontarget Insect Testing

Data and other information provide sufficient 

information to determine that 

toxicity/pathogenicity to honey bees and

nontarget insects is not expected as a result of the 

proposed pesticidal uses.

Classification: Acceptable

481517-17

885.4380 Honey Bee Testing

Tiers II, III, and IV

Not required for Pasteuria nishizawae – Pn1 based on the acceptability of the data and other information provided for Tier I.
Pasteuria nishizawae – Pn1 Page 34 of 34

Biopesticides Registration Action Document

APPENDIX B. PESTICIDE PRODUCTS

EPA 

Registration 

Number

Registration 

Name

Percentage 

Active 

Ingredient

Formulation 

Type

Use 

Site(s)

Method(s) of 

Application

Application Rate(s) Target Pest

85004-6 Soyacyst 

Tech 99.88% Technical N/A N/A N/A N/A

85004-7 Soyacyst 

Tech+ 99.88% End Use –

Liquid Soybean Seed Treatment 

(Preplant or Commercial)

2–5 fluid ounces of 

Soyacyst Tech+ per 100 

pounds of soybean seed

Soybean cyst 

nematode

(Heterodera glycines)

85004-9 Soyacyst LF 33.29% End Use –

Liquid Soybean

Seed Treatment 

(Preplant or Commercial)

2–5 fluid ounces of 

Soyacyst LF per 100 

pounds of soybean seed

Soybean cyst 

nematode

(Heterodera glycines) Ground application 

equipment or 

chemigation

1.3–64 fluid ounces of 

Soyacyst LF per 1,000 

square feet of soybean