BIOPESTICIDES REGISTRATION ACTION DOCUMENT
Pasteuria usgae
PC Code: 006545
U.S. Environmental Protection Agency
Office of Pesticide Programs
Biopesticides and Pollution Prevention Division
Last Updated - July 14, 2009
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Pasteuria usgae
Biopesticides Registration Action Document (BRAD) July 2009
Table of Contents
I. EXECUTIVE SUMMARY 4
II. ACTIVE INGREDIENT OVERVIEW 6
III. REGULATORY BACKGROUND 6
IV. RISK ASSESSMENT 6
A. PRODUCT ANALYSIS ASSESSMENT 7
1. Product Chemistry and Composition 7
2. Analysis and Certified Limits 8
3. Physical and Chemical Characteristics 9
B. HUMAN HEALTH ASSESSMENT 9
1. Toxicology 9
2. Dietary Exposure and Risk Characterization 12
3. Drinking Water Exposure and Risk Characterization 12
4. Acute and Chronic Dietary Exposure and Risks for Sensitive
Subpopulations, Particularly Infants and Children 13
5. Occupational, Residential, School, and Day care Exposure
and Risk Characterization 13
6. Aggregate Exposure from Multiple Routes Including
V.
VI.
VII.
VIII.
Dermal, Oral, and Inhalation
7. Cumulative Effects
8. Risk Characterization
C. ENVIRONMENTAL ASSESSMENT
1. Summarv of Non-Target Organism Waiver Rationales
2. Environmental Effects Conclusions
3. Threatened and Endangered Species Assessment
ENVIRONMENTAL JUSTICE
PUBLIC INTEREST FINDING
RISK MANAGEMENT AND REGISTRATION DECISIONS
A. DETERMINATION OF ELIGIBILITY
B. REGISTRATION REVIEW
C. REGULATORY DECISION
D. LABELING
ACTIONS REQUIRED BY THE REGISTRANT
A. FINAL PRINTED LABELING
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14
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16
18
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B. CONDITIONAL, TIME-LIMITED REGISTRATION REQUIRED
DATA/INFORMATION 21
C. REPORTING OF ADVERSE EFFECTS AND
HYPERSENSITIVITY INCIDENTS 22
IX. GLOSSARY OF ACRONYMS AND ABBREVIATIONS 23
X. BIBLIOGRAPHY 24
A. STUDIES SUBMITTED IN SUPPORT OF THIS REGISTRATION 24
B. EPA RISK ASSESSMENT MEMORANDUMS 26
C. OTHER REFERENCES 26
APPENDIX A 28
APPENDIX B 35
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Pasteuria usgae
Biopesticides Registration Action Document (BRAD) July 2009
BIOPESTICIDES REGISTRATION ACTION DOCUMENT TEAM
Office of Pesticide Programs
Biopesticides and Pollution Prevention Division
Microbial Pesticides Branch
Science Reviews
Stuart Schiissel, B.S. Product Analysis, Human Health
John Kough, Ph.D. Product Analysis, Human Health
Shannon Borges, M.S. Environmental Effects
Zigfridas Vaituzis, Ph.D. Environmental Effects
Regulations
Sheryl Reilly, Ph.D. Chief, Microbial Pesticides Branch
Jeannine Kausch, M.S.E.L. Regulatory Action Leader
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Pasteuria usgae
Biopesticides Registration Action Document (BRAD) July 2009
I. EXECUTIVE SUMMARY
Pasteuria spp. are gram-positive, mycelial, endospore-forming bacteria that are
endoparasitic to nematodes and water fleas. Pasteuria usgae, a recently discovered strain,
is host-specific to the sting nematode (Belonolaimus longicaudatus), which can be
damaging to a wide variety of crops, particularly turf. The active agent of Pasteuria
usgae is an endospore that attaches and infects the host nematode during all life stages
(except eggs). After attachment of the endospore, a germ tube penetrates the nematode
cuticle and mycelial microcolonies are formed in the pseudocoelom, leading to eventual
death of the host. The endospores formed inside the host are released into the soil when
the infected nematode decomposes. The spores are non-motile and stable in the soil
environment for several years.
To decide whether to grant a conditional, time-limited registration under Section
3(c)(7)(C) of the Federal Insecticide, Fungicide, and Rodenticide Act (FIFRA) to
Pasteuria usgae - BL1, a proposed manufacturing-use product containing Pasteuria
usgae as an active ingredient at 0.01% by weight, the Biopesticides and Pollution
Prevention Division (BPPD) reviewed microbial pesticide product analysis, toxicology,
and non-target organism and environmental fate data requirements [40 Code of Federal
Regulations (CFR) §§ 158.2120, 158.2140, and 158.2150, respectively]. It was
determined that the data/information submitted adequately satisfy current guideline
requirements for the purposes of this type of registration only.
For the purposes of a conditional, time-limited FIFRA section 3(c)(7)(C) registration
only, product analysis data requirements (to include product chemistry and composition,
analysis and certified limits, and physical and chemical characteristics) for the technical
grade of the active ingredient (TGAI)/manufacturing-use product (MP), Pasteuria usgae
- BL1, were satisfied by acceptable guideline studies and waiver rationales. Additional
product analysis data that the Agency is requiring by specific dates falling within two
years of registration include the following:
• A written description of quality control measures taken during the manufacturing
process to screen for Pasteuria species that parasitize saprophytic nematodes.
• A new five-batch analysis with all batches from production level.
• Results from an ongoing 12-month storage stability study.
Lastly, documentation indicating official recognition of Pasteuria usgae by the Judicial
Commission of the International Committee for Systematic Bacteriology must be
submitted to the Agency when it becomes available.
Adequate mammalian toxicology data/information were submitted to support the
registration of Pasteuria usgae - BL1. Acceptable acute toxicity guideline studies were
submitted, and data waivers were granted by the Agency to fulfill the remaining Tier I
acute toxicity data requirements based on the lack of toxicity and/or pathogenicity of
Pasteuria usgae, because the manufacturing-use product is equivalent to the technical
grade of the active ingredient, and/or the low potential for worker exposure attributed to
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Biopesticides Registration Action Document (BRAD) July 2009
appropriate precautionary statements and requirements for personal protective equipment
on the label.
Dietary, drinking water, and non-dietary, non-occupational exposures to Pasteuria usgae
are unlikely to occur because the use of the proposed product is limited to manufacturing
of end-use products intended for eventual application to turf. The associated end-use
products are still undergoing Agency review. Despite the low toxicological profile of
Pasteuria usgae, personal protective equipment (PPE) is required for handlers that may
be exposed to the active ingredient, due to their occupation, for prolonged periods.
Handlers working with Pasteuria usgae in manufacturing facilities must 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. Overall, a determination has been made that no
unreasonable adverse effects to the United States population in general, and to infants and
children in particular, will result from the use of Pasteuria usgae when used in
accordance with Environmental Protection Agency (EPA)-approved labeling.
Data waiver rationales were submitted in response to data requirements for avian,
freshwater fish and invertebrate, insect, and honey bee non-target organism testing
requirements. The information provided is sufficient to satisfy the Tier I non-target
organism data requirements for the technical product and proposed end-use products
containing Pasteuria usgae as an active ingredient. Further testing of non-target
organisms at higher tier levels is not required. Based on the rationales submitted, adverse
effects to terrestrial animals and plants or freshwater and marine/estuarine fish,
invertebrates, and plants are not expected as a result of exposure to proposed labeled
applications of Pasteuria usgae. Furthermore, BPPD makes "No Effect" (NE)
determinations for direct and indirect effects to listed species and their habitat as a result
of the proposed uses of Pasteuria usgae.
The Agency determines whether conditional registration of a pesticide is in the public
interest in accordance with the criteria set forth in the Federal Register dated March 5,
1986 (58 Federal Register 7268). There is a presumption that registration of a pesticide is
in the public interest if (1) the use is for a minor crop, (2) the use is a replacement for
another pesticide that is of continuing concern to the Agency, (3) the use is one for which
an emergency exemption under FIFRA section 18 has been granted (i.e., the basis for the
exemption was lack of a registered alternative product), or (4) the use is against a pest of
public health significance. Pasteuria usgae - BL1 will be used to create end-use
products, that once reviewed and determined to be eligible for registration under FIFRA,
will serve as partial replacements for conventional nematicides of continuing concern to
the Agency [e.g., methyl bromide (ozone-depleting substance) and 1,3-dichloropropene
(probable human carcinogen)]. Based on this information, this registration is presumed to
be in the public interest.
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Pasteuria usgae
Biopesticides Registration Action Document (BRAD) July 2009
II. ACTIVE INGREDIENT OVERVIEW
Biological Name: Pasteuria usgae
Culture Deposit: American Type Culture Collection in Manassas,
Virginia under Accession Number SD-5835
Office of Pesticide
Programs (OPP)
Chemical Code: 006545
Type of Pesticide: Microbial Pesticide - Nematicide
See Appendix B for specific information (i.e.,
use sites, application rate, method of application,
formulation type, and target pests) regarding
the manufacturing-use product, Pasteuria usgae -
BL1, containing this active ingredient.
III. REGULATORY BACKGROUND
On May 5, 2008, Macintosh and Associates, Incorporated (address: 1203 Hartford
Avenue, Saint Paul, Minnesota 55116-1622), acting as the authorized agent for Pasteuria
Bioscience, Incorporated (address: 12085 Research Drive, Suite 185, Alachua, Florida
32615), submitted an application to register Pasteuria usgae - BL1 (EPA File Symbol
85004-R) under Section 3 of the Federal Insecticide, Fungicide, and Rodenticide Act. On
August 13, 2008, the EPA announced receipt of this application to register a pesticide
product containing a new active ingredient [73 Federal Register (FR) 47166] and opened
a 60-day public comment period. No comments were received following this publication.
Pursuant to FIFRA section 3(c)(7)(C), a conditional, time-limited registration was issued
for Pasteuria usgae - BL1 on June 2, 2009 (EPA Registration Number 85004-1). The
Agency announced the approval to conditionally register Pasteuria usgae - BL1, which
does not contain an active ingredient included in any previously registered product, in the
Federal Register of July 1, 2009 (74 FR 31426).
IV. RISK ASSESSMENT
On October 26, 2007, the Agency issued a Final Rule in the Federal Register on the data
requirements to support registration of biochemical and microbial pesticides, and updated
the definitions for biochemical and microbial pesticides (72 FR 61002). The rule became
effective on December 26, 2007. The data and information evaluated for this
Biopesticides Registration Action Document (BRAD) were considered in light of these
requirements.
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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: UPGRADABLE"). If a study is rated as "SUPPLEMENTAL:
UPGRADABLE," the Environmental Protection Agency 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 the current 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 need to be submitted.
For the acute toxicity data requirements, toxicity categories are assigned based on the
hazard(s) identified from studies and/or information submitted to the Agency. 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.
A. Product Analysis Assessment (40 CFR § 158.2120)
Although there are some product analysis data that are lacking, a reasonable period of
time sufficient for generation of these data has not elapsed since the Agency first imposed
the relevant data requirements. Therefore, all product analysis data requirements for a
conditional, time-limited FIFRA section 3(c)(7)(C) registration of Pasteuria usgae -
BL1, containing Pasteuria usgae as an active ingredient, have been satisfied by either
acceptable guideline studies or waiver rationales. The Agency will allow for generation
and submission of the lacking product analysis data within two years of registration as the
use of the manufacturing-use Pasteuria usgae product will not cause any unreasonable
adverse effects on the environment and registration of such a product is presumed to be in
the public interest.
For a comprehensive guideline-by-guideline summary of the product analysis data
requirements described in sections IV(A)(1), IV(A)(2), and IV(A)(3), refer to Table 1 in
Appendix A.
1. Product Chemistry and Composition [Office of Prevention,
Pesticides, and Toxic Substances (OPPTS) Guidelines 885.1100, 885.1200,
and 885.1300]
Pasteuria spp. are gram-positive, mycelial, endospore-forming bacteria that are
endoparasitic to nematodes and water fleas. The endospores of Pasteuria spp. can be
identified and counted microscopically, but it is difficult to distinguish between species
unless high magnification electron microscopy is used so that size and shape of the spores
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Biopesticides Registration Action Document (BRAD) July 2009
can be visualized and measured. Molecular techniques such as polymerase chain reaction
(PCR) methods have been reported for Pasteuriapenetrans but have not been developed
for the other species. Therefore, species identification via host specificity assays,
requiring the establishment of nematode cultures and a bioassay, is the most direct and
reliable technique for species identification.
Pasteuria usgae, a recently discovered strain, is host-specific to the sting nematode
(Belonolaimus longicaudatus), which can be damaging to a variety of crops, particularly
turf. Currently, full recognition of Pasteuria usgae is still pending with the Judicial
Commission of the International Committee for Systematic Bacteriology. Once Pasteuria
usgae has been recognized and removed from the category Candidates, documentation
indicating official recognition must be provided to the Agency.
The active agent of Pasteuria usgae is an endospore that attaches and infects the host
nematode during all life stages (except eggs). Increased moisture, neutral pH,
temperatures above 10°C, and sandy soil seem to provide the best environments for spore
attachment to the host. In laboratory studies, Pasteuria spp. were able to attach to
nematodes after exposure to high temperatures and wide ranges of pH; however, at
extremes, the number of attached spores per nematode was reduced and, in some cases,
no infection occurred after attachment. After attachment of the endospore, a germ tube
penetrates the nematode cuticle and mycelial microcolonies are formed in the
pseudocoelom, leading to eventual death of the host. The endospores formed inside the
host are released into the soil when the infected nematode decomposes. The spores are
non-motile and stable in the soil environment for several years.
Submitted data also adequately describe the production process and potential microbial
contaminants and these requirements have been satisfied for the purposes of a
conditional, time-limited FIFRA section 3(c)(7)(C) registration only. The original
manufacturing process did not describe quality control measures taken to confirm that
Pasteuria usgae was the only Pasteuria species present in the master stock. A follow-up
explanation, which demonstrated that batches are screened for the presence of other
microbes and screened against sting nematodes to confirm the presence of Pasteuria
usgae, still did not provide sufficient information to indicate that screening for other
Pasteuria species is conducted during the manufacturing process. Other Pasteuria
species, specifically those that parasitize saprophytic (non-plant pathogenic) nematodes,
must be screened for during the manufacturing process. Pasteuria Bioscience,
Incorporated must integrate quality control measures that screen for Pasteuria species
that parasitize saprophytic nematodes, either immediately after freezing or after making
seed stock, into their current manufacturing process and provide a written description of
these measures to the Agency on or before October 1, 2009.
2. Analysis and Certified Limits (OPPTS Guidelines 885.1400 and 885.1500)
Results of a preliminary five-batch analysis were provided and the requirement for
analysis of samples has been satisfied for purposes of a conditional, time-limited FIFRA
section 3(c)(7)(C) registration only. The preliminary analysis of samples provided to the
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Agency revealed a wide range of concentrations, with many replicates containing 1-2
orders of magnitude fewer Pasteuria usgae spores than the minimum specified on the
Confidential Statement of Formula (CSF) and product label (i.e., 1 x 106 spores per
milliliter). A follow-up explanation, attempting to address this variability, mentioned
several factors that contributed to this inconsistency and discussed how the concentration
of spores could be adjusted during the manufacturing process, if necessary. Despite this
additional information, a new five-batch analysis, with all batches from production level,
must be submitted to the Agency on or before May 1, 2011.
The certified limits for the active and inert ingredients fall within the OPPTS Guideline
830.1750 specified ranges; therefore, the requirement for certified limits has been
satisfied.
3. Physical and Chemical Characteristics (OPPTS Guidelines 830.6302,
830.6303, 830.6304, 830.6313, 830.6317, 830.6319, 830.6320, 830.7000,
830.7100, and 830.7300)
Submitted data adequately describe the physical and chemical characteristics and waiver
rationales provided are acceptable; however, a 12-month storage stability study is cited to
as ongoing and miscibility is not required because Pasteuria usgae - BL1 is not an
emulsifiable form of microbial pesticide. Therefore, the requirements for color, physical
state, odor, stability to normal and elevated temperatures, metals, and metal ions,
corrosion characteristics, pH, viscosity, and density/relative density/bulk density (specific
gravity) have been satisfied. When the storage stability study has been completed and the
results have been compiled, these data must be submitted to the Agency on or before June
1,2010.
B. Human Health Assessment
1. Toxicology
Adequate mammalian toxicology data/information are available to support the
registration of Pasteuria usgae - BL1, which contains Pasteuria usgae. All Tier I
toxicology data requirements for the technical grade of the active
ingredient/manufacturing-use product have been satisfied by guideline studies or waiver
rationales. Tier II and Tier III studies were not required for Pasteuria usgae based on the
lack of acute toxicity/pathogenicity in the Tier I studies.
For a comprehensive guideline-by-guideline summary of the toxicology data
requirements described in sections IV(B)(l)(a) and IV(B)(l)(b), refer to Table 2 in
Appendix A.
a. Acute Toxicity/Pathogenicity - Tier I (40 CFR § 158.2140)
Acute Oral Toxicity andPathogenicity - Rat [OPPTS Guideline 885.3050; Master
Record Identification (MPJD) Number (No.) 474267-091: There were no treatment-
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related clinical signs or necropsy findings in rats receiving a single oral dose of 1 x 108
Pasteuria usgae spores. Three males in the microbial pest control agent (MPCA)-treated
group gained weight through day 14 but lost weight by day 21. All other animals gained
weight prior to scheduled sacrifice. Pasteuria usgae does not appear to be toxic and/or
pathogenic in rats when dosed at 1 x 108 spores/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 sting nematode,
infectivity is unlikely to be a concern. This study was rated "ACCEPTABLE" and
Pasteuria usgae was classified as TOXICITY CATEGORY IV.
Acute Dermal Toxicity - Rat (OPPTS Guideline 885.3100; MRID No. 474267-12): There
were no treatment-related significant adverse effects seen in the dosed rats. Two males
and one female had very slight erythema on day 1 with clearance by day 4. One male lost
weight slightly during the second week and one male and two females lost weight during
the first week, but all gained weight by the end of the study. All other animals gained
weight throughout the study. Based on the results of this study, Pasteuria usgae does not
appear to be toxic in rats when treated with 2,000 milligrams (mg)/kilogram (kg) at 108
spores/milliliter (mL). The acute dermal LD50 is greater than 2,000 mg/kg for 108
spores/mL in male and female rats. This study was rated "ACCEPTABLE" and Pasteuria
usgae was classified as TOXICITY CATEGORY IV.
Acute Pulmonary Toxicity andPathogenicity - Rat (OPPTS Guideline 885.3150; MRID
No. 474267-10): In an acute pulmonary toxicity and pathogenicity assessment, there were
no test substance-related significant adverse effects seen in rats receiving a single dose of
approximately 1-3 x 108 spores of Pasteuria usgae. 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 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 does not
appear to be toxic and/or pathogenic in rats when dosed at approximately 1-3 x 108
spores/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 sting nematode, infectivity is unlikely to be a concern. This
study was rated "ACCEPTABLE" and Pasteuria usgae was classified as TOXICITY
CATEGORY IV.
Acute Injection Toxicity andPathosenicity - Rat (OPPTS Guideline 885.3200; MRID
No. 474267-11): There were no treatment-related significant adverse effects seen in rats
receiving a single intravenous dose of 108 Pasteuria usgae 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 LDso of Pasteuria usgae is greater than 1 x 108 spores/animal in male
and female rats. Pasteuria usgae does not appear to be toxic and/or pathogenic in rats
when dosed at 108 spores/animal. Since microbial enumeration was not performed
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Biopesticides Registration Action Document (BRAD) July 2009
because the test material would not grow on agar media, the infectivity was uncertain.
However, because the spores are highly specific to sting nematode, infectivity is unlikely
to be a concern. This study was rated "ACCEPTABLE" and Pasteuria usgae was
classified as TOXICITY CATEGORY IV.
HvversensitivityIncidents (OPPTS Guideline 885.3400; MRIDNo. 474350-02): No
hypersensitivity incidents, including immediate-type or delayed-type reactions of humans
and domestic animals, occurring during the testing or production of the TGAI/MP were
reported by Pasteuria Bioscience, Incorporated. Any future hypersensitivity incidents
must be reported per OPPTS Guideline 885.3400.
Cell Culture (OPPTS Guideline 885.3500): This study is not required because Pasteuria
usgae is not a virus (refer to test note #4 of 40 CFR § 158.2140(d)).
Acute Oral Toxicity (OPPTS Guideline 870.1100; MRID No. 474979-01): Waived based
on the results of MRID No. 474267-09, which showed that Pasteuria usgae was not toxic
and/or pathogenic by oral route of exposure, and because the MP is equivalent to the
TGAI. This waiver request was rated "ACCEPTABLE" and Pasteuria usgae - BL1 was
classified as TOXICITY CATEGORY IV.
Acute Dermal Toxicity (OPPTS Guideline 870.1200; MRID No. 474979-01): Waived
based on the results of MRID No. 474267-12, which showed that Pasteuria usgae was
not toxic by dermal route of exposure, and because the MP is equivalent to the TGAI.
This waiver request was rated "ACCEPTABLE" and Pasteuria usgae - BL1 was
classified as TOXICITY CATEGORY IV.
Acute Inhalation Toxicity (OPPTS Guideline 870.1300; MRID No. 474979-01): Waived
based on the results of MRID No. 474267-10, which showed that Pasteuria usgae was
not toxic and/or pathogenic by pulmonary route of exposure, and because the MP is
equivalent to the TGAI. This waiver request was rated "ACCEPTABLE" and Pasteuria
usgae - BL1 was classified as TOXICITY CATEGORY IV.
Acute Eve Irritation (OPPTS Guideline 870.2400; MRID No. 474350-03): Waived based
on Pasteuria usgae'?, lack of toxicity in the other toxicology studies and precautionary
statements on the label that mitigate potential for worker exposure. This waiver request
was rated "ACCEPTABLE" and Pasteuria usgae - BL1 was classified as TOXICITY
CATEGORY III.
Primary Dermal Irritation (OPPTS Guideline 870.2500; MRID No. 474350-03): Waived
based on the results of MRID No. 474267-12, which showed that Pasteuria usgae was
not toxic by dermal route of exposure, and precautionary statements/personal protective
equipment on the label that mitigate potential for worker exposure. This waiver request
was rated "ACCEPTABLE" and Pasteuria usgae - BL1 was classified as TOXICITY
CATEGORY III.
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b. Acute Toxicology and Subchronic Toxicity/Pathogenicity - Tier II;
Reproductive Fertility Effects, Carcinogenicity, Immunotoxicity, and
Infectivity/Pathogenicity Analysis - Tier III (40 CFR $ 158.2140)
Tier II and Tier III studies were not required for Pasteuria usgae based on the lack of
acute toxicity/pathogenicity in the Tier I studies.
c. Effects on the Endocrine System
Section 408(p) of the Federal Food, Drug, and Cosmetic Act (FFDCA) requires EPA to
develop a screening program to determine whether certain substances (including all
pesticide active and other ingredients) "may have an effect in humans that is similar to an
effect produced by a naturally occurring estrogen, or such other endocrine effect as the
Administrator may designate." Following the recommendations of its Endocrine
Disrupter Screening and Testing Advisory Committee (EDSTAC), EPA determined that
there were scientific bases for including, as part of its program, androgen and thyroid
hormone systems, in addition to the estrogen hormone system. The Environmental
Protection Agency also adopted EDSTAC's recommendation that the Program include
evaluations of potential effects on wildlife.
The Agency has no knowledge of Pasteuria usgae being an endocrine disrupter, nor is
this microbe related to any class of known endocrine disrupters. Due to the unique mode
of action of Pasteuria usgae, the narrow host range, and lack of mammalian toxicity (see
section IV(A)Q) and section IV(B)(l)(a)X no effects on the immune or endocrine
systems are anticipated. Additional data, specifically on the endocrine effects of this
microbial pesticide, are not required at this time. When the appropriate screening and/or
testing protocols being considered under the Agency's Endocrine Disrupter Screening
Program (EDSP) have been developed and vetted, Pasteuria usgae may be subjected to
additional screening and/or testing to better characterize effects related to endocrine
disruption.
2. Dietary Exposure and Risk Characterization
Dietary exposure to Pasteuria usgae is not expected because there are no approved food
uses for this active ingredient at this time.
3. Drinking Water Exposure and Risk Characterization
Exposure of humans to residues of Pasteuria usgae in drinking water is unlikely. The
current use pattern for Pasteuria usgae (i.e., manufacturing into nematicide end-use
products, which are still undergoing Agency review) does not include direct application
to aquatic environments. In the unlikely event that Pasteuria usgae is transferred to
surface or ground water intended for eventual human consumption, the microbe would
not survive the conditions water is subjected to in a drinking water treatment facility,
including chlorination, pH adjustments, and/or filtration. Even if oral exposure should
occur through drinking water, the Agency concludes that there is a reasonable certainty
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that no harm will result from the exposure to the residues of Pasteuria usgae in all the
anticipated drinking water exposures because of the lack of acute oral
toxicity/pathogenicity to mammals (see section IV(B)(l)(a)) and the host specific nature
of the microbe.
4. Acute and Chronic Dietary Exposure and Risks for Sensitive
Subpopulations, Particularly Infants and Children
Acute and chronic dietary exposure to Pasteuria usgae is not expected for sensitive
subpopulations, particularly infants and children, because there are no approved food
uses for this active ingredient at this time.
5. Occupational, Residential, School, and Daycare Exposure and Risk
Characterization
Significant additional human exposure to Pasteuria usgae from its use as a
manufacturing product to produce nematicide end-use products is not expected in
occupational, residential, school, or daycare areas.
a. Occupational Exposure and Risk Characterization
In light of the Tier I acute toxicity/pathogenicity studies, which did not show any toxic
and/or pathogenic effects to rats via oral, pulmonary, dermal, and intravenous routes of
exposure (see section IV(B)(l)(a)X handler exposure to Pasteuria usgae is not expected
to pose any undue risk. Regardless, appropriate personal protective equipment and
precautionary statements are required on the product label to mitigate any potential risks
to pesticide handlers due to prolonged exposure. Handlers working with Pasteuria usgae
in manufacturing facilities must wear a long-sleeved shirt, long pants, socks, shoes,
waterproof gloves, and a dust/mist filtering respirator meeting NIOSH standards of at
least N-95, R-95, orP-95.
b. Residential, School, and Daycare Exposure and Risk Characterization
According to the label, Pasteuria usgae - BL1 is only to be used to manufacture
nematicide end-use products. No indoor residential, school, or daycare uses currently
appear on the label; thus, human exposure to Pasteuria usgae should not occur in these
areas.
6. Aggregate Exposure from Multiple Routes Including Dermal, Oral, and
Inhalation
Potential non-occupational dermal, oral, and inhalation exposure to Pasteuria usgae is
not expected as there are no approved food uses for this active ingredient at this time and
the active ingredient is to be used only to manufacture nematicide end-use products,
which are still undergoing Agency review.
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7. Cumulative Effects
Cumulative effects of exposure to Pasteuria usgae and to other substances that might
have a common mechanism of toxicity have not been considered because there are no
approved food uses for this active ingredient at this time.
8. Risk Characterization
The Agency considered human exposure to Pasteuria usgae in light of the standard for
registration in FIFRA, as amended by the Food Quality Protection Act (FQPA) of 1996.
A determination has been made that no unreasonable adverse effects to the United States
population in general, and to infants and children in particular, will result from the use of
Pasteuria usgae when used in accordance with EPA-approved labeling.
C. Environmental Assessment
Exposure of non-target organisms is possible with application of end-use products
containing Pasteuria usgae to turf. Pasteuria Bioscience, Incorporated has submitted data
waiver rationales containing information cited in published literature to satisfy data
requirements for non-target organism testing with the technical grade of the active
ingredient (TGAI). The following is a review of the rationales submitted to support the
registration of the manufacturing-use product and proposed end-use products, as well as
resulting conclusions regarding environmental risks based on the submitted waiver
justifications.
For a comprehensive guideline-by-guideline summary of the non-target toxicity data
requirements, refer to Table 3 in Appendix A.
1. Summary of Non-Target Organism Waiver Rationales (40 CFR §
158.2150)
Data waiver rationales were submitted in response to data requirements for avian,
freshwater fish and invertebrate, insect, and honey bee non-target organism testing
requirements. Justification for these waivers is described in sections IV(C)(l)(a),
IV(C)(l)(c), and IV(C)(l)(f). Some data requirements were not required based on the test
notes described in 40 CFR § 158.2150 (e) and explanations are provided in sections
IV(C)(l)(b), IV(C)(l)(d), and IV(C)(l)(e). Overall, the information provided is sufficient
to satisfy the Tier I non-target organism data requirements for the manufacturing-use
product and proposed end-use products. Further testing of non-target organisms at higher
tier levels is not required.
a. Avian Oral Toxicity/Pathogenicity (OPPTS Guideline 885.4050)
The request to waive avian oral toxicity/pathogenicity testing is based on the rationale
that exposure of birds to Pasteuria usgae as a result of its proposed applications will be
minimal, and that no records are available in which toxicity or pathogenicity of Pasteuria
usgae or other Pasteuria spp. to birds is reported. Pasteuria spp. are ubiquitous in the
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environment and are found in soil environments where plant-parasitic nematodes exist
(Chen and Dickson 1998; Hewlett etal. 1994), and their population dynamics are
strongly linked to parasite-prey interactions with their nematode hosts (Ciancio and
Queneherve 2000; Darban et al. 2005). Pasteuria usgae is very specific to its host (Bekal
etal. 2001; Giblin-Davis etal. 2001, 2003).
Extensive literature searches were performed within Agricola and PubMed databases, as
well as several relevant journals, and no information was found relating exposure to
naturally occurring Pasteuria usgae or other Pasteuria spp. to toxic or pathogenic effects
in birds. The searches also did not result in finding any reports of genotoxic,
carcinogenic, allergenic, mutagenic, or teratogenic effects.
The rationale that exposure of birds to Pasteuria usgae would be minimal is acceptable.
An argument could be made that the absence of a record of toxicity/pathogenicity to birds
is not evidence that such effects could not occur. However, Pasteuria usgae is a naturally
occurring and ubiquitous bacterium and would be recorded as an agent of avian disease if
it were toxic or pathogenic to birds. Therefore, the rationale presented is acceptable and
fulfills the data requirement for avian oral toxicity/pathogenicity testing. Based on this
information, adverse effects to avian wildlife resulting from exposure to Pasteuria usgae
are not anticipated.
b. Avian Inhalation Toxicity/Pathogenicity (OPPTS Guideline 885.4100)
This data requirement is 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)).
c. Freshwater Fish Toxicity/Pathogenicity (OPPTS Guideline 885. 4200) and
Freshwater Invertebrate Toxicity/Pathogenicity (OPPTS Guideline
885.4240)
The requests to waive requirements for freshwater fish toxicity/pathogenicity and
freshwater invertebrate toxicity/pathogenicity were combined. The rationale is similar to
that for birds described above, wherein exposure to freshwater fish and invertebrates is
expected to be limited and no records are available in which toxicity or pathogenicity of
Pasteuria usgae to freshwater fish or invertebrates is reported. Additionally, Pasteuria
usgae and other Pasteuria spp. do not produce crystalline insect toxins as seen in other
bacteria that are known to be toxic to insects (e.g., Bacillus thuringiensis). Pasteuria
ramosa is a closely related species to Pasteuria usgae that is known to parasitize
Daphnia magna and other Daphnia species. However, Pasteuria ramosa does not infect
nematode hosts typical to other Pasteuria spp. (Ebert et al. 1996). Pasteuria usgae is
host-specific and it is unique in its spore ultrastructural characteristics that contributes to
its specificity and separates it from other closely related Pasteuria spp. (Ebert et al. 1996;
Giblin-Davis etal. 2001). Furthermore, neither Pasteuria usgae nor Pasteuria ramosa are
included in published fish pathogen listings.
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This rationale is acceptable and sufficient to fulfill the requirements for freshwater fish
toxicity/pathogenicity and freshwater invertebrate toxicity/pathogenicity testing. Based
on this information, adverse effects to freshwater fish or invertebrates resulting from
proposed label applications of Pasteuria usgae are not expected.
d. Estuarine/Marine Fish Testing and Estuarine/Marine Invertebrate
Testing (OPPTS Guideline 885.4280)
This data requirement is not required because Pasteuria usgae will not be applied to
water and is not expected to enter marine/estuarine environments in amounts that are
significantly higher than naturally occurring concentrations (refer to test note #6 of 40
CFR§ 158.2150(e)).
e. Non-Target Plant Testing (OPPTS Guideline 885.4300)
This data requirement is not required because Pasteuria usgae is not related to known
plant pathogens, and adverse effects to plants are not expected (refer to test note #7 of 40
CFR§ 158.2150(e)).
/ Non-Target Insect Testing (OPPTS Guideline 885.4340) and Honey Bee
Testing (OPPTS Guideline 885.4380)
The requests to waive requirements for non-target insect testing and honey bee testing
were combined. The rationale is similar to that given for other taxa above, wherein
exposure to non-target insects and honey bees is expected to be limited, and neither
Pasteuria usgae nor other Pasteuria spp. are included in published insect or honey bee
pathogen listings. Pasteuria usgae and other Pasteuria spp. do not produce crystalline
insect toxins as seen in other bacteria that are known to be toxic to insects.
This rationale is acceptable and sufficient to fulfill the requirements for non-target insect
testing and honey bee testing. Based on this information, adverse effects to non-target
insects or honey bees resulting from proposed labeled applications of Pasteuria usgae are
not expected.
2. Environmental Effects Conclusions
a. Terrestrial A nimals and Plants
Data waiver rationales provide sufficient information to conclude that adverse effects are
not expected in birds, non-target insects, and honey bees as a result of exposure to
Pasteuria usgae (MRTD No. 474267-13). These rationales address exposure via
applications of granules containing Pasteuria usgae to turf; however, the rationales
presented are also applicable to liquid applications to turf. Pasteuria spp. are ubiquitous
in soils worldwide, and Pasteuria usgae is highly host-specific, parasitizing exclusively
the plant-parasitic sting nematode (Belonolaimus longicaudatus). None of the proposed
applications are expected to increase the concentrations of Pasteuria usgae in the soil
above naturally occurring levels. Soil applications of granules to turf are expected to limit
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exposure in the environment. While liquid applications would result in exposure on foliar
surfaces of turf, there is no available literature documenting reports of terrestrial animal
pathogenicity or toxicity as a result of exposure to Pasteuria usgae.
An acute oral toxicity and pathogenicity study with laboratory rats (MRID No. 474267-
09) is available with which to determine the potential effects of Pasteuria usgae on wild
mammals. Laboratory rats were dosed with 108 spores/mL, and were observed for 21
days. No signs of toxic or pathogenic effects were observed throughout the test or at
necropsy. This study demonstrates that toxicity/pathogenicity to wild mammals is not
expected from labeled applications of Pasteuria usgae.
Non-target plant testing is not required because Pasteuria usgae is not related to any
known plant pathogen. Adverse effects on plants are not expected to result from labeled
applications of Pasteuria usgae.
Based on the above rationales, adverse effects are not expected to occur to terrestrial
animals or plants as a result of proposed labeled applications of Pasteuria usgae.
b. Aquatic Animals and Plants
Data waiver rationales were submitted to fulfill data requirements for and support effects
conclusions for freshwater aquatic organisms (MRID No. 474267-13). These rationales
address exposure via applications of granules containing Pasteuria usgae to turf;
however, the rationales presented are also applicable to liquid applications to turf. The
rationales provide sufficient information to conclude that adverse effects are not expected
in freshwater fish or freshwater invertebrates as a result of exposure to Pasteuria usgae.
The rationales were similar to those presented for terrestrial organisms in that Pasteuria
spp. are ubiquitous in soils worldwide, and Pasteuria usgae is highly host-specific,
parasitizing exclusively the plant-parasitic sting nematode (Belonolaimus longicaudatus}.
Soil applications in turf only are expected to limit exposure in the aquatic environment,
and runoff from both soil and liquid applications is not expected to increase the
concentrations of Pasteuria usgae in freshwater environments above naturally occurring
levels. There is no available literature documenting reports of pathogenicity or toxicity to
freshwater fish or invertebrates as a result of exposure to Pasteuria usgae. Pasteuria
ramosa is a closely related species to Pasteuria usgae that does parasitize Daphnia
magna and other Daphnia spp. However, Pasteuria ramosa does not infect nematode
hosts typical to other Pasteuria spp., and Pasteuria usgae is very host-specific and is not
known to parasitize aquatic invertebrates.
Because the use of Pasteuria usgae is limited to applications to turf and is not intended
for application directly to water, exposure is not expected to be significant in aquatic
environments, including marine or estuarine systems. Therefore, testing with
marine/estuarine animals and testing with aquatic plants is not necessary.
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Based on the rationales submitted, adverse effects to freshwater and marine/estuarine
fish, invertebrates, and plants are not expected as a result of exposure to proposed labeled
applications of Pasteuria usgae.
3. Threatened and Endangered Species Assessment
There are no listed endangered or threatened species related to the target pest. Since it is
concluded that effects are not anticipated for non-target species exposed to Pasteuria
usgae as a result of proposed labeled applications, direct and indirect effects to listed
species or their habitat are also not expected. Therefore, BPPD makes "No Effect" (NE)
determinations for direct and indirect effects to listed species and their habitat as a result
of the proposed uses of Pasteuria usgae.
V. ENVIRONMENTAL JUSTICE
The Environmental Protection Agency 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.
At this time, the Environmental Protection Agency does not believe the use of Pasteuria
usgae - BL1, which contains Pasteuria usgae as an active ingredient, will cause harm or
a disproportionate impact on at-risk communities.
For additional information regarding environmental justice issues, please visit EPA's web
site at http://www.epa.sov/compliance/environmentaliustice/index.html.
VI. PUBLIC INTEREST FINDING
EPA determines whether conditional registration of a pesticide is in the public interest in
accordance with the criteria set forth in the Federal Register dated March 5, 1986 (58 FR
7268). There is a presumption that registration of a pesticide is in the public interest if (1)
the use is for a minor crop, (2) the use is a replacement for another pesticide that is of
continuing concern to the Agency, (3) the use is one for which an emergency exemption
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under FIFRA section 18 has been granted (i.e., the basis for the exemption was lack of a
registered alternative product), or (4) the use is against a pest of public health
significance. Pasteuria usgae - BL1 will be used to create end-use products, that once
reviewed and determined to be eligible for registration under FIFRA, will serve as partial
replacements for conventional nematicides of continuing concern to the Agency [e.g.,
methyl bromide (ozone-depleting substance) and 1,3-dichloropropene (probable human
carcinogen)]. Based on this information, this registration is presumed to be in the public
interest.
VII. RISK MANAGEMENT AND REGISTRATION DECISIONS
A. Determination of Eligibility
Section 3(c)(7)(C) of the Federal Insecticide, Fungicide, and Rodenticide Act provides
for the conditional registration of a pesticide containing an active ingredient not
contained in any currently registered pesticide for a period reasonably sufficient for the
generation and submission of required data if the following are determined:
(1) Use of the pesticide during a defined period will not cause any
unreasonable adverse effect on the environment;
(2) Use of the pesticide is in the public interest; AND
(3) For data that are lacking, a reasonable period of time sufficient for
generation of that data has not elapsed since the Agency first imposed the data
requirements.
As discussed in this document, use of the manufacturing-use product containing
Pasteuria usgae is not likely to result in any unreasonable adverse effects to human
health or the environment, fulfilling criterion 1. Furthermore, as mentioned in section VI,
the registration of Pasteuria usgae - BL1 is presumed to be in the public interest because
it will be used to create end-use products that will eventually serve as partial
replacements for particular conventional nematicides of continuing concern to the
Agency; thus, criterion 2 has been satisfied. To satisfy criterion 3, insufficient time has
elapsed since the initial imposition of the data requirements outlined in section VIII(B).
B. Registration Review
The Federal Insecticide, Fungicide, and Rodenticide Act, as amended by the FQPA of
1996, mandated the continuous review of existing pesticides. All pesticides distributed
and sold in the United States must generally be registered by the EPA, based on scientific
data showing that they will not cause unreasonable risks to human health, workers, or the
environment when used as directed in product labeling. The registration review program
is intended to make sure that, as the ability to assess risk evolves and as policies and
practices change, all registered pesticides continue to meet the statutory standard of no
unreasonable adverse effects to human health or the environment. Changes in science,
public policy, and pesticide use practices will occur over time. Through the registration
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review program, the Agency periodically reevaluates pesticides to make sure that as
change occurs, products in the marketplace can be used safely. Information on this
program is provided at http://www.epa.gov/oppsrrdl/registration review/.
The Agency has implemented the registration review program pursuant to FIFRA section
3(g) and will review each registered pesticide every 15 years to determine whether it
continues to meet the FIFRA standard for registration. A registration review decision is
the Agency's determination whether a pesticide meets, or does not meet, the standard for
registration in the Federal Insecticide, Fungicide, and Rodenticide Act. Pasteuria usgae
will be included in the schedule for registration review at the end of the Fiscal Year when
schedules are updated.
C. Regulatory Decision
Based on the data submitted and under FIFRA section 3(c)(7)(C), BPPD recommends
conditional, time-limited registration of the manufacturing-use product, Pasteuria usgae
- BL1, containing Pasteuria usgae as a new active ingredient. Although the data are
satisfactory with respect to this particular new active ingredient in this pesticide, they are
not sufficient to support an unconditional registration under FIFRA section 3(c)(5).
Additional data, as outlined in section VIII(B), are necessary for a finding of
registrability under FIFRA section 3(c)(5) and are considered terms or conditions for the
purposes of this registration.
Handler exposure to Pasteuria usgae is not expected to pose any undue risk. Regardless,
appropriate personal protective equipment and precautionary statements are required on
the product label to mitigate any potential risks to pesticide handlers due to prolonged
exposure. Handlers working with Pasteuria usgae in manufacturing facilities must wear a
long-sleeved shirt, long pants, socks, shoes, waterproof gloves, and a dust/mist filtering
respirator meeting NIOSH standards of at least N-95, R-95, or P-95.
D. Labeling
The label for the registered manufacturing-use product containing the active ingredient,
Pasteuria usgae, is available at http://oaspub.epa.gov/pestlabl/ppls.home.
VIII. ACTIONS REQUIRED BY THE REGISTRANT
A. Final Printed Labeling
Before releasing pesticide products containing Pasteuria usgae for shipment, the
registrant is required to provide appropriate final printed labeling to the Agency.
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B. Conditional, Time-Limited Registration Required Data/Information
July 2009
The Agency evaluated the data submitted in connection with the initial registration of the
manufacturing-use product, Pasteuria usgae - BL1, containing Pasteuria usgae as a new
active ingredient. Although it was determined that these data fulfill current guideline
requirements for a conditional, time-limited FIFRA section 3(c)(7)(C) registration, the
Agency is requiring Pasteuria Bioscience, Incorporated to submit the following additional
data and/or information in the time frames listed:
Study Type
Required Data/Information
Due Date
Analysis of Samples
(OPPTS Guideline
885.1400)
The preliminary analysis of samples provided to the Agency revealed a
wide range of concentrations, with many replicates containing 1-2 orders
of magnitude fewer Pasteuria usgae spores than the minimum specified
on the Confidential Statement of Formula and product label (i.e., 1 x 106
spores per milliliter). A follow-up explanation, attempting to address this
variability, mentioned several factors that contributed to this
inconsistency and discussed how the concentration of spores could be
adjusted during the manufacturing process, if necessary. Despite this
additional information, a new five-batch analysis, with all batches from
production level, must be submitted to the Agency.
May 1,2011
Manufacturing Process
(OPPTS Guideline
885.1200)
The original manufacturing process did not describe quality control
measures taken to confirm that Pasteuria usgae was the only Pasteuria
species present in the master stock. A follow-up explanation, which
demonstrated that batches are screened for the presence of other microbes
and screened against sting nematodes to confirm the presence of
Pasteuria usgae, still did not provide sufficient information to indicate
that screening for other Pasteuria species is conducted during the
manufacturing process. Other Pasteuria species, specifically those that
parasitize saprophytic (non-plant pathogenic) nematodes, must be
screened for during the manufacturing process. Pasteuria Bioscience,
Incorporated must integrate quality control measures that screen for
Pasteuria species that parasitize saprophytic nematodes, either
immediately after freezing or after making seed stock, into their current
manufacturing process and provide a written description of these
measures to the Agency.
October 1, 2009
Product Identity (OPPTS
Guideline 885.1100)
Currently, full recognition of Pasteuria usgae is still pending with the
Judicial Commission of the International Committee for Systematic
Bacteriology. Once Pasteuria usgae has been recognized and removed
from the category Candidatus, documentation indicating official
recognition must be provided to the Agency.
As soon as the
specified
information becomes
available
Storage Stability (OPPTS
Guideline 830.6317)
The storage stability study for the technical grade of the active ingredient
(TGAI)/manufacturing-use product (MP) has been cited as ongoing.
When the study has been completed and the results have been compiled,
these data must be submitted to the Agency. Within the discussion of
these results, the test substance must be described to ensure that this data
requirement has been fulfilled for both the TGAI and the MP (i.e.,
reference that the TGAI is equivalent to the MP for Pasteuria usgae -
BL1).
June 1, 2010
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C. Reporting of Adverse Effects and Hypersensitivity Incidents
Notwithstanding the information stated in the previous section, it should be clearly
understood that certain, specific data are required to be reported to the Agency 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
Agency 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 Agency under the provisions of 40 CFR § 158.2140(d).
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IX. GLOSSARY OF ACRONYMS AND ABBREVIATIONS
July 2009
BPPD Biopesticides and Pollution Prevention Division
BRAD Biopesticides Registration Action Document
CFR Code of Federal Regulations
cm3 cubic centimeter
CSF Confidential Statement of Formula
°C degrees Celsius
EDSP Endocrine Disrupter Screening Program
EDSTAC Endocrine Disrupter Screening and Testing Advisory Committee
EPA Environmental Protection Agency (the "Agency")
FFDCA Federal Food, Drug, and Cosmetic Act
FIFRA Federal Insecticide, Fungicide, and Rodenticide Act
FQPA Food Quality Protection Act
FR Federal Register
g gram
kg kilogram
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).
MRID No. Master Record Identification Number
mg milligram
mL milliliter
MP manufacturing-use product
MPCA microbial pest control agent
NE "No Effect"
NIOSH National Institute for Occupational Safety and Health
OPP Office of Pesticide Programs
OPPTS Office of Prevention, Pesticides, and Toxic Substances
PCR polymerase chain reaction
PPE personal protective equipment
TGAI technical grade of the active ingredient
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X. BIBLIOGRAPHY
A. Studies Submitted in Support of This Registration
474267-01 Macintosh S, Smith K. 2008. Product Identity: Pasteuria usgae Spores-
Liquid Suspension. Project Number: PBI-2008-001. Unpublished report
prepared by Pasteuria Bioscience, Incorporated and Macintosh &
Associates, Incorporated, 17 pages.
474267-02 Smith K. 2008. Manufacturing Process for Pasteuria usgae - Liquid
Suspension. Project Number: PBI-2008-003. Unpublished report prepared
by Pasteuria Bioscience, Incorporated, 17 pages.
474267-03 Macintosh S. 2008. Discussion of Formation of Unintentional Ingredients
(Pasteuria usgae). Project Number: RES-32244301-0, 11467-07.
Unpublished report prepared by Pasteuria Bioscience, Incorporated, 17
pages.
474267-04 Smith K. 2008. Analysis of Samples/Enforcement Methods for Pasteuria
usgae Spores - Liquid Suspension. Unpublished report prepared by
Pasteuria Bioscience, Incorporated, 77 pages.
474267-05 Smith K. 2008. Certification of Limits for Pasteuria usgae - Liquid
Suspension. Project Number: PB1-2008-004. Unpublished report prepared
by Pasteuria Bioscience, Incorporated, 13 pages.
474267-06 Kaminsky M. 2008. Pasteuria usgae Spores - Liquid Suspension (TGAI):
Final Report. Project Number: 11565-08. Unpublished report prepared by
Stillmeadow, Incorporated, 17 pages.
474267-07 Smith K. 2008. Product Stability to Elevated Temperatures (Pasteuria
usgae). Project Number: PB 1-2008-005. Unpublished report prepared by
Pasteuria Bioscience, Incorporated, 8 pages.
474267-08 Smith K. 2008. Interim Report: Storage Stability (Pasteuria usgae).
Project Number: PB 1-2008-006. Unpublished report prepared by Pasteuria
Bioscience, Incorporated, 8 pages.
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 report prepared by Stillmeadow,
Incorporated, 15 pages.
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July 2009
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 report prepared by Stillmeadow,
Incorporated, 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 report prepared by Stillmeadow,
Incorporated, 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 report prepared by Stillmeadow,
Incorporated, 12 pages.
474267-13 Macintosh S, Smith K, Hewlett T. 2008. Request for Waivers from the
Requirement to Conduct Guideline Studies: Avian Oral Toxicity,
Freshwater Fish Toxicity/Pathogenicity, Freshwater Invertebrate Toxicity/
Pathogenicity, Non-Target Insect Testing, and Honey Bee Testing
(Pasteuria usgae). Project Number: PBI-2008-002. Unpublished report
prepared by Pasteuria Bioscience, Incorporated, 29 pages.
474267-14 Pasteuria Bioscience, Incorporated. 2008. Copies of Publications
(Pasteuria usgae). Unpublished report prepared by Pasteuria Bioscience,
Incorporated, 334 pages.
474350-01 Macintosh S. 2008. Request for Waivers from the Requirement to
Conduct Physical and Chemical Characteristics Guideline Studies:
Corrosion Characteristics - 830.6320 and Viscosity - 830.1700. Project
Number: PBI-2008-008. Unpublished report prepared by Macintosh &
Associates, Incorporated, 43 pages.
474350-02 Macintosh S. 2008. Hypersensitivity Incidents - 885.3400. Project
Number: PBI-2008-009. Unpublished report prepared by Macintosh &
Associates, Incorporated, 7 pages.
474350-03 Macintosh S. 2008. Request for Waivers from the Requirement to
Conduct Toxicology Guideline Studies: Acute Eye Irritation - 870.2400
and Primary Dermal Irritation - 870.2500. Project Number:
PBI-2008-007. Unpublished report prepared by Pasteuria Bioscience,
Incorporated, 8 pages.
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474979-01 Macintosh S. 2008. Request for Waivers from the Requirement to
Conduct Toxicology Guideline Studies: Acute Oral Toxicity - 870.1100,
Acute Dermal Toxicity - 870.1200, and Acute Inhalation Toxicity -
870.1300. Project Number: PBI-2008-012. Unpublished report prepared
by Macintosh & Associates, Incorporated, 9 pages.
B. Environmental Protection Agency Risk Assessment Memorandums
Borges S, Vaituzis Z. 2009. 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. U.S. EPA,
Washington, D.C. Memorandum dated April 8, 2009.
Schiissel S, Kough JL. 2009. Review of Product Chemistry and Human Health Studies
for Pasteuria usgae (AF254387) - Liquid Suspension to Support the Registration
of Technical Grade Active Ingredient (EPA Reg. No. 85004-R). U.S. EPA,
Washington, D.C. Memorandum dated April 9, 2009.
C. Other References
Bekal S, Borneman J, Springer MS, Giblin-Davis RM, Becker JO. 2001. Phenotypic and
molecular analysis of a Pasteuria strain parasitic to the sting nematode.
Journal of Nematology 33:110-115.
Chen ZX, Dickson DW. 1998. Review of Pasteuria penetrans: biology, ecology, and
biological control potential. Journal of Nematology 30:313-340.
Ciancio A, Queneherve P. 2000. Populations dynamics of Meloidogyne incognita and
infestation levels by Pasteuria penetrans in a naturally infested field in
Martinique. Nematropica 30:77-86.
Darban DA, Pathan MA, Bhatti AG, Maitelo SA. 2005. The effect of different initial
densities of nematode (Meloidogyne javanica) on the build-up of Pasteuria
penetrans population. Journal ofZhejiang University 6B:113-118.
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 3 51:1689-1701.
26
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Biopesticides Registration Action Document (BRAD) July 2009
Giblin-Davis RM, Williams DS, Wergin WP, Dickson DW, Hewlett TE, Bekal S, Becker
JO. 2001. infrastructure and development of Pasteuria sp. (S-l strain), an
obligate endoparasite of Belonolaimus longicaudatus (Nemata: Tyenchida).
Journal ofNematology 3 3:227-23 8.
Giblin-Davis RM, Williams DS, Bekal S, Dickson DW, Brito JA, Becker JO, Preston JF.
2003. ' Candidates Pasteuria usgae' sp. Nov., an obligate endoparasite of the
phytoparasitic nematode, Belonolaimus longicaudatus. International Journal of
Systematic and Evolutionary Microbiology 53:197-200.
Hewlett TE, Cox R, Dickson DW, Dunn RA. 1994. Occurrence of Pasteuria spp. in
Florida. Journal ofNematology 26:616-619.
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Pasteuria usgae
Biopesticides Registration Action Document (BRAD) July 2009
APPENDIX A - MICROBIAL PESTICIDE DATA REQUIREMENTS
TABLE 1. Product Analysis Data Requirements for the Technical Grade of the Active Ingredient (TGAI)/
Manufacturing-Use Product (MP), Pasteuria usgae - BL1 (40 CFR § 158.2120)
Data Requirement
(OPPTS Guideline)
Results
TGAI
MP
MRID
Number
Product Chemistry and Composition
Product Identity
(885.1100)
Not
applicable
Pasteuria spp. are gram-positive, mycelial,
endospore-forming bacteria that are endoparasitic to
nematodes and water fleas. The endospores of
Pasteuria spp. can be identified and counted
microscopically, but it is difficult to distinguish
between species unless high magnification electron
microscopy is used so that size and shape of the
spores can be visualized and measured. Molecular
techniques such as PCR methods have been reported
for Pasteuria penetrans but have not been developed
for the other species. Therefore, species
identification via host specificity assays, requiring
the establishment of nematode cultures and a
bioassay, is the most direct and reliable technique for
species identification.
Pasteuria usgae, a recently discovered strain, is host-
specific to the sting nematode (Belonolaimus
longicaudatus), which can be damaging to a variety
of crops, particularly turf. The active agent of
Pasteuria usgae is an endospore that attaches and
infects the host nematode during all life stages
(except eggs). Increased moisture, neutral pH,
temperatures above 10°C, and sandy soil seem to
provide the best environments for spore attachment
to the host. In laboratory studies, Pasteuria spp. were
able to attach to nematodes after exposure to high
temperatures and wide ranges of pH; however, at
extremes, the number of attached spores per
nematode was reduced and, in some cases, no
infection occurred after attachment.
After attachment of the endospore, a germ tube
penetrates the nematode cuticle and mycelial
microcolonies are formed in the pseudocoelom,
leading to eventual death of the host. The endospores
formed inside the host are released into the soil when
the infected nematode decomposes. The spores are
non-motile and stable in the soil environment for
several years.
Classification: Acceptable. Currently, full
recognition of Pasteuria usgae is still pending with
the Judicial Commission of the International
Committee for Systematic Bacteriology. Once
Pasteuria usgae has been recognized and removed
474267-01
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Pasteuria usgae
Biopesticides Registration Action Document (BRAD)
July 2009
Data Requirement
(OPPTS Guideline)
Manufacturing Process
(885.1200)
Deposition of a Sample in a
Nationally Recognized
Culture Collection
(Not applicable)
Discussion of Formation of
Unintentional Ingredients
(885.1300)
Results
TGAI
MP
from the category Candidates, documentation
indicating official recognition must be provided to
the Agency.
Submitted data satisfy the requirements of manufacturing process
for the TGAI/MP for the purposes of a conditional, time-limited
FIFRA section 3(c)(7)(C) registration only.
Classification: Supplemental: Upgradeable. Pasteuria
Bioscience, Incorporated must integrate quality control
measures that screen for Pasteuria species that parasitize
saprophytic nematodes, either immediately after freezing or
after making seed stock, into their current manufacturing
process and provide a written description of these measures to
the Agency on or before October 1, 2009.
Pasteuria usgae is on deposit
with the American Type
Culture Collection in Manassas,
Virginia under Accession
Number SD-5835.
Not applicable
Submitted data satisfy the requirements of discussion of formation
of unintentional ingredients for the TGAI/MP.
Classification: Acceptable
MRID
Number
474267-02
Not applicable
474267-03
Analysis and Certified Limits
Analysis of Samples
(885.1400)
Certification of Limits
(885.1500)
Submitted data satisfy the requirements of analysis of samples for
the TGAI/MP for the purposes of a conditional, time-limited
FIFRA section 3(c)(7)(C) registration only.
Classification: Unacceptable. A new five-batch analysis, with
all batches from production level, must be submitted to the
Agency on or before May 1, 2011.
Not applicable
The certified limits for the
active and inert ingredients fall
within the OPPTS Guideline
830.1750 specified ranges.
Classification: Acceptable
474267-04
474267-05
Physical and Chemical Characteristics
Color
(830.6302)
Physical State
(830.6303)
Odor
(830.6304)
Stability to Normal and
Elevated Temperatures,
Metals, and Metal Ions
(830.6313)
Storage Stability
(830.6317)
Clear to very faint yellow-
brown or cloudy at 24°C (5
batches)
Liquid at 20°C
No odor
Stable after exposure to 54°C
for 14 days.
Not applicable
Not applicable
Not applicable
Not applicable
Preliminary results indicate that after 1 month of storage at 4°C,
the TGAI/MP had a 5% loss of attachment and a 15% loss of
infectivity.
A 12-month storage stability study is ongoing and results must
be submitted to the Agency upon its completion (on or before
June 1, 2010).
474267-06
474267-06
474267-06
474267-07
474267-08
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Pasteuria usgae
Biopesticides Registration Action Document (BRAD)
July 2009
Data Requirement
(OPPTS Guideline)
Miscibility
(830.6319)
Corrosion Characteristics
(830.6320)
pH
(830.7000)
Viscosity
(830.7100)
Density /Relative Density /Bulk
Density (Specific Gravity)
(830.7300)
Results
TGAI
Not applicable
Not applicable
6.80-6.96 (5 batches)*
6-7.3**
Not applicable
Specific gravity = 1.006-1.008
gram (g)/cubic centimeter
(cm3)(5 batches)*
Specific gravity = 1.0 g/cm3**
MP
Not required because Pasteuria
usgae - BL1 is not an
emulsifiable form of microbial
pesticide (refer to test note #2
of40CFR§ 158.2120(d)).
Waived because Pasteuria
usgae - BL1 will be packaged
for very short time periods in
plastic as it moves from the
manufacturing facility to the
formulation facility for end-use
products. There will be no
long-term exposure to the
packaging.
Classification: Acceptable
Not applicable
Waived because Pasteuria
usgae - BL 1 is not viscous and
will not be applied as a product.
There will be no environmental
exposure.
Classification: Acceptable
Not applicable
MRID
Number
Not applicable
474350-01
474267-06*
CSF**
474350-01
474267-06*
CSF**
TABLE 2. Toxicology Data Requirements for the Technical Grade of the Active Ingredient (TGAI)/
Manufacturing-Use Product (MP), Pasteuria usgae- BL1 (40 CFR § 158.2140)
Data Requirement
(OPPTS Guideline)
Results
TGAI
MP
MRID
Number
Tier I
Acute Oral
Toxicity/Pathogenicity
(885.3050)
Not toxic and/or pathogenic to
rats by oral dose of 1 x 108
spores/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 sting
nematode, infectivity is unlikely
to be a concern.
Classification: Acceptable
TOXICITY CATEGORY IV
Not applicable
474267-09
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Pasteuria usgae
Biopesticides Registration Action Document (BRAD)
July 2009
Data Requirement
(OPPTS Guideline)
Results
TGAI
MP
MRID
Number
Acute Dermal Toxicity
(885.3100)
Not toxic to rats when treated
with 2,000 mg/kg at 108
spores/mL. The acute dermal
LD50 is greater than 2,000 mg/kg
for 108 spores/mL.
Classification: Acceptable
TOXICITY CATEGORY IV
Not applicable
474267-12
Acute Pulmonary
Toxicity/Pathogenicity
(885.3150)
Not toxic and/or pathogenic to
rats by pulmonary dose of 1-3 x
108 spores/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
sting nematode, infectivity is
unlikely to be a concern.
Classification: Acceptable
TOXICITY CATEGORY IV
Not applicable
474267-10
Acute Injection
Toxicity/Pathogenicity
(885.3200)
Not toxic and/or pathogenic to
rats by intravenous dose of Ix
108 spores/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
sting nematode, infectivity is
unlikely to be a concern.
Classification: Acceptable
TOXICITY CATEGORY IV
Not applicable
474267-11
Hypersensitivity Incidents
(885.3400)
No hypersensitivity incidents, including immediate-type or
delayed-type reactions of humans and domestic animals, occurring
during the testing or production of the TGAI/MP were reported by
the registrant. Any future hypersensitivity incidents must be
reported per OPPTS Guideline 885.3400.
474350-02
Cell Culture
(885.3500)
Not required because Pasteuria
usgae is not a virus (refer to test
note#4of40CFR§
158.2140(d)).
Not applicable
Not applicable
Acute Oral Toxicity
(870.1100)
Not applicable
Waived based on the results of
MRID Number 474267-09 and
because the MP is equivalent
to the TGAI.
Classification: Acceptable
TOXICITY CATEGORY
IV
474979-01
Acute Dermal Toxicity
(870.1200)
Not applicable
Waived based on the results of
MRID Number 474267-12 and
because the MP is equivalent
to the TGAI.
474979-01
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Pasteuria usgae
Biopesticides Registration Action Document (BRAD)
July 2009
Data Requirement
(OPPTS Guideline)
Acute Inhalation Toxicity
(870.1300)
Acute Eye Irritation
(870.2400)
Primary Dermal Irritation
(870.2500)
Results
TGAI
Not applicable
Not applicable
Not applicable
MP
Classification: Acceptable
TOXICITY CATEGORY
IV
Waived based on the results of
MRID Number 474267-10 and
because the MP is equivalent
to the TGAI.
Classification: Acceptable
TOXICITY CATEGORY
IV
Waived based on Pasteuria
usgae' s lack of toxicity in the
other toxicology studies and
precautionary statements that
mitigate potential for worker
exposure.
Classification: Acceptable
TOXICITY CATEGORY
III
Waived based on the results of
MRID Number 474267-12 and
precautionary
statements/personal protective
equipment that mitigate
potential for worker exposure.
Classification: Acceptable
TOXICITY CATEGORY
III
MRID
Number
474979-01
474350-03
474350-03
Tiers II and III
Not required for Pasteuria usgae based on the lack of acute toxicity/pathogenicity in the Tier I studies.
TABLE 3. Non-Target Organisms and Environmental Fate Data Requirements for the Technical Grade of the
Active Ingredient (TGAI), Pasteuria usgae (40 CFR § 158.2150)
Data Requirement
(OPPTS Guideline)
Results
Toxicity
Category/Description
MRID
Number
Tier I
Avian Oral Toxicity
(885.4050)
Avian Inhalation
Toxicity /Pathogenicity
(885.4100)
Data waiver rationale provides
sufficient information to determine that
toxicity/pathogenicity to avian wildlife
is not expected.
Classification: Acceptable
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)).
Not applicable
Not applicable
474267-13
Not applicable
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Pasteuria usgae
Biopesticides Registration Action Document (BRAD)
July 2009
Data Requirement
(OPPTS Guideline)
Wild Mammal
Toxicity/Pathogenicity
(885.4150)
Freshwater Fish
Toxicity/Pathogenicity
(885.4200)
Freshwater Invertebrate
Toxicity/Pathogenicity
(885.4240)
Estuarine/Marine Fish Testing
Estuarine and Marine
Invertebrate Testing
(885.4280)
Non-Target Plant Testing
(885.4300)
Non-Target Insect Testing
(885.4340)
Honey Bee Testing
(885.4380)
Results
Tests required by 40 CFR § 158.2140
are adequate and appropriate for
assessment of hazards to wild
mammals (refer to test note #4 of 40
CFR § 158.2150(e)).
Classification: Acceptable for wild
mammal risk assessment
Pasteuria usgae will not be applied
directly to water and is not expected to
enter freshwater environments in
amounts that are significantly higher
than naturally occurring concentrations.
Data waiver rationale provides
sufficient information to determine that
toxicity/pathogenicity or substantial
exposure to freshwater fish is not
expected.
Classification: Acceptable
Pasteuria usgae will not be applied
directly to water and is not expected to
enter freshwater environments in
amounts that are significantly higher
than naturally occurring concentrations.
Data waiver rationale provides
sufficient information to determine that
toxicity/pathogenicity or substantial
exposure to freshwater invertebrates is
not expected.
Classification: Acceptable
Not required because Pasteuria usgae
will not be applied to water and is not
expected to enter marine/estuarine
environments in amounts that are
significantly higher than naturally
occurring concentrations (refer to test
note #6 of 40 CFR § 158.2150(e)).
Not required because Pasteuria usgae
is not related to known plant pathogens,
and adverse effects to plants are not
expected (refer to test note #7 of 40
CFR § 158.2150(e)).
Data waiver rationale provides
sufficient information to determine that
toxicity/pathogenicity to non-target
insects is not expected.
Classification: Acceptable
Data waiver rationale provides
sufficient information to determine that
toxicity/pathogenicity to honey bees is
not expected.
Classification: Acceptable
Toxicity
Category/Description
Testing indicates no
adverse effects to
laboratory rats at 1 x
108 spores/mL when
dosed orally.
Not applicable
Not applicable
Not applicable
Not applicable
Not applicable
Not applicable
MRID
Number
474267-09
474267-13
474267-13
Not applicable
Not applicable
474267-13
474267-13
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Pasteuria usgae
Biopesticides Registration Action Document (BRAD)
July 2009
Data Requirement 1
(OPPTS Guideline) |
Results
Toxicity
Category/Description
MRID
Number
Tiers II, III, and IV
Not required for Pasteuria
usgae based
on the acceptability of the waiver rationales provided for Tier I.
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Pasteuria usgae
Biopesticides Registration Action Document (BRAD)
July 2009
APPENDIX B - PASTEURIA USGAE MANUFACTURING-USE AND END-USE
PRODUCTS
EPA
Registration
Number
85004-1
Registration
Name
Pasteuria
usgae - BL1
Percentage
Active
Ingredient
0.01%
Formulation
Type
Technical
Use Site
N/A
Method of
Application
N/A
Application
Rate
N/A
Target Pests
N/A
35
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