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BIOPESTICIDES REGISTRATION ACTION DOCUMENT
T
Chromobacterium subtsugae strain PRAA4-1
Pesticide Chemical (PC) Code: 016329
U.S. Environmental Protection Agency
Office of Pesticide Programs
Biopesticides and Pollution Prevention Division
September 27, 2011
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Chromobacterium subtsugae strain PRAA4-1T Page 2 of 36
Biopesticides Registration Action Document
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 6
IV. RISK ASSESSMENT 7
A. Product Analysis Assessment (40 CFR § 158.2120) 7
B. Human Health Assessment (40 CFR § 158.2140) 7
C. Environmental Assessment (40 CFR § 158.2150) 13
V. ENVIRONMENTAL JUSTICE 17
VI. RISK MANAGEMENT DECISION 18
VII. ACTIONS REQUIRED BY THE REGISTRANT 18
A. Final Printed Labeling 18
B. Terms of Registration 18
C. Reporting of Adverse Effects and Hypersensitivity Incidents 19
VIII. GLOSSARY OF ACRONYMS AND ABBREVIATIONS 20
IX. BIBLIOGRAPHY 21
A. Studies Submitted to Support the Chromobacterium subtsugae strain PRAA4-1T
Pesticide Product Registrations 21
B. Environmental Protection Agency Risk Assessment Memoranda and Other Documents 24
C. Other References 24
APPENDIX A. MICROBIAL PESTICIDES DATA REQUIREMENTS 26
APPENDIX B. PESTICIDE PRODUCTS 36
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Biopesticides Registration Action Document
BIOPESTICIDES REGISTRATION ACTION DOCUMENT TEAM
Office of Pesticide Programs (OPP)
Biopesticides and Pollution Prevention Division
Microbial Pesticides Branch
Science Reviews
Joel V. Gagliardi, Ph.D.
John L. Kough, Ph.D.
Anna Gross
Zigfridas Vaituzis, Ph.D
Shannon Borges
Regulations
Product Analysis, Human Health
Product Analysis, Human Health
Environmental Effects
Environmental Effects
Environmental Effects
Sheryl K. Reilly, Ph.D.
Jeannine Kausch
Chief, Microbial Pesticides Branch
Regulatory Action Leader
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I. EXECUTIVE SUMMARY
Chromobacterium subtsugae strain PRAA4-11 is a gram-negative, violet-pigmented bacterium
that was isolated from soil under an eastern hemlock (Tsuga canadensis) in the Catoctin
Mountain region of central Maryland. The United States Department of Agriculture found this
isolate of Chromobacterium subtsugae to be orally toxic to Colorado potato beetle (Leptinotarsa
decemlineata) larvae, small hive beetle (Aethina tumida) larvae, southern corn rootworm
(Diabrotica undecimpunctata) larvae and adults, and southern green stink bug (Nezara viridula)
adults. Additional testing has shown that Chromobacterium subtsugae strain PRAA4-lT-treated
diet resulted in reduced feeding in beet armyworm (Spodoptera exigua), cabbage looper
(.Trichoplusia ni), tobacco budworm (Heliothis virescens), diamondback moth (Plutella
xylostella), and southern corn rootworm, suggesting this microbe's insecticidal activity is due to
reduction in weight or inhibition of feeding. In light of the demonstrated insecticidal and
miticidal capabilities of Chromobacterium subtsugae strain PRAA4-11, Marrone Bio Innovations
proposed to register both a manufacturing-use pesticide product, MBI-203 TGAI, and an end-use
pesticide product, MBI-203 EP Bioinsecticide (formerly MBI-203 EP), containing this
bacterium. MBI-203 EP Bioinsecticide will be used to control label-specified insect and mite
pests (e.g., European corn borer, citrus rust mite, and stink bugs) on agricultural and greenhouse
crops, including vegetables, fruit, flowers, bedding plants, ornamentals, and turf.
Environmental Protection Agency (EPA) scientists reviewed product analysis, toxicology, and
nontarget organism data and information (40 Code of Federal Regulations (CFR) §§ 158.2120,
158.2140, and 158.2150, respectively) submitted to support the registration of the two
aforementioned Chromobacterium subtsugae strain PRAA4-11 pesticide products. Overall, such
data and information are adequate for risk assessment purposes, fulfill current data requirements,
and allow for registration under Section 3(c)(5) of the Federal Insecticide, Fungicide, and
Rodenticide Act (FIFRA).
Product analysis data requirements for Chromobacterium subtsugae strain PRAA4-11, including
product chemistry and composition, analysis of samples, and physical and chemical
characteristics, were fulfilled by acceptable guideline studies.
Adequate mammalian toxicology data and information were submitted to support the
Chromobacterium subtsugae strain PRAA4-11 pesticide products. The acute injection
toxicity/pathogenicity study showed that, at a single high dose, Chromobacterium subtsugae
strain PRAA4-11 is not toxic, infective, and/or pathogenic via the intravenous route of exposure.
Acute pulmonary and oral toxicity/pathogenicity tests were waived after considering the results
of the intravenous test and MBI-203 TGAI-specific toxicity tests (acute oral and inhalation).
Moreover, no hypersensitivity incidents, occurring during research, development, or testing of
Chromobacterium subtsugae strain PRAA4-11, were reported by the applicant. In light of the
results of the intravenous study, the acceptable rationale provided to waive the acute pulmonary
and oral toxicity/pathogenicity tests, and the absence of hypersensitivity incidents, testing at
higher tiers (i.e., Tiers II and III) was not required.
EPA concluded that there is a reasonable certainty that no harm will result to the United States
population, including infants and children, from aggregate exposure to residues of
Chromobacterium subtsugae strain PRAA4-1'. No dietary risks are expected from use of
Chromobacterium subtsugae strain PRAA4-11 as an active ingredient in pesticide products.
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Significant dietary exposure—through food and drinking water—is not anticipated given the
potential for this bacterium to degrade due to both predation by other biological organisms and
exposure to environmental factors (e.g., sunlight and varying temperatures) in the phyllosphere,
the filtering effect of many particulate soil types, and conditions (e.g., filtration and pH
adjustments) water is subjected to in wastewater treatment systems and drinking water facilities.
Should Chromobacterium subtsugae strain PRAA4-11 be present on food or in drinking water,
the acute oral toxicity and pathogenicity data/information demonstrated no toxicity, infectivity,
and/or pathogenicity is likely to occur with any exposure level of this microbial pest control
agent.
Despite the low toxicological profile of Chromobacterium subtsugae strain PRAA4-11, 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 Chromobacterium subtsugae strain PRAA4-11 in agricultural settings are directed to wear a
long-sleeved shirt, long pants, socks, shoes, waterproof gloves, and a dust/mist filtering respirator
meeting National Institute for Occupational Safety and Health (NIOSH) standards of at least N-
95, R-95, or P-95. EPA may require additional PPE, other than the standard described above, on
a product-specific basis.
The data and data waiver rationale, submitted by the applicant to support the pesticide products
containing Chromobacterium subtsugae strain PRAA4-11, are sufficient to fulfill the Tier I
nontarget organism data requirements and for risk assessment purposes. Further testing of
nontarget organisms at higher tier levels (i.e., Tiers n, m, and IV) is not required for the current
uses and application methods. EPA performed an environmental risk assessment based on the
data and data waiver rationale provided by the applicant and determined that the uses of
Chromobacterium subtsugae strain PRAA4-11 do not pose significant risk to nontarget
organisms when used according to label directions. Based on study results, there are concerns of
toxicity to terrestrial arthropods, aquatic invertebrates, and honey bees. Use sites and application
methods, in combination with mitigating label language (see Table 7 in Appendix A), however,
will limit exposure. EPA has made "no effect" determinations for direct effects, indirect effects,
and effects to habitat (including designated critical habitat) to listed species for all foliar and soil-
directed applications to crop plants treated within greenhouses. On the other hand, EPA
concludes that all outdoor foliar applications made by chemigation and aerial spray equipment
have the potential to cause direct effects, indirect effects, and effects to habitat (including
designated critical habitat) to listed species.
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.
Consistent with the policy of making registration actions more transparent, the pesticide products
containing Chromobacterium subtsugae strain PRAA4-11, 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 Chromobacterium subtsugae strain PRAA4-11 pesticide products, MBI-203 TGAI
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and MBI-203 EP Bioinsecticide. Therefore, EPA maintained that, based upon the risk assessment
and information submitted in support of registration of such pesticide products, it was in the best
interest of the public and the environment to issue the MBI-203 TGAI and MBI-203 EP
Bioinsecticide registrations. The basis for this decision can be found in the risk assessment for
Chromobacterium subtsugae strain PRAA4-11, which is characterized throughout this BRAD.
II. ACTIVE INGREDIENT OVERVIEW
Biological Name: Chromobacterium subtsugae strain PRAA4-11
Culture Deposit: Agricultural Research Service Culture Collection (also
known as the Northern Regional Research Laboratory
(NRRL) Collection) in Peoria, Illinois under Accession
Number NRRLB-3 065 5
OPP Chemical Code: 016329
Type of Pesticide: Microbial Pesticide - Insecticide
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
On December 22, 2009, Marrone Bio Innovations (address: 2121 Second Street, Suite B-107;
Davis, California 95618) submitted applications to register a manufacturing-use pesticide
product, MBI-203 TGAI (EPA File Symbol 84059-0), and an end-use pesticide product, MBI-
203 EP (EPA File Symbol 84059-RN), under FIFRA section 3. On March 10, 2010, EPA
announced receipt of these applications to register pesticide products containing a new active
ingredient (75 Federal Register (FR) 11175) and opened a 30-day public comment period
pursuant to the provisions of FIFRA section 3(c)(4). No comments were received following this
publication.
B. Food Tolerance Exemption
Concurrent with its registration applications and under Federal Food, Drug, and Cosmetic Act
(FFDCA) section 408(d), Marrone Bio Innovations submitted a petition to establish an
exemption from the requirement of a tolerance for Chromobacterium subtsugae strain PRAA4-11
(Pesticide Petition (PP) 9F7674). In the Federal Register of March 10, 2010 (75 FR 11171). EPA
announced that Marrone Bio Innovations proposed to establish an exemption from the
requirement of a tolerance for residues of the insecticide, Chromobacterium subtsugae strain
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PRAA4-1T, in or on all food commodities and opened a 30-day comment period. No comments
were received following this publication.
On September 7, 2011, EPA established an exemption from the requirement of a tolerance for
residues of Chromobacterium subtsugae strain PRAA4-11 in or on all food commodities when
applied as an insecticide or miticide and used in accordance with good agricultural practices (40
CFR § 180.1305; 76 FR 552721
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, n, HI, or IV, where Toxicity Category I indicates the highest toxicity and Toxicity
Category IV indicates the lowest toxicity (see 40 CFR § 156.62).
A. Product Analysis Assessment (40 CFR § 158.2120)
All product analysis data requirements for Chromobacterium subtsugae strain PRAA4-11 have
been fulfilled. Refer to Tables 1, 2, 3, and 4 in Appendix A for a brief 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 Chromobacterium subtsugae strain PRAA4-11 have been
fulfilled. Acceptable Tier I mammalian toxicology data and information support registration of
the Chromobacterium subtsugae strain PRAA4-11 pesticide products. Furthermore, Tier II and
Tier in studies were not required for Chromobacterium subtsugae strain PRAA4-11 based on the
lack of acute toxicity/pathogenicity in the Tier I studies.
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For a comprehensive summary of the generic toxicology data requirements described in sections
IV(B)(l)(a) and IV(B)(l)(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 (Office of Chemical Safety and Pollution Prevention (OCSPP)
Guideline 885.3050; Master Record Identification Number (MRID No.) 479450-23): Upon
consideration of results of other definitive toxicological data submitted by the applicant, EPA
waived acute oral toxicity/pathogenicity testing for Chromobacterium subtsugae strain PRAA4-
1T. In an acute up and down oral toxicity study (MRID No. 479450-03), three female rats were
gavaged once with 5,000 milligrams per kilogram (mg/kg) of the test material. All animals
survived and gained weight, and there were no signs of gross toxicity, adverse effects, or
abnormal behavior. The acute oral median lethal dose (LD50) was greater than 5,000 mg/kg. In an
acute intravenous study (MRID No. 479450-11), groups of rats received a single intravenous
injection of inactive test material or active test material (2.62 x 108 colony-forming units per
milliliter (cfu/mL)). Three males and three females from the active test material group were
sacrificed and necropsied on days 0, 3, 7, 14, and 21 after treatment. Tissue and blood samples
were collected at necropsy and cultured to provide qualitative and quantitative measurements of
the test organism. There was no mortality during the study, and all rats appeared normal
throughout. The test organism had cleared completely from blood, kidneys, mesenteric lymph
nodes, lungs, brain, liver, spleen, and cecum contents by day 3. The test material was nontoxic
and noninfective. EPA believes these data, when taken together, clearly indicate that
Chromobacterium subtsugae strain PRAA4-11 would not be toxic, infective, and/or pathogenic
through the oral route of exposure and that further testing is not necessary.
Acute Pulmonary Toxicity/Pathogenicity (OCSPP Guideline 885.3150; MRID No. 479450-23):
Upon consideration of results of other definitive toxicological data submitted by the applicant,
EPA waived acute pulmonary toxicity/pathogenicity testing for Chromobacterium subtsugae
strain PR.AA4-1'. In an acute inhalation study (MRID No. 479450-05), male and female rats
were exposed to an aerosol concentration of 2.12 milligrams per liter (mg/L) of the test material
for 4 hours in a nose-only inhalation chamber. No deaths occurred during exposure or during the
14-day observation period. Clinical signs included decreased activity and piloerection, which
resolved by day 5. Necropsy was unremarkable. The acute inhalation median lethal concentration
(LC50) was greater than 2.12 mg/L. In an acute intravenous study (MRID No. 479450-11), groups
of rats received a single intravenous injection of inactive test material or active test material (2.62
x 108 cfu/mL). Three males and three females from the active test material group were sacrificed
and necropsied on days 0, 3, 7, 14, and 21 after treatment. Tissue and blood samples were
collected at necropsy and cultured to provide qualitative and quantitative measurements of the
test organism. There was no mortality during the study, and all rats appeared normal throughout.
The test organism had cleared completely from blood, kidneys, mesenteric lymph nodes, lungs,
brain, liver, spleen, and cecum contents by day 3. The test material was nontoxic and
noninfective. EPA believes these data, when taken together, clearly indicate that
Chromobacterium subtsugae strain PRAA4-11 would not be toxic, infective, and/or pathogenic
through the inhalation route of exposure and that further testing is not necessary.
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Biopesticides Registration Action Document
Acute Injection Toxicity/Pathogenicity (Intravenous) - Rat (OCSPP Guideline 885.3200; MRID
No. 479450-11): In an acute intravenous injection toxicity and pathogenicity study, groups of
young adult Sprague-Dawley rats were injected at 3.1 x 106 colony-forming units (cfu)/animal.
Animals were observed up to 21 days. Five males and five females were treated with inactive
MOI203 [MBI203] TGAI, while five males and five females were not treated and served as
controls. All animals survived, gained weight, and appeared normal during the study. No
observable abnormalities were noted in any animal at necropsy. There were no significant
variations among organ weights. MOI 203 [MBI 203] organism cleared from the blood, brain,
lungs, spleen, liver, kidneys, mesenteric lymph nodes, and cecum contents of animals by day 3.
MOI 203 [MBI 203] was not detected in the tissues/organs of animals treated with autoclaved
test material or the untreated control group. MOI 203 [MBI 203] does not appear to be toxic,
infective, and/or pathogenic to rats when dosed intravenously at 3.1 x 106 cfu per animal. This
study was rated acceptable.
Hypersensitivity Incidents (OCSPP Guideline 885.3400; MRID No. 479450-23) : No
hypersensitivity incidents, including immediate-type or delayed-type reactions of humans and
domestic animals that occurred during research, development, or testing of Chromobacterium
subtsugae strain PRAA4-11, were reported by the applicant. Any future hypersensitivity incidents
must be reported to EPA (refer to test note #3 of 40 CFR § 158.2140(d)).
Cell Culture (OCSPP Guideline 885.3500): This study is not required because Chromobacterium
subtsugae strain PRAA4-11 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 HI studies were not required for Chromobacterium subtsugae strain PRAA4-11
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.
Chromobacterium subtsugae strain PRAA4-11 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 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 Table 5 in Appendix A. the data required for a FFDCA risk assessment for
Chromobacterium subtsugae strain PRAA4-11 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: Any exposure to this naturally occurring soil
bacterium is anticipated to be negligible. Although Chromobacterium subtsugae strain PRAA4-
1T may be applied directly to food, it is not expected to persist or accumulate in any reservoirs on
plants or food commodities (the phyllosphere) because, as a soil microorganism, it is best
adapted to more favorable conditions underground. Rather, after application, it likely will
degrade due to predation by other biological organisms (e.g., protists) and exposure to particular
environmental factors (e.g., sunlight and varying temperatures) (Lindow and Brandl 2003; U.S.
EPA 1996). Should this microbial pesticide be present on food, the acute oral toxicity and
pathogenicity data/information demonstrated no toxicity, infectivity, and/or pathogenicity is
likely to occur with any exposure level of Chromobacterium subtsugae strain PRAA4-11 (see
section IV(B)(l)(a) and Table 5 in Appendix A).
Drinking Water Exposure and Risk Characterization: Exposure of humans to residues of
Chromobacterium subtsugae strain PRAA4-11 in consumed drinking water is unlikely. The
proposed use patterns for Chromobacterium subtsugae strain PRAA4-11 do not include direct
application to aquatic environments, thereby limiting contact with surface water. Furthermore,
ground water is not expected to have significant exposure to Chromobacterium subtsugae strain
PRAA4-11 since, like other microorganisms, this bacterium 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)
and is not known to survive in water or deep soil. If Chromobacterium subtsugae strain PRAA4-
1T were to be transferred to surface or ground waters that are intended for eventual human
consumption (e.g., through spray drift or runoff) and directed to wastewater treatment systems or
drinking water facilities, it likely would not survive the conditions water is subjected to in such
systems or facilities, including high temperatures, chlorination, pH adjustments, and/or filtration
(Centers for Disease Control and Prevention 2009; U.S. EPA 2004). In the remote likelihood that
this microbial pesticide is present in drinking water (e.g., in water not subject to treatment
systems or facilities), the acute oral toxicity and pathogenicity data/information demonstrated no
toxicity, infectivity, and/or pathogenicity is likely to occur with any exposure level of
Chromobacterium subtsugae strain PRAA4-11 (see section IV(B)(l)(a) and Table 5 in Appendix
A).
Non-occupational, Residential Risk Characterization: Dermal and inhalation non-occupational
exposure to Chromobacterium subtsugae strain PRAA4-11 is not expected as all proposed
pesticide applications will take place in distinct agricultural settings. Even if dermal and
inhalation non-occupational exposures were to occur, such exposures would not exceed EPA's
level of concern given testing that indicated that Chromobacterium subtsugae strain PRAA4-11
is not toxic (acute inhalation and dermal toxicity), is only slightly irritating (primary dermal
irritation), is not a sensitizer (dermal sensitization), and is not pathogenic or infective (acute
injection toxicity/pathogenicity) (see section IV(B)(l)(a) and Table 5 in Appendix A).
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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."
EPA has not found Chromobacterium subtsugae strain PRAA4-11 to share a common
mechanism of toxicity with any other substances, and Chromobacterium subtsugae strain
PRAA4-11 does not appear to produce a toxic metabolite produced by other substances. For the
purposes of this tolerance action, therefore, EPA has assumed that Chromobacterium subtsugae
strain PRAA4-11 does not have a common mechanism of toxicity with other substances.
Following from this, therefore, EPA concludes that there are no cumulative effects associated
with Chromobacterium subtsugae strain PRAA4-11 that need to be considered. 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. sov/pesticides/cumulative.
c. Determination of Safety for the United States (U.S.) Population, Infants and
Children
FFDCA section 408(b)(2)(C) provides that 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 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 safety will be safe for infants and children. This additional margin of 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/information summarized in section
IV(B)(l)(a) and Table 5 in Appendix A. 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 the residues of Chromobacterium subtsugae strain PRAA4-1'. 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 (e.g.,
lack of toxicity noted for oral, dermal, and inhalation routes of exposure) available on
Chromobacterium subtsugae strain PRAA4-11 do not demonstrate toxic, pathogenic, and/or
infective potential to sensitive populations from exposure to this microbial pest control agent.
Thus, there are no threshold effects of concern and, as a result, the provision requiring an
additional margin of safety is not necessary.
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3. Occupational Exposure and Risk Characterization
Handler exposure to Chromobacterium subtsugae strain PRAA4-11 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 Chromobacterium subtsugae strain
PRAA4-11 end-use pesticide products in agricultural settings must wear a long-sleeved shirt,
long pants, socks, shoes, waterproof gloves, and a dust/mist filtering respirator meeting NIOSH
standards of at least N-95, R-95, or P-95. Additional PPE, other than the standard described
above, may be required on a product-specific basis.
4. Human Health Risk Characterization
EPA considered human exposure to Chromobacterium subtsugae strain PRAA4-11 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 Chromobacterium subtsugae strain PRAA4-11
pesticide products are used in accordance with EPA-approved labeling.
C. Environmental Assessment (40 CFR § 158.2150)
The data and data waiver rationale, submitted by the applicant to support the pesticide products
containing Chromobacterium subtsugae strain PRAA4-11, fulfill the Tier I nontarget organism
data requirements and are sufficient for risk assessment purposes. Further testing of nontarget
organisms at higher tier levels (i.e., Tiers II, IE, and IV) is not required for the current uses and
application methods. EPA has performed an environmental risk assessment based on the data and
data waiver rationale provided by the applicant and has determined that the use of
Chromobacterium subtsugae strain PRAA4-11 does not pose significant risk to nontarget
organisms when used according to label directions. Based on study results, there are concerns of
toxicity to terrestrial arthropods, aquatic invertebrates, and honey bees. Use sites and application
methods, in combination with mitigating label language, however, will limit exposure.
For a comprehensive summary of the generic data requirements described in sections IV(C)(1),
refer to Table 7 in Appendix A.
1. Ecological Exposure and Risk Characterization
a. Terrestrial Animals and Plants
The end-use product (EP) is a concentrated liquid that is labeled as a foliar spray with ground or
aerial applications at 3-10 day intervals. Aerial applications would increase exposure to nontarget
insects and honey bees and, due to potential toxicity concerns, were not recommended.
Terrestrial animals and plants may be exposed to Chromobacterium subtsugae strain PRAA4-1 T
through direct sprays and exposure on foliar surfaces.
Birds (OCSPP Guideline 885.4050) and Mammals (OCSPP Guideline 885.4150): A study
(MRID No. 479450-13) was conducted to determine the acute toxicity and pathogenicity of MBI-
203 TGAI (active ingredient, 100.00% Chromobacterium subtsugae strain PRAA4-P cells and
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spent fermentation media) to northern bobwhite (Colinus virginianus). Fourteen-day-old chicks
were dosed with the test material (4.0 x 1011 colony-forming units per kilogram (cfu/kg) body
weight), attenuated test material, or sterile saline once a day for five days and observed for an
additional 25 days. No signs of toxic or pathogenic effects were observed throughout the test or
at necropsy. This study demonstrates that toxicity/pathogenicity to birds is not expected from
labeled applications of Chromobacterium subtsugae strain PRAA4- P.
In an acute oral toxicity study (MRID No. 479450-03), three fasted young adult female Sprague-
Dawley rats were given a single oral gavage dose of MOI203 [MBI203] TGAI at a
concentration of 5,000 mg/kg body weight. The test material was allowed to thaw at room
temperature prior to dosing. The animals were observed for 14 days. All animals survived,
gained weight, and appeared active and healthy throughout the study. No gross abnormalities
were found in any animal at necropsy. This study demonstrates that toxicity/pathogenicity to wild
mammals is not expected from labeled applications of Chromobacterium subtsugae strain
PRAA4-11.
Nontarset Insects (OCSPP Guideline 885.4340) and Honey Bees (OCSPP Guideline 885.4380) :
Nontarget insect studies were submitted for ladybird beetles, green lacewing larvae and parasitic
wasps. Two studies were submitted for ladybird beetles, both of which indicated toxicity. An
acute toxicity study (MRID No. 479450-16) was of insufficient duration but supplemental for the
purposes of the risk assessment, and a feeding study (MRID No. 479450-17) fulfilled the testing
guidelines but also indicated toxicity. A green lacewing study (MRID No. 479450-18) that
fulfilled testing requirements was submitted; there were no observed effects from test
concentrations of Chromobacterium subtsugae strain PRAA4-P, and the LC50 for MBI TGAI
was greater than 70,000 parts per million (ppm) in a moth egg diet. A parasitic wasp study
(MRID No. 481129-05) was also submitted and was supplemental for the purposes of the risk
assessment. While there were no statistically significant differences in the control and test
substance groups, the dose of Chromobacterium subtsugae strain PRAA4-11 to which the wasps
were exposed was not adequately characterized and the duration of the test was insufficient.
Three honey bee studies were submitted, all of which were sufficient for the purposes of the risk
assessment. Results indicated toxicity to honey bees with the use of Chromobacterium subtsugae
strain PRAA4-1T (MRID Nos. 479450-19, 479450-20 and 479450-21).
In conclusion, according to studies conducted on nontarget insects and honey bees, there are
toxicity concerns with the use of Chromobacterium subtsugae strain PRAA4-11. To address
these concerns and to reduce exposure to nontarget insects and honey bees that may occur
through outdoor ground and aerial applications, EPA is requiring particular aerial drift reduction
information, as well as the following mitigation language, on the MB1-203 EP Bioinsecticide
label:
Environmental Hazards Statements
This product is highly toxic to bees exposed to direct treatment or residues on blooming crops or
weeds. Do not apply this product or allow it to drift to blooming crops or weeds if bees are
visiting the treatment area.
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This product is toxic to certain nontarget terrestrial arthropods. Minimize spray drift away from
target area to reduce effects to nontarget insects.
Nontarget Plants (OCSPP Guideline 885.4300): The applicant submitted both data waiver
rationale (MRID No. 479450-23) and a study (MRID No. 479450-15) to fulfill the nontarget
plant testing requirement. While Chromobacterium subtsugae strain PRAA4-P occurs naturally
in soils and is not related to any known plant pathogen, effects of mild phytotoxicity were
observed in cucumbers at 5X and 10X the field application rate, and in chrysanthemum flowers
at all application rates. Even though effects in cucumbers and chrysanthemum flowers were mild,
the MB1-203 EP Bioinsecticide label will still require all users to test for phytotoxicity prior to
pesticide application.
b. Aquatic Animals and Plants
Freshwater Fish (OCSPP Guideline 885.4200), Freshwater Invertebrates (OCSPP Guideline
885.4240). and Estuarine/Marine Fish and Invertebrates (OCSPP Guideline 885.4280): MBI-
203 EP Bioinsecticide (94.50% Chromobacterium subtsugae strain PRAA4-11) is not labeled for
aquatic uses, and the label includes language to prevent application to or contamination of water.
Due to demonstrated toxicity to aquatic invertebrates, aquatic uses should not be added to the
label without further testing. The applicant submitted studies to fulfill the freshwater fish and
freshwater invertebrate testing requirements, as well as data waiver rationale for testing on
estuarine/marine organisms. The freshwater fish study (MRID No. 479450-14) fulfills the testing
requirements and did not show any adverse effects to fathead minnows (Pimephalespromelas).
Toxicity was shown in two supplemental studies (MRID Nos. 479450-12 and 481129-04)
submitted by the applicant that tested the effects of Chromobacterium subtsugae strain
PRAA4-P on Daphnia magna. The studies were sufficient for the purposes of this registration,
but do not support aquatic uses. The data waiver rationale submitted for estuarine/marine
organisms was acceptable for the labeled use patterns, as MBI-203 EP Bioinsecticide is not
intended for direct application into the estuarine or marine environment or expected to enter such
environment in significant concentrations.
Even though MBI-203 EP Bioinsecticide is not applied directly to aquatic environments, EPA is
requiring that the maximum labeled application rate not exceed 12 quarts of product per acre and
that specific aerial drift reduction text, as well the following mitigation language, be included on
the label to reduce the potential for exposure to aquatic invertebrates:
Environmental Hazards Statements
This product is toxic to aquatic invertebrates. Drift and runoff may be hazardous to aquatic
organisms in water adjacent to treated areas.
Buffer Zone for Aerial Applications (in the Directions for Use) (see U.S. EPA 201 lc. 201 Id)
Do not apply within 75 feet of aquatic habitats (such as, but not limited to, lakes, reservoirs,
rivers, streams, marshes, ponds, estuaries, and commercial fish ponds).
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Information on Droplet Size (in the Directions for Use) (see U.S. EPA 201 lc. 201 Id)
Use only medium or coarser spray nozzles according to ASAE (S572) definition for standard
nozzles.
c. Open Literature Information
Literature searches were performed to determine if other effects have been reported that were not
contained within the information provided by the applicant. Both the Science Citation Index
Expanded database (1970-present) and the Environmental Information Search service (1964-
present) were searched using the terms "Chromobacterium" and "Chromobacterium subtsugae
The Environmental Information Search service includes the Agricola; Biosis Previews; CAB
Abstracts; Energy, Science, and Technology; General Science Abstracts; National Technical
Information Service; and Waternet databases. None of the records found in any of the searches
described here reported effects on any nontarget species. The only results relevant to the active
ingredient were two documents published by the United States Department of Agriculture -
Agricultural Research Service concerning the toxicity of Chromobacterium subtsugae to
Colorado potato beetle and other insect pests (Martin et al. 2007a), as well as to southern green
stink bug and corn rootworm (Martin et al. 2007b). Neither document provided additional
information on environmental or nontarget effects useful for the purposes of this risk assessment.
2. Environmental Fate Data
As the information provided, for the current uses and application methods, is sufficient to both
satisfy the Tier I nontarget organism data requirements and for nontarget organism risk
assessment for Chromobacterium subtsugae strain PRAA4-11, further testing at higher tier levels
(i.e., Tiers n, IE, and IV) is not required.
3. Threatened and Endangered Species Assessment
EPA has determined that there is a potential for effects to nontarget organisms, including
endangered and threatened species as a result of the proposed labeled applications (aerial sprays
and chemigation) of Chromobacterium subtsugae strain PRAA4-11. There are toxicity concerns
for certain terrestrial arthropods and honey bees, as well as for aquatic invertebrates. Further
analysis is needed to characterize the specific effects and affected species. For this reason, aerial
applications were not recommended. Terrestrial applications through sprinkler irrigation systems
(chemigation) are expected to result in exposure to nontarget insects, though to a lesser extent
than aerial applications. MBI-203 EP Bioinsecticide is also labeled for greenhouse use, which
will not result in significant exposure to nontarget organisms. Aquatic applications are not
permitted for this product, which will limit exposure to aquatic invertebrates and other aquatic
organisms. Mitigation language (e.g., requirement for a buffer between aquatic and treatment
areas and use of only medium to coarser spray nozzles for aerial applications), addressing
concerns of Chromobacterium subtsugae strain PRAA4-lT's toxicity to honey bees, terrestrial
arthropods, and aquatic invertebrates, has been added to the MBI-203 EP Bioinsecticide label.
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The Biopesticides and Pollution Prevention Division (BPPD) makes "no effect" determinations
for direct effects, indirect effects, and affects to habitat (including designated critical habitat) for
listed species for the following labeled applications of Chromobacterium subtsugae strain
PRAA4-P to the proposed crops:
• All foliar and soil directed liquid applications to the proposed crop plants treated within
enclosed greenhouses
BPPD concludes that the following types of labeled applications of Chromobacterium subtsugae
strain PRAA4-P to the proposed crops have the potential to cause direct effects, indirect effects,
and effects to habitat (including designated critical habitat) to listed species. Further analyses are
needed to characterize the effects that are likely to occur and the species potentially affected.
• All outdoor foliar applications made by chemigation and aerial spray equipment.
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
result their location, cultural practices, or other factors, may have atypical, unusually high
exposure to Chromobacterium subtsugae strain PRAA4-11, 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/environmentaliustice/index. html.
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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 Chromobacterium subtsugae strain PRAA4-11 pesticide products have
well-known properties. EPA has no knowledge that would contradict the claims made on the
MBI-203 TGAI and MBI-203 EP Bioinsecticide labels, and such products are not expected to
cause unreasonable adverse effects on the environment when used according to the 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 Chromobacterium subtsugae strain
PRAA4-11 pesticide products will not cause any unreasonable adverse effects on the
environment, and MBI-203 EP Bioinsecticide (end-use pesticide product), in particular, is likely
to provide protection against insect and mite pests as claimed, satisfying criterion 3. Criterion 4 is
satisfied in that the Chromobacterium subtsugae strain PRAA4-11 pesticide products are not
expected to cause unreasonable adverse effects when used according to label instructions.
Therefore, MBI-203 TGAI and MBI-203 EP Bioinsecticide, containing Chromobacterium
subtsugae strain PRAA4-11 as a new active ingredient, are eligible for registration under FIFRA
section 3(c)(5) for the labeled uses. If the applicant proposes uses that are more extensive in the
future (e.g., aquatic uses), EPA will likely require that additional data, in conjunction with a
registration amendment request, be submitted.
VII. ACTIONS REQUIRED BY THE REGISTRANT
A. Final Printed Labeling
Before releasing pesticide products containing Chromobacterium subtsugae strain PRAA4-11 for
shipment, the registrant is required to provide appropriate final printed labeling to EPA.
B. Terms of Registration
As terms of the MBI-203 EP Bioinsecticide registration, the registrant must submit the following
data within one year of this product's registration:
(1) Analysis of Samples (OCSPP Guideline 885.1400): A five-batch analysis, addressing all
quality control parameters (i.e., dry weight, standard plate count, viscosity, density, and
pH).
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(2) Storage Stability (OCSPP Guideline 830.6317) and Corrosion Characteristics (OCSPP
Guideline 830.6320): The results of a two-year storage stability and corrosion
characteristics study (ongoing).
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
ASAE
BPPD
BRAD
CFR
cfu
cfu/kg
cfu/mL
cP
EDSP
EP
EPA
FFDCA
FIFRA
FQPA
FR
g/mL
LC50
LD
50
mg/kg
mg/L
MP
MRIDNo.
NIOSH
NRRL
OCSPP
OPP
PC Code
PP
PPE
ppm
TGAI
U.S.
American Society of Agricultural Engineers
Biopesticides and Pollution Prevention Division
Biopesticides Registration Action Document
Code of Federal Regulations
colony-forming unit(s)
colony-forming units per kilogram
colony-forming units per milliliter
centipoise
Endocrine Disruptor Screening Program
end-use product
Environmental Protection Agency (the "Agency")
Federal Food, Drug, and Cosmetic Act
Federal Insecticide, Fungicide, and Rodenticide Act
Food Quality Protection Act
Federal Register
gram per milliliter
median lethal concentration. A statistically derived concentration of a
substance that can be expected to cause death in 50% of test animals. It is
usually expressed as the weight of substance per weight or volume of water,
air, or feed (e.g., mg/L, mg/kg, or ppm).
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).
milligrams per kilogram
milligrams per liter
manufacturing-use product
Master Record Identification Number
National Institute for Occupational Safety and Health
Northern Regional Research Laboratory
Office of Chemical Safety and Pollution Prevention
Office of Pesticide Programs
Pesticide Chemical Code
Pesticide Petition
personal protective equipment
parts per million
technical grade of the active ingredient
United States
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IX. BIBLIOGRAPHY
A. Studies Submitted to Support the Chromobacterium subtsugae strain PRAA4-11
Pesticide Product Registrations
479450-01 Koivunen M. 2009. Product Chemistry for MB1-203 TGAI. Unpublished study
prepared by Marrone Bio Innovations, 190 pages.
479450-02 Koivunen M. 2009. Product Chemistry for MBI-203 EP. Unpublished study
prepared by Marrone Bio Innovations, 143 pages.
479450-03 Oley S. 2009. MOI203 Chromobacteria: Acute Oral Toxicity Up and Down
Procedure in Rats. Project Number: 25936, P320/UDP. Unpublished study
prepared by Eurofins/Product Safety Laboratories, 15 pages.
479450-04 Kuhn J. 2009. MOI 203 Chromobacteria: Acute Dermal Toxicity Study in Rats -
Final Report. Project Number: 12780/09. Unpublished study prepared by
Stillmeadow, Incorporated, 11 pages.
479450-05 Younger C. 2009. MOI 203 Chromobacteria: Acute Inhalation Toxicity Study in
Rats - Final Report. Project Number: 12781/09. Unpublished study prepared by
Stillmeadow, Incorporated, 17 pages.
479450-06 Kuhn J. 2009. MOI 203 Chromobacteria: Acute Eye Irritation Study in Rabbits -
Final Report. Project Number: 12782/09. Unpublished study prepared by
Stillmeadow, Incorporated, 16 pages.
479450-07 Kuhn J. 2009. MOI 203 Chromobacteria: Acute Dermal Irritation Study in
Rabbits - Final Report. Project Number: 12783/09. Unpublished study prepared
by Stillmeadow, Incorporated, 11 pages.
479450-08 Kuhn J. 2009. MOI 203 Chromobacteria: Skin Sensitization Study in Guinea Pigs
- Final Report. Project Number: 12784/09. Unpublished study prepared by
Stillmeadow, Incorporated, 16 pages.
479450-09 Monds K. 2009. MOI 203 Chromobacteria: MPCA Mouse Safety Study - Final
Report. Project Number: 12446/08. Unpublished study prepared by Stillmeadow,
Incorporated, 10 pages.
479450-10 Monds K. 2009. MOI 203 Chromobacterium: Acute Injection
Toxicity/Pathogenicity Pilot Study in Mice - Final Report. Project Number:
12661/08. Unpublished study prepared by Stillmeadow, Incorporated, 11 pages.
479450-11 Monds K. 2009. MOI 203 Chromobacterium: Acute Injection Toxicity/Infectivity
Study in Rats - Final Report. Project Number: 13056/09. Unpublished study
prepared by Stillmeadow, Incorporated, 19 pages.
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479450-12 Gallagher S, Krueger H. 2009. MBI-203: A 48-Hour Static Acute Toxicity Test
with the Cladoceran (Daphnia magna) - Final Report. Project Number:
664A/103A. Unpublished study prepared by Wildlife International, Limited, 38
pages.
479450-13 Hubbard P, Beavers J. 2009. MBI-203: An Avian Oral Pathogenicity and Toxicity
Study in the Northern Bobwhite. Project Number: 664/101. Unpublished study
prepared by Wildlife International, Limited, 50 pages.
479450-14 Gallagher S, Krueger H. 2009. MBI-203: A Five-Concentration Toxicity and
Pathogenicity Test with the Fathead Minnow (Pimephalespromelas) - Final
Report. Project Number: 664A/102. Unpublished study prepared by Wildlife
International, Limited, 61 pages.
479450-15 Morgan C. 2009. MBI-203: Nontarget Plants. Project Number: 4401/11, 1503/18,
4501/16. Unpublished study prepared by Marrone Bio Innovations, 8 pages.
479450-16 Chanbusarakum L. 2009. MBI-203: Ladybird Beetle (.Hippodamia convergens)
Contact Assay. Project Number: 2702/145. Unpublished study prepared by
Marrone Bio Innovations, 9 pages.
479450-17 Chanbusarakum L. 2009. MBI-203: Ladybird Beetle (Hippodamia convergens)
Feeding Assay. Project Number: 104/47. Unpublished study prepared by Marrone
Bio Innovations, 9 pages.
479450-18 Porch J, Krueger H. 2009. MBI-203: A Dietary Pathogenicity and Toxicity Study
with Green Lacewing Larvae (Chrysoperla rufilabris) - Final Report. Project
Number: 664/103B. Unpublished study prepared by Wildlife International,
Limited, 46 pages.
479450-19 Richards K. 2009. Evaluation of the Dietary Effect(s) of MB1203 on Adult Honey
Bees {Apis mellifera!^). Project Number: CAR/140/09. Unpublished study
prepared by California Agricultural Research, Incorporated, 45 pages.
479450-20 Richards K. 2009. Evaluation of the Dietary Effect(s) of MB1203 on Adult Honey
Bees (Apis mellifera L.) - Final Report. Project Number: CAR/140/09B.
Unpublished study prepared by California Agricultural Research, Incorporated, 12
pages.
479450-21 Richards K. 2009. Evaluation of the Dietary Effect(s) of MB1203 on Adult Honey
Bees (Apis mellifera L.) - Final Report. Project Number: CAR/140/09A.
Unpublished study prepared by California Agricultural Research, Incorporated, 11
pages.
479450-22 Mileson B. 2009. Endangered Species Evaluation for MBI-203. Unpublished
study prepared by Technology Sciences Group, Incorporated, 23 pages.
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479450-23 Mileson B. 2009. Response to Tier 1 Microbial Pesticide Data Requirements for
MBI-203 TGAI. Unpublished study prepared by Technology Sciences Group,
Incorporated, 29 pages.
479450-24 Mileson B. 2009. Response to Tier 1 Microbial Pesticide Data Requirements for
MBI-203 EP. Unpublished study prepared by Technology Sciences Group,
Incorporated, 77 pages.
479450-25 Pitts K. 2009. Published Literature to Support the Registration of MB1203.
Unpublished study prepared by Marrone Bio Innovations, 275 pages.
481129-02 Koivunen M. 2010. Supplemental Product Identity Study, Secondary Metabolites
(MBI-203). Unpublished study prepared by Marrone Bio Innovations, 136 pages.
481129-03 Chanbusarakum L. 2010. Supplemental Manufacturing Process Study, Cabbage
Looper Potency Unit Justification (MBI-203). Unpublished study prepared by
Marrone Bio Innovations, 25 pages.
481129-04 Gallagher S, Krueger H. 2009. MBI-203: A 21-Day Life Cycle Toxicity and
Pathogenicity Test with the Cladoceran (Daphnia magna). Project Number:
664A/101. Unpublished study prepared by Wildlife International, Limited, 33
pages.
481129-05 Younger C. 2010. MBI203: Parasitic Wasp (Aphidius matricariae), Nontarget
Insect Microbial Testing - Final Report. Project Number: 13902/10. Unpublished
study prepared by Stillmeadow, Incorporated, 16 pages.
481129-06 Koivunen M. 2010. Supplemental Product Identity and Manufacturing Study,
Replacement of Inert Ingredient for MBI-203 EP. Unpublished study prepared by
Marrone Bio Innovations, 24 pages.
481129-07 Chanbusarakum L. 2010. Supplemental Manufacturing Process, Cabbage Looper
Potency Unit Justification for MBI-203 EP. Unpublished study prepared by
Marrone Bio Innovations, 27 pages.
481129-08 Pitts K. 2010. Response to Tier 1 Microbial Pesticide Data Requirements for
MBI-203 EP: Supplemental Study for Revised EP Formulation. Unpublished
study prepared by Marrone Bio Innovations, 123 pages.
481307-01 Chanbusarakum L. 2010. MBI 203 TGAI: Supplemental Product Identity Study,
Mode of Action. Unpublished study prepared by Marrone Bio Innovations, 30
pages.
484406-01 Pitts K. 2011. MBI 203 TGAI: 12-Month Storage Stability and Corrosion
Characteristics Study. Unpublished study prepared by Marrone Bio Innovations,
21 pages.
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484407-01 Pitts K. 2011. MBI203 EP: 7-Month Storage Stability and Corrosion
Characteristics Study. Unpublished study prepared by Marrone Bio Innovations,
21 pages.
B. Environmental Protection Agency Risk Assessment Memoranda and Other Documents
U.S. EPA. 201 la. Chromobacterium subtsugae strain PRAA4-1'. Memorandum from
J.V. Gagliardi, Ph.D. and J.L. Kough, Ph.D. to J. Kausch dated April 21, 2011.
U.S. EPA. 201 lb. Environmental Risk Assessment of Chromobacterium subtsugae strain
PRAA4-11 (PC Code: 016329) for Section 3 Registrations of the Technical Product
(MBI-203 TGAI; EPA File Symbol 84059-0) and End Use Product (MBI-203 EP; EPA
File Symbol 84059-RN) for Control of Insects and Mites in Ornamental Plants, Turf and
Edible Crops, Memorandum from A. Gross and Z. Vaituzis, Ph.D. to J. Kausch dated
July 7,2011.
U.S. EPA. 201 lc. Spray Drift Buffers for Chromobacterium subtsugae strain PRAA4-I. Analysis
drafted by S. Borges and dated August 11, 2011.
U.S. EPA. 201 Id. Spray Buffers Revised C. subtsugae. Spreadsheet assembled by S. Borges and
dated August 17, 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.
Centers for Disease Control and Prevention. 2009. Drinking Water - Water Treatment. Available
from http://www.cdc.sov/healthywater/drinkins/public/water treatment.html.
DeFelice K, Wollenhaupt N, Buchholz D. 1993. Aquifers and Soil Filter Effect. Available from
http://extension, missouri. edu/p/W024.
Lindow SE, Brandl MT. 2003. Microbiology of the phyllosphere. Applied and Environmental
Microbiology 69:1875-1883.
Martin PAW, Gundersen-Rindal D, Blackburn M, Buyer J. 2007a. Chromobacterium subtsugae
sp. nov., a betaproteobacterium toxic to Colorado potato beetle and other insect pests.
International Journal of Systematic and Evolutionary Microbiology 57:993-999.
Martin PAW, Hirose E, Aldrich JR. 2007b. Toxicity of Chromobacterium subtsugae to southern
green stink bug (Heteroptera: Pentatomidae) and corn rootworm (Coleoptera:
Chrysomelidae). Journal of Economic Entomology 100(3):680-684.
Pang L, McLeod M, Aislabie J, Simunek J, Close M, Hector R. 2008. Modeling transport of
microbes in ten undisturbed soils under effluent irrigation. Vadose Zone Journal 7:97-
111.
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Chromobacterium subtsugae strain PRAA4-11
Biopesticides Registration Action Document
Page 25 of 36
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.
U.S. EPA. 1996. Microbial Pesticide Test Guidelines - Background for Residue Analysis of
Microbial Pest Control Agents (OPPTS 885.2000). Available from
http://www. epa. gov/ocspp/pubs/frs/publications/Test Guidelines/series885. htm.
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Chromobacterium subtsugae strain PRAA4-11
Biopesticides Registration Action Document
Page 26 of 36
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)/Manufacturing-Use Product (MP), MBI-203 TGAI (40 CFR § 158.2120)
OCSPP
Guideline
Number
Data Requirement
Results
MRID No.
TGAI
MP
885.1100
Product Identity
Not applicable
Submitted data fulfill the
requirement for product
identity. MBI-203 TGAI
contains 100.00% by weight
Chromobacterium subtsugae
strain PRAA4-11 cells and
spent fermentation media
(minimum of 1,500 Cabbage
Looper Potency (or Killing)
Units per milligram).
479450-01
481129-02
481307-01
885.1200
Manufacturing Process
Submitted data fulfill the requirement for manufacturing
process.
479450-01
481129-03
Not applicable
Deposition of a Sample
in a Nationally
Recognized Culture
Collection
Chromobacterium subtsugae
strain PRAA4-11 is on
deposit with the Agricultural
Research Service Culture
Collection (also known as the
Northern Regional Research
Laboratory Collection) in
Peoria, Illinois under
Accession Number NRRL B-
30655.
Not applicable
479450-01
885.1300
Discussion of Formation
of Unintentional
Ingredients
Submitted data fulfill the requirement for discussion of
formation of unintentional ingredients.
479450-01
885.1400
Analysis of Samples
Submitted data fulfill the requirement for analysis of samples.
479450-01
885.1500
Certification of Limits
Not applicable
Limits listed on the
confidential statement of
formula are
adequate/acceptable.
479450-01
Additional Studies
830.1800
Enforcement Analytical
Method
Not applicable
Submitted data fulfill the
requirement for an
enforcement analytical
method.
479450-01
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Chromobacterium subtsugae strain PRAA4-11
Biopesticides Registration Action Document
Page 27 of 36
TABLE 2. Product Analysis Data Requirements for
(40 CFR \
the End-Use Product (EP), MBI-203 EP Bioinsecticide
158.2120)
OCSPP Guideline Number
Data Requirement
Results
MRU) No.
885.1100
Product Identity
Submitted data fulfill the requirement for
product identity. MBI-203 EP
Bioinsecticide contains 94.50% by weight
Chromobacterium subtsugae strain
PRAA4-11 cells and spent fermentation
media (minimum of 1,000 Cabbage Looper
Potency (or Killing) Units per milligram).
479450-02
481129-06
885.1200
Manufacturing Process
Submitted data fulfill the requirement for
manufacturing process.
479450-02
481129-06
481129-07
Not applicable
Deposition of a Sample in a
Nationally Recognized Culture
Collection
Not applicable
Not applicable
885.1300
Discussion of Formation of
Unintentional Ingredients
Submitted data fulfill the requirement for
discussion of formation of unintentional
ingredients.
479450-02
885.1400
Analysis of Samples
Submitted date fulfill the requirement for
analysis of samples for purposes of FIFRA
section 3(c)(5) registration. As a term of
the MBI-203 EP Bioinsecticide
registration, EPA is requiring a five-
batch analysis, addressing all quality
control parameters (i.e., dry weight,
standard plate count, viscosity, density,
and pH), be submitted within one year.
479450-02
885.1500
Certification of Limits
Limits listed on the confidential statement
of formula are adequate/acceptable.
479450-02
Additional Studies
830.1800
Enforcement Analytical
Method
Submitted data fulfill the requirement for
an enforcement analytical method.
479450-02
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Chromobacterium subtsugae strain PRAA4-11
Biopesticides Registration Action Document
Page 28 of 36
TABLE 3. Physical and Chemical Characteristics for the Technical Grade of the Active Ingredient
(TGAI)/Manufacturing-Use Product (MP), MBI-203 TGAI (40 CFR § 158.2120)
OCSPP
Guideline
Number
Data Requirement
Results
MRU) No.
TGAI
MP
830.6302
Color
Purple-blue
Not applicable
479450-01
830.6303
Physical State
Liquid
Not applicable
479450-01
830.6304
Odor
Grass-like
Not applicable
479450-01
830.6313
Stability to Normal and
Elevated
Temperatures, Metals,
and Metal Ions
Expected to be stable
Not applicable
479450-01
830.6317
Storage Stability
Stable for one year when stored per label directions.
479450-01
484406-01
830.6319
Miscibility
Not applicable
Not required because MBI-203
TGAI is not an emulsifiable form
of microbial pesticide (refer to test
note #2 of 40 CFR § 158.2120(d)).
Not
applicable
830.6320
Corrosion
Characteristics
Not applicable
Not corrosive to packaging
materials in a one-year study.
479450-01
484406-01
830.7000
pH
4.1 (l%w/w)
Not applicable
479450-01
830.7100
Viscosity
Not applicable
5.4 cP
479450-01
830.7300
Density/Relative
Density/Bulk Density
(Specific Gravity)
1.01 g/mL
Not applicable
479450-01
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Chromobacterium subtsugae strain PRAA4-11
Biopesticides Registration Action Document
Page 29 of 36
TABLE 4. Physical and Chemical Characteristics for the End-Use Product (EP), MBI-203 EP Bioinsecticide
(40 CFR § 158.2120)
OCSPP Guideline Number
Data Requirement
Results
MRU) No.
830.6302*
Color
Purple-blue
479450-02
830.6303*
Physical State
Liquid
479450-02
830.6304*
Odor
Herbaceous
479450-02
830.6313
Stability to Normal and
Elevated Temperatures,
Metals, and Metal Ions
Not applicable
Not applicable
830.6317
Storage Stability
Expected to be stable for two years when
stored per label instructions. As a term of
the MBI-203 EP Bioinsecticide
registration, EPA is requiring the
results of a two-year storage stability
and corrosion characteristics study
(ongoing) be submitted within one
year.
479450-02
484407-01
830.6319
Miscibility
Miscible with water
479450-02
830.6320
Corrosion Characteristics
Not expected to be corrosive to packaging
materials. As a term of the MBI-203 EP
Bioinsecticide registration, EPA is
requiring the results of a two-year
storage stability and corrosion
characteristics study (ongoing) be
submitted within one year.
479450-02
484407-01
830.7000*
pH
6.2
479450-02
830.7100
Viscosity
21.8 cP
479450-02
830.7300*
Density/Relative Density/Bulk
Density (Specific Gravity)
1.02 g/mL
479450-02
* According to 40 CFR § 158.2120, these data are only required for the technical grade of the active
ingredient. Since Marrone Bio Innovations included this information with its application for MBI-203 EP
Bioinsecticide, it is summarized appropriately in this table.
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Chromobacterium subtsugae strain PRAA4-11
Biopesticides Registration Action Document
Page 30 of 36
TABLE 5. Toxicology Data Requirements for the Technical Grade of the Active Ingredient (TGAI)/Manufacturing-Use
Product (MP), MBI-203 TGAI (40 CFR § 158.2140)
OCSPP Guideline
Number
Data Requirement
Results
MRID No.
TGAI
MP
Tier I
885.3050
Acute Oral
T oxicity/Pathogenicity
Waived based on the results of
MRID Nos. 479450-11 and
479450-03.
Classification: Acceptable
Not applicable
479450-23
885.3150
Acute Pulmonary
Toxicity/Pathogenicity
Waived based on the results of
MRID Nos. 479450-11 and
479450-05.
Classification: Acceptable
Not applicable
479450-23
885.3200
Acute Injection
Toxicity/Pathogenicity
(Intravenous)
Not toxic, infective, and/or
pathogenic to rats when dosed
intravenously at 3.1 x 106 cfu
per animal.
Classification: Acceptable
Not applicable
479450-11
885.3400
Hypersensitivity
Incidents
No hypersensitivity incidents, including immediate-type or
delayed-type reactions of humans and domestic animals that
occurred during research, development, or testing of the
TGAI/MP, were reported by the applicant. Any future
hypersensitivity incidents must be reported to EPA (refer to test
note #3 of 40 CFR § 158.2140(d)).
479450-23
885.3500
Cell Culture
Not required because
Chromobacterium subtsugae
strain PRAA4-11 is not a virus
(refer to test note #4 of 40
CFR § 158.2140(d)).
Not applicable
Not
applicable
870.1100
Acute Oral Toxicity
Not applicable
Oral LD50 female rats > 5,000
mg/kg
Classification: Acceptable
TOXICITY CATEGORY IV
479450-03
870.1200
Acute Dermal Toxicity
Not applicable
Dermal LD50 combined (male
and female) rats > 5,050 mg/kg
Classification: Acceptable
TOXICITY CATEGORY IV
479450-04
870.1300
Acute Inhalation
Toxicity
Not applicable
Inhalation LC50 combined
(male and female) rats >2.12
mg/L
Classification: Acceptable
TOXICITY CATEGORY IV
479450-05
870.2400
Acute Eye Irritation
Not applicable
MBI-203 TGAI was minimally
irritating to the eyes of rabbits.
Classification: Acceptable
TOXICITY CATEGORY IV
479450-06
870.2500
Primary Dermal
Irritation
Not applicable
MBI-203 TGAI was slightly
irritating to the skin of rabbits.
Classification: Acceptable
TOXICITY CATEGORY IV
479450-07
Tiers II and III
Not required for Chromobacterium subtsugae strain PRAA4-11 based on the lack of acute toxicity/pathogenicity in the Tier I
studies.
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Chromobacterium subtsugae strain PRAA4-11
Biopesticides Registration Action Document
Page 31 of 36
TABLE 5. Toxicology Data Requirements for the Technical Grade of the Active Ingredient (TGAI)/Manufacturing-Use
Product (MP), MBI-203 TGAI (40 CFR § 158.2140)
OCSPP Guideline
Number
Data Requirement
Results
MRU) No.
TGAI
MP
Additional Studies
870.2600
Skin Sensitization
Not a dermal sensitizer
Classification: Acceptable
479450-08
Not applicable
Safety Study
(Subcutaneous
Injection)
Not toxic or pathogenic at > 106 cfu/mouse subcutaneously
Classification: Supplemental - Not a required study
479450-09
885.3200
Acute Injection
Toxicity/Pathogenicity
(Intravenous) Pilot
Study
Provides no useful data as to the mechanism of toxicity observed
Classification: Supplemental - Not a required study
479450-10
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Biopesticides Registration Action Document
Page 32 of 36
TABLE 6. Toxicology Data Requirements for the End-Use Product (EP), MBI-203 EP Bioinsecticide
(40 CFR § 158.2140)
OCSPP Guideline Number
Data Requirement
Results
MRID No.
885.3050
Acute Oral
T oxicity/Pathogenicity
Not applicable
Not applicable
885.3150
Acute Pulmonary
T oxicity/Pathogenicity
Not applicable
Not applicable
885.3200
Acute Injection
T oxicity/Pathogenicity
Not applicable
Not applicable
885.3400
Hypersensitivity Incidents
No hypersensitivity incidents, including
immediate-type or delayed-type
reactions of humans and domestic
animals that occurred during research,
development, or testing of the EP, were
reported by the applicant. Any future
hypersensitivity incidents must be
reported to EPA (refer to test note #3 of
40 CFR § 158.2140(d)).
Not applicable
885.3500
Cell Culture
Not applicable
Not applicable
870.1100
Acute Oral Toxicity
Waived based on the results of MRID
No. 479450-03 and because this
formulation contains inert ingredients
that are present in small amounts, are not
expected to be of toxicological concern,
and are cleared for food use.
Classification: Acceptable
TOXICITY CATEGORY III
479450-24
481129-08
870.1200
Acute Dermal Toxicity
Waived based on the results of MRID
No. 479450-04 and because this
formulation contains inert ingredients
that are present in small amounts, are not
expected to be of toxicological concern,
and are cleared for food use.
Classification: Acceptable
TOXICITY CATEGORY III
479450-24
481129-08
870.1300
Acute Inhalation Toxicity
Waived based on the results of MRID
No. 479450-05 and because this
formulation contains inert ingredients
that are present in small amounts, are not
expected to be of toxicological concern,
and are cleared for food use.
Classification: Acceptable
TOXICITY CATEGORY III
479450-24
481129-08
870.2400
Acute Eye Irritation
Waived based on the results of MRID
No. 479450-06 and because this
formulation contains inert ingredients
that are present in small amounts, are not
expected to be of toxicological concern,
and are cleared for food use.
Classification: Acceptable
TOXICITY CATEGORY III
479450-24
481129-08
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Biopesticides Registration Action Document
Page 33 of 36
TABLE 6. Toxicology Data Requirements for the End-Use Product (EP), MBI-203 EP Bioinsecticide
(40 CFR § 158.2140)
OCSPP Guideline Number
Data Requirement
Results
MRID No.
870.2500
Primary Dermal Irritation
Waived based on the results of MRID
No. 479450-07 and because this
formulation contains inert ingredients
that are present in small amounts, are not
expected to be of toxicological concern,
and are cleared for food use.
Classification: Acceptable
TOXICITY CATEGORY III
479450-24
481129-08
TABLE 7. Nontarget Organism Toxicity and Environmental Fate Data Requirements for the Technical Grade of the
Active Ingredient (TGAI), Chromobacterium subtsugae strain PRAA4-1T (40 CFR § 158.2150)
OCSPP Guideline
Number
Data Requirement
Results
MRID No.
Tier I
885.4050
Avian Oral Toxicity
The no-observed-effect dosage of MBI-203 TGAI
in this study was 4.0 x 1011 cfu/kgbody
weight/day for 5 days.
Classification: Acceptable
479450-13
885.4100
Avian Inhalation
T oxicity/Pathogenicity
Data waiver rationale provides sufficient
information to determine that pathogenicity to
birds is not expected.
Classification: Acceptable
479450-23
885.4150
Wild Mammal
T oxicity/Pathogenicity
Tests required by 40 CFR § 158.2140 are
adequate and appropriate for assessment of
hazards to wild mammals. Testing indicates no
adverse effects to laboratory rats when dosed
orally at 5,000 mg MBI-203 TGAI/kg body
weight.
Classification: Acceptable
479450-03
885.4200
Freshwater Fish
T oxicity/Pathogenicity
The study indicates no adverse effects to fathead
minnow (Pimephales promelas) when dosed with
Chromobacterium subtsugae at a rate of 8.24 x
107 cfu/mL.
Classification: Acceptable
479450-14
885.4240
Freshwater Invertebrate
T oxicity/Pathogenicity
Tests indicate toxicity to Daphnia magna, and the
addition of aquatic uses would require further
testing.
Classification: Supplemental but acceptable
for risk assessment
479450-12
481129-04
The following mitigating language is required in
the Environmental Hazards section of the MBI-
203 EP Bioinsecticide label: "This product is
toxic to aquatic invertebrates. Drift and runoff
may be hazardous to aquatic organisms in water
adjacent to treated areas." Additionally, the MBI-
203 EP Bioinsecticide label will (1) bear a
maximum application rate of 12 quarts of product
per acre, (2) include a restriction that aerial
applications not occur within 75 feet of aquatic
habitats, (3) require aerial applicators to use
medium or coarser spray nozzles, and (4) list
certain requirements and instructions to further
reduce potential for spray drift to offsite areas.
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Chromobacterium subtsugae strain PRAA4-11
Biopesticides Registration Action Document
Page 34 of 36
TABLE 7. Nontarget Organism Toxicity and Environmental Fate Data Requirements for the Technical Grade of the
Active Ingredient (TGAI), Chromobacterium subtsugae strain PRAA4-1T (40 CFR § 158.2150)
OCSPP Guideline
Number
Data Requirement
Results
MRU) No.
885.4280
Estuarine/Marine Fish and
Invertebrate Testing
Not required because Chromobacterium
subtsugae strain PRAA4-11 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 applicable
885.4300
Nontarget Plant Testing
Effects of mild phytotoxicity were observed in
cucumbers at 5X and 10X the field application
rates and in chrysanthemum flowers at all
application rates.
Classification: Supplemental but acceptable
for risk assessment
479450-15
Even though effects in cucumbers and
chrysanthemum flowers were mild, the MBI-203
EP Bioinsecticide label will still require all users
to test for phytotoxicity prior to pesticide
application.
885.4340
Nontarget Insect Testing
Tests indicate toxicity to ladybird beetles. There
were no observed effects to green lacewing larvae
or parasitic wasps. The dosing and duration of the
parasitic wasp study, however, were insufficient
for a conclusive determination of no effects.
Classification: Supplemental but acceptable
for risk assessment
479450-16
479450-17
479450-18
481129-05
The following mitigating language is required in
the Environmental Hazards section of the MBI-
203 EP Bioinsecticide label: "This product is
toxic to certain nontarget terrestrial arthropods.
Minimize spray drift away from target area to
reduce effects to nontarget insects." Further
testing would be required for the removal of this
mitigating language from the MBI-203 EP
Bioinsecticide label. Additionally, the MBI-203
EP Bioinsecticide label will list certain
requirements and instructions to further reduce
potential for spray drift to offsite areas.
885.4380
Honey Bee Testing
Tests indicate toxicity to Apis mellifera.
Classification: Supplemental but acceptable
for risk assessment
479450-19
479450-20
479450-21
The following mitigating language is required in
the Environmental Hazards section of the MBI-
203 EP Bioinsecticide label: "This product is
highly toxic to bees exposed to direct treatment or
residues on blooming crops or weeds. Do not
apply this product or allow it to drift to blooming
crops or weeds if bees are visiting the treatment
area." Further testing would be required for the
removal of this mitigating language from the
MBI-203 EP Bioinsecticide label. Additionally,
the MBI-203 EP Bioinsecticide label will list
certain requirements and instructions to further
reduce potential for spray drift to offsite areas.
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Chromobacterium subtsugae strain PRAA4-11
Biopesticides Registration Action Document
Page 35 of 36
TABLE 7. Nontarget Organism Toxicity and Environmental Fate Data Requirements for the Technical Grade of the
Active Ingredient (TGAI), Chromobacterium subtsugae strain PRAA4-1T (40 CFR § 158.2150)
OCSPP Guideline
Number
Data Requirement
Results
MRU) No.
Tiers II, III, and IV
Not required for Chromobacterium subtsugae strain PRAA4-11 based on the current uses and application methods.
Additional Studies
Not applicable
Endangered Species
Evaluation
Sufficient information was provided to determine
that any potential risk from use of MBI-203
TGAI to endangered mammalian, avian, or fish
species would likely be below the level of
concern, and that adverse effects to endangered
plant species are unlikely. Sufficient information
was not provided to determine the potential risk
to endangered nontarget insects or aquatic
invertebrates. BPPD will make an independent
endangered species determination.
Classification: Supplemental - not a required
study
479450-22
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Page 36 of 36
APPENDIX B. PESTICIDE PRODUCTS
EPA
Registration
Number
Registration
Name
Percentage
Active
Ingredient
Formulation
Type
Use Site(s)
Method(s)
of
Application
Application
Rate
Target
Pest
84059-9
MBI-203
TGAI
100.00%
Technical
N/A
N/A
N/A
N/A
84059-10
MBI-203 EP
Bioinsecticide
94.50%
End Use -
Liquid
Various
agricultural
and
greenhouse
crops (e.g.,
vegetables,
ornamentals,
and turf)
Standard
ground or
aerial spray
application
equipment
and
chemigation
2-12 quarts
per acre
Various
insects and
mites
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