OFFICE OF CHEMICAL SAFETY
AND POLLUTION PREVENTION
UNITED STATES ENVIRONMENTAL PROTECTION AGENCY
WASHINGTON, D.C. 20460
MEMORANDUM
SUBJECT: Human Health Risk Assessment for the New Active Ingredients Bacillus
subtilis strain RTI477 and Bacillus velezensis strain RTI301 in the Proposed
Manufacturing-use Products 279-OAUT, 279-OAUI and End-use Products 279-
OAUO, 279-OALN and 279-OALR for FIFRA Section 3 Registration with
Tolerance Exemption Petitions
Decision Numbers: 549769, 549770, 549772, 549771, 549767, 549773, 549774
DP Numbers: 451944, 451948, 451958, 452003, 452028, 457332, 457333
Submission Numbers: 1032814, 1032816, 1032837, 1032818, 1032828, 1032815,
1032817
EPA File Symbols: 279-OAUT, 279-OAUI, 279-OALR, 279-OALN, 279-OAUO
Active Ingredient Names: Bacillus subtilis strain RTI477 and Bacillus velezensis strain
RTI301
PC Codes: 006552, 006553
Tolerance Exemption
Petitions: 9F8749 (B. subtilis strain RTI477),
9F8750 (B. velezensis strain RTI301)
Associated MRIDs: 50354402, 50354403, 50354404, 50354405, 50354406, 50354415,
50354501, 50354503, 50354504, 50354505, 50354506, 50354515,
50354601, 50354602, 50354603, 50354701, 50354702, 50354703,
50354801, 50354802, 50354803, 50354804, 50354805, 50354414,
50925702, 51255601, 50354411, 50354412, 50354413, 50354418,
50354514, 50925701, 51255501, 50354511, 50354512, 50354513,
50354518, 50354705, 51391802, 50354707, 50354706, 50354704,
50354708, 50354605, 51391801, 50354607, 50354606, 50354604,
50354608, 50354808, 51391803, 50354809, 50354810, 50354811,
50354807, 50644801, 50923401, 50644901, 50923601, 50354812
FROM: Mark Perry, Risk Assessment Process Leader
Risk Assessment Branch
Biopesticides and Pollution Prevention Division
THRU: Shannon Borges, Branch Chief
John Kough, Ph.D., Senior Scientist
Risk Assessment Branch
Biopesticides and Pollution Prevention Division
Active Ingredients: B. subtilis strain RTI477 / B. velezensis strain RTI301 PC Codes: 006552, 006553
Action: Human Health Risk Assessment
2
TO: Monica Thapa, Risk Manager
Microbial Pesticides Branch
Biopesticides and Pollution Prevention Division
I. Action Requested
Under Section 3 of the Federal Insecticide, Fungicide, and Rodenticide Act (FIFRA), FMC
Corporation has requested registration of two new food-use active ingredients, Bacillus subtilis
strain RTI477 (hereafter “Bacillus subtilis RTI477”) and Bacillus velezensis strain RTI301
(hereafter “Bacillus velezensis RTI301”). This application includes two proposed manufacturing-
use products, Bacillus subtilis RTI477 Technical (EPA File Symbol 279-OAUT) and Bacillus
velezensis RTI301 Technical (EPA File Symbol 279-OAUI), as well as three end-use products.
The proposed end-use products are F4034-5 (EPA File Symbol 279-OALN) containing Bacillus
subtilis RTI477 and Bacillus velezensis RTI301, F4007-9 (EPA File Symbol 279-OAUO)
containing Bacillus velezensis RTI301, and F4092-3 (EPA File Symbol 279-OALR) containing
Bacillus subtilis RTI477, Bacillus velezensis RTI301 and bifenthrin as the active ingredients.
The three end-use products (F4034-5, F4007-9 and F4092-3) are proposed for use on a wide
variety of crops to protect seedlings against fungal diseases primarily caused by Rhizoctonia,
Fusarium, and Phytophthora. Since F4092-3 contains bifenthrin as an active ingredient, it also
has insecticidal uses. In support of registration, the applicant has submitted product analysis data,
mammalian toxicology data, Confidential Statements of Formula (CSFs) and data matrices for
the proposed manufacturing-use and end-use products. Since FMC Corporation has requested the
establishment of tolerance exemptions for Bacillus subtilis RTI477 and Bacillus velezensis
RTI301 in or on all food commodities, tolerance exemption petitions under 9F8749 (Bacillus
subtilis RTI477) and 9F8750 (Bacillus velezensis RTI301) have also been submitted.
II. Executive Summary
FMC Corporation has applied for registration of the new food-use active ingredients Bacillus
subtilis strain RTI477 and Bacillus velezensis strain RTI301. The application includes two
proposed manufacturing-use products; Bacillus subtilis strain RTI477 Technical (EPA File
Symbol 279-OAUT) and Bacillus velezensis strain RTI301 Technical (EPA File Symbol 279-
OAUI), as well as three end-use products; F4034-5 (EPA File Symbol 279-OALN), F4007-9
(EPA File Symbol 279-OAUO) and F4092-3 (EPA File Symbol 279-OALR). Bacillus subtilis
and Bacillus velezensis are environmentally ubiquitous bacteria that produce endospores under
certain environmental conditions and are naturally resident in soil and on plant undergrowth.
Both Bacillus subtilis RTI477 and Bacillus velezensis RTI301 were isolated from environmental
sources within the United States.
Bacillus subtilis RTI477 colonizes plant seed and root surfaces and obstructs development of
pathogenic organisms through competition for space and nutrients. There are multiple modes of
action for how this is achieved, including the ability of Bacillus subtilis RTI477 to produce
Active Ingredients: B. subtilis strain RTI477 / B. velezensis strain RTI301 PC Codes: 006552, 006553
Action: Human Health Risk Assessment
3
volatile compounds and metabolites that enhance plant growth, as well as secondary metabolites
that are antagonistic against certain fungal pathogens. Bacillus subtilis RTI477 is also capable of
stimulating the host plant’s immune system, which results in Induced Systemic Resistance (ISR)
and Systemic Acquired Resistance (SAR) responses (MRID 50925702). The pesticidal modes of
action for Bacillus velezensis RTI301 are very similar to those for Bacillus subtilis RTI477, as
this microbe is also known to produce volatile compounds and metabolites that enhance plant
growth, in addition to secondary metabolites that are antagonistic against fungal pathogens.
Bacillus velezensis RTI301 is also capable of increasing ISR and SAR responses in the host plant
(MRID 50925701).
The end-use product, F4034-5 is proposed for use as a bio-fungicide to protect seedlings against
diseases caused by Rhizoctonia, Fusarium, and Phytophthora. F4034-5 contains 2.5% B. subtilis
RTI477 and 7.5% B. velezensis RTI301. The minimum viability of Bacillus subtilis RTI477 and
Bacillus velezensis RTI301 in this product is 1.15 x 109 CFU/mL and 1.5 x 1010 CFU/mL,
respectively. This product is intended for application to a variety of crops/seed including corn,
dried bean/pea, soybean, fruiting vegetables, cucurbit vegetable, cereal grains, oil seed,
root/tuber vegetables, cotton and brassica vegetables. The application methods include in-furrow,
at-transplant drench water treatment and seed treatment. Seed treatment application are
performed by mechanical, slurry, or mist-type seed treating equipment. The application rates
given for in-furrow applications and transplant drench water applications for all crops on the
proposed label are 4 to 10 ounces of product per acre diluted in a minimum of five gallons of
water. The application rates for seed treatment applications are given on a per seed weight basis
or on a per seed unit basis depending on the crop being treated, with a maximum rate of 0.27 fl.
oz./100,000 seeds.
F4007-9 is an end-use product intended only for use as a seed treatment to protect seedlings
against diseases caused by Rhizoctonia, Fusarium, and Phytophthora. This product contains
5.5% Bacillus velezensis RTI301 (minimum viability of 1.1 x 1010 CFU/mL) as the sole active
ingredient. F4007-9 is intended for application to seed for corn, dried bean/pea, soybean, fruiting
vegetables, cucurbit vegetable, cereal grains, oil seed, root/tuber vegetables and cotton.
Application rates for this product are given on a per seed weight basis or on a per seed unit basis
depending on the crop being treated, with a maximum rate of 0.34 fl. oz./100,000 seeds. The
application instructions indicate that the product may be applied to the seeds with mechanical,
slurry, or mist-type seed treating equipment.
F4092-3 is an end-use product intended for use as combination bio-fungicide and insecticide to
protect seedlings against nematodes as well as diseases caused by Rhizoctonia solani, Fusarium
spp., Pythium spp. and Phytophthora spp. F4092-3 contains Bacillus subtilis RTI477 (2.5%),
Bacillus velezensis RTI301 (2.5%) and bifenthrin (15.7%) as the active ingredients. The
minimum viability of Bacillus subtilis RTI477 and Bacillus velezensis RTI301 in this product is
5.0 x 109 CFU/mL for each microbe. The product is proposed for use on a variety of crops
including artichoke, brassica vegetables, berries, citrus, cucurbit, beans/peas, eggplant, grapes
and many tuber/root vegetables. Application methods include in-furrow at plant, pre-plant
incorporated, pre-emergent, lay-by in-furrow (with seed), as a T-band (over the open furrow), as
Active Ingredients: B. subtilis strain RTI477 / B. velezensis strain RTI301 PC Codes: 006552, 006553
Action: Human Health Risk Assessment
4
a broadcast (to soil) and in transplant drench water during setting. The field application rates
range from 2.9 to 42.5 fl. oz./acre. The product is applied in a minimum of five gallons of water
per acre for all field applications except for citrus which must be applied in a minimum of 40
gallons of water per acre. The directions for use on citrus crops instruct the applicator to apply
F4092-3 by ground equipment to bare soil beneath citrus trees from the trunk to the drip line of
the trees.
The generic and product-specific mammalian toxicology data performed with Bacillus subtilis
RTI477 and/or Bacillus velezensis RTI301 as the only active ingredients did not exhibit any
toxicity, infectivity or pathogenicity. While moderate toxicity/irritation was observed in the acute
oral, primary eye irritation and primary dermal irritation testing for the end-use product F4092-3
(EPA File Symbol 279-OALR), these results are very likely due to the presence of bifenthrin as
an active ingredient in this product.
Food and drinking water exposures to Bacillus subtilis RTI477 and Bacillus velezensis RTI301
are expected to be negligible as significant residues are not likely due to the low application rates
and the use of primarily soil-directed applications early in the growing season. There is potential
for occupational exposure, however, no toxicological endpoints have been identified in guideline
mammalian toxicology studies except for one end-use product containing bifenthrin as an active
ingredient (F4092-3; EPA File Symbol 279-OALR). Potential occupational risks resulting from
the use of F4092-3 should be adequately addressed through product label restrictions/warnings
and the use of required personal protective equipment (PPE). Residential exposure resulting from
the pesticidal use of these active ingredients is expected to be minimal. While residential use of
treated turf seed is allowed on two of the end-use product labels, the likelihood of significant
exposure from this use is very low. Considering the results of the mammalian toxicology data
review and the available exposure data/information, the Agency has determined that no further
studies are needed at this time on Bacillus subtilis RTI477 and Bacillus velezensis RTI301.
Based on the available toxicology and exposure information, no unreasonable adverse effects to
humans are expected from the use of Bacillus subtilis strain RTI477 and Bacillus velezensis
strain RTI301 as pesticides when used in accordance with the product label instructions and
requirements. Further, there is a reasonable certainty that no harm will result to the U.S.
population, including infants and children, from aggregate exposure to residues of Bacillus
subtilis strain RTI477 and Bacillus velezensis strain RTI301 resulting from the proposed
pesticidal uses.
III. Background
Bacillus subtilis RTI477 and Bacillus velezensis RTI301 are naturally occurring microorganisms
that have been discovered to have fungicidal and plant health promoting characteristics. Bacillus
subtilis and Bacillus velezensis are found in many environmental habitats including in/on organic
material/plant undergrowth, plant roots and in soil samples. These active ingredients are
proposed for use primarily in agricultural settings as food use bio-fungicides. Exposure to
Bacillus subtilis and Bacillus velezensis occurs naturally and is likely a common component of
Active Ingredients: B. subtilis strain RTI477 / B. velezensis strain RTI301 PC Codes: 006552, 006553
Action: Human Health Risk Assessment
5
the human diet via its presence on surfaces of plant-based foods. When used as pesticidal active
ingredients Bacillus subtilis RTI477 and Bacillus velezensis RTI301 are not expected to result in
significant residues in/on food or in water due to the specific use patterns and application
methods employed. According to the results of toxicity/infectivity and acute toxicity studies,
Bacillus subtilis RTI477 and Bacillus velezensis RTI301 have a low toxicity profile and no
toxicological endpoints were identified in studies containing these microorganisms as the only
active ingredients.
IV. Product Identity and Analysis Review
The applicant submitted data to comply with product identity and analysis data requirements
published in 40 CFR § 158.2120 in support of the registration of Bacillus subtilis RTI477 and
Bacillus velezensis RTI301, as well as the products containing these active ingredients. In
addition, certain product identity and analysis data requirements published in 40 CFR § 158.310
were provided to support EPA File Symbol 279-OALR which contains bifenthrin. The product
identity, manufacturing process, discussion of formation of unintentional ingredients, analysis of
samples, certification of limits, storage stability/corrosion characteristics, and physical/chemical
characteristics data submitted for these products are acceptable. See Appendix I for additional
details on the data supporting these requirements.
V. Summary of Toxicology Data
Tables 1 and 2 below provide the status of the generic or technical-grade active ingredient
(TGAI) data requirements for human health risk assessment as published in 40 CFR § 158.2140
for Bacillus subtilis RTI477 and Bacillus velezensis RTI301, respectively. Table 3 provides the
status of the product-specific data requirements for human health risk assessment as published in
40 CFR § 158.2140. Specifically, Table 3 summarizes the manufacturing-use (EPA File Symbols
279-OAUT and 279-OAUI) and end-use (EPA File Symbols 279-OALN, 279-OAUO and 279-
OALR) products containing Bacillus subtilis RTI477 and/or Bacillus velezensis RTI301 as active
ingredients. Note that scientific rationales were submitted to satisfy the generic acute oral
toxicity/pathogenicity and acute injection toxicity/pathogenicity data requirements for Bacillus
subtilis RTI477, and the acute injection toxicity/pathogenicity data requirement for Bacillus
velezensis RTI301. Additionally, scientific rationales were submitted to satisfy several of the
manufacturing-use product (MP) and end-use product (EP) data requirements.
The information provided satisfies the Tier I toxicology data requirements for the human health
risk assessment for Bacillus subtilis RTI477 and Bacillus velezensis RTI301, as well as the
associated pesticide products. Further testing at higher tiers is not required for the current uses
identified in the proposed pesticide labels. While moderate toxicity/irritation was observed in the
acute oral (870.1100), primary eye irritation (870.2400) and primary dermal irritation (870.2500)
testing for the end-use product F4092-3 (EPA File Symbol 279-OALR), these findings are very
likely due to the presence of bifenthrin as an active ingredient in this product. This conclusion is
supported by the fact that the testing for the MPs and EPs containing only Bacillus subtilis
Active Ingredients: B. subtilis strain RTI477 / B. velezensis strain RTI301 PC Codes: 006552, 006553
Action: Human Health Risk Assessment
6
RTI477 and/or Bacillus velezensis RTI301 as active ingredients did not demonstrate significant
acute oral toxicity or eye/dermal irritation.
Table 1. Summary of generic data submitted to comply with toxicology data requirements
published in 40 CFR § 158.2140 for support of the registration of products containing Bacillus
subtilis strain RTI477.
Data Requirement
OCSPP
Guideline No.
Summary, Results and Classification
MRID
Generic Bacillus subtilis RTI477 Toxicology Data
Acute Oral
Toxicity/Pathogenicity
885.3050 Testing waived based on the applicant’s scientific
rationale which cited the lack of toxicity and/or
pathogenicity in MRID Nos. 50354403 (acute oral
toxicity [870.1100] study conducted with the B. subtilis
MP) and 50354402 (acute pulmonary
toxicity/pathogenicity study conducted with the TGAI).
In addition, the applicant referenced supporting
literature which indicates that B. subtilis is very
unlikely to cause toxicity/pathogenicity in humans.
Classification: Acceptable
50354415
Acute Pulmonary
Toxicity/Pathogenicity
885.3150
A study was provided that demonstrated a pattern of
clearance for B. subtilis RTI477 from the organs of
treated animals. Not toxic, infective, and/or pathogenic
to rats when administered by intra-tracheal instillation
in a single dose of 3.08 x 108 CFU/animal.
Classification: Acceptable
50354402
Acute Injection
Toxicity/Pathogenicity
885.3200
Testing waived based on the applicant’s scientific
rationale which cited the lack of toxicity and/or
pathogenicity in MRID Nos. 50354403 (acute oral
toxicity [870.1100] study conducted with the B. subtilis
MP) and 50354402 (acute pulmonary
toxicity/pathogenicity study conducted with the TGAI).
In addition, the applicant referenced supporting
literature which indicates that B. subtilis is very
unlikely to cause toxicity/pathogenicity in humans.
Classification: Acceptable
50354415
Hypersensitivity
Incidents
885.3400
The applicant reported that no hypersensitivity incidents
or other adverse effects occurred during research,
development, manufacturing or handling of the TGAI.
Any future hypersensitivity incidents must be reported
to the EPA (refer to test note #3 of 40 CFR §
158.2140(d)).
Classification: Acceptable
50354415
Cell Culture 885.3500
Not required because B. subtilis RTI477 is not a virus
(refer to test note #4 of 40 CFR § 158.2140(d)).
Classification: Acceptable
50354415
Table 2. Summary of generic data submitted to comply with toxicology data requirements
published in 40 CFR § 158.2140 for support of the registration of products containing Bacillus
velezensis strain RTI301.
Active Ingredients: B. subtilis strain RTI477 / B. velezensis strain RTI301 PC Codes: 006552, 006553
Action: Human Health Risk Assessment
7
Data Requirement
OCSPP
Guideline No.
Summary, Results and Classification
MRID
Generic Bacillus velezensis RTI301 Toxicology Data
Acute Oral
Toxicity/Pathogenicity
885.3050 A study was provided that showed a pattern of
clearance for B. velezensis RTI301 from the organs of
treated animals. Not toxic, infective, and/or pathogenic
to rats when administered orally in a single dose of 2.1
x 108 CFU/animal.
Classification: Acceptable
50354506
Acute Pulmonary
Toxicity/Pathogenicity
885.3150
A study was provided that showed a pattern of
clearance for B. velezensis RTI301 from the organs of
treated animals. Not toxic, infective, and/or pathogenic
to rats when administered by intra-tracheal instillation
in a single dose of 5.27 x 108 CFU/animal.
Classification: Acceptable
50354501
Acute Injection
Toxicity/Pathogenicity
885.3200
Testing waived based on the applicant’s scientific
rationale which cited the lack of toxicity/pathogenicity
in MRID Nos. 50354506 (acute oral toxicity/
pathogenicity study conducted with the TGAI),
50354501 (acute pulmonary toxicity/pathogenicity
study conducted with the TGAI) and 50354503 (acute
dermal toxicity on the B. velezensis MP). In addition,
the applicant referenced supporting literature which
indicates that B. velezensis (which is closely related to
B. subtilis) is very unlikely to cause
toxicity/pathogenicity in humans.
Classification: Acceptable
50354515
Hypersensitivity
Incidents
885.3400
The applicant reported that no hypersensitivity incidents
or other adverse effects occurred during research,
development, manufacturing or handling of the TGAI.
Any future hypersensitivity incidents must be reported
to the EPA (refer to test note #3 of 40 CFR §
158.2140(d)).
Classification: Acceptable
50354515
Cell Culture 885.3500
Not required because B. velezensis RTI301 is not a
virus (refer to test note #4 of 40 CFR § 158.2140(d)).
Classification: Acceptable
50354515
Table 3. Summary of product-specific data submitted to comply with toxicology data
requirements (40 CFR § 158.2140) for support of the registration of manufacturing-use products
and end-use products containing B. subtilis strain RTI477 and/or B. velezensis strain RTI301.
Data Requirement
OCSPP
Guideline No.
Results Summary, Classification and Toxicity
Category
MRID
Product-specific (MP) Toxicology Data - Bacillus subtilis strain RTI477 Technical (EPA File Symbol
279-OAUT)
Acute Oral Toxicity 870.1100
Oral LD50 (female rats) > 5,000 mg/kg
Classification: Acceptable
Toxicity Category: IV
50354403
Active Ingredients: B. subtilis strain RTI477 / B. velezensis strain RTI301 PC Codes: 006552, 006553
Action: Human Health Risk Assessment
8
Data Requirement
OCSPP
Guideline No.
Results Summary, Classification and Toxicity
Category
MRID
Acute Dermal
Toxicity
870.1200
Dermal LD50 (male and female rats) > 5,000 mg/kg
Classification: Acceptable
Toxicity Category: IV
50354404
Acute Inhalation
Toxicity
870.1300 Testing waived based on oral study with TGAI and
because this formulation contains inert ingredients that
are not expected to be of toxicological concern (refer to
test note #5 of 40 CFR § 158.2140(d)).
Classification: Acceptable
Toxicity Category: IV
50354415
Primary Eye Irritation 870.2400 The product was mildly irritating to the eyes of rabbits.
No corneal opacity, iritis, conjunctivitis, or other signs
of ocular irritation were noted at 24 hours post
installation.
Classification: Acceptable
Toxicity Category: IV
50354405
Primary Dermal
Irritation
870.2500 The product was slightly irritating to the skin of rabbits.
All animals were free of slight erythema or edema at 72
hours.
Classification: Acceptable
Toxicity Category: IV
50354406
Hypersensitivity
Incidents
885.3400 The applicant reported that no hypersensitivity incidents
or other adverse effects occurred during research,
development, manufacturing or handling of the TGAI.
Any future hypersensitivity incidents must be reported
to the EPA (refer to test note #3 of 40 CFR §
158.2140(d)).
Classification: Acceptable
50354415
Product-specific (MP) Toxicology Data – Bacillus velezensis strain RTI301 Technical (EPA File Symbol
279-OAUI)
Acute Oral Toxicity 870.1100
Testing waived based on the applicant’s scientific
rationale which cited the lack of toxicity/pathogenicity
in MRID Nos. 50354506 (acute oral toxicity/
pathogenicity study conducted with the TGAI),
50354501 (acute pulmonary toxicity/pathogenicity
study conducted with the TGAI) and 50354503 (acute
dermal toxicity on the B. velezensis MP). In addition,
the applicant referenced supporting literature which
indicates that B. velezensis is very unlikely to cause
toxicity/pathogenicity in humans, and inert ingredients
are not expected to be of toxicological significance.
Classification: Acceptable
Toxicity Category: IV
50354515
Acute Dermal
Toxicity
870.1200
Dermal LD50 (male and female rats) > 5,000 mg/kg
Classification: Acceptable
Toxicity Category: IV
50354503
Active Ingredients: B. subtilis strain RTI477 / B. velezensis strain RTI301 PC Codes: 006552, 006553
Action: Human Health Risk Assessment
9
Data Requirement
OCSPP
Guideline No.
Results Summary, Classification and Toxicity
Category
MRID
Acute Inhalation
Toxicity
870.1300 Testing waived based on the applicant’s scientific
rationale which cited the lack of toxicity/pathogenicity
in MRID Nos. 50354501 (acute pulmonary
toxicity/pathogenicity study conducted with the TGAI)
and 50354503 (acute dermal toxicity on the B.
velezensis MP). In addition, the applicant referenced
supporting literature which indicates that B. velezensis
is very unlikely to cause toxicity/pathogenicity in
humans, and inert ingredients are not expected to be of
toxicological significance.
Classification: Acceptable
Toxicity Category: IV
50354515
Primary Eye Irritation 870.2400 The product was mildly irritating to the eyes of rabbits.
No positive effects (corneal opacity, iritis,
conjunctivitis, or other signs of ocular irritation) were
noted at any time point in the study.
Classification: Acceptable
Toxicity Category: IV
50354504
Primary Dermal
Irritation
870.2500 The product was slightly irritating to the skin of rabbits.
All animals were free of significant erythema or edema
by 72 hours.
Classification: Acceptable
Toxicity Category: IV
50354505
Hypersensitivity
Incidents
885.3400 The applicant reported that no hypersensitivity incidents
or other adverse effects occurred during research,
development, manufacturing or handling of the TGAI.
Any future hypersensitivity incidents must be reported
to the EPA (refer to test note #3 of 40 CFR §
158.2140(d)).
Classification: Acceptable
50354515
Product-specific (EP) Toxicology Data - F4034-5 (EPA File Symbol 279-OALN)
Acute Oral Toxicity 870.1100
Testing waived based on the applicant’s scientific
rationale which cited studies performed with Bacillus
subtilis RTI477 (MRIDs 50354403, 50354404,
50354402) and Bacillus velezensis RTI301 (MRIDs
50354503, 50354506, 50354501), and because this
formulation contains inert ingredients that are not
expected to be of toxicological concern (refer to test
note #5 of 40 CFR § 158.2140(d)).
Classification: Acceptable
Toxicity Category: IV
50354703
Active Ingredients: B. subtilis strain RTI477 / B. velezensis strain RTI301 PC Codes: 006552, 006553
Action: Human Health Risk Assessment
10
Data Requirement
OCSPP
Guideline No.
Results Summary, Classification and Toxicity
Category
MRID
Acute Dermal
Toxicity
870.1200
Testing waived based on the applicant’s scientific
rationale which cited studies performed with Bacillus
subtilis RTI477 (MRIDs 50354403, 50354404,
50354402, 50354406) and Bacillus velezensis RTI301
(MRIDs 50354503, 50354506, 50354501, 50354505),
and because this formulation contains inert ingredients
that are not expected to be of toxicological concern
(refer to test note #5 of 40 CFR § 158.2140(d)).
Classification: Acceptable
Toxicity Category: IV
50354703
Acute Inhalation
Toxicity
870.1300 Testing waived based on the applicant’s scientific
rationale which cited studies performed with Bacillus
subtilis RTI477 (MRIDs 50354403, 50354404,
50354402) and Bacillus velezensis RTI301 (MRIDs
50354503, 50354506, 50354501), and because this
formulation contains inert ingredients that are not
expected to be of toxicological concern (refer to test
note #5 of 40 CFR § 158.2140(d)).
Classification: Acceptable
Toxicity Category: IV
50354703
Primary Eye Irritation 870.2400 Treated eyes were free of all irritation by 48 hours. The
product was minimally irritating to the rabbit eye.
Classification: Acceptable
Toxicity Category: IV
50354701
Primary Dermal
Irritation
870.2500 The product elicited well-defined irritation on rabbit
skin at 72 hours.
Classification: Acceptable
Toxicity Category: IV
50354702
Hypersensitivity
Incidents
885.3400 The applicant reported that no hypersensitivity incidents
or other adverse effects occurred during research,
development, manufacturing or handling of these
TGAIs. Any future hypersensitivity incidents must be
reported to the EPA (refer to test note #3 of 40 CFR §
158.2140(d)).
Classification: Acceptable
50354703
Product-specific (EP) Toxicology Data - F4007-9 (EPA File Symbol 279-OAUO)
Acute Oral Toxicity 870.1100
Testing waived based on the applicant’s scientific
rationale which cited studies performed with Bacillus
velezensis RTI301 (MRIDs 50354503, 50354506,
50354501), and because this formulation contains inert
ingredients that are not expected to be of toxicological
concern (refer to test note #5 of 40 CFR § 158.2140(d)).
Classification: Acceptable
Toxicity Category: IV
50354603
Active Ingredients: B. subtilis strain RTI477 / B. velezensis strain RTI301 PC Codes: 006552, 006553
Action: Human Health Risk Assessment
11
Data Requirement
OCSPP
Guideline No.
Results Summary, Classification and Toxicity
Category
MRID
Acute Dermal
Toxicity
870.1200
Testing waived based on the applicant’s scientific
rationale which cited studies performed with Bacillus
velezensis RTI301 (MRIDs 50354503, 50354506,
50354501, 50354505), and because this formulation
contains inert ingredients that are not expected to be of
toxicological concern (refer to test note #5 of 40 CFR §
158.2140(d)).
Classification: Acceptable
Toxicity Category: IV
50354603
Acute Inhalation
Toxicity
870.1300 Testing waived based on the applicant’s scientific
rationale which cited studies performed with Bacillus
velezensis RTI301 (MRIDs 50354503, 50354506,
50354501), and because this formulation contains inert
ingredients that are not expected to be of toxicological
concern (refer to test note #5 of 40 CFR § 158.2140(d)).
Classification: Acceptable
Toxicity Category: IV
50354603
Primary Eye Irritation 870.2400 The product elicited very mild irritation in rabbit eyes at
24-hours after treatment. No corneal opacity, iritis,
conjunctivitis, or other signs of ocular irritation was
noted at 48-hours after treatment.
Classification: Acceptable
Toxicity Category: IV
50354601
Primary Dermal
Irritation
870.2500 The product was a slight irritant to the skin of rabbits.
All animals were free of slight erythema at 7 days after
treatment.
Classification: Acceptable
Toxicity Category: IV
50354602
Hypersensitivity
Incidents
885.3400 The applicant reported that no hypersensitivity incidents
or other adverse effects occurred during research,
development, manufacturing or handling of the TGAI
(B. velezensis RTI301). Any future hypersensitivity
incidents must be reported to the EPA (refer to test note
#3 of 40 CFR § 158.2140(d)).
Classification: Acceptable
50354603
Product-specific (EP) Toxicology Data - F4092-3 (EPA File Symbol 279-OALR)
Acute Oral Toxicity 870.1100
Oral LD50 (female rats) 748.8 mg/kg
Classification: Acceptable
Toxicity Category: III
50354801
Acute Dermal
Toxicity
870.1200
This study requirement is waived; the acute toxicity
category is based on the acute oral toxicity study
results. Classification: Acceptable
Toxicity Category: III
50354806
Acute Inhalation
Toxicity
870.1300 Inhalation LC50 (male and female rats) > 2.04 mg/L
Classification: Acceptable
Toxicity Category: IV
50354802
Active Ingredients: B. subtilis strain RTI477 / B. velezensis strain RTI301 PC Codes: 006552, 006553
Action: Human Health Risk Assessment
12
Data Requirement
OCSPP
Guideline No.
Results Summary, Classification and Toxicity
Category
MRID
Primary Eye Irritation 870.2400 The product elicited redness and chemosis in all rabbit
eyes at 24 hours. No corneal opacity, iritis,
conjunctivitis, or other signs of ocular irritation were
noted by 72 hours after treatment.
Classification: Acceptable
Toxicity Category: III
50354804
Primary Dermal
Irritation
870.2500 The product elicited moderate to severe erythema and
moderate edema at 72 hours. All animals were free of
abnormalities by day 14.
Classification: Acceptable
Toxicity Category: II
50354803
Dermal Sensitization
870.2600 The study results indicate that the product is a dermal
sensitizer under the test parameters of the Local Lymph
Node Assay (LLNA).
Classification: Acceptable
Toxicity Category: Sensitizer (Positive)
50354805
A. Toxicology Study Summaries
1. Generic (TGAI) Toxicology Data for Bacillus subtilis RTI477
Study Title: 885.3150; Acute Pulmonary Toxicity/Pathogenicity (B. subtilis RTI477)
MRID No.: 50354402
Classification: Acceptable
Study Summary: In this study, rats received a dose of 3.08 x 108 viable spores in 0.1 ml
of dosing solution by intra-tracheal administration, while a control group remained untreated.
Lung, brain, liver, spleen, kidney, lymph nodes, cecum contents and blood were collected 4,
8, 15, 22, 36, 57, and 85/86 days after dosing, and lung samples were collected soon after
dosing on Day 1. There was no mortality in the study animals and no significant effect on
body weight or body weight gain associated with strain RTI477. A pattern of microbial clearance
from the organs of treated animals was demonstrated during the study. Although this study
demonstrated low level persistence in the lungs on days 85/86, this finding is not uncommon for
spore-forming microbes. Further, with the absence of adverse clinical signs and the
establishment of a pattern of clearance, this persistence is not considered significant.
2. Generic (TGAI) Toxicology Data for Bacillus velezensis RTI301
Study Title: 885.3050; Acute Oral Toxicity/Pathogenicity (B. velezensis RTI301)
MRID No.: 50354506
Classification: Acceptable
Study Summary: In this study, male and female rats received an oral dose of 2.1 x 108
CFU/animal. Various tissue samples were collected 2, 3, 7, 14, and 21 days after dosing and
cultured for detecting Bacillus velezensis strain RTI301. There were no mortalities or test
substance-related clinical signs observed during the study. No macroscopic findings at scheduled
Active Ingredients: B. subtilis strain RTI477 / B. velezensis strain RTI301 PC Codes: 006552, 006553
Action: Human Health Risk Assessment
13
sacrifice. Bacillus velezensis strain RTI301 was completely cleared from all cultured tissue
samples evaluated during the study.
Study Title: 885.3150; Acute Pulmonary Toxicity/Pathogenicity (B. velezensis RTI301)
MRID No.: 50354501
Classification: Acceptable
Study Summary: In this study, male and female rats received a dose of 5.27 x 108 spores
inactivated (Group 2) or viable (Group 3) in 0.1 ml of dosing solution by intra-tracheal
administration, while a third group (Group 1) remained untreated. No mortality was observed in
the study animals and no test substance-related effects on body weight or body weight gain for
either male or female rats during the study. A pattern of microbial clearance from the organs of
treated animals was demonstrated for B. velezensis RTI301during the study. Although this study
demonstrated low level persistence in the lungs on days 85/86, this finding is not uncommon for
spore-forming microbes. Further, with the absence of adverse clinical signs and the
establishment of a pattern of clearance, this persistence is not considered significant.
3. Product-specific (MP) Toxicology Data – Bacillus subtilis strain RTI477
Technical (EPA File Symbol 279-OAUT)
Study Title: 870.1100; Acute Oral Toxicity
MRID No.: 50354403
Classification: Acceptable
Toxicity Category: IV
Study Summary: Three fasted Sprague-Dawley-derived female rats were treated by oral gavage
with Bacillus subtilis strain RTI477 as a 20% w/w mixture in distilled water at a dose level of
5000 mg/Kg bw (24.20 mL/Kg volume) divided into two approximately equal portions and
administered two hours apart. There was no perceptible change in viable test substance during
the observation period. There were no deaths, abnormal clinical signs, or abnormal gross
necropsy findings. All the animals gained weight during both weeks of the study. The acute oral
lethal dose of the test substance is greater than 5000 mg/Kg bw.
Study Title: 870.1200; Acute Dermal Toxicity
MRID No.: 50354404
Classification: Acceptable
Toxicity Category: IV
Study Summary: Five male and five female fasted young adult Sprague-Dawley derived albino
rats were dermally exposed for 24 hours to Bacillus subtilis strain RTI477 as a 55% w/w mixture
with distilled water applied to a 6-square-inch area of previously shaved, intact skin (~9.6% to
12.8% of the body surface area) at a dose of 5000 mg/Kg bw. Following exposure on Day 0, the
animals were observed for a period of 14 days. There were no deaths, abnormal clinical signs, or
abnormal gross necropsy findings. All the animals gained weight during both weeks of the study.
The acute dermal lethal dose of Bacillus subtilis strain RTI477 is greater than 5000 mg/Kg bw.
Study Title: 870.2400; Primary Eye Irritation
Active Ingredients: B. subtilis strain RTI477 / B. velezensis strain RTI301 PC Codes: 006552, 006553
Action: Human Health Risk Assessment
14
MRID No.: 50354405
Classification: Acceptable
Toxicity Category: IV
Study Summary: A 0.1 mL volume (0.056 g) of Bacillus subtilis strain RTI477 was instilled
into the conjunctival sac of the unanesthetized right eye of three 17-week-old female New
Zealand White rabbits, and the upper and lower lids were held shut for approximately one
second, while the untreated left eye of each animal served as control. Eyes were scored for ocular
irritation according to the Draize method at 1, 24, 48, and 72 hours after instillation, with
fluorescein staining done at 24 hours. There were no observations of corneal opacity, iritis, or
“positive” conjunctival findings at any time during the study. Based on the MMTS of 6.0 (and
non-zero scores at 48 hours), Bacillus subtilis strain RTI477 is mildly irritating to the eye (Kay
and Calandra, 1962). Bacillus subtilis strain RTI477 caused minimal effects that cleared within
24 hours.
Study Title: 870.2500; Primary Dermal Irritation
MRID No.: 50354406
Classification: Acceptable
Toxicity Category: IV
Study Summary: Three young adult female New Zealand albino rabbits were dermally exposed
for 4 hours to 500 mg of Bacillus subtilis strain RTI477 in distilled water (60% w/w; 0.83 g of
mixture) applied via a 4-ply gauze pad to intact, previously clipped application sites measuring 6
cm2 and then covered by a semi-occlusive dressing. The application sites were scored for
irritation according to the Draize method at 30-60 minutes and at 1, 24, 48, and 72 hours after
patch removal. At 30-60 minutes following patch removal, all three sites showed very slight,
barely perceptible erythema (score=1), and two sites also had very slight, barely perceptible
edema (score=1). At 24 hours, the three sites had very slight erythema without edema. At 48 and
72 hours, all sites were free of erythema, edema, and other signs of dermal irritation. The
Primary Dermal Irritation Index was 0.67. Based on the PDII, the test material is a slight irritant.
4. Product-specific (MP) Toxicology Data – Bacillus velezensis strain RTI301
Technical (EPA File Symbol 279-OAUI)
Study Title: 870.1200; Acute Dermal Toxicity
MRID No.: 50354503
Classification: Acceptable
Toxicity Category: IV
Study Summary: Five male and five female fasted young adult Sprague-Dawley derived albino
rats were dermally exposed for 24 hours to Bacillus velezensis Strain RTI301 manufacturing-use
product as an 80% w/w mixture with distilled water applied to a 6-squareinch area of previously
shaved, intact skin (~8.4% to 12.9% of the body surface area) at a dose of 5000 mg/Kg bw.
Following exposure on Day 0, the animals were observed for a period of 14 days. There were no
deaths, abnormal clinical signs, or abnormal gross necropsy findings. All animals gained weight
during both weeks of the study. The acute dermal lethal dose of Bacillus velezensis Strain
RTI301 is greater than 5000 mg/Kg bw.
Active Ingredients: B. subtilis strain RTI477 / B. velezensis strain RTI301 PC Codes: 006552, 006553
Action: Human Health Risk Assessment
15
Study Title: 870.2400; Primary Eye Irritation
MRID No.: 50354504
Classification: Acceptable
Toxicity Category: IV
Study Summary: A 0.1 mL volume (0.044 g) of Bacillus velezensis Strain RTI301 was instilled
into the conjunctival sac of the unanesthetized right eye of three 11-week-old female New
Zealand White rabbits, and the upper and lower lids were held shut for approximately one
second, while the untreated left eye of each animal served as control. Eyes were scored for ocular
irritation according to the Draize method at 1, 24, 48, and 72 hours after instillation, with
fluorescein staining done at 24 hours. There were no observations of corneal opacity, iritis, or
“positive” conjunctivitis at any time during the study. “Non-positive” conjunctivitis was noted in
the three treated eyes: one-hour post instillation, conjunctival redness and discharge were seen in
all three treated eyes, and one treated eye also had chemosis (all scores=1); at 24 hours, chemosis
was noted in all three treated eyes, with redness and discharge each noted in 2/3 eyes (all
scores=1). All eyes were clear of abnormal findings at 48 and 72 hours. Bacillus velezensis
Strain RTI301 is minimally irritating to the eye.
Study Title: 870.2500; Primary Dermal Irritation
MRID No.: 50354505
Classification: Acceptable
Toxicity Category: IV
Study Summary: Three young adult female New Zealand albino rabbits were dermally exposed
for 4 hours to 0.5 g of Bacillus velezensis Strain RTI301 as an 80% w/w mixture with distilled
water applied to intact, previously clipped application sites measuring 6 cm2 then covered by a
semi-occlusive dressing. Following patch removal, any residual test substance was removed via
gentle cleansing with a 3% soap solution followed by tap water and a clean paper towel. The
application sites were scored for irritation according to the Draize method at 30-60 minutes, at 1,
24, 48, and 72 hours, and at 7 and 10 days after patch removal. At 30-60 minutes following patch
removal, all three sites had very slight erythema, and one site also had very slight edema. At 24,
48, and 72 hours, two sites exhibited both very slight erythema and very slight edema, and the
third site was normal. On Day 7, one site had very slight erythema without edema, and two sites
were normal. All sites were free of erythema, edema, and/or other signs of skin irritation on Day
10. The Primary Dermal Irritation Index was 1.33.
5. Product-specific (EP) Toxicology Data - F4034-5 (EPA File Symbol 279-OALN)
Study Title: 870.2400; Primary Eye Irritation
MRID No.: 50354701
Classification: Acceptable
Toxicity Category: IV
Study Summary: A 0.1 mL volume of undiluted F4034-5 was instilled into the conjunctival sac
of the unanesthetized right eye of three 17-week-old female New Zealand White rabbits, and the
upper and lower lids were held shut for approximately one second, while the untreated left eye of
Active Ingredients: B. subtilis strain RTI477 / B. velezensis strain RTI301 PC Codes: 006552, 006553
Action: Human Health Risk Assessment
16
each animal served as control. Eyes were scored for ocular irritation according to the Draize
method at 1, 24, 48, and 72 hours after instillation, with fluorescein staining done at 24 hours.
There were no observations of corneal opacity or iritis at any time during the study. Fluorescein
staining at 24 hours verified the absence of corneal opacity. The Maximum Mean Total Score
(MMTS) of F4034-5 was 6.7, noted one-hour post treatment.
Study Title: 870.2500; Primary Dermal Irritation
MRID No.: 50354702
Classification: Acceptable
Toxicity Category: IV
Study Summary: Three young adult female New Zealand albino rabbits were dermally exposed
for 4 hours to 0.5 mL of undiluted F4034-5 applied to intact, previously clipped application sites
measuring 6 cm2 and covered by a semi-occlusive dressing. Following patch removal, any
residual test substance was removed via gentle cleansing with a 3% soap solution followed by
tap water and a clean paper towel. The application sites were scored for irritation according to
the Draize method at 30-60 minutes, at 1, 24, 48, and 72 hours, and at 7 days after patch
removal. At 30-60 minutes following patch removal, all three sites showed very slight or well-
defined erythema with very slight or slight edema. The erythema persisted, unchanged, on the
three application sites through 72 hours and resolved by Day 7. At 24 and 48 hours, one site was
free of edema, and the other two sites had very slight edema or slight edema. At 72 hours, very
slight edema was present on two sites, and all sites were free of edema by Day 7. The Primary
Dermal Irritation Index was 2.7.
6. Product-specific (EP) Toxicology Data – F4007-9 (EPA File Symbol 279-OAUO)
Study Title: 870.2400; Primary Eye Irritation
MRID No.: 50354601
Classification: Acceptable
Toxicity Category: IV
Study Summary: A 0.1 mL volume of undiluted F4007-9 was instilled into the conjunctival sac
of the right eye of three young adult female New Zealand White rabbits, and the upper and lower
lids were held shut for approximately one second, while left eye of each animal was not treated
with the test material and served as a control. Topical anesthetic was not used for the first animal
treated. Because the initial treated animal exhibited signs of distress following dosing, an
unspecified topical anesthetic was used for the remaining two animals; it is unknown whether the
contralateral eyes of these two animals were also anesthetized. Eyes were scored for ocular
irritation according to the Draize method at 1, 24, 48, and 72 hours after instillation, with
fluorescein staining done at 24 hours. There were no observations of corneal opacity, iritis, or
“positive” conjunctivitis at any time during the study. Fluorescein staining at 24 hours verified
the absence of opacity. The Maximum Mean Total Score (MMTS) of F4007-9 was 4.0, noted
one-hour post treatment.
Study Title: 870.2500; Primary Dermal Irritation
MRID No.: 50354602
Active Ingredients: B. subtilis strain RTI477 / B. velezensis strain RTI301 PC Codes: 006552, 006553
Action: Human Health Risk Assessment
17
Classification: Acceptable
Toxicity Category: IV
Study Summary: Three young adult female New Zealand albino rabbits dermally exposed for 4
hours to 0.5 mL of undiluted F4007-9 applied to intact, previously clipped application sites
measuring 6 cm2 and covered by a semi-occlusive dressing. Following patch removal, any
residual test substance was removed via gentle cleansing with a 3% soap solution followed by
tap water and a clean paper towel. The application sites were scored for irritation according to
the Draize method at 30-60 minutes, at 1, 24, 48, and 72 hours, and at 7 days after patch
removal. At 30-60 minutes following patch removal, all three sites showed very slight or well-
defined erythema with very slight or slight edema. At 24 hours, one site had well-defined
erythema with very slight edema, one site had very slight erythema without edema, and one site
was free of abnormal findings. At 48 hours, very slight erythema was present without edema on
two sites, and the third site remained free of abnormal findings. At 72 hours, one site had very
slight erythema without edema, one site had desquamation without other abnormalities, and one
site was normal. All sites were free of erythema, edema, and other signs of skin irritation by Day
7. The Primary Dermal Irritation Index was 1.3.
7. Product-specific (EP) Toxicology Data - F4092-3 (EPA File Symbol 279-OALR)
Study Title: 870.1100; Acute Oral Toxicity
MRID No.: 50354801
Classification: Acceptable
Toxicity Category: III
Study Summary: Eight fasted 8-10-week-old female Sprague-Dawley-derived rats were given
single oral gavage doses of undiluted F4092-3 at dose levels of 110 mg/Kg bw (1 animal), 350
mg/Kg bw (2 animals), 1100 mg/Kg bw (3 animals), or 2000 mg/Kg bw (2 animals) with dose
selection, progression, and stopping criteria in accordance with AOT 425 (limit dose: 2000
mg/Kg bw; assumed LD50: 1100 mg/Kg bw; sigma: 0.5). Dosing was on Day 0, and the animals
were observed for up to 14 days. The animal treated with 110 mg/Kg bw remained active and
healthy throughout the study duration. One 350-mg/Kg animal was found dead on Day 1 after
exhibiting irregular respiration and tremors on Day 0. The surviving 350-mg/Kg animal
exhibited hypoactivity and irregular respiration following dosing on Day 0, had irregular
respiration, slight tremors, and reduced fecal volume on Day 1, and appeared active and healthy
on Day 2 and thereafter. One 1100-mg/Kg animal was found dead on Day 1 after exhibiting
irregular respiration and tremors on Day 0. The two surviving 1100-mg/Kg animals exhibited
hypoactivity and irregular respiration on Days 0 and/or 1, with reduced fecal volume and
piloerection or hypersensitivity on Day 1; both recovered by Day 3 and appeared active and
healthy thereafter. Both 2000-mg/Kg animals died: one was found dead on Day 1 after exhibiting
irregular respiration and hypoactivity on Day 0; and the other was found dead on Day 2 without
any preceding abnormal clinical signs. The surviving animals gained weight during both weeks
of the study.
Study Title: 870.1300; Acute Inhalation Toxicity
MRID No.: 50354802
Active Ingredients: B. subtilis strain RTI477 / B. velezensis strain RTI301 PC Codes: 006552, 006553
Action: Human Health Risk Assessment
18
Classification: Acceptable
Toxicity Category: IV
Study Summary: Five male and five female 9-10-week-old Sprague-Dawley-derived rats were
exposed by the nose-only inhalation route to aerosolized undiluted F4092-3 for four hours at a
mean gravimetric concentration of 2.04 mg/L (mean MMAD 2.98 μm and GSD of 2.23).
Exposure was on Day 0, and the animals were observed for 14 days. There were no deaths or
abnormal gross necropsy findings. Upon removal from the chamber and/or beginning on Day 0,
all the animals had moist rales, irregular respiration, and hypoactivity, with abnormal gait and
tremors noted from each of two animals (four animals in total). All the animals subsequently
exhibited anogenital staining, with or without red nasal discharge, beginning on Day 1. Tremors
were noted in four additional animals with onset on Day 3, and one female exhibited
hypersensitivity to sound on Days 3-4. Most of the animals recovered by Days 5-8, but one
female exhibited anogenital staining through Day 13. All the animals lost weight during Days 0-
1, and one female lost additional weight during Days 1-3. All the animals had net body weight
gains for Days 0-14.
Study Title: 870.2400; Primary Eye Irritation
MRID No.: 50354804
Classification: Acceptable
Toxicity Category: III
Study Summary: A 0.1 mL volume of F4092-3 was instilled into the conjunctival sac of the
unanesthetized right eye of three 17-week-old female New Zealand White rabbits, and the upper
and lower lids were held shut for approximately one second, while the untreated left eye of each
animal served as a control. Eyes were scored for ocular irritation according to the Draize method
at 1, 24, 48, and 72 hours after instillation, with fluorescein staining done at 24 hours. There
were no observations of corneal opacity or iritis at any time during the study. One hour after
instillation, the three treated eyes exhibited conjunctival redness (scores of 1-2) and chemosis
(scores of 1-2). At 24 hours, 3/3 eyes had chemosis (scores of 1-2), with redness noted in 2/3
(scores of 1) and discharge noted in 2/3 (scores of 1). At 48 hours, abnormalities were limited to
chemosis (score=1) in 3/3 eyes. All eyes were free of abnormal findings (all scores of zero) at 72
hours.
Study Title: 870.2500; Primary Dermal Irritation
MRID No.: 50354803
Classification: Acceptable
Toxicity Category: II
Study Summary: Three young adult female New Zealand albino rabbits were dermally exposed
for 4 hours to 0.5 mL of undiluted F4092-3 applied to intact, previously clipped application sites
measuring 6 cm2 and covered by a semi-occlusive dressing. The application sites were scored
for irritation according to the Draize method at 30-60 minutes, at 1, 24, 48, and 72 hours, and at
7 and 10 days after patch removal. At 30-60 minutes and at 24 hours following patch removal,
all three sites showed well-defined or moderate-to-severe erythema with slight edema. One site
had well-defined erythema with very slight edema at 48 and 72 hours; on this site, the edema
resolved by Day 7, and erythema resolved by Day 10. The other two sites had moderate-to-
Active Ingredients: B. subtilis strain RTI477 / B. velezensis strain RTI301 PC Codes: 006552, 006553
Action: Human Health Risk Assessment
19
severe erythema at 48 and 72 hours, with slight edema on both sites at 48 hours and slight or
moderate edema at 72 hours. For the latter two sites, edema resolved by Day 10, and erythema
resolved by Day 10 or Day 14. The three sites were free of abnormalities on Day 14. The
Primary Dermal Irritation Index was 4.58.
Study Title: 870.2600; Dermal Sensitization
MRID No.: 50354805
Classification: Acceptable
Toxicity Category: Positive Sensitizer
Study Summary: Groups of five female CBA/J mice were tested using the Local Lymph Node
Assay (individual animal approach). Concentrations of 0% (vehicle only), 25%, 50%, or 100%
were evaluated. An additional positive control group of five animals was treated with Alpha-
Hexylcinnamaldehyde (≥95% a.i.) administered at a 25% concentration in 1% Pluronic® L92.
The stimulation indices (SIs) of the groups treated with 25%, 50%, and 100% concentrations of
the test material were 5.04, 8.38, and 7.63, respectively, while the SI of the positive control
group was 8.63. The mean radioactive DPM (disintegrations per minute) values of the groups
treated with the 50% and 100% concentrations of the test material were significantly greater than
control (p<0.05), and the mean DPM of the positive controls also was significantly greater
(p<0.01).
VI. Human Exposure and Risk Characterization Assessment
A. Description of Uses
1. F4034-5; EPA File Symbol 279-OALN - The application methods for F4034-5 include
in-furrow, at-transplant drench water treatment and seed treatment. Seed treatment
application are performed by mechanical, slurry, or mist-type seed treating equipment in
commercial facilities or on-farm treatments. The application rates given for in-furrow
applications and transplant drench water applications for all crops on the proposed label
are 4 to 10 ounces of product per acre diluted in a minimum of five gallons of water. The
application rates for seed treatment applications are given on a per seed weight basis or
on a per seed unit basis depending on the crop being treated, with a maximum rate of
0.27 fl. oz./100,000 seeds.
2. F4007-9; EPA File Symbol 279-OAUO - F4007-9 is intended for seed treatment only.
Application rates for this product are given on a per seed weight basis or on a per seed
unit basis depending on the crop being treated, with a maximum rate of 0.34 fl.
oz./100,000 seeds. The application instructions indicate that the product may be applied
to the seeds with mechanical, slurry, or mist-type seed treating equipment in commercial
facilities or on-farm treatments.
3. F4092-3; EPA File Symbol 279-OALR - The application methods for F4092-3 include
in-furrow at plant, pre-plant incorporated, pre-emergent, and lay-by in-furrow (with the
seed), as a T-band (over the open furrow), as a broadcast application, as a band over the
Active Ingredients: B. subtilis strain RTI477 / B. velezensis strain RTI301 PC Codes: 006552, 006553
Action: Human Health Risk Assessment
20
row or in transplant drench water during setting. The product application rates range from
2.9 to 42.5 fl. oz./acre. The directions for citrus crops instruct the applicator to apply
F4092-3 by ground equipment to bare soil beneath citrus trees from the trunk to the drip
line of the trees.
B. 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
of 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 FFDCA section
408(c)(2)(B), in establishing or maintaining in effect an exemption from the requirement of a
tolerance, EPA must take into account the factors set forth in FFDCA section 408(b)(2)(C) and
(D), which require EPA to give special consideration to exposure of infants and children to the
pesticide chemical residue in establishing a tolerance or tolerance exemption and to “ensure that
there is a reasonable certainty that no harm will result to infants and children from aggregate
exposure to the pesticide chemical residue…” Additionally, FFDCA section 408(b)(2)(D)
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.”
As a result, EPA performs analyses to determine the risks from aggregate exposure to pesticide
residues. First, for microbial pesticides, EPA determines the pathogenicity and toxicity of the
pesticide. Second, EPA examines exposure to the pesticide through food, drinking water, and
other exposures that occur as a result of pesticide use in residential settings, as well as other non-
occupational exposure to the substance or substances.
1. Aggregate Exposure and Risk Characterization
In examining aggregate exposure, FFDCA section 408 directs EPA to consider available
information concerning exposures from the pesticide residue in food and all other non-
occupational 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). However, since no adverse effects of concern were observed in toxicological tests
performed with Bacillus subtilis RTI477 and Bacillus velezensis RTI301, the EPA did not
conduct a quantitative exposure assessment.
a. Food Exposure and Risk Characterization
Although Bacillus subtilis RTI477 and Bacillus velezensis RTI301 are intended for direct food
use, significant dietary exposure to these organisms resulting from these uses is not expected.
Active Ingredients: B. subtilis strain RTI477 / B. velezensis strain RTI301 PC Codes: 006552, 006553
Action: Human Health Risk Assessment
21
Both Bacillus subtilis and Bacillus velezensis are ubiquitous environmental microbes that are
common in soils and on plant surfaces. As pesticides, Bacillus subtilis RTI477 and Bacillus
velezensis RTI301 will be applied by seed treatment and soil-directed applications early in the
growing season. Due to the relatively low application rates proposed for the end-use products,
the pesticidal use of these microbes would not be expected to significantly increase the levels of
B. subtilis and B. velezensis naturally present in the treated environment. Further, since food
crops undergo a post-harvest washing process to remove soil and surface residues, it is unlikely
that significant amounts of these organisms would remain on treated crops. Should these
organisms happen to remain on some food commodities at substantial levels, the results of the
guideline mammalian toxicology testing performed with these microorganisms demonstrated a
lack of significant adverse effects (toxicity, pathogenicity or infectivity). Therefore, as a result of
the minimal likelihood of significant food exposure and low hazard findings, quantitative food
exposure assessments were not performed for these microbial active ingredients.
b. Drinking Water Exposure and Risk Characterization
Although Bacillus subtilis RTI477 and Bacillus velezensis RTI301 are intended for agricultural
use, significant drinking water exposure to these organisms resulting from these uses is not
expected to occur. Both B. subtilis and B. velezensis are ubiquitous environmental microbes that
are common in soils and on plant surfaces. As pesticides, Bacillus subtilis RTI477 and Bacillus
velezensis RTI301 will be applied by seed treatment and direct soil applications which reduce the
likelihood of off-site airborne movement of these active ingredients. If these active ingredients
were to enter surface or ground water to some degree, considering the relatively low application
rates they would likely be present at levels below that of the naturally occurring B. subtilis and
B. velezensis organisms. Further, if these active ingredients were to ultimately enter drinking
water systems, they would be even further reduced in numbers by way of standard drinking
water treatment processes. In addition, the results of the guideline mammalian toxicology testing
performed with these organisms demonstrated a lack of significant adverse effects (toxicity,
pathogenicity or infectivity). Therefore, as a result of these low exposure and hazard findings,
quantitative drinking water exposure assessments were not performed for these microbial active
ingredients.
c. Residential, Non-occupational Exposure and Risk Characterization
As previously noted, Bacillus subtilis RTI477 and Bacillus velezensis RTI301 are intended for
agricultural use. However, there is some potential for indirect residential exposure as F4034-5
(EPA File Symbol 279-OALN) and F4007-9 (EPA File Symbol 279-OAUO) allow for treatment
of turf seed that could be used in residential locations. Residential exposures resulting from this
use would likely be infrequent and would pose minimal concern due to the small amount of
active ingredient involved. As a result, this use scenario is not expected to result in significant
residential exposure. Further, since Bacillus subtilis RTI477 and Bacillus velezensis RTI301 will
be applied agriculturally by seed treatment and direct soil applications, other non-occupational
exposures through drift or other means are also considered very unlikely. Should significant
residential or non-occupational exposures occur, the results of the guideline mammalian
toxicology testing performed with these active ingredients demonstrated a lack of significant
Active Ingredients: B. subtilis strain RTI477 / B. velezensis strain RTI301 PC Codes: 006552, 006553
Action: Human Health Risk Assessment
22
adverse effects (toxicity, pathogenicity or infectivity). Therefore, quantitative non-
occupational/residential exposure assessments were not performed for these microbial active
ingredients.
2. Cumulative Effects
Section 408(b)(2)(D)(v) of FFDCA requires that, when considering whether to establish, modify,
or revoke a tolerance, EPA consider “available information concerning the cumulative effects of
[a particular pesticide’s] . . . residues and other substances that have a common mechanism of
toxicity.”
Bacillus subtilis RTI477 and Bacillus velezensis RTI301are not toxic and do not have a common
mechanism of toxicity with other substances. Consequently, FFDCA section 408(b)(2)(D)(v)
does not apply.
3. Determination of Safety for U.S. Population, Infants and Children
a. U.S. Population
For all the reasons discussed previously, EPA concludes that there is reasonable certainty that no
harm will result to the U.S. population, including infants and children, from aggregate exposure
to residues of Bacillus subtilis RTI477 and Bacillus velezensis RTI301. This includes all
anticipated dietary exposures and all other exposures for which there is reliable information.
b. Infants and Children
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 based on reliable data 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. As discussed previously, EPA has concluded that Bacillus subtilis RTI477 and
Bacillus velezensis RTI301 are not toxic, pathogenic, or infective to mammals, including infants
and children. Because there are no threshold levels of concern to infants, children, and adults
when Bacillus subtilis RTI477 and Bacillus velezensis RTI301 are used in accordance with label
directions and good agricultural practices, EPA concludes that no additional margin of safety is
necessary to protect infants and children.
c. Occupational Exposure and Risk Characterization
Based on the agricultural use pattern proposed for Bacillus subtilis RTI477 and Bacillus
velezensis RTI301, there is potential for occupational exposure to applicators and other handlers
Active Ingredients: B. subtilis strain RTI477 / B. velezensis strain RTI301 PC Codes: 006552, 006553
Action: Human Health Risk Assessment
23
through the dermal, ocular, and inhalation routes. However, the supporting mammalian
toxicological testing has shown a lack of irritation, toxicity, infectivity, or pathogenicity
associated with these active ingredients. Although one of the proposed end-use products, F4092-
3 (EPA File Symbol 279-OALR), did demonstrate significant eye and skin irritation, the irritant
effects were likely due to the presence of bifenthrin in the product formulation. As a result, the
product labeling for F4092-3 will require the appropriate precautions and protective equipment
deemed necessary to address these hazards. In addition, all three end-use products proposed for
registration with this application include labeling that specifies use of a National Institute for
Occupational Safety and Health-approved respirator, long-sleeved shirt, long pants, gloves,
socks, and shoes while handling these products. Therefore, since occupational exposure to
Bacillus subtilis RTI477 and Bacillus velezensis RTI301 are not expected to exceed any toxicity
thresholds when pesticide handlers follow the precautions and requirements identified on the
product label, quantitative occupational assessments were not performed.
4. Human Health Conclusions
EPA does not expect significant dietary (food and drinking water) or other non-occupational
risks from use of Bacillus subtilis RTI477 and Bacillus velezensis RTI301 as active ingredients
in the proposed pesticide products. The required toxicology data demonstrated that Bacillus
subtilis RTI477 and Bacillus velezensis RTI301 are not toxic, pathogenic or infective. Any risks
to handlers resulting from exposure to Bacillus subtilis RTI477, Bacillus velezensis RTI301 or
the proposed end-use product containing bifenthrin are expected to be minimized by use of the
required personal protective equipment. Based on the available toxicology and exposure
information, EPA concludes that no unreasonable adverse effects to humans are expected from
the use of Bacillus subtilis strain RTI477 and/or Bacillus velezensis strain RTI301 as pesticides,
when EPA-approved product label instructions are followed. Further, there is a reasonable
certainty that no harm will result to the U.S. population, including infants and children, from
aggregate exposure to residues of Bacillus subtilis strain RTI477 and/or Bacillus velezensis strain
RTI301 resulting from the proposed pesticidal uses. In addition, residues of Bacillus subtilis
strain RTI477 and Bacillus velezensis strain RTI301 will be covered by exemptions from the
requirement of a tolerance in or on food commodities.
Active Ingredients: B. subtilis strain RTI477 / B. velezensis strain RTI301 PC Codes: 006552, 006553
Action: Human Health Risk Assessment
24
Appendix I
Table 4. Summary of data submitted to comply with product analysis data requirements
published in 40 CFR § 158.2120 for support of the registration of products containing [Microbial
active ingredient].
Data Requirement
OCSPP
Guideline No.
Results and Classification (As Applicable)
MRID No.
Product Analysis Data for Bacillus subtilis RTI477 Technical (EPA File Symbol 279-OAUT)
Product Identity 885.1100 Submitted data fulfill the requirement for product
identity.
Classification: Acceptable
50354414,
50925702,
51255601
Manufacturing
Process
885.1200 Submitted data fulfill the requirement for
manufacturing process.
Classification: Acceptable
51255601*
Deposition of a
Sample in a
Nationally
Recognized Culture
Collection
885.1250
Bacillus subtilis RTI477 is on deposit with the ATCC;
PTA-121167 (Deposit at American Type Culture
Collection) and DSM 32474 (Deposit at Deutche
Sammlung von Mikroorganismen und Zellkulturen)
Classification: Acceptable
50354414,
50925702,
51255601
Discussion of
Formation of
Unintentional
Ingredients
885.1300
Submitted data fulfill the requirement for discussion of
formation of unintentional ingredients.
Classification: Acceptable
50354414,
50925702,
51255601
Analysis of Samples 885.1400
Submitted data fulfill the requirement for analysis of
samples.
Classification: Acceptable
50354411,
50354412,
51255601
Certification of Limits 885.1500 Submitted data fulfill the requirement for certification
of limits.
Classification: Acceptable
50354412,
51255601
Color 830.6302
Brownish beige 50354413,
51255601
Physical State 830.6303 Solid powder 50354413,
51255601
Odor 830.6304 Slightly sweet, musty oder 50354413,
51255601
Stability to Normal
and Elevated
Temperatures, Metals,
and Metal Ions
830.6313 The TGAI product was stable for at least 12 months at
25°C. It is expected to be stable at elevated
temperatures since the optimal growth temperature
range for Bacillus subtilis strain RTI477 is 28°C - 30°C.
51255601
Storage Stability 830.6317 The two-year storage stability study is ongoing.
The interim results showed that the TGAI
product was stable when stored in foil pockets at
25°C for at least 12 months.
50354418,
51255601
Miscibility 830.6319 Not applicable NA
Active Ingredients: B. subtilis strain RTI477 / B. velezensis strain RTI301 PC Codes: 006552, 006553
Action: Human Health Risk Assessment
25
Data Requirement
OCSPP
Guideline No.
Results and Classification (As Applicable)
MRID No.
Corrosion
Characteristics
830.6320 No signs of corrosion were noted on the commercial
packaging (foil
pockets) during the 12-month storage stability study at
25°C.
50354418,
51255601
pH 830.7000 8.09 (1% w/v aqueous suspension) at 20°C 50354413,
51255601
Viscosity 830.7100 Not applicable NA
Density/Relative
Density/Bulk Density
830.7300 0.60 g/mL (pour density), 0.63 g/mL (tap density) 50354413,
51255601
Product Analysis Data for Bacillus velezensis RTI301 Technical (EPA File Symbol 279-OAUI)
Product Identity 885.1100 Submitted data fulfill the requirement for product
identity.
Classification: Acceptable
50354514,
50925701,
51255501
Manufacturing
Process
885.1200 Submitted data fulfill the requirement for
manufacturing process.
Classification: Acceptable
51255501*
Deposition of a
Sample in a
Nationally
Recognized Culture
Collection
885.1250
Bacillus velezensis RTI301 is on deposit with the
ATCC; PTA-121165 (Deposit at American Type
Culture Collection) and DSM 32473 (Deposit at
Deutche Sammlung von Mikroorganismen und
Zellkulturen) Classification: Acceptable
50354514,
50925701,
51255501
Discussion of
Formation of
Unintentional
Ingredients
885.1300
Submitted data fulfill the requirement for discussion of
formation of unintentional ingredients.
Classification: Acceptable
50354514,
50925701,
51255501
Analysis of Samples 885.1400
Submitted data fulfill the requirement for analysis of
samples.
Classification: Acceptable
50354511,
50354512,
51255501
Certification of Limits 885.1500 Submitted data fulfill the requirement for certification
of limits.
Classification: Acceptable
50354512,
51255501
Color 830.6302
Brownish beige 50354513,
51255501
Physical State 830.6303 Solid powder 50354513,
51255501
Odor 830.6304 Sweet, yeast-like odor 50354513,
51255501
Stability to Normal
and Elevated
Temperatures, Metals,
and Metal Ions
830.6313 The TGAI product was found to be stable for at
least 12 months at 25°C. It is expected to be
stable at elevated temperatures since the optimal
growth temperature range for Bacillus velezensis
strain RTI301 is 30°C - 40°C.
51255501
Storage Stability 830.6317 The TGAI product was found to be stable for at
least 12 months at 25°C. It is expected to be
stable at elevated temperatures since the optimal
growth temperature range for Bacillus velezensis
strain RTI301 is 30°C - 40°C.
50354518,
51255501
Miscibility 830.6319 Not applicable NA
Active Ingredients: B. subtilis strain RTI477 / B. velezensis strain RTI301 PC Codes: 006552, 006553
Action: Human Health Risk Assessment
26
Data Requirement
OCSPP
Guideline No.
Results and Classification (As Applicable)
MRID No.
Corrosion
Characteristics
830.6320 No signs of corrosion were noted on the
commercial packaging (foil pockets) during the
12-month storage stability study at 25°C.
50354518,
51255501
pH 830.7000 8.01 (1% w/v aqueous suspension) at 20°C 50354513,
51255501
Viscosity 830.7100 Not applicable NA
Density/Relative
Density/Bulk Density
830.7300 0.34 g/mL (pour density), 0.44 g/mL (tap
density)
50354513,
51255501
Product Analysis Data for F4034-5 (EPA File Symbol 279-OALN)
Product Identity 885.1100 Submitted data fulfill the requirement for product
identity.
Classification: Acceptable
50354705
51391802
Manufacturing
Process
885.1200
Submitted data fulfill the requirement for
manufacturing process.
Classification: Acceptable
50354705
51391802*
Discussion of
Formation of
Unintentional
Ingredients
885.1300
Submitted data fulfill the requirement for discussion of
formation of unintentional ingredients.
Classification: Acceptable
50354705
51391802
Analysis of Samples 885.1400 Submitted data fulfill the requirement for analysis of
samples.
Classification: Acceptable
50354707
50354706
Certification of Limits 885.1500 Submitted data fulfill the requirement for certification
of limits.
Classification: Acceptable
50354705
51391802
Color 830.6302 Brownish beige 50354704
Physical state 830.6303 Liquid suspension 50354704
Odor 830.6304 Slightly sweet musty odor 50354704
Storage Stability 830.6317 Product was stable when stored in HDPE screw top
bottles at 25°C for at least 12 months.
50354708
Miscibility 830.6319 Not applicable NA
Corrosion
Characteristics
830.6320 No signs of corrosion were noted during the 12-month
storage stability study at 25°C.
50354708
pH 830.7000 6.81 (1% w/v aqueous suspension) at 20°C;
6.29 (neat sample) at 20°C
50354704
Viscosity 830.7100 20°C: 604.9 mPa·s (30 rpm), 446.3 mPa·s (50 rpm),
97.1 mPa·s (60 rpm); 40°C: 454.0 mPa·s (30 rpm),
350.4 mPa·s (50 rpm), 316.9 mPa·s (60 rpm)
50354704
Density/Relative
Density/Bulk Density
830.7300 1.16 g/mL at 20.5°C 50354704
Product Analysis Data for F4007-9 (EPA File Symbol 279-OAUO)
Product Identity 885.1100 Submitted data fulfill the requirement for product
identity.
Classification: Acceptable
50354605
51391801
Manufacturing
Process
885.1200
Submitted data fulfill the requirement for
manufacturing process.
Classification: Acceptable
50354605
51391801*
Active Ingredients: B. subtilis strain RTI477 / B. velezensis strain RTI301 PC Codes: 006552, 006553
Action: Human Health Risk Assessment
27
Data Requirement
OCSPP
Guideline No.
Results and Classification (As Applicable)
MRID No.
Discussion of
Formation of
Unintentional
Ingredients
885.1300
Submitted data fulfill the requirement for discussion of
formation of unintentional ingredients.
Classification: Acceptable
50354605
51391801
Analysis of Samples 885.1400 Submitted data fulfill the requirement for analysis of
samples.
Classification: Acceptable
50354607
50354606
Certification of Limits 885.1500 Submitted data fulfill the requirement for certification
of limits.
Classification: Acceptable
50354605
51391801
Color 830.6302 Brownish beige 50354604
Physical state 830.6303 Liquid suspension 50354604
Odor 830.6304 Sweet musty 50354604
Storage Stability 830.6317 Stable at least 12 months in commercial packaging at
25°C
50354608
Miscibility 830.6319 Not applicable NA
Corrosion
Characteristics
830.6320 Stable via visual inspection at least 12 months in
commercial packaging at 25°C
50354608
pH 830.7000 6.76 (neat); 7.98 (1% in deionized water) 50354604
Viscosity 830.7100 609.6 mPa s 20°C 30 rpm; 451.3 mPa s 20oC 50 rpm;
408.6 mPa s 20°C 60 rpm; 625.7 mPa s 40oC 30 rpm;
465.0 mPa s 40°C 50 rpm; 414.8 mPa s 40oC 60 rpm
50354604
Density/Relative
Density/Bulk Density
830.7300 1.14 g/mL at 20°C 50354604
Product Analysis Data for F4092-3 (EPA File Symbol 279-OALR)
Product Identity 885.1100,
830.1550
Submitted data fulfill the requirement for product
identity.
Classification: Acceptable
50354808
51391803
Materials used to
Product the Product
830.1600 Submitted data fulfill the requirement for materials used
to produce the product.
Classification: Acceptable
50354808
51391803
Manufacturing
Process,
Production Process,
Formulation Process
885.1200,
830.1620,
830.1650
Submitted data fulfill the requirement for
manufacturing/production process.
Classification: Acceptable
50354808
51391803*
Discussion of
Formation of
Unintentional
Ingredients/ Impurities
885.1300,
830.1670
Submitted data fulfill the requirement for discussion of
formation of unintentional ingredients/impurities.
Classification: Acceptable
50354808
51391803
Analysis of Samples 885.1400 Submitted data fulfill the requirement for analysis of
samples.
Classification: Acceptable
50354809
50354810
Certification of Limits 885.1500,
830.1750
Submitted data fulfill the requirement for certification
of limits.
Classification: Acceptable
50354808
51391803
Enforcement
Analytical Method
830.1800 Submitted data fulfill the requirement for enforcement
analytical method.
Classification: Acceptable
50354809
50354811
Active Ingredients: B. subtilis strain RTI477 / B. velezensis strain RTI301 PC Codes: 006552, 006553
Action: Human Health Risk Assessment
28
Data Requirement
OCSPP
Guideline No.
Results and Classification (As Applicable)
MRID No.
Color 830.6302 Brownish beige 50354807
Physical state 830.6303 Liquid suspension 50354807
Odor 830.6304 Not applicable NA
Stability to normal
and elevated temps,
metals, etc.
830.6313 Product is expected to be stable under the storage and
use conditions
50644801,
50923401,
50644901,
50923601
Oxidation/Reduction 830.6314 Product does not contain strong oxidizing/reducing
agents
50354807
Flammability 830.6315 Product does not contain combustible materials 50354807
Explodability 830.6316 Product does not contain high-energy materials 50354807
Storage Stability 830.6317 Product was stable when stored in HDPE screw top
bottles at 25°C for at least 12 months.
50354811,
50354812
Miscibility 830.6319 Not applicable 50354807
Corrosion
Characteristics
830.6320 No signs of corrosion were noted during the 12-month
storage stability study at 25°C.
50354811,
50354812
Dielectric breakdown
voltage
830.6321 Not applicable 50354807
pH 830.7000 5.97 (1% w/v aqueous suspension) at 20°C; 5.97 (neat
sample) at 20°C
50354807
Viscosity 830.7100 Viscosity at 20°C: 668.1 mPa·s (30 rpm), 533.3 mPa·s
(50 rpm), 488.3 mPa·s (60 rpm); Viscosity at 40°C:
577.1 mPa·s (30 rpm), 420.9 mPa·s (50 rpm), 371.4
mPa·s (60 rpm)
50354807
Density/Relative
Density/Bulk Density
(Specific Gravity)
830.7300 1.16 g/mL at 20.5°C
50354807
* See reference 1
REFERENCES
1. Memorandum: Addendum to Data Evaluation Records for the Active Ingredients Bacillus
subtilis strain RTI477 and Bacillus velezensis strain RTI301 contained in the Proposed
Manufacturing-use Products (File Symbols 279-OAUT and 279-OAUI); M. Perry
(BPPD, RAB) to M. Thapa (BPPD, MPB), 2/9/2021.