Document ID: EPA-HQ-OPP-2015-0420-0002
Agency: epa
Document Type: Supporting & Related Material
Title: 
Posted Date: 2015-08-26T04:00Z

EPA BIOPESTICIDES AND POLLUTION PREVENTION DIVISION COMPANY 

NOTICE OF FILING FOR PESTICIDE PETITIONS PUBLISHED IN THE FEDERAL
REGISTER

EPA Biopesticides and Pollution Prevention Division contact: Shannon
Borges (703.305.7175)

LidoChem, Inc. 

20 Village Court

Hazlet, NJ  07730

Petition Number: _______________

EPA has received a pesticide petition [Petition Number:
____________________] from LidoChem, Inc. (hereinafter “LidoChem”),
20 Village Court; Hazlet, NJ 07730 (submitted by RegWest Company, LLC:
8203 West 20th St., Suite A; Greeley, CO 80634-4696) requesting,
pursuant to section 408(d) of the Federal Food, Drug, and Cosmetic Act
(FFDCA), 21 U.S.C. 346a(d), to amend 40 CFR part 180

To establish an exemption from the requirement of a tolerance for:

	

Microbial pesticide:  Bacillus amyloliquefaciens strain PTA-4838

	

Pursuant to section 408(d)(2)(A)(i) of  FFDCA, as amended, LidoChem has
submitted the following summary of information, data and arguments in
support of their pesticide petition. This summary was prepared by
RegWest Company, LLC for LidoChem and EPA has not fully evaluated the
merits of the pesticide petition. The summary may have been edited by
EPA if the terminology used was unclear, the summary contained
extraneous material, or the summary unintentionally made the reader
conclude that the findings reflected EPA’s position and not the
position of the petitioner.

LidoChem Petition Summary

	Petition Number: _______________

Product Name and Proposed Use Practices

The microbial pest control product, Bacillus amyloliquefaciens strain
PTA-4838, powder formulation is a chitinase-, protease- and
lipase-producing soil bacterium that is efficacious as a biocontrol
agent against certain pathogenic fungi, nematodes and to help establish
a vigorous root system, resulting in more uniform foliage and overall
enhanced plant health. 

Commercial/Agricultural Uses

Varnimo WSP may be applied alone, in combination and/or rotation with
chemical fungicides as a tool for integrated disease management in
bushes, crops, herbs, spices, ornamentals, shrubs, trees (forest and
shade), turf, vegetables and vines. It offers a valuable tool for the
management of chemical pesticides’ resistance through its multiple and
unique modes of action. 

The proposed uses are as follows:

Varnimo WSP, as a fungicide and nematode suppressant, may be used on all
crops, ornamentals and turf at the rate of 3-32 ounces of product in a
minimum of 25 gallons of water (drench and drip applications) and the
same dilution  applied per acre (foliar application). 

Varnimo WSP, as a plant growth promoter, may be used in the following
methods:

Transplants (including plugs): Apply Varnimo WSP to transplants by
dipping or drenching, making sure the root system is thoroughly soaked.
For dipping, follow the instructions for "Cutting and Root Dips" before
planting transplants into soil medium. For drenching, first plant the
transplants into soil medium and then follow instructions for
"Drenching". 

Drenching:  Apply Varnimo WSP to seedlings or to newly rooted cuttings.
Drench plants with Varnimo WSP’s suspension making sure the root
system is thoroughly soaked, allowing Varnimo WSP to work into the root
zone.

Cutting and Root Dips: Stir suspension for several minutes to ensure
complete mixture and to eliminate clumps. Place rootstock in the
suspension for 5-10 minutes, allowing time for Varnimo WSP to penetrate
the root zone. For ornamentals, apply at least one follow-up drench
treatment 2-3 weeks following initial treatment.

Tree Injection: Refer to manufacturers’ instructions for individual
tree injection methods/devices.

New Turf Grow-Ins: Optimal results are obtained by making the first
application 2-7 days prior to seeding, sprigging, laying sod or at the
time of seeding, sprigging or sodding. Maintain soil moisture until
seeds, sprigs or sod are installed.

Cool Season and Warm Season Turf: Optimal results are obtained by making
the first application before the first flush of roots.

Hydroponics: Prepare a stock solution by adding 0.25–0.75 ounces of
Varnimo WSP, for every 50 feet of irrigation tubing, in one gallon of
water. Stir product for several minutes to ensure complete suspension.
Add solution to circulating water system and circulate through 3 to 5
watering cycles before clearing the system. For best results, make 2 or
3 weekly applications.

Soil Incorporation: Mix Varnimo WSP into soil or soilless growing media
at a rate of 0.5-2 ounces per cubic yard. Thoroughly mix media, using
mechanical mixing equipment, to ensure uniform distribution of the
product. Incorporated into soil, Varnimo WSP may be raked into growing
beds prior to planting. 

Residential Uses

Varnimo WSP is a fungicide used for suppression of fungal plant diseases
on home garden plants, lawn and other turfgrass areas.  Application
methods are limited to drench and spray (4-8 ounces of product to 10
gallons of water with 1 gallon per every 10 square feet).

Varnimo WSP is a Plant Growth Promoter used for enhancing root
development and crop production on bushes, crops, herbs, spices,
ornamentals, shrubs, trees (forest and shade), turf, vegetables and
vines.  Varnimo WSP helps establish a vigorous root system, which often
results in more uniform foliage and overall enhanced plant health. Apply
Varnimo WSP to field soil, to planting substrate, to solid absorbent
soil amendments and directly on propagation material.

Commercial/Agricultural Seed Treatment Uses

Varnimo ST is a biological seed treatment that provides early-season
protection against listed plant parasitic nematodes and helps to
establish a vigorous root system resulting in greater yields, more
uniform foliage and overall enhanced plant health. Varnimo ST is applied
as a water-based suspension of adequate volume to uniformly distribute
the product over the seed surface area. The volume of suspension varies
according to seed type and whether companion seed treatments are used.
Application rates, depending upon the type of seed, range from 0.01 to
0.09 ounce per 100 pounds of seed; 0.02 to 0.12 ounce per 140,000
soybean seeds; and 0.01 to 0.03 ounce per 100,000 sugar beet seeds.

Product Identity/Chemistry

Identity of the pesticide and corresponding residues. 

Bacillus amyloliquefaciens strain PTA-4838 is a gram-positive,
nonpathogenic and harmless bacterium, and is ubiquitous in the
environment; it may be isolated from the air, streams, soils and oceans
and a variety of plants. 

Bacillus amyloliquefaciens strain PTA-4838 has been described as
irregular, beige, crateriform with undulate margin. Bacillus
amyloliquefaciens are gram-positive rods with peritrichous flagella
allowing motility. The cells often appear as long chains unlike many
other Bacillus species that form as single cells. The optimal
temperature for cellular growth is between 30-40°C. Similar to other
Bacillus species, Bacillus amyloliquefaciens forms endosphores allowing
survival for a long period of time. Endospores appear centrally in the
cells which do not have a swollen appearance. 

The culture is deposited in the American Type Culture Collection’s
(“ATCC”) Patent Collection and identified as “New strain of
Bacillus, APM-1 (ATCC® PTA-4838TM).

Bacillus amyloliquefaciens strain PTA-4838 is not considered to be a
human pathogen. Bacillus subtilis species, including Bacillus subtilis
variety amyloliquefaciens PTA-4838, produce proteolytic enzymes and are
widely used as additives in laundry detergents. Bacillus
amyloliquefaciens is used in broiler feed to improve poultry
productivity by its actions at different levels.

Magnitude of residues at the time of harvest and method used to
determine the residue. 

Bacillus amyloliquefaciens strain PTA-4838 is ubiquitous in the
environment, especially in soils and agricultural environments all over
the world. As a result, dietary exposure to background levels of the
naturally-occurring microbe may already be occurring and will likely
continue to occur. Because of its ubiquitous presence in the
environment, no increase in exposure to Bacillus amyloliquefaciens
strain PTA-4838 resulting from the proposed pesticidal uses, when
compared to existing exposure to background levels of Bacillus
amyloliquefaciens strain PTA-4838, is expected to occur.

Any potentially occurring residual deposits on crops will not harm
humans because the strain shows no toxicity, infectivity or
pathogenicity in appropriate tests.

A statement of why an analytical method of detecting and measuring the
levels of the pesticide residue are not needed. 

The acute toxicity studies discussed in Section C of this document are
sufficient to show that there are no foreseeable human or domestic
health hazards likely to arise from the use of the product as proposed.
LidoChem is requesting an exemption from the requirement for a tolerance
for any residues remaining in/on all agricultural commodities. Since
enforcement of residue levels would not be needed, an enforcement
analytical method is not required.

Mammalian Toxicological Profile

LidoChem conducted and submitted the required toxicology studies to
support its petition for an exemption from the requirement of a
tolerance and associated registration. Toxicology data submitted in
support of the exemption from the requirement of a tolerance for
Bacillus amyloliquefaciens strain PTA-4838 included studies on the
technical form and formulated product as follows:

Acute Toxicity and/or Pathogenicity

Varnimo® Technical

OCSPP Guideline 885.3050: The acute oral toxicity/pathology of Varnimo
Technical was evaluated when administered to rats by gavage in a single
high dose exposure and given an adequate post-exposure observation
period. The only observed general health finding was one male (17-M) in
Group IV – MCPA with a slight decrease in activity on Day 0. There
were no other abnormal general health findings. There were no observed
abnormalities during necropsy. The test substance appeared to clear
completely from brain and kidneys by Day 7, from the blood by Day 14 and
from the cecum by Day 21. The lungs, spleen, liver and mesenteric lymph
nodes (“MLN”) showed patterns of clearance through Day 21. There
were no indications of infectivity or pathogenicity by the test
substance. Varnimo Technical was determined to be non-toxic to rats when
administered by oral gavage in a single dose of 3.5 x 108 CFU/rat.

OCSPP Guideline Number 885.3150: The pulmonary toxicity, infectivity and
pathogenicity of Varnimo Technical were evaluated when administered to
rats at a single high dose exposure and an adequate post-exposure
observation period. There were no abnormal general health observations.
There were no observed abnormalities during necropsy. The test organism
was considered completely clear from the blood and cecum by Day 7;
considered completely clear from the kidneys and MLN by Day 14;
considered completely cleared from the brain, lungs and spleen by Day
21; and was present in the liver on Day 14 in one out of six animals,
but was clear on Day 21. Varnimo Technical was determined to be
non-toxic to rats when administered by tracheal injection in a single
dose of 3.1 x 109 CFU/g or 3.1 x 108 CFU/rat.

OCSPP Guideline Number 885.3200: Varnimo Technical was evaluated to
assess the acute intravenous toxicity and infectivity using a single
high dose exposure and an adequate post-exposure observation period. No
abnormalities were seen during daily observations or at necropsy. There
was no mortality in any group during the study. The test organism had
cleared completely from the Group III (MPCA) blood by Day 3 and from the
brain and the cecum by Day 7. The test organism showed a pattern of
clearance from the Group III lungs, liver, spleen and MLN. Varnimo
Technical was determined to be non-toxic to rats when injected
intravenously at a dose of 2.7 x 108 CFU/mL or 2.7 x 107 CFU/rat.

Varnimo® WSP

OCSPP Guideline Number 870.1100: Varnimo WSP was evaluated for acute
oral toxicity potential in female albino rats when administered as a
gavage dose at 5000 mg/kg. No mortality occurred during the study. There
were no clinical signs of toxicity during the study. Animals exhibited
weekly weight gain during the study. Gross necropsy conducted at
termination revealed no observable abnormalities. The acute oral LD50
was determined to be greater than 5000 mg/kg.

OCSPP Guideline Number 870.1200: Varnimo WSP was evaluated for dermal
toxicity potential and relative skin irritancy when a single dose
moistened with 0.5 mL of deionized (“DI”) water/g test substance, at
5050 mg/kg, was applied to the intact skin of albino rats. No mortality
occurred during the study. There were no clinical signs of toxicity at
any time throughout the study. Signs of dermal irritation included
erythema, atonia, desquamation and alopecia on Days 7-14. Animals
exhibited weekly weight gain, except for one that failed to gain weight
between Days 0 and 7. Gross necropsy conducted at study termination
revealed no observable abnormalities. The LD50 was determined to be
greater than 5050 mg/kg.

OCSPP Guideline Number 870.1300: Varnimo WSP was evaluated for acute
inhalation toxicity potential in albino rats. Pre-study trial assays
were conducted to determine if aerosolizing the test substance into the
exposure chamber would produce an acceptable concentration of at least 2
mg/L with mass median aerodynamic diameter (“MMAD”) between 1 and 4
μm. Prior to aerosolization in some assays, the test substance was
heat-dried and/or ground. An aerosol was generated using a Venturi
Aspirator connected to a 500 L exposure chamber. No attempt resulted in
an appropriate exposure concentration with an appropriate correspond
MMAD. Due to the nature of the test substance, an exposure concentration
could not be achieved that would provide meaningful acute inhalation
toxicity data. Based on the laboratory report, LidoChem submitted a
waiver request for this data requirement.

OCSPP Guideline Number 870.2400: An acute eye irritation study was
conducted on three albino rabbits using Varnimo WSP. The test substance,
100 mg, was placed into the conjunctival sac of the right eye of each
animal selected for testing. All treated eyes were washed with room
temperature deionized (“DI”) water for one minute immediately after
recording the 24-hour observation. There were no positive effects
exhibited in any eyes after treatment. The test substance is rated
non-irritating.

OCSPP Guideline Number 870.2500: A primary dermal irritation study was
conducted on three albino rabbits using Varnimo WSP. There was one
intact test site per animal. Each test site was treated with 500 mg of
Varnimo WSP moistened with 0.5 mL of deionized (“DI”) water and
covered with semi-permeable dressing. Test substance was maintained in
contact with the skin for 4 hours. Observations for dermal irritation
and defects were made at 1, 24, 48 and 72 hours after removal of the
dressings. Based on 0.2 PII, the test substance is rated slightly
irritating. Based on scores at only the 72-hour observation, the test
substance is assigned Toxicity Category IV. The death of one animal on
Day 3 of the study is considered unrelated to administration of the test
substance.

Aggregate Exposure

Dietary exposure

 Food 

Bacillus amyloliquefaciens strain PTA-4838 is ubiquitous in the
environment, especially in soils and agricultural environments. As a
result, dietary exposure to background levels of the naturally-occurring
microbe may already be occurring and will likely continue to occur.
Because of its ubiquitous presence in the environment, no increase in
exposure to Bacillus amyloliquefaciens strain PTA-4838, resulting from
the proposed pesticidal uses, is expected when compare to existing
exposure to background levels of Bacillus amyloliquefaciens strain
PTA-4838.

As previously discussed, dietary exposure to the naturally-occurring
microbe may already be occurring and will likely continue to occur.
Notably, similar Bacillus subtilis strains are used internationally in
the production of food-grade products and in fermented foods in Japan
and Thailand. Reports in the literature implicating Bacillus subtilis
sp. in food-borne illness do not describe any pathogen or toxin
production, but rather simple spoilage from Bacillus subtilis growth in
dough. Such low-quality dough would not be suitable for bread production
by commercial bakeries, therefore this particular food exposure scenario
is unlikely and the human risk negligible. 

An extensive literature search yielded no reliable reports of adverse
effects on non-target terrestrial plants. One on-line report identified
Bacillus amyloliquefaciens as infecting an onion in cold storage. The
temperature of the storage unit was 0-1°C. It has been established that
Bacillus amyloliquefaciens remains inactive at temperatures below 15°C
or above 50°C. Eight bacterial isolates were found all to infect
longitudinally-cut onions at 30°C. LidoChem concludes this report is
not reliable as the bacteria would have remained dormant during cold
storage and the isolation and identification of the Bacillus
amyloliquefaciens was incidental or incorrect.

The risk posed to adults, infants and children from food-related
exposure to Bacillus amyloliquefaciens strain PTA-4838 is minimal due to
the demonstrated lack of acute oral toxicity/pathogenicity associated
with the microbial pesticide. Based on the submitted data, there are no
dietary risks exceeding regulatory levels of concern.

Drinking water

Bacillus amyloliquefaciens strain PTA-4838 is ubiquitous in the
environment and exposure to the microbe through drinking water may
already be occurring and will likely continue to occur. While the
proposed and existing use sites do not include direct application to
aquatic environments, the intended use of Bacillus subtilis treatment of
growing crops or seed for the control of plant disease might result in
the potential for drift or runoff that might reach surface or ground
waters. Bacillus subtilis species are not recognized aquatic bacteria
and are not expected to grow in aquatic environments.  Municipal
drinking water treatment processes and deep water wells both further
reduce any such residues. Even if oral exposure to this ubiquitous
microbe should occur through drinking water, due to its demonstrated
lack of acute oral toxicity/pathogenicity, it is likely that no harm
will result from such exposure.

Non-dietary exposure 

The pesticide uses of Bacillus amyloliquefaciens strain PTA-4838, as
proposed, do not increase in a significant way the potential for
non-dietary, non-occupational exposure to its residues for the general
population, including infants and children, because Bacillus
amyloliquefaciens strain PTA-4838 is naturally-occurring and ubiquitous
in the environment and because it is likely populations have been
previously exposed, and will likely continue to be exposed, to
background levels of the microbe. Neither such common human exposure to
Bacillus amyloliquefaciens strain PTA-4838 naturally present in soils,
waters and plants, nor exposures associated with similar Bacillus
subtilis strains used internationally in producing food-grade products
and fermented foods, have resulted in reports of infections in humans
(which consistently are reported only in otherwise-compromised
individuals) those reports are most notable for their rare and
exceptional nature.

We believe evaluation of the high-dose Tier 1 acute toxicity and
pathogenicity tests will result in the assignment of Toxicity Category
IV (least toxic) and determinations of not infective and not pathogenic
will be concluded for all exposure routes. No toxicological end points
of concern have been identified. There are no dietary endpoints that
exceed regulatory levels of concern. Any additional exposure to the
microbe resulting from residues attributable to Bacillus subtilis var.
amyloliquefaciens strain PTA-4838 pesticide use, will not result in
additional aggregate non-occupational risk from dermal and inhalation
exposures. Based solely on non-occupation exposure scenarios, regular
occupational exposures associated with this active ingredient pose
negligible risk. 

Cumulative Effects

No mechanism of toxicity in mammals has been identified for Bacillus
amyloliquefaciens strain PTA-4838. Therefore, no cumulative effect with
other related organisms is anticipated, because the available data
demonstrate a lack of toxicity/pathogenicity potential for the active
ingredient, adverse dietary effects are unlikely. 

Safety Determination

U.S. population

Based on the acute toxicity information discussed in 3.C., there is a
reasonable certainty that no harm will result to the United States
population from aggregate exposure to residues of Bacillus
amyloliquefaciens strain PTA-4838. See “2. Infants and Children” for
rationale support.

Infants and children

Based on the acute toxicity information discussed in 3.C., 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 Bacillus amyloliquefaciens strain PTA-4838. This includes
all anticipated dietary exposures and all other exposures for which
there is reliable information. This is based on the available data on
Bacillus subtilis species, including Bacillus amyloliquefaciens strain
PTA-4838, that demonstrate a lack of toxicity/pathogenicity potential.
There are no threshold effects of concern and therefore, the additional
tenfold margin of safety for infants and children is unnecessary. The
need to consider consumption patterns, special susceptibility and
cumulative effects is unnecessary when dealing with pesticides with no
demonstrated significant adverse effects.

Effects on the Immune and Endocrine Systems

Bacillus amyloliquefaciens strain PTA-4838 is a ubiquitous organism in
the environment that is non-toxic to mammals. To date, there is no
evidence to suggest that Bacillus amyloliquefaciens strain PTA-4838
affects the immune system, functions in a manner similar to any known
hormone, or that it acts as an endocrine disruptor. The submitted
toxicity/pathogenicity studies in rodents indicate that, following
several routes of exposure, the immune system is intact and able to
process and clear the active ingredient. Therefore, it is unlikely that
this organism will have estrogenic or endocrine effects.

Existing Tolerances

There are no existing tolerance exemptions or tolerances established for
Bacillus amyloliquefaciens strain PTA-4838.

International Tolerances

No Codex maximum residue level (“MRL”) exists for Bacillus
amyloliquefaciens strain PTA-4838.

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