### BIOPESTICIDES REGISTRATION ACTION DOCUMENT



## Bacillus thuringiensis subspecies galleriae strain SDS

- 502

Pesticide Chemical (PC) Code: 0 0 6399

U.S. Environmental Protection Agency Office of Pesticide Programs Biopesticides and Pollution Prevention Division

July 24, 2013Bacillus thuringiensis subspecies galleriae strain SDS

- 502 Page 2 of 42 Biopesticides Registration Action Document



### TABLE OF CONTENTS

I. EXECUTIVE SUMMARY

................................ ................................ ................................ ......................... 4

II. ACTIVE INGREDIENT OV ERVIEW

................................ ................................ ................................ ..... 6

III. REGULATORY BACKGROUN D

................................ ................................ ................................ ............ 6

A. Applications for Pesticide Product Registration

................................ ................................ ........... 6

B. Food Tolerance Exemption

................................ ................................ ................................ ............. 7

C

. Request for an Experimental Use Permit

................................ ................................ ...................... 8

IV. RISK ASSESSMENT

................................ ................................ ................................ ................................ .. 8

A. Product Analysis Assessment (40 C.F.R. § 158.2120)

................................ ................................ ... 8

B. Human Health Assessment (40 C.F.R. § 158.2140)

................................ ................................ ....... 8

C. Environmental Assessment (40 C.F.R. § 158.2150)

................................ ................................ ..... 15

V. ENVIRONMENTAL JUSTIC E

................................ ................................ ................................ ............... 18

VI. RISK MANAGEMENT DECI SION

................................ ................................ ................................ ....... 19

VII. ACTIONS REQUIRED OF THE REGISTRANT

................................ ................................ ................. 19

VIII. GLOSSARY OF ACRONYMS AND ABBREVIATIONS

................................ ................................ .... 22

IX. BIBLIOGRAPHY

................................ ................................ ................................ ................................ ..... 23

A. Studies Submitted to Support the Bacillus thuringiensis subspecies galleriae strain SDS

- 502 Pesticide Product Registrations

................................ ................................ .................... 23

B. Environmental Protection Agency Risk Assessment Memoranda

................................ ............ 29

C. Other References

................................ ................................ ................................ ............................ 29

APPENDIX A. MICROBI AL PESTICIDES DATA R EQUIREMENTS

................................ ........................ 32

APPENDIX B. PESTICI DE PRODUCTS

................................ ................................ ................................ .......... 42Bacillus thuringiensis subspecies galleriae strain SDS

- 502 Page 3 of 42 Biopesticides Registration Action Document



### BIOPESTICIDES REGISTRATION ACTION DOCUMENT TEAM

Office of Pesticide Programs (OPP) Biopesticides and Pollution Prevention Division Microbial Pesticides Branch

Science Reviews

Joel V. Gagliardi , Ph.D. Product Analysis, Human Health John L. Kough, Ph.D. Product Analysis, Human Health Gail Tomimatsu, Ph.D. Environmental Effects Shannon Borges Environmental Effects

Regulations

Kimberly Nesci Chief, Microbial Pesticides Branch Jeannine Kausch Regulatory Action LeaderBacillus thuringiensis subspecies galleriae strain SDS

- 502 Page 4 of 42 Biopesticides Registration Action Document

I. EXECUTIVE SUMMARY

Background In July and August 2011

, Phyllom

, LLC, submitted applications for a manufacturing

- use p esticide product, Phyllom SDS

- 502 MP ( EPA File Symbol : 88347

- R )

, and an end

- use pesticide product

, Phyllom grubGONE! G ( EPA File Symbol : 88 347

- E )

, to the U .S. Environmental Protection Agen cy (EPA) under section 3 of the Federal Insecticide, Fungicide, and Rodenticide Act (FIFRA)

. In November 2012, Phyllom, LLC, submitted another application for an end

- use pesticide product, Phyllom beetleGONE ! TM biological insecticide (EPA File Symbol: 88347

- G)

. These pesticide products contain a new active ingredient, Bacillus thuringiensis subspecies galleriae strain SDS

- 502

.
Bacillus thuringiensis is a naturally occurring organism commonly found in soil. It forms parasporal crystalline bodies, which are toxic for certain invertebrates, especially species of insect larvae belonging to the insect orders Coleoptera (beetles), Diptera (flies), and Lepid optera (butterflies and moths) (Bravo 2007). The toxicity of the crystalline bodies is strain

- specific; that is, not all Bacillus thuringiensis strains are toxic to all insects.

Bacillus thuringiensis was first discovered in Japan (1901) by Ishiwata durin g his study of a bacterial disease of silkworms. Later, in 1915, a German scientist named Berliner found a similar bacillus killing the larvae of flour moths; he published a description of the bacterium and its properties, naming it Bacillus thuringiensis

. 1 In the 1930 ’ s, French researchers recognized its potential for use in agriculture and develo ped a product known as Sporeine (Federici 2005)

. In 1961, Bacillus thuringiensis was registered as a pesticide in the United States (U.S. EPA 1998)

. Today

, there are more than 100 registered pesticide products containing Bacillus thuringiensis as an active ingredient.

Bacillus thuringiensis subspecies galleriae strain SDS

- 502 was isolated from a soil sample in Japan, and data demonstrate that it is highly active against scarab beetles, such as the cupreous chafer ( Anomala
cuprea ) (Asano et al . 2003)

. Bacillus thuringiensis pesticide products used against coleopteran insects are not as common as those used against lepidopteran insects (e.g., Bacillus thuringiensis subspecies tenebrionis and Bacillus thuringiensis subspecies israelensis ). Given Bacillus thuringiensis subspecies galleriae strain SDS

- 502 ’s ability to control certain coleopteran insects, Phyllom, LLC, proposed to register a manufacturing

- use pesticide product and two end

- use pesticide product s that are intended to be applied to turf, ornamentals, and food crops, as well as in poultry premises or houses , to control white grubs of beetles and adult beetles

.

The EPA ’s s cientists reviewed product analysis, toxicology, and nontarget organism data and information ( 40 C

. F

. R

. §§ 158.2120, 158.2140, and 158.2150, respectively) submitted to support the registration of the three proposed Bacillus thuringiensis subspecies galleriae strain SDS

- 502 pesticide products

. Overall, such data and information are adequate for risk assessment purposes , fulfill the
current microbial pesticide data requirements

, and support registration of the proposed products under FIFRA section 3(c)(5)

.

1 The species name “thuringiensis” is derived from Thuringia, the region in Germany in which the disease occurred (Federici 2005).Bacillus thuringiensis subspecies galleriae strain SDS

- 502 Page 5 of 42 Biopesticides Registration Action Document

Product Analysis T he applicant fulfilled most of the product analysis data requirements for Bacillus thuringiensis subspecies galleriae strain SDS

- 502 , including product chemistry and composition, analysis of samples, and physical and chemical characteristics, by acceptable guideline studies. As terms of registration, the EPA will require the applicant to submit storage stability, corrosion characteristics, and pH studies.

Toxicology The applicant submitted a dequate mamm alian toxicology data and information to support the Ba cillus thuringiensis subspecies galleriae strain SDS

- 502 pesticide products. A cute oral and injection toxicity/pathogenicity studies and an acute inhalation toxicity study showed t hat, at a single high dose, Bacillus thuringiensis subspecies galleriae strain SDS

- 502 is not toxic , infective, and/or pathogenic via th ese route s of exposure

. Moreover, the applicant reported that no hypersensitivity incidents occurred during research, development, or testing of Bacillus thuringiensis subspecies galleriae st rain SDS

- 502

. In l ight of the results of the acute toxicity/pathogenicity data and the absence of hypersensitivity incidents, the EPA is not requiring testing at higher tiers (i.e., Tiers II and III)

.
Tolerance Exemption The EPA concludes that there is a reasonable certainty that no harm will result to the U.S. population, including infants and children, from aggregate exposure to residues of Bacillus thuringiensis subspecies galleriae strain SDS

- 502

. The EPA expects no dietar y risks are expected from use of Bacillus thuringiensis subspecies galleriae strain SDS

- 502 as an active ingredient in pesticide products. Although exposure to Bacillus thuringiensis subspecies galleriae strain SDS

- 502 through consumed food and d rinking water may be possible, the EPA anticipates these types of exposure s not to be of concern because of supporting toxicological data

. Bacillus thuringiensis subspecies galleriae strain SDS

- 502 currently meets the criteria of 40 C.F.R. § 180.1011.

Occ upational Exposure Despite the low toxicological profile of Bacillus thuringiensis subspecies galleriae strain SDS

- 502

, the EPA is requiring baseline personal protective equipment (PPE) for handlers who, due to their occupation, may be exposed to the active ingredient for prolonged periods and/ or numerous times

. Handlers working with Bacillus thuringiensis subspecies galleriae strain SDS

- 502 in agricultural or commercial settings are directed to wear a long

- sleeved shirt, long pants, socks, shoes, wate rproof gloves, protective eyewear and /or a dust/mist filtering respirator meeting National Institute for Occupational Safety and Health (NIOSH) standards of at least N

- 95, R

- 95, or P

- 95. For future products, the EPA may require additional /different PPE (e. g., chemical

- resistant gloves ) , other than the standard described above, on a product

- specific basis.

Nontarget Organisms The EPA found that guideline studies and scientific rationale submitted by the applicant are sufficient to fulfill the nontarget organism data requirements and for risk assessment purposes. Further testing of nontarget organisms at higher tier levels (i.e. , Tiers II, III, and IV) is not required. The EPA performed an environmental risk assessment, based on information provided by the applicant, and determined that no effects are anticipated for most nontarget species exposed to Bacillus thuringiensis subspe cies galleriae strain SDS

- 502 as a result of the proposed labeled applications.Bacillus thuringiensis subspecies galleriae strain SDS

- 502 Page 6 of 42 Biopesticides Registration Action Document

Public Participation On Octob er 1, 2009, the EPA announced a policy to provide a more meaningful opportunity for the public to participate in major registration decisions before they occur. According to this policy, the EPA provides a public comment period prior to making a registration decision for the following types of applications: new active ingredients; first food uses; first outdoor uses; first residential uses; or a ny other registration actions for which the EPA believes there may be significant public interest.

Consistent with the policy of making registration actions more transparent, the proposed pesticide products containing Bacillus thuringiensis subspecies galleriae strain SDS

- 502, a new active ingredient, and allowing for this active ingredient’s first food, outdoor, and residential uses were subject to a 15

- day comment period. During this comment period, the EPA received no comments on its preliminary dec ision to register the Bacillus thuringiensis subspecies galleriae strain SDS

- 502 pesticide products, Phyllom SDS

- 502 MP, Phyllom grubGONE! G, and Phyllom beetleGONE! TM biological insecticide

. Therefore, the EPA maintains that, based upon the risk assessment and information submitted in support of such pesticide products , it is appropriate to issue these registrations. The basis for this decision can be found in the risk assessment for Bacillus thuringiensis subspecies galleriae strain SDS

- 502, which is chara cterized throughout this Biopesticides Registration Action Document ( BRAD )

.

II. ACTIVE INGREDIENT OVERVIEW

Biological Name: Bacillus thuringiensis subspecies galleriae strain SDS

- 502

Culture Deposit: Japan International Patent Organism Depository, National Institute of Technology and Evaluation in Tsukuba

- shi, Japan under Accession Number FERM BP

- 7667

OPP Chemical Code: 0 0 6399

Type of Pesticide: Microbial Pesticide

– Insecticide

See Appendix B for specific information (e.g., use sites, application rates, methods of application, formulation types, and target pests) regarding the pesticide products containing this active ingredient.

III. REGULATORY BACKGROUND

A. Applications for Pesticide Product Registration

In July and August 2011

, Phyllom, LLC (address: 922 San Leandro Avenue ; Suite F ; Mountain View, California 94043 )

, submitted application s to register a manufacturing

- use pesticide product, Phyllom SDS

- 502 MP (EPA File Symbol: 88347

- R )

, and an end

- use pesticide product, Phyllom grubGONE! G (EPA File Symbol: 88347

- E )

, under FIFRA section 3. On February 29, 2012 ( 77 FR 12295 )

, the EPA announced receipt of these application s to register pesticide product s containing a new active ingredient and opened a 3 0

- day public comment period pursuant to the provisions of FIFRA section 3(c)(4). No

comme nts were received following th is publication.Bacillus thuringiensis subspecies galleriae strain SDS

- 502 Page 7 of 42 Biopesticides Registration Action Document

In November 2012, Phyllom, LLC, submitted an application to register an end

- use pesticide product, Phyllom beetleGONE! TM biological insecticide (EPA File Symbol : 88347

- G) under FIFRA section 3. On April 19, 2013, the EPA announced receipt of this application to register a pesticide product containing a new active ingredient ( 7 8 FR 23558 ) and opened a 30

- day public comment period pursuant to the provisions of FIFRA s ection 3(c)(4). In response to this notice of receipt, the EPA received one comment generally supportive of the EPA’s process for notifying the public of receipt of pesticide applications and of the Phyllom beetleGONE! TM biological insecticide application, specifically. The EPA appreciates this comment and intends to continue to meet the statutory requirement, as indicated above, to inform the public of applications for pesticide products containing a new active ingredient, such as Phyllom beetleGONE! TM biolo gical insecticide, or new uses.

B. Food Tolerance Exemption

An exemption from the requirement of a tolerance is established for viable spores of the microorganism Bacillus thuringiensis Berliner in or on honey and honeycomb and all other raw agricultural commodities when it is applied either to growing crops, or when it is applied after harvest in accordance with good agricultural practices, provided the specifications set forth in the exemption are met ( 40 C.F.R. § 180.1011 ). The specifications are listed below:

(1) The microorganism shall be an authentic strain of Bacillus thuringiensis Berliner c onforming to the morphological and biochemical characteristics of Bacillus thuringiensis as described in Bergey's Manual of Determinative Bacteriology, Eighth Edition.

(2) Spore preparations of Bacillus thuringiensis Berliner shall be produced by pure culture fermentation procedures with adequate control measures during production to detect any changes from the characteristics of the parent strain or contamination by other microorganisms.

(3) Each lot of spore preparation, prior to the addition of other materials , shall be tested by subcutaneous injection of at least 1 million spores into each of five laboratory test mice weighing 17 grams to 23 grams. Such test shall show no evidence of infection or injury in the test animals when observed for 7 days following in jection.

(4) Spore preparations shall be free of the Bacillus thuringiensis β

- exotoxin when tested with the fly larvae toxicity test (“Microbial Control of Insects and Mites,” R.P.M. Bond et al ., p. 280 ff., 1971). This specification can be satisfied either b y determining that each master seed lot brought into production is a Bacillus thuringiensis strain that does not produce β

- exotoxin under standard manufacturing conditions or by periodically determining that β

- exotoxin synthesized during spore production i s eliminated by the subsequent spore

- harvesting procedure.

The EPA determined that Bacillus thuringiensis subspecies galleriae strain SDS

- 502 currently meet s the criteria of 40 C

. F

. R . § 180.1011. To ensure that producers of Phyllom SDS

- 502 MP ( the source of active ingredient for the Bacillus thuringiensis subspecies galleriae strain SDS

- 502 end

- use pesticide products) continue to manufacture product in accordance with the tolerance exemption criteria, the EPA will also be integrating these quali ty control measures into the text of the Phyllom SDS

- 502 MP registration notice as terms.Bacillus thuringiensis subspecies galleriae strain SDS

- 502 Page 8 of 42 Biopesticides Registration Action Document

C. Request for an Experimental Use Permit

The EPA received an experimental use permit (EUP) application from Phyllom, LLC, on January 31, 2012 and issued the EU P on May 8, 2013. This EUP allows the use of 2,448 pounds of the insecticide, Bacillus thuringiensis subspecies galleriae strain SDS

- 502, on 1,400 acres of forested area to evaluate the control of emerald ash borer ( Agrilus planipennis ). The program is aut horized only in the states of Illinois, Minnesota, Pennsylvania, and Wisconsin and is effective from May 8, 2013 to August 31, 2014. In accordance with 40 C.F.R. § 172.11(c) , the EPA announced issuance of this EUP in the Federal Register of July 8, 2013 ( 78 FR 40737 ).

IV. RISK ASSESSMENT

In the Federal Register s of October 26, 2007 ( 72 FR 60988 ) and August 30, 2012 ( 77 FR 52610 )

, the EPA issued Final Rule s on the data requirements to support registration of microbial pesticides and updated the definition for microbial pesticides. The data and information evaluated for this BRAD were considered in light of these requirements.

The classifications that are found for each data submission are assigned by the EPA ’s science reviewers and are an indication of the usefulness of the information contained in the documents for risk assessment. A rating of “ acceptable ” indicates the study is scientifically sound and is useful for risk assessment. A “ supplemental ” rating indicates the data provide some information that can be useful for risk assessment. The studies may have certain aspects determined not to be scientifically acceptable (“ supple mental : upgradeable ”). If a study is rated as “supplemental: upgradeable ,” the EPA always provides an indication of what is lacking or what can be provided to change the rating to “ acceptable .” If there is simply a “ supplemental ” rating, the reviewer will often state that t he study is not required by 40 C

. F

. R

. P art 158. Both “ acceptable” and “supplemental ” studies may be used in the risk assessment
process as appropriate. An “unacceptable ” rating indicates that new data must be submitted.

For the acu te tox icity data requirements, Toxicity C ategories are assigned based on the hazard(s) identified from studies and/or other information submitted to the EPA in support of a pesticide registration. The active ingredient or particular product is classified into Toxicity Category I, II, III, or IV, where Toxicity Category I indicates the highest toxicity and Toxicity Category IV indicates the lowest toxicity (se e section II of U.S. EPA ( 2012 b ) )

.
A. Product Analysis Assessment ( 40 C

. F

. R

. § 158.2120 )

With the exception of a one

- year storage stability and corrosion characteristics study and a pH study, t he applicant fulfilled a ll product analysis data requirements for Bacillus thuringiensis subspecies galleriae strain SDS

- 502 . Refer to Tables 1, 2, 3, a nd 4 in Appendix A for a summary of the data requirements, including both generic and product

- specific information.

B. Human Health Assessment ( 40 C

. F

. R

. § 158.2140 )

1. Toxicity

The applicant fulfilled a ll toxicology data requirements for Bacillus thu ringiensis subspecies galleriae strain SDS

- 502

. Acceptable Tier I mammalian toxicology data and information support registration ofBacillus thuringiensis subspecies galleriae strain SDS

- 502 Page 9 of 42 Biopesticides Registration Action Document

the Bacillus thuringiensis subspecies galleriae strain SDS

- 502 pesticide products

. The EPA is not requiring Tier II and Tier III studies for Bacillus thuringiensis subspecies galleriae strain SDS

- 502 based on the lack of acute toxicity/pathogenicity in the Tier I studies attributed to the bacterium

.
For a summary of the generic toxicology data requirements desc ribed in sections IV(B)(1)(a) and IV(B)(1)(b), as well as additional product

- specific data submitted to support the individual registrations, refer to Tables 5 and 6 in Appendix A

.

a. Acute Toxicity/Pathogenicity

– Tier I

Acute Oral Toxicity/Pathogenicity

– Rat ( Harmonized Guideline 885.3050; Master Record Identification Number s (MRID No s .) 485532

- 08, 485532

- 11, and 489401

- 08 ) : In an acute oral toxicity and pathogenicity study

, groups of fasted, 9

- week

- old rats (2 or 3 per sex) were given a single oral dose of Bacillus thuringiensis subspecies galleriae strain SDS

- 502 in sterile phosphate buffered saline at a dose of 2.4 x 10 8 cfu per animal. The animals were observed for a period of up to 21 days, with interim scheduled sac rifices on days 3, 7, and 14. Three males and three females were treated with heat

inactivated Bacillus thuringiensis subspecies galleriae strain SDS

- 502 as inactivated controls, two males and two females were untreated shelf controls, and two males and tw o females were untreated, non

- shelf controls. Based on the results of this study, Bacillus thuringiensis subspecies galleriae strain SDS

- 502 does not appear to be toxic, infective, or pathogenic in rats, when dosed at 2.4 x 10 8 cfu per animal. There were no treatment

- related deaths, clinical signs, necropsy findings, or changes in body weight or body weight gain. Only samples from day 0 and day 21 were analyzed; high levels for test organism were present in feces on day 0, while no test organism was recovered from the cecum contents or any organ or blood from treated animals sacrificed on day 21. The EPA rated this study acceptable.

Acute Pulmonary Toxicity/Pathogenicity

– Rat ( Harmonized Gui deline 885.3150; MRID No s

. 485532

- 09, 485532

- 11, and 489401

- 08 ) : In an acute pulmonary toxicity and pathogenicity study, groups of 9

- week

- old rats (3 or 5 per sex per group) were exposed by the intratracheal route to Bacillus thuringiensis subspecies galleriae strain SDS

- 502 in sterile phosph ate buffered saline. Six males and five females were treated with heat

- inactivated Bacillus thuringiensis subspecies galleriae strain SDS

- 502 , and two males and two females served as untreated controls. The inactive

- and active

- treated animals were initial ly mis

- dosed at approximately 0.2 x 10 8 cfu per animal, and three male and three female active

treated animals were killed (on schedule) before the mis

- dosing was recognized. The following day, three male inactive controls were treated with a dose approximately equivalent to 2 x 10 8 cfu per ani mal with immediate mortality. The remaining animals scheduled to be treated, as well as three male and three female replacement animals, were treated at a dose of 1.2 x 10 8 cfu per animal in a 0.5 milliliter dosing volume, and the surviving inactive

- treate d animals were treated at an approximately equivalent dose. The animals were observed for 21 days after the second dose. In addition to the three males treated with the initial formula tion of the inactive control, 2 of 3 males and 3 of 5 females treated wi th the further diluted and homogenized inactive suspension were found dead on days 0

– 1, and an additional female in this group was sacrificed in extremis on day 0. One female treated with the active suspension was found dead on day 1 after exhibiting letha rgy, labored breathing, gasping, and inappetence. Red, mottled discoloration of the lungs and red staining of the face were found on this animal at necropsy. Additional treatment

- related clinical signs among the surviving active

- treated animals included in appetence, ocular and/or nasal discharge, lethargy, noisy breathing, and staining of the fur and skin. The study author attributed these signs to the physical properties of the dosing formulations. Body weight losses were noted for the active

- treated anima ls of both sexes killed on day 3, and the active

treated males had decreased body weight gain during days 0

– 7. Abnormal gross necropsy findingsBacillus thuringiensis subspecies galleriae strain SDS

- 502 Page 10 of 42 Biopesticides Registration Action Document

among animals killed on schedule were limited to tan, mottled discoloration of the lungs in the three males and three females that were killed on day 7. Average cfu counts decreased over time in all of the evaluated tissues or samples, but the test organisms did remain present through day 21 with the highest counts seen in lung tissue (126 cfu per gram). No conclusi ons can be made regarding infectivity, pathogenicity, or toxicity based on these study results. Attributing lethal effects to dosing was not followed up with a lternate dosing methods. The EPA rated this study unacceptable and concluded that a replacement p ulmonary toxicity/pathogenicity study or an inhalation toxicity study was required to address pulmonary effects. (See review summary for MRID No. 489731

- 01 , directly below .)

Acute Inhalation Toxicity

– Rat (Harmonized Guideline 870.1300; MRID No. 489731

- 01) : All animals survived and gained weight with exception of 1 male that lost 7 grams after day 7. Symptoms noted between 4.5 and 6 hours after exposure included red nasal discharge, red crust around eyes, decreased activity and piloerection noted as sli ght in 2

– 5 of the males and very slight red crust around the nose of 1 female at 4.5 hours. No other symptoms were reported through day 14 of the study. No observable abnormalities were noted at necropsy. The inhalation median lethal concentration (LC 50 ) w as greater than 4.30 mg/L, and the EPA rated this study acceptable.

Acute Injection Toxicity/Pathogenicity ( Intraperitoneal )

– Rat (Harmonized Guideline 885.3200; MRID No s

. 485532

- 10, 485532

- 11, and 489401

- 08 ) : In an acute intraperitoneal injection toxicity and pathogenicity study, 15 male and 15 female 11

- week

- old rats were injected with Bacillus thuringiensis subspecies galleriae strain SDS

- 502 in sterile phosphate buffered saline at a dose of 2.02 x 10 7 cfu pe r animal and observed for up to 21 days with interim sacrifices of 3 animals per sex done on days 0, 3, 7, and 14. Two additional groups of two males and two females were either left untreated or treated with a suspension of the heat

- inactivated test mater ial. Bacillus thuringiensis subspecies galleriae strain SDS

502 was not pathogenic or overtly toxic when dosed intraperitoneally; however, the test organism persisted in the animals through day 21. There were no treatment

- related deaths, clinical signs, or necropsy findings, and body weight gain was not significantly affected. The t est organism was recovered from cecum contents, organ and blood samples from active

- treated animals sacrificed on day 21 and was most prevalent in lymph node samples (33.3 cfu per gram). The EPA rated this study acceptable.

Hypersensitivity Incidents ( Harmonized Guideline 885.3400; MRID No. 485532

- 12 ) : The applicant reported that n o hypersensitivity incidents, including immediate

- type or delayed

- type reactions of humans and domestic animals

, occurred during research, development, or testing of Bacillus thuringiensis subspecies galleriae strain SDS

- 502 . Any future hypersensitivity incidents must be reported to the EPA (refer to test note #3 of 40 C

. F

. R

. § 158.2140(d))

.
Cell Culture ( Harmonized Guideline 885.3500 ; MRID No. 485532

- 13 ) : This study is not required because Bacillus thuringiensis subspecies galleriae strain SDS

- 502 is not a virus (refer to test note #4 of 40 C

. F

. R

. § 158.2140(d)).

b. Acute Toxicology and Subchronic Toxicity/Pathogenicity

– Tier II; Reproductive Fertility Effects, Carcinogenicity, Immunotoxicity, and Infectivity/Pathogenicity Analysis

– Tier III

The EPA is not requiring Tier II and Tier III studies for Bacillus thuringiensis subspecies galleriae strain SDS

- 502 based on the lack of acute toxicity/pathogenicity in the Tier I studies attributed to the bacterium

. Any uncertainties about the toxicity , pathogenicity, or infectivity of Bacillus thuringiensis subspecies galleriae strain SDS

- 502 raised in the acute p ul m o nary toxicity/pathogenicity study (MRIDBacillus thuringiensis subspecies galleriae strain SDS

- 502 Page 11 of 42 Biopesticides Registration Action Document

No. 485532

- 09 ) were addressed by a subsequent acute inhalation toxicity study (MRID No. 489731

- 01 ) and information to support lack of infectivity of Bacillus thuringiensis subspecies galleriae strain SDS

502 (MRID No. 48 9401

- 08).

c. Endocrine Disruptors

As required by the Administrator under FFDCA section 408(p), the EPA has developed the Endocrine Disru ptor Screening Program (EDSP) and has begun to implement the screening program that is to be used to test all pesticides in order to determine whether certain substances (including pesticide active and other ingredients) may have an effect in humans or wil dlife similar to an effect produced by a “naturally occurring estrogen, or other such endocrine effects as the Administrator may designate.”

FFDCA section 408(p)(4) authorizes the Administrator, by order, to exempt from the requirements of the Estrogenic Substances Screening Program a biologic substance or other substance if a determination is made that the substance is not anticipated to produce any effect in humans similar to an effect produced by a naturally occurring estrogenic substance.

Between Oct ober 2009 and February 2010, the EPA issued test orders/data call

- ins for the first group of 67 chemicals, which contains 58 pesticide active ingredients and 9 inert ingredients. Bacillus thuringiensis subspecies galleriae strain SDS

- 502 is not among the g roup of 58 pesticide active ingredients on the initial list to be screened under the EDSP.

The EPA believes that Bacillus thuringiensis subspecies galleriae strain SDS

- 502 , the active ingredient described in this document, likely is a substance that woul d not produce any effect in humans similar to an effect produced by a naturally occurring estrogenic substance. As such, pursuant to Section 408(p)(4), the EPA will determine in the future whether it can exempt Bacillus thuringiensis subspecies galleriae s train SDS

- 502 from the requirements of the Section 408(p) EDSP. In the event the EPA does determine to exempt this substance from the EDSP, an order will be issued.
For further information on the status of the EDSP, the policies and procedures, the list of 67 chemicals, the test guidelines and the Tier 1 screening battery, please visit our websit e at http://www.epa.gov/endo/

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

Section 408(c)(2)(A)(i) of FFDCA allows the EPA to establish an exemption from the requirement for a tolerance (the legal limit for a pesticide chemical residue in or on a food) only if the 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 in formation.” 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 tolera nce, the EPA must take into account the factors set forth in FFDCA section 408(b)(2)(C), which require the EPA to give special consideration to exposure of infants and children to the pesticide chemical residue in establishing a tolerance exemption, and to “ensure that there is a reasonable certainty that no harm will result to infants and children from aggregate exposure to the pesticide chemical residue....” Additionally, FFDCA section 408(b)(2)(D) requires that the EPA consider “available information con cerning the cumulative effects of [a particular pesticide's] . . . residues and other substances that have a common mechanism of toxicity.”Bacillus thuringiensis subspecies galleriae strain SDS

- 502 Page 12 of 42 Biopesticides Registration Action Document

The EPA performs a number of analyses to determine the risks from aggregate exposure to pesticide residues. First, the EPA determines the toxicity of pesticides. Second, the EPA examines exposure to the pesticide through food, drinking water, and through other exposures that occur as a result of pesticide use in residential settings.

Consistent with FFDCA section 408(b )(2)(D), the EPA reviewed the available scientific data and other relevant information on Bacillus thuringiensis subspecies galleriae strain SDS

- 502 , and considered its validity, completeness, and reliability, as well as the relationship of this informatio n to human risk. The EPA also considered available information concerning the variability of the sensitivities of major identifiable subgroups of consumers, including infants and children.

Based on the acute toxicity/pathogenicity data and information disc ussed previously and presented in Tables 5 and 6 in Appendix A , t he data required for a FFDCA risk assessment for Bacillus thuringiensis subspecies galleriae strain SDS

- 502 have been fulfilled.

a. Aggregate Exposure

In examining aggregate exposure, FFDCA section 408 directs the 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 wat er and exposure through pesticide use in gardens, lawns, or buildings (residential and other indoor uses).

Food Exposure and Risk Characterization : Exposure to residues of Bacillus thuringiensis subspecies galleriae strain SDS

- 502 in food may occur throug h the consumption of food commodities that have been treated with microbial pesticides containing this bacterium. The EPA expects the exposure to food residues of Bacillus thuringiensis subspecies galleriae strain SDS

- 502 to be negligible when compared to the ordinary, everyday levels to which individuals are already exposed in the natural environment

— Bacillus thuringiensis is commonly found in soil and on plant s (Damgaard et al . 1998 ; Gao et al . 2008; Mizuki et al . 1999 )

. Although residues from the use of pesticides containing Bacillus thuringiensis subspecies galleriae strain SDS

- 502 may be present on certain food commodities, processing procedures (e.g., peeling and washing) may reduce such residues ( U.S. EPA 1996 ). Rega rdless of the scenario, should Bacillus thuringiensis subspecies galleriae strain SDS

- 502 be present on food, supporting toxicological data indicate that no toxicity, pathogenicity, and/or infectivity is likely to occur with any food exposure resulting fro m the use of Bacillus thuringiensis subspecies galleriae strain SDS

- 502 as a pesticide when manufactured to meet the specifications of 40 C.F.R. § 180.1011 and applied in accordance with label directions and good agricultural practices (see section IV(B)(1)(a) and Tables 5 and 6 in Appendix A ).

Drinking Water Exposure and Risk Characterization : Ex posure to residues of Bacillus thuringiensis subspecies galleriae strain SDS

- 502 , a naturally occurring bacterium (D amgaard et al . 1998; Gao et al

. 2008; Mizuki et al . 1999) , in consumed drinking water is possible. End

- use pesticide products containing Bacillus thuringiensis subspecies galleriae strain SDS

- 502 can be applied with ground and aerial equipment to many diff erent plants. Runoff from treated areas and spray drift are definite possibilities. If Bacillus thuringiensis subspecies galleriae strain SDS

- 502 were to be transferred to ground or surface waters as a result of runoff or spray drift, this bacterium could survive the conditions water is subjected to in wastewater treatment systems and drinking facilities (e.g., pH adjustments andBacillus thuringiensis subspecies galleriae strain SDS

- 502 Page 13 of 42 Biopesticides Registration Action Document

chlorination). According to the World Health Organization, Bacillus species, such as thuringiensis , are ofte n detected in drinking water even after going through acceptable water treatment processes, largely because the spores are resistant to these disinfection processes (World Health Organization 2011). Should Bacillus thuringiensis subspecies galleriae strain SDS

- 502 be present in drinking water, supporting toxicological data indicate that no toxicity, pathogenicity, and/or infectivity is likely to occur with any drinking water exposure resulting from the use of Bacillus thuringiensis subspecies galleriae
stra in SDS

- 502 as a pesticide when manufactured to meet the specifications of 40 C.F.R. § 180.1011 and applied in accordance with label directions and good agricultural practices (see section IV(B)(1)(a) and Tables 5 and 6 in Appendi x A ).

Non

- occupational, Residential Risk Characterization : Given Bacillus thuringiensis ’ natural presence in the environment (Damgaard et al . 1998; Gao et al . 2008; Mizuki et al . 1999) , dermal and inhalation

non

- occupational exposures to Bacillus thuringiensis subspecies galleriae strain SDS

- 502 almost certainly are already occurring. Additional exposure to Bacillus thuringiensis subspecies galleriae strain SDS

- 502 may occur because the end

- use pesticide products are labeled for applicatio n to sites like athletic fields, residential lawns, and playgrounds . Even if additional non

- occup ational exposures were to occur, no adverse effects attributed to Bacillus thuringiensis subspecies galleriae strain SDS

- 502 are anticipated, particularly in l ight of testing results that indicated this bacterium is not toxic , pathogenic, or infective and is essentially non

- irritating to skin at high doses ( see section IV(B)(1)(a) and Tables 5 and 6 in Appendix A ).

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

Section 408(b)(2)(D)(v) of FFDCA requires that, when considering whether to establish, modify, or revoke a tolerance, the EPA consider “available information concerning the cumulative effects of [a particular pesticide’s] . . . residues and other substances that have a common mechanism of toxicity.”

The EPA has not found Bacillus thuringiensis subspecies galleriae strain SDS

- 502 to share a common mechanism of toxicity with any other substances, a nd Bacillus thuringiensis subspecies galleriae strain SDS

- 502 does not appear to produce a toxic metabolite produced by other substances. For the purposes of this tolerance action, therefore, the EPA has assumed that Bacillus thuringiensis subspecies galle riae strain SDS

- 502 does not have a common mechanism of toxicity with other substances. Following from this, therefore, the EPA concludes that there are no cumulative effects associated with Bacillus thuringiensis subspecies galleriae strain SDS

- 502 that n eed to be considered. For information regarding the EPA’s efforts to determine chemicals that have a common mechanism of toxicity and to evaluate the cumulative effects of such chemicals, see the EPA’s website at http://www.epa.gov/pesticides/cumulative

.

c. Determination of Safety for the United States Population, Infants and Children

FFDCA section 408(b)(2)(C) provides that, in considering the establishment of a tolerance or tolerance exemption for a pesticide chemical residue, the EPA shall assess the available information about consumption patterns among infants and children, special susceptibility of infants and children to pesticide chemical residues, and the cumulative effects on infants and children of the residues and other substances with a common mechanism of toxicity. In addition, FFDCA section 408(b)(2)(C) provides that the E PA 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 the EPA determines th at a different margin of safety will be safe forBacillus thuringiensis subspecies galleriae strain SDS

- 502 Page 14 of 42 Biopesticides Registration Action Document

infants and children. This additional margin of safety is commonly referred to as the Food Quality Protection Act Safety Factor. In applying this provision, the EPA either retains the default value of 10X,

o r uses a different additional or no safety factor when reliable data are available to support a different additional or no safety factor.

Based on the acute toxicity and pathogenicity data/information discussed in section IV(B)(1)(a) and Tables 5 and 6 i n Appendix A

, the EPA concludes that there are no threshold effects of concern to infants, children, or adults when Bacillus thuringiensis subspecies galleriae strain SDS

- 502 is manufactured according to the criteria set forth in 40 C.F.R. § 180.1011, is used as labeled, and good agricultural practices are followed . As a result, the EPA concludes that no additional margin of exposure (safety) is necessary.

Moreover , based on the same data and analysis as presented directly above, the EPA conclude s that there is a reasonable certainty that no harm will result to the U.S. population, including infants and children, from aggregate exposure to the residues of Bacillus thuringiensis subspecies galleriae strain SDS

- 502 when it is manufactured according to the criteria set forth in 40 C.F.R. § 180.1011 and used as labeled as an insecticide . Such exposure includes all anticipated dietary exposures and all other exposures for which there is reliable information. The EPA has arrived at this conclusion because, considered collectively, the data and information available on Bacillus thuringiensis subspecies galleriae strain SDS

- 502 do not demonstrate toxic, pathogenic, and/or infective potential to mammals, including infants and children.

3 . Occupational Exposure and Risk Characterization

Handler exposure to Bacillus thuringiensis subspecies galleriae strain SDS

- 502 is not expected to pose any undue risk. Regardless, the EPA is requiring appropriate personal protective equipment and precautionary statements on pesticide product labels to mitigate any potential risks to pesticide handlers due to prolonged and/ or numerous exposures. Handlers working with Bacillus thuringiensis subspecies galleriae strain SDS

- 502 in agricultural or commercial settings are directed to wear a long

- sleeved shirt, long pants, socks, shoes, waterproof gloves, protective eyewear, and /or a dust/mist filtering respirator meeting NIOSH standards of at least N

- 95, R

- 95, or P

- 95. For future products, the EPA may require additional /different PPE (e.g., chemical

- resistant gloves ), other than the standard described above, on a product

- specific basis.

4 . Human Health Risk Characterization

The EPA considered human expo sure to Bacillus thuringiensis subspecies galleriae strain SDS

- 502 in light of the standard for registration in FIFRA and the relevant safety factors in FFDCA. A determination has been made that no unreasonable adverse effects to the U.S. population in gen eral, and to infants and children in particular, will result from incidental exposures when Bacillus thuringiensis subspecies galleriae strain SDS

- 502 pesticide products are applied to turf, to ornamentals, to various crops, and in poultry premises and hou ses in accordance with the EPA’s approved labeling.Bacillus thuringiensis subspecies galleriae strain SDS

- 502 Page 15 of 42 Biopesticides Registration Action Document

C. Environmental Assessment ( 40 C

. F

. R

. § 158.2150 )

The EPA found that guideline studies and scientific rationale submitted by the applicant are sufficient to fulfill the nontarget organism data requirements and for risk assessment purposes. Further testing of nontarget organisms at higher tier levels (i .e., Tiers II, III, and IV) is not required. The EPA performed an environmental risk assessment, based on information provided by the applicant, and determined that no effects are anticipated for most nontarget species exposed to Bacillus thuringiensis sub species galleriae strain SDS

- 502 as a result of the proposed labeled applications.

For a summary of the generic data requirements described in sections IV(C)(1), refer to Table 7 in Appendix A

.

1. Ecological Exposure and Risk Characterization

The end

- use pesticide product (EP) , Phyllom beetleGONE! TM biological insecticide, is proposed for application to many types of grain, vegetable and small fruit crops, including fruit and nut trees, greenhouse /shadehouse and ornamental plants and trees, and poultry premises. Proposed application methods include ground and aerial sprays, as well as chemigation. The EP , Phyllom grubGONE! G, is proposed for use on turfgrass and ornamentals plants and trees, and is intended to be applied by ground application equipment. The turfgrass sites include golf course greens, reside ntial lawns, commercial grounds and sod farms, and the ornamentals sites include landscapes, interiorscapes, nurseries, fields and greenhouses. S ince the proposed applications are expected to result in contact of the active ingredient (a.i.) with foliar surfaces and soil, and spray drift is expected to occur, exposure to terrestrial and aquatic nontarget organisms is possible.

Based on current in formation concerning the amount of active ingredient 2 in the EP, Phyllom beetleGONE! TM biological insecticide , the EPA estimates terrestrial maximum environmental concentrations (EC) of Bacillus thuringiensis subspecies galleriae strain SDS

- 502 as approxima tely 6,078 g/A. An aquatic maximum EC was estimated at 10.0 mg a.i./L of water.

a. Birds (Harmonized Guidelines 885.4050 and 885.4100) and Wild Mammals (Harmonized Guideline 885.4150) (MRID Nos. 485532

- 19, 489401

- 07, 485532

- 20 , and 485532

- 21 ; Rose 2013b )

Birds and mammals may be exposed to Bacillus thuringiensis subspecies galleriae strain SDS

- 502, primarily through ingestion or contact with treated surfaces. Information presented to the EPA indicates low risk to birds and wild mammals as a result of the proposed applications. An acute oral toxicity study with birds at the maximum hazard dose (MRID No. 485532

- 19) showed that the acute oral median lethal dose (LD 50 ) is 3.0 x 10 11 cfu/kg of body weight per day. A study with laborat ory rats exposed orally (MRID No. 485532

- 08 ) showed that Bacillus thuringiensis subspecies galleriae strain SDS

- 502 is not toxic, infective, or pathogenic when administered at maximum hazard doses. Other acute toxicological data volumes (MRID No s

. 485532

- 1 1 and 489401

- 08 ) provide additional information to conclude that toxicity to mammals is not expected.

2 Terrestrial maximum EC calculation : 17.5 lb/A * 454 g/lb * 1 acre * 76.5% a.i. in EP ; aquatic maximum EC calculation: 17.5 lb /A * 454,000 g/lb * 1A/40,468,564 cm 2 = 0. 1963 mg/cm 2 ÷ 15 cm 2 = 0.013 mg/mL * 1000 mL/L = 13.08 mg/L * 76.5 % = 10.0 mg a.i./L.Bacillus thuringiensis subspecies galleriae strain SDS

- 502 Page 16 of 42 Biopesticides Registration Action Document

b. Nontarget Plants (Harmonized Guideline 885.4300) (MRID No. 485532

- 25 )

The applicant submitted a scientific rationale to satisfy the data requirement for nontarget plants. Bacillus thuringiensis is well characterized wi th respect to effects on plants and is not taxonomically related to known plant pathogens. Therefore, adverse effects on nontarget pl ants (terrestrial and aquatic) are not expected to result from the proposed applications of Bacillus thuringiensis subspecies galleriae strain SDS

- 502

.

c. Nontarget Insects (Harmonized Guideline 885.4340) and Honey Bees (Harmonized Guideline 885.4380) (MRID No s

. 485532

- 04, 485532

- 26, 485532

- 27, 485532

- 28, 485532

- 29, 485532

- 30, 487168

- 03, and 489401

- 07 ; Rose 2013b )

The EPA reviewed submitted studies and information from the open literature. Most of the submitted insect studies (bioassays) provided limited information regarding potential dietary toxicity of the Cry8Da protein, extracted and purified from Bacillus thur ingiensis subspecies galleriae strain SDS

- 502 (Table 7 in Appendix A ). T hese studies may have been of insufficient duration to note potential activity by an insecticide that must be ingested by susceptible insect larvae for toxic effects. Insect bioassays may be influenced by relative proportions of the different proteins within the crystal, thereby obscuring the role an individual toxin may play in insect toxicity . Variability in the composition and potency of crystals within the same strain may be caused by factors such as mobile genetic elements and nutriti onal requirements (Masson et al . 1998) . At least seven different delta

- endotoxins were identified in Bacillus thuringiensis subspecies galleriae (Chestukhina et al . 1994)

.

It is not possible to determine whether these exposures were conducted at maximum hazard concentrations, based on submitted information about the protein levels in the test materials for some of the studies (MRID No. 485532

- 04). A couple of submitted studies used different protein extraction, purification and quantification methods and did not report the percentage of the Cry8Da protein.

The EPA determined that the i temized studies below ( studies b through f) are supplemental ; however

, all studies are usefu l for the risk assessment of Bacillus thuringiensis subspecies galleriae strain SDS

- 502 because they provide a spectrum of the tested protein’s (Cry8Da) activity. The conclusions of these insect bioassays/studies were as follows :

a. No sustained adverse effe cts are anticipated for populations of H ippodamia convergens (Order Coleoptera)

. b. Potential Cry8Da toxicities were demonstrated for Plutella xylostella (Order Lepidoptera); defined as “weakly toxic” (cited by Asano et al . (2003)

, see MRID No. 485532

- 27).

c. No Cry8Da toxicity was demonstrated for Bombyx mori (Oriental silkworm moth) or Spodoptera litura (Oriental leafworm moth)

. d. No Cry8Da toxicity was demonstrated for Leptinotarsa decemlineata (Colorado p otato beetle)

. e. No Cry8Da toxicity to 4 species of adult parasitic Hymenoptera

. f. High toxicity of purified Cry8Da from Bacillus thuringiensis subspecies galleriae strain SDS

502 to the Emerald Ash Borer ( Agrilus planipennis or Agrilus marcopoli ) and to Scarabaeidae beetle “white grubs” ( Anomala cuprea ) w as demonstrated i n another submitted study.

Asano et al

. (2003) noted that other species of the Scaraba eid ae (e.g., cupreous chafer, Anom a la cuprea ; Japanese beetle, Popillia japonica Newman) are affected by other strains or subspecies of BacillusBacillus thuringiensis subspecies galleriae strain SDS

- 502 Page 17 of 42 Biopesticides Registration Action Document

thuringiensis (e.g., Bacillus thuringiensis subspecies galleriae NRRL B

- 4027 and Bacillus thuringiensis subspecies japonensis ) and other C ry 8 genes were active against Cotin i s sp p

. (June beetle).

Although the Cry8Da protein was reported as weakly toxic to a lepidopteran pest, Plutella xylostella

, another formulation with Bacillus thuringiensis subspecies galleriae was developed to control the wax moth ( Galleriae sp p

. ) in honeybee hives (Melin and Cozzi 1990)

.

Based on these studies and the informatio n presented above, effects to some insect species in the orders Coleoptera and Lepidoptera as a result of Bacillus thuringiensis subspecies galleriae strain SDS

- 502 applications cannot be precluded. More r efined estimates of the potential risk are not possible because (1) the majority of the studies were not conducted at maximum hazard concentrations and (2) there are uncertainties associated with the bioassays because of variation in the protein extraction methods used in those studies. T hese bi oassays were conducted primarily to determine the potential insecticidal activity of Cry8Da protein.

The applicant also submitted a study with honey bee larvae ( Apis mellifera ) exposed to Bacillus thuringiensis subspecies galleriae strain SDS

- 502 Cry8Da protein. Based on this study, the no observed effect concentration ( NOEC ) was 500 nanograms Bacillus thuringiensis subspecies galleriae strain SDS

- 502 Cry8Da protein/honeybee larval cell; however, the study was supplemental because of the uncertainty in do sing. However, a commercial formulation containing Bacillus thuringiensis subspecies galleriae (strain not identified) with Lepidopteran activity was discussed in a review paper by Melin and Cozzi (1990) . The formulation (no potency information provided) was developed to control the wax moth ( Galleriae spp.) in bee hives, and the bacterium can be sprayed on the surface of the honeycomb or impregnated into the wax. Hives treated with Bacillus thuringiensis subspecies galleriae (strain and viability not prov ided) in a 40% sucrose solution showed no adverse effect on adult workers or colony life as determined by egg laying, brood production, brood capping or honey production (Melin and Cozzi 1990) . Therefore, EPA does not have concerns as a result of exposure of this new active ingredient to honeybees

d. Freshwater Fish (Harmonized Guideline 885.4200), Freshwater Invertebrates (Harmonized Guideline 885.4240), and Estuarine/Marine Fish and Invertebrates (Harmonized Guideline 885.4280) (MRID No s

. 485532

- 04, 485532

- 22, 485532

- 23, 485532

- 24, and 489401

- 07; Rose 2013b )

The applicant submitted scientific rationale s in lieu of studies for freshwater fish and estuarine/marine organisms . Resubmitted information and data confirmed the taxonomic classification as Bacillus thuringiensis subspecies galleriae , with no known pathogenesis to fish species (U.S. EPA 1998) and a limited host range to certain insect orders.

The applicant also sub mitted a study with Daphnia magna in which daphnids were exposed to water containing a range of nominal concentrations of the technical grade of the technical grade of the active ingredient (TGAI) for 21 days. Based on the percentage of the active ingredie nt in the EP, a direct application to a body of water equivalent to the EPA Standard Wetland would be approximately 10.0 mg a.i./L, which is less than the NOEC values for reproduction and growth (i.e., 50 mg/L) and for survival (i.e., 100 mg/L). Although high daphnid mortality and low neonate production were noted in the filtered treatment, the adverse effects did not affect the overall conclusions of the study and were attributed to low dissolved oxygen content, and anomalous values for conductivity and water hardness. Since exposure resulting from the proposed applications is well below the defined toxicity endpoints,Bacillus thuringiensis subspecies galleriae strain SDS

- 502 Page 18 of 42 Biopesticides Registration Action Document

adverse effects to freshwater invertebrates are not expected.

2. Environmental Fate Data

Microbial pesticides containing Bacillus thu ringiensis readily break down in the environment, degrade within 48 hrs under ultraviolet light exposures (Myasnik et al . 2001), and have a half

- life of 3.8 hours under normal sunlit conditions (USDA 1994) . The microorganism can persist for several months in soil (Melin and Cozzi 1990) , and certain Cry proteins may degrade within 32 days. Half

- life values (DT 50 ) ranged from 8.3 to 31.3 days (Mendelsohn et al . 2003) . A spray

- dried powder of Bacillus thuringiensis subspecies galleriae strain SDS

- 502 provided nearly 100% control of Anomala cuprea larvae as long as 56 days after an initial application of 0.12 g/m 2 (Asano et al . 2003)

.

3

. Threatened and Endangered Species Assessment

The EPA has determined that no effects are anticipated for most nontarget species exposed to Bacillus thuringiensis subspecies galleriae strain SDS

- 502 as a result of the proposed labeled applications, including mammals, birds, terrestrial and aquatic plan ts, and freshwater and marine/estuarine animals. Therefore, a “No Effect” determination is made for direct and indirect effects to federally listed endangered and threatened (“listed”) mammals, birds, terrestrial and aquatic plants, and freshwater and mari ne/estuarine animals and for their designated critical habitat.

Sustained population

- level effects to nontarget insects are not anticipated; however, it is known that Bacillus thuringiensis subspecies galleriae can affect insects in the orders Coleoptera and Lepidoptera. Direct effects to individuals of listed insect species (USFWS 2013) cannot be precluded, nor can indirect effects to listed species that depend specifically on a listed insect. Additional analysis of the potential effects to these is nece ssary to make a determination for these species.

V. ENVIRONMENTAL JUSTICE

The EPA seeks to achieve environmental justice

— the fair treatment and meaningful involvement of all people regardless of race, color, national origin, or income

— with respect to the development, implementation, and enforcement of environmental laws, regulations , and policies. Fair treatment means that no group of people, including racial, ethnic, or socioeconomic groups, should bear a disproportionate share of the negative environmental consequences resulting from industrial, municipal, and commercial operations or the execution of federal, state, local, and tribal environmental programs and policies. Meaningful involvement means that (1) potentially affected community residents have an appropriate opportunity to participate in decisions about a proposed activity that will affect their environment and/or health; (2) the public’s contribution can influence the regulatory agency’s decision; (3) the concerns of all participants involved will be considered in the decision

- making process; and (4) the decision

- makers se ek out and facilitate the involvement of those potentially affected. The EPA has this goal for all communities and persons across the United States.

To help address potential environmental justice issues, during the 15

- day public participation comment
pe riod, the EPA sought information on any groups or segments of the population who, as a result their location, cultural practices, or other factors, may have atypical, unusually high exposure to Bacillus thuringiensis subspecies galleriae strain SDS

- 502 , co mpared to the general population. The EPA received no public comments on this matter.Bacillus thuringiensis subspecies galleriae strain SDS

- 502 Page 19 of 42 Biopesticides Registration Action Document

For additional information regarding environmental justice issues, please visit the EPA’s web site at http://www.epa.gov/compliance/environmentaljustice/index.html

.

V I

. RISK MANAGEMENT DECISION

Section 3(c)(5) of FIFRA permits for the registration of a pesticide provided that all the following determinations are made:

(1) Its composition is such as to warrant the proposed claims for it; (2) Its labeling and other material required to be submitted comply with the requirements of FIFRA; (3) It will perform its intended function without unreasonable adverse effects on the environ ment; AND (4) When used in accordance with widespread and commonly recognized practice, it will not generally cause unreasonable adverse effects on the environment.

To satisfy criterion 1, the EPA understands that Bacillus thuringiensis subs pecies galleriae strain SDS

502 is well

- known to have insecticidal capabilities

. The EPA has no knowledge that would contradict the claims made on the Phyllom SDS

- 502 MP, Phyllom grubGONE! G, and Phyllom beetleGONE! TM biological insecticide labels, and such pesticide products are not expected to cause unreasonable adverse effects on the environment when used according to the ir respective label instructions. Criterio n 2 is satisfied by the product labels, as well as the data and information presented in this document. It is believed that the Bacillus thuringiensis subspecies galleriae strain SDS

- 502 pesticide products will not cause any unreasonable adverse effects on the environment, and Phyllom grubGONE! G and Phyllom beetleGONE! TM biological insecticide ( end

- use pesticide product s ) , in particular, are likel y to control certain beetle pests as claimed, satisfying criterion 3. Criterion 4 is satisfied in that the Bacillus thuringiensis subspecies galleriae strain SDS

- 502 pesticide products are not expected to cause unreasonable adverse effects when used according to label instructions. Therefore, the EPA believes Phyllom SDS

- 502 MP, Phyllom grubGONE! G, and Phyllom beetleGONE! TM biological insecticide

, containing Bacillus thuringiensis subspecie s galleriae strain SDS

- 502 as a new active ingredient, are eligible for registration under FIFRA section 3(c)(5) for the labeled uses.

V I I

. ACTIONS REQUIRED OF THE REGISTRANT

A. Final Printed Labeling

Before releasing pesticide products containing Bacillus thuringiensis subspecies galleriae strain SDS

502 for shipment, the registrant is required to provide appropriate final printed labeling to the EPA

.Bacillus thuringiensis subspecies galleriae strain SDS

- 502 Page 20 of 42 Biopesticides Registration Action Document

B. Terms of Registration

The following items will be terms of the Phyllom SDS

- 502 MP (EPA File Symbol: 88347

- R) registration:

(1) Storage Stability (Harmonized Guideline 830.6317) : By June 6, 2014 , Phyllom, LLC, must provide the EPA with the results of a one

- year storage stability study on Phyllom SDS

- 502 MP

.
(2) Corrosion Characteristics (Harmonized Guideline 830.6320) : By June 6, 2014 , Phyllom, LLC, must provide the EPA with the results of a one

- year corrosion characteristics study on Phyllom SDS

- 502 MP

.
(3) pH (Harmonized Guideline 830.7000) : By December 6, 2013 , Phyllom, LLC, must provide the EPA wit h the results of a pH study on Phyllom SDS

- 502 MP

.
(4) Producers of Phyllom SDS

- 502 MP will conduct the following production quality control measures to detect contamination or changes in the parent strain:

• Spore preparations of Bacillus thuringiensis subsp. galleriae strain SDS

- 502 shall be produced by pure culture fermentation procedures with adequate quality control measures during production to detect any changes from the characteristics of the parent strain or contamination by other microorganisms.

• Each lot of spore preparation, prior to the addition of other materials, shall be tested by subcutaneous injection of at least 1 million spores into each of 5 laboratory mice weighing 17 grams to 23 grams. Such test shall show no evidence of infection or injur y in the test animals when observed for 7 days following injection.

• Spore preparations shall be free of the Bacillus thuringiensis β

- exotoxin when tested with the fly larvae toxicity test (“Microbial Control of Insects and Mites,” R.P.M. Bond et al ., p. 2 80 ff., 1971). This specification can be satisfied either by determining that each master seed lot brought into production is a Bacillus thuringiensis strain that does not produce β

- exotoxin under standardized manufacturing conditions or by periodically de termining that β

- exotoxin synthesized during spore production is eliminated by the subsequent spore

- harvesting procedure.

(5) Producers of Phyllom SDS

- 502 MP will maintain records of the quality control tests described directly above (in #4) for a period of at least five years from the date each batch is released for production.Bacillus thuringiensis subspecies galleriae strain SDS

- 502 Page 21 of 42 Biopesticides Registration Action Document

The following item s will be term s of the Phyllom grubGONE! G (EPA File Symbol: 88347

- E) registration :

(1) Storage Stability (Harmonized Guideline 830.6317) : By June 6, 2014 , Phyllom, LLC, must provide the EPA with the results of a one

- year storage stability study on Phyllom grubGONE! G.

(2) Corrosion Characteristics (Harmonized Guideline 830.6320) : By June 6, 2014 , Phyllom, LLC, must provide the EPA with the results of a one

- year corrosi on characteristics study on Phyllom grubGONE! G.

The following items will be terms of the Phyllom beetleGONE! TM biological insecticide (EPA File Symbol: 88347

- G) registration:

(1) Storage Stability (Harmonized Guideline 830.6317) : By July 11, 2014 , Phyllom, LLC, must provide the EPA with the results of a one

- year storage stability study on Phyllom beetleGONE! TM biological insecticide.

(2) Corrosion Characteristics (Harmonized Guideline 830.6320) : By July 11, 2014 , Phyllom, LLC, must provide the EPA with the results of a one

- year corrosion characteristics study on Phyllom beetleGONE! TM biological insecticide.

C. Reporting of Adverse Effects and Hypersensitivity Incidents

Notwithstanding the information stated in the previous sections, it should be clearly un derstood that certain specific data are required to be reported to the EPA as a requirement for maintaining the federal registration for a pesticide product. A brief summary of these types of data are described below.

Reports of all incidents of adverse e ffects to the environment must be submitted to the EPA under the provisions stated in FIFRA section 6(a)(2). Additionally, all incidents of hypersensitivity (including both suspected and confirmed incidents) must be reported to the EPA under the provisions of 40 C

. F

. R

. § 158.2140(d).Bacillus thuringiensis subspecies galleriae strain SDS

- 502 Page 22 of 42 Biopesticides Registration Action Document

VIII. GLOSSARY OF ACRONYMS AND ABBREVIATIONS

A /cm 2 acre per square centimeter a.i. active ingredient BRAD Biopesticides Registration Action Document C

. F

. R

. Code of Federal Regulations cfu colony

- forming units cfu/kg colony

- forming units per kilogram cfu/mL colony

- forming units per milliliter

cm centimeter EC environmental concentration EC 50 half maximal effective concentration EDSP Endocrine Disruptor Screening Program EP end

- use pesticide product EPA United States Envi ronmental Protection Agency EUP experimental use permit FFDCA Federal Food, Drug, and Cosmetic Act FIFRA Federal Insecticide, Fungicide, and Rodenticide Act FR Federal Register g/A grams per acre g/lb grams per pound g/m 2 grams per square meter lb/A pounds per acre lb/ft 3 pounds per cubic foot LC 50 median lethal concentration. A statistically derived concentration of a substance that can be expected to cause death in 50% of test animals. It is usually expressed as the weight of substance per

we ight or volume of water, air, or feed (e.g., mg/L, mg/kg, or ppm). LD 50 median lethal dose. A statistically derived single dose that can be expected to cause death in 50% of the test animals when administered by the route indicated (oral, dermal, or inhalation). It is expressed as a weight of substance per unit weight of animal (e.g., mg/kg). μg/μL micrograms per microliter μg/mL micrograms per milliliter mg/cm 2 milligrams per square centimeter mg/kg milligram s per kilogram mg/L milligrams per liter mg/mL milligrams per milliliter mL/L milliliters per liter

mm millimeter MP manufacturing

- use pesticide product MPCA microbial pest control agent MRID No. Master Record Identification Number
ng nanogram NIOSH National Institute for Occupational Safety and Health NOEC no observed effect concentration OPP Office of Pesticide Programs PC Code Pesticide Chemical Code PPE personal protective equipment
ppm parts per million QC quality control TGAI technical grade of the active ingredient U.S. United States USDA United States Department of Agriculture USFWS United States Fish and Wildlife ServiceBacillus thuringiensis subspecies galleriae strain SDS

- 502 Page 23 of 42 Biopesticides Registration Action Document

IX. BIBLIOGRAPHY

A. Studies Submitted to Support the Bacillus thuringiensis subspecies galleriae strain SD S

- 502 Pesticide Product Registrations

MRID No. Study Information

485532

- 01 Libs J, Rose R. 2010

. Phyllom SDS

- 502 MP, Product Identity and Characterization. Unpublished study pr epared by Phyllom, LLC, 10 pages.

485532

- 02 Libs J , Rose R . 2010

. Phyllom SDS

- 502 MP, Manufacturing Process

. Unpublished study prepared by Phyllom

, LLC, 6 pages.

485532

- 03 Libs J, Rose R. 2010. Phyllom SDS

- 502 MP, Discussion of Formation of Unintentional Ingredients. Unpublished study prepared by Phyllom, LLC, 6 pages.

485532

- 04 Libs J, Rose R. 2010

. Phyllom SDS

- 502 MP, Analysis of Samples

. Unpublished study prepared by Phyllom

, LLC, 14 pages.

485532

- 05 Libs J, Rose R. 2011. Phyllom SDS

- 502 MP, Certification of Limits. Unpublished study prepared by Phyllom, LLC, 2 pages.

485532

- 08 Munley S. 2011. Bacillus thuringiensis subsp. galleriae , Strain SDS

- 502: Acute Oral Toxicity/Pathogenicity Study in Rats. Project Number: DUPONT/20580 16248 1013. Unpublished study prepared by E.I. du Pont de Nemours and Company and Phyllom, LLC, 51 pages.

485532

- 09 Munley S. 2011. Bacillus thuringiensis subsp. galleriae , Strain SDS

- 502: Acute Pulmonary Toxicity/Pathogenicity Study in Rats. Project Number: DUPONT/20579 16248 1014. Unpublished study prepared by E.I. du Pont de Nemours and Company and Phyllom, LLC, 56 pages.

485532

- 10 Munley S. 2011. Bacillus thuringiensis subsp. galleriae , Strain SDS

- 502: Acute Intraperitoneal Injection Toxicity/Pathogenicity Study in Rats. Project Number: DUPONT/20581 16248 1015. Unpublished study prepared by E.I. du Pont de Nemours and Company and Phyllom, LLC, 49 pages.

485532

- 11 Libs J, Rose R. 2008. Phyllom SDS

- 502 MP, Spore Clearance. Unpublished study prepared by Phyllom, LLC, 27 pages.Bacillus thuringiensis subspecies galleriae strain SDS

- 502 Page 24 of 42 Biopesticides Registration Action Document

MRID No. Study Information

485532

- 12 Libs J, Rose R. 2010. Phyllom SDS

- 502 MP, Hypersensitivity Incidents. Unpublished study prepared by Phyllom, LLC, 2 pages.

485532

- 13 Libs J, Rose R. 2010. Phyllom SDS

- 502 MP, Cell Culture (Waiver Request). Unpublished study prepared by Phyllom, LLC, 2 pages.

485532

- 14 Libs J, Rose R. 2010. Phyllom SDS

- 502 MP, Acute Oral Toxicity (Waiver Request). Unpublished study prepared by Phyllom, LLC, 3 pages.

485532

- 15 Libs J, Rose R. 2010. Phyllom SDS

- 502 MP, Acute Dermal Toxicity (Waiver Request). Unpublished study prepared by Phyllom, LLC, 3 pages.

485532

- 17 Kuhn J. 2006. Acute Eye Irritation Study in Rabbits

– SDS

- 502 TGAI (Final Report). Project Number: 9645/06. Unpublished study prepared by Stillmeadow, Inc., 16 pages.

485532

- 18 Kuhn J. 2006. SDS

- 502 TGAI

- Acute Dermal Toxicity Study in Rabbits with a Microbial Pest Control Agent (MPCA) (Final Report). Project Number: 9644/06. Unpublished study prepared by Stillmeadow, Inc., 12 pages.

485532

- 19 Borchert J. 2006. Avian Oral Limit Toxicity Test with Bacillus thuringiensis subsp. galleriae strain SDS

- 502 in Mallard Ducks ( Anas platyrhynchos ) (Final Report). Project Number: 06001. Unpublished study prepared by Genesis Laboratories, Inc., 58 pages.

485532

- 20 Libs J, Rose R. 2010. Phyllom SDS

- 502 MP, Avian Inhalation Toxicity/Pathogenicity (Waiver Request). Unpublished study by Phyllom, LLC, 3 pages.

485532

- 21 Libs J, Rose R. 2010. Phyllom SDS

- 502 MP, Wild Mammal Toxicity/Pathogenicity (Waiver Request). Unpublished study prepared by Phyllom, LLC, 3 pages.

485532

- 22 Libs J, Rose R. 2010. Phyllom SDS

- 502 MP, Freshwater Fish Toxicity/Pathogenicity (Waiver Request). Unpublished study prepared by Phyllom, LLC, 3 pages.Bacillus thuringiensis subspecies galleriae strain SDS

- 502 Page 25 of 42 Biopesticides Registration Action Document

MRID No. Study Information

485532

- 23 Hartwell T. 2009. Bacillus thuringiensis subsp. galleriae Strain SDS

- 502

– Microbial Pest Control Agent (MPCA) Freshwater Aquatic Invertebrate Test with Daphnia magna (Final Report). Project Number: 13048/09. Unpublished study prepared by Stillmeadow, Inc., 27
pages.

485532

- 24 Libs J, Rose R. 2011. Phyllom SDS

- 502 MP, Estuarine/Marine Fish Testing and Estuarine/Marine Invertebrate Testing (Waiver Request). Unpublished study prepared by Phyllom, LLC, 3 pages.

485532

- 25 Libs J, Rose R. 2011. Phyllom SDS

- 502 MP, Nontarget Plant Testing (Waiver Request). Unpublished study prepared by Phyllom, LLC, 3 pages.

485532

- 26 Porch J, Krueger H. 2011. Bacillus thuringiensis subsp. galleriae Strain SDS

- 502 TGAI

– A Dietary Pathogenicity and Toxicity Study with Ladybird Beetle ( Hippodamia
convergens ) (Final Report) . Project Number: 702/101. Unpublished study prepared by Wildlife International, Ltd., 31 pages.

485532

- 27 Asano S, Yamashita C, Iizuka T, Takeuchi K, Yamanaka S, Cerf D, Yamamoto T. 2003. A Strain of Bacillus thuringiensis subsp. galleriae Containing a Novel Cry8 Gene Highly Toxic to Anomala cuprea (Coleoptera: Scarabaeidae). Biological Control 28:191

– 196.

485532

- 28 Libs J, Rose R. 2010. Phyllom SDS

- 502 MP, Bacillus thuringiensis subsp. galleriae

, Strain SDS

- 502 Toxicity to the Colorado Potato Beetle ( Leptinotarsa decemlineata ) (Coleoptera: Chrysomelidae), Acute Nontarget Insect Testing, Tier I. Project Number: PHY/SDS502/GRUB. Unpublished study prepared by the University of Maryland, 5
pages.

485532

- 29 Libs J. 2010. Phyllom SDS

- 502 MP, Bacillus thuringiensis subsp. galleriae , Strain SDS

502 Toxicity to the Spotted Cucumber Beetle/Southern Corn Rootworm ( Diabrotica undecimpunctata howardi ) (Coleoptera: Chrysomelidae), Acute Nontarget Insect Testing, Tier I. Unpublished study prepared by Phyllom, LLC, 4 pages.

485532

- 30 Richards R. 2006. Evaluation of Dietary Effects of SDS

- 502 Cry8Da on Larval Honeybee Development ( Apis mellifera L.). Project Number: CAR/152/05. Unpublished study prepared by California Agricultural Research, Inc., 37 pages.Bacillus thuringiensis subspecies galleriae strain SDS

- 502 Page 26 of 42 Biopesticides Registration Action Document

MRID No. Study Information

485698

- 01 Libs J, Rose R. 2011. Phyllom GrubGone EP, Product Identity and Characterization. Unpublished study prepared by Phyllom, LLC, 2 pages.

485698

- 02 Libs J, Rose R. 2011. Phyllom GrubGone EP, Manufacturing Process. Unpublished study prepared by Phyllom, LLC, 4 pages.

485698

- 03 Libs J, Rose R. 2011. Phyllom GrubGone EP, Discussion of Formation of Unintentional Ingredients. Unpublished study prepared by Phyllom, LLC, 4 pages.

485698

- 04 Libs J, Rose R. 2011. Phyllom GrubGone EP, Analysis of Samples. Unpublished study prepared by Phyllom, LLC, 11 pages.

485698

- 05 Libs J, Rose R. 2011. Phyllom GrubGone EP, Certification of Limits. Unpublished study prepared by Phyllom, LLC, 2 pages.

485698

- 08 Libs J, Rose R. 2011. Phyllom GrubGone EP, Hypersensitivity Incidents. Unpublished study prepared by Phyllom, LLC, 2 pages.

485698

- 09 Libs J, Rose R. 2011. Phyllom GrubGone EP, Acute Oral Toxicity (Waiver Request). Unpublished study prepared by Phyllom, LLC, 4 pages.

485698

- 10 Libs J, Rose R. 2011. Phyllom GrubGone EP, Acute Dermal Toxicity (Waiver Request). Unpublished study prepared by Phyllom, LLC, 4 pages.

485698

- 12 Kuhn J. 2006. SDS

- 502 End Use: Acute Eye Irritation Study in Rabbits (Final Report). Project Number: 9643/06. Unpublished study prepared by Stillmeadow, Inc., 16 pages.

485698

- 13 Kuhn J. 2006. SDS

- 502 End Use: Acute Dermal Toxicity Study in Rabbits with a Microbial Pest Control Agent (MPCA) (Final Report). Project Number: 9642/06. Unpublished study prepared by Stillmeadow, Inc., 12 pages.

487168

- 01 Libs J. 2012. Physical and Chemical Characteristics for Phyllom SDS

- 502 MP, Bacillus thuringiensis subsp. galleriae . Unpublished study prepared by Phyllom, LLC, 15 pages.Bacillus thuringiensis subspecies galleriae strain SDS

- 502 Page 27 of 42 Biopesticides Registration Action Document

MRID No. Study Information

487168

- 02 Libs J. 2012. Physical and Chemical Characteristics for Phyllom GrubGONE Granular (G). Unpublished study prepared by Phyllom, LLC, 15 pages.

487168

- 03 Bauer L, Londono D. 2012. Acute Nontarget Insect Testing, Tier I: Parasitic Hymenoptera Wasps: Tetrastichus planipennisi

, Oobius agrili

, Spathius agrili , and Atanycolus cappaerti . Unpublished study prepared by the U.S. Forest Service, 9 pages.

48 9401

- 01 Libs J. 2012 . Phyllom boreGONE! EP, Product Identity and Characterization. Unpublished s tudy prepared by Phyllom, LLC, 3 pages.

489401

- 02 Libs J. 2012. Phyllom boreGONE! Biological Insecticide, Manufacturing Process. Unpublished study prepared by Phyllom, LLC, 4 pages.

489401

- 03 Libs J. 2012. Phyllom boreGONE! Biological Insecticide, Discussion of Formation of Unintentional Ingredients. Unpublished study prepared by Phyllom, LLC, 3 pages.

489401

- 04 Libs J. 2012. Phyllom boreGONE! EP, Analysis of Samples. Unpublished study prepared by Phyllom, LLC, 10 pages.

489401

- 05 Libs J. 2012. Phyllom boreGONE! EP, Certification of Limits. Unpublished study prepared by Phyllom, LLC, 2 pages.

489401

- 06 Libs J. 2012. Physical and Chemical Properties for Phyllom boreGONE! Biological Insecticide Data Requirements. Unpublished study prepared by Phyllom, LLC, 12 pages.

489401

- 07 Rose R. 2012. Product

- Specific Data for the End

- Use Pesticide Product, Phyllom grubGONE! Granular (G), and the Manufacturing

- Use Pesticide Product, Phyllom SDS

502 MP. Unpublished study prepared by Robert I. Rose, 8 pages.

489401

- 08 Libs J. 2012. Phyll om Bacillus thuringiensis subsp./var. galleriae , Strain SDS

- 502, Primary Powder TGAI/MUP/MP

– Rationale/Clearance Demonstration of Lack of, or No Infectivity in Studies Conducted for Registration of Phyllom SDS

- 502 MP. Unpublished study prepared by Phyllom, LLC, 9 pages.Bacillus thuringiensis subspecies galleriae strain SDS

- 502 Page 28 of 42 Biopesticides Registration Action Document

MRID No. Study Information

489731

- 01 Doig A. 2012. Acute In halation Toxicity in Rats: Bacillus thuringiensis serovar galleriae SDS

- 502 (BTG SDS

- 502) TGAI Powder. Project Number: 16757/12. Unpublished study prepared by Stillmeadow, Inc., 17 pages.

489731

- 02 Libs J. 2012. Phyllom boreGONE! Biological Insecticide, Hypersensitivity Incidents. Unpublished study prepared by Phyllom, LLC, 3 pages.

489731

- 03 Libs J. 2012. Phyllom boreGONE! Biological Insecticide End

- Use Product, Acute Oral Toxicity. Unpublished study prepared by Phyllom, LLC, 3 pages.

489731

- 04 Libs J. 2012. Phyllom boreGONE! Biological Insecticide End

- Use Product, Acute Dermal Toxicity. Unpublished s tudy prepared by Phyllom, LLC, 4 pages.

489731

- 05 Libs J. 2012. Phyllom boreGONE! Biological Insecticide End

- Use Product, Acute Inhalation Toxicity. Unpublished study prepared by Phyllom, LLC, 4 pages.

489731

- 06 Libs J. 2012. Phyllom boreGONE! Biological Insecticide End

- Use Product, Acute Eye Irritation. Unpublished study p repared by Phyllom, LLC, 3 pages.

489731

- 07 Libs J. 2012. Phyllom boreGONE! Biological Insecticide End

- Use Product, Primary Dermal Irritation. Unpublished study prepared by Phyllom, LLC, 3 pages.

489731

- 08 Libs J. 2012. Phyllom grubGONE! Granular (G) End

- Use Product, Acute Inhalation Toxicity. Unpublished study prepared by Phyllom, LLC, 4 pages.

490940

- 01 Benzon G, Finkenbiner C. 2013. Phyllom Bacillus thuringiensis subsp. galleriae , Strain SDS

- 502 MP (TGAI) Primary Powder Beta

- Exotoxin Bioassay on Musca domestica (House Fly Larvae), Diptera: Muscidae. Project Number: PH13001, PH13002. Unpublished study prepared by Benzon Research, 14 pages.

490940

- 02 Libs J. 2013. Phyllom Manufacturing Process Quality Control Supplement to MRID No. 485532

- 02 for Phyllom’s Bacillus thuringiensis subsp. galleriae , Strain SDS

- 502, MP (TGAI). Unpublished study prepared by Phyllom LLC, 5 pages.Bacillus thuringiensis subspecies galleriae strain SDS

- 502 Page 29 of 42 Biopesticides Registration Action Document

MRID No. Study Information

490940

- 03 Doig A. 2013. Phyllom, LLC Bacillus thuringiensis subsp. galleriae , Strain SDS

- 502, MP (TGAI) Primary Powder MPCA Subcutaneous Injection Safety Test in Mice. Project Number: 17193/13. Unpublished study prepared by Stillmeadow, Inc., 16 pages.

490940

- 04 Matthews T. 2013

. Phyllom Bacillus thuringiensis subsp. galleriae , Strain SDS

- 502, MP (TGAI) Primary Colony Forming Units (CFU) and Culture Purity Quality Control (QC) Analyses

. Project Number: PHY/6/13/1. Unpublished study prepared by San Diego State Univ ersity, 12 pages.

490940

- 05 Matthews T. 2013. Phyllom Bacillus thuringiensis subsp. galleriae , Strain SDS

- 502, grubGONE! Granular (G) Colony Forming Units (CFU) and Culture Purity Quality Control (QC) Analyses. Project Number: PHY/6/13/3. Unpublished study prepared by San Diego State University, 9 pages.

490940

- 06 Matthews T. 2013. Phyllom Bacillus thuringiensis subsp. galleriae , Strain SDS

- 502, boreGONE!/beetleGONE! Colony Forming Units (CFU) and Culture Purity Quality Control (QC) Analyses. Project Number: PHY/6/13/2. Unpublished study prepared by San Diego State University, 13 pages.

B. Environmental Protection Agency Risk Assessment Memoranda

U.S. EPA. 201 3a

. Bacillus thuringiensis subsp

. galleriae strain SDS

- 502. Memorandum from J.V. Gagliardi , Ph.D. through J.L. Kough, Ph.D. to J. Kausch

, dated April 30 , 2013

.
U.S. EPA. 2013b. Evaluation of a new microbial pesticide, Phyllom boreGONE! Biological Insecticide (a.i.: Bacillus thuringiensis subspecies galleriae strain SDS

- 502) for use in experimental testing to control Emerald Ash Borer ( Agrilus planipennis ) (EPA File Symbol: 88347

- EUP

- R). Memorandum from G. Tomimatsu, Ph.D. through S. Borges to J. Kausch, dated May 7, 2013.

U.S. EPA. 2013c. Environmental risk assessment for the FIFRA section 3 registration of Bacillus thuringiensis subsp. galleriae strain SDS

- 502 (EPA File Symbols: 88347

- R, 88347

- E, and 88347

- G). Memorandum from G. Tomimatsu, Ph.D. through S. Borges to J. Kausch, dated May 14, 2013.

U.S. EPA. 2012 a

. Bacillus thuringiensis serotype galleria strain SDS

- 502. Memorandum from J.V. Gagliardi, Ph.D. through J.L. Kough, Ph.D. to J. Kausch, dated December 18, 2012.

C. Other References

Asano S, Yamashita C, Iizuka T, Takeuchi K, Yamanaka S, Cerf D, Yamamoto T. 2003. A strain of Bacillus thuringiensis subsp. galleriae containing a novel cry8 gene highly toxic to Anomala
cuprea (Coleoptera: Scarabaeidae). Biological Control 28:191

– 19 6 (also see MRID No. 485532

- 27).Bacillus thuringiensis subspecies galleriae strain SDS

- 502 Page 30 of 42 Biopesticides Registration Action Document

Bravo A. 2007. Mode of action of Bacillus thuringiensis Cry and Cyt toxins and their potential for insect control. Toxicon 49(4):423

– 435.

Chestukhina GG, Kostina LI, Zalunin IA, Revina LP, Mikhailova AL, Stepanov VM. 1994. Production of multiple delta

- endotoxins by Bacillus thuringiensis : delta

- endotoxins produced by strains of the subspecies galleriae and wuhanensis

. Canadian Journal of Microbiology 40:1026

– 1034.

Damgaard PH, Abdel

- Hameed A, Eilenberg J, Smits PH. 199 8 . Natural occurrence of Bacillus thuringiensis on grass foliage. World Journal of Microbiology & Technology 14:239

– 242.

Federici BA. 2005. Insecticidal bacteria: An overwhelming success for invertebrate pathology. Journal of Invertebrate Pathology 8 9(2005):30

– 38.

Gao M, Li R, Dai S, Wu Y, Yi D. 2008. Diversity of Bacillus thuringiensis strains from soils in China and their pesticidal activities. Biological Control 44(3):380

– 388.

Masson L, Erlandson M, Puzstai

- Carey M, Brousseau R, Juárez

- Pérez V, Frutos R. 1998. A holistic approach for determining the entomopathogenic potential of Bacillus thuringiensis strains. Applied and Environmental Microbiology 64(12):4782

– 4788.

Melin BE, Cozzi EM. 1990. Safety to nontarget invertebrates of Lepidopteran strains of Bacillus thuringiensis and their delta

- endotoxins. Pp. 149

– 167 In Laird M, Lacey LA, Davidson EWD, Eds., Safety of Microbial Insecticides . CRC Press, Boca Raton, Florida.

Mend elsohn M, Kough J, Vaituzis Z, Matthews K. 2003. Are Bt crops safe? Nature Biotechnology 21(9):1003

– 1009.

Mizuki E, Ichimatsu T, Hwang S

- H, Park YS, Saitoh H, Higuchi K, Ohba M. 1999. Ubiquity of Bacillus thuringiensis on phylloplanes or arboreous and herbaceous plants in Japan. Journal of Applied Microbiology 86:979

– 984.

Myasnik M, Manasherob R, Ben

- Dov E, Zaritsky A, Margalith Y, Barak Z. 2001. Comparative sensitivity to UV

- B radiation of two Bacillus thuringiensis subspecies and other Bacillus sp

. Current Microbiology 43:140

– 143.

Rose RI. 2013a. January 11, 2013 Letter from R.I. Rose, Ph.D., Authorized Agent for Phyllom, LLC , to the EPA (re: Culture Collection Certificate from the Japan International Patent Organism Depository, National Institute of Technology and Evaluation; Additional Product Identity Information on Bacillus thuringiensis subspecies galleriae strain SDS

- 502; and Revised Experimental Use Permit Description Forms).

Rose RI. 2013b. March 30, 2013 Le tter from R.I. Rose, Ph.D., Authorized Agent for Phyllom, LLC, to the EPA (re: Response to the Meeting Minutes/Action Items from the February 28, 2013 Meeting between Phyllom. LLC, and the EPA and the Deficiencies Described in a March 30, 2013 90

- day Pr eliminary Technical Screening Letter from the EPA).

USDA. 1994. Extension Service, National Agricultural Pesticide Impact Assessment Program.Bacillus thuringiensis subspecies galleriae strain SDS

- 502 Page 31 of 42 Biopesticides Registration Action Document

U.S. EPA. 2012 b . Chapter 7 of the Label Review Manual (Precautionary Statements). Available from http://www.epa.gov/oppfead1/labeling/lrm/chap

- 07.pdf

.
U.S. EPA. 1998. Reregistration Eligibility Decision for Bacillus thuringiensis (List D

– Case 0247). Available from http://www.epa.gov/oppsrrd1/REDs/0247.pdf

.

U.S. EPA. 1996. Microbial Pesticide Test Guidelines

– Background for Residue Analysis of Microbial Pest Control Agents (OPPTS 885.2000). Available from http://www.epa.gov/ocspp/pubs/frs/publications/Test_Guidelines/series885.htm

.
USFWS. 2013. Endangered Species. Available from http://www.fws.gov/endangered/

.
World Health Organization. 2011. Guidelines for Drinking

- water Quality (Fourth Edition). Available from http://whqlibdoc.who.int/publicati ons/2011/9789241548151_eng.pdf

.Bacillus thuringiensis subspecies galleriae strain SDS

- 502 Page 32 of 42 Biopesticides Registration Action Document

A PPENDIX A. MICROBIAL PESTICIDES DATA REQUIREMENTS (40 C

. F

. R

. PART 158

– SUBPART V )

TABLE 1. Product Analysis Data Requirements for the Technical Grade of the Active Ingredient (TGAI) , Bacillus thuringiensis subspecies galleriae strain SDS

- 502 , and Its Manufacturing

- Use Pesticide Product (MP) , Phyllom SDS

- 502 MP (40 C

. F

. R

. § 158.2 120 )
Harmonized

Guideline Number Data Requirement Results MRID No. or Other Reference TGAI MP

885.1100 Product Identity N/A Submitted data fulfill the requirement for product identity. Phyllom SDS

- 502 MP contains 9 0 .0% by weight Bacillus thuringiensis subspecies galleriae strain SDS

- 502 fermentation solids, spores, and insecticidal toxins (contains a minimum of 1 x 10 1 0 cfu per gram ). 485532

- 01 Rose (2013a) Rose (2013b)

885.1200 Manufacturing Process Submitted data fulfill the requirement for manufacturing process. As a term of the Phyllom SDS

- 502 MP registration, all producers of Phyllom SDS

502 MP will conduct certain production quality control measures to detect contamination or changes to the parent strain and will maintain records of these quality control tests for a period of at least five years from the date each batch is rel eased. 485532

- 02 490940

- 02 490940

- 03 Rose (2013b)

885.1250 Deposition of a Sample in a Nationally Recognized Culture Collection Bacillus thuringiensis subspecies galleriae strain SDS

- 502 is on deposit with the Japan International Patent Organism Depository, National Institute of Technology and Evaluation in Tsukuba

- shi, Japan under Accession Number FERM BP

7667. N/A 485532

- 01 Rose (2013a) Rose (2013b)

885.1300 Discussion of Formation of Unintentional Ingredients Submitted data fulfill the requirement for discussion of formation of unintentional ingredients. As a term of the Phyllom SDS

- 502 MP registration, all producers of Phyllom SDS

- 502 MP will conduct certain production quality control measures to detect contamination or changes to the p arent strain and will maintain records of these quality control tests for a period of at least five years from the date each batch is released. 485532

- 03 490940

- 02 490940

- 03 Rose (2013b)

885.1400 Analysis of Samples Submitted data fulfill the requirement for analysis of samples. 485532

- 04 490940

- 04

885.1500 Certification of Limits N/A Limits listed on the confidential statement of formula are adequate/acceptable. 485532

- 05 Rose (2013b)Bacillus thuringiensis subspecies galleriae strain SDS

- 502 Page 33 of 42 Biopesticides Registration Action Document

TABLE 2. Product Analysis Data Requirements for the End

- Use Pesticide Product s ( E P s ), Phyllom grubGONE! G and beetleGONE! TM biological insecticide (40 C

. F

. R

. § 158.2120)

Harmonized

Guideline Number Data Requirement Results MRID No. or Other Reference Phyllom grubGONE! G Phyllom beetleGONE! TM biological insecticide

885.1100 Product Identity Submitted data fulfill the requirement for product identity. Phyllom grubGONE ! G contains 9.0 % by

weight Bacillus thuringiensis subspecies galleriae strain SDS

- 502

fermentation solids, spores, and insecticidal toxins (contains a minimum of 1 x 10 9 cfu per gram). Submitted data fulfill the requirement for product identity. Phyllom beetleGONE! TM biological insecticide

contains 76.5% by weight Bacillus thuringiensis subspecies galleriae strain SDS

- 502 f ermentation solids, spores, and insecticidal toxins (contains a minimum of 0.85 x 10 10 cfu per gram). 485698

- 01 1
489401

- 01 2 Rose (2013b) 1,2

885.1200 Manufacturing Process Submitted data fulfill the requirement for manufacturing process. 485698

- 02 1
489401

- 02 2 Rose (2013b) 1,2

885.1250 Deposition of a Sample in a Nationally

Recognized Culture Collection N/A N/A

885.1300 Discussion of Formation of Unintentional

Ingredients Submitted data fulfill the requirement for discussion of formation of unintentional ingredients. 485698

- 03 1
489401

- 03 2 Rose (2013b) 1,2

885.1400 Analysis of Samples Submitted data fulfill the requirement for analysis of samples. 485698

- 04 1
490940

- 05 1
489401

- 04 2
490940

- 06 2

885.1500 Certification of Limits Limits listed on the confidential statement of formula are adequate/acceptable. 485698

- 05 1
489401

- 05 2 Rose (2013b) 1,2

1 Study supporting Phyllom grubGONE! G (EPA Reg. No. 88347

- 2)

. 2 Study supporting Phyllom beetleGONE! TM biological insecticide ( EPA Reg. No. 88347

- 3)

.Bacillus thuringiensis subspecies galleriae strain SDS

- 502 Page 34 of 42 Biopesticides Registration Action Document

TABLE 3. Physical and Chemical Characteristics for the Technical Grade of the Active Ingredient (TGAI), Bacillus thuringiensis subspecies galleriae strain SDS

- 502 , and Its Manufacturing

- Use Pesticide Product (MP), Phyllom SDS

- 502 MP (40 C.F.R. § 158.2120)

Harmonized

Guideline Number Data Requirement Results MRID No. or Other Reference TGAI MP

830.6302 Color Light brown, beige, or tan

N/A 487168

- 01

830.6303 Physical State Flour

- like powder 487168

- 01 Rose (2013b)

830.6304 Odor Mild earthy, slightly yeast

like 487168

- 01 830.6313 Stability to Normal and Elevated Temperatures, Metals, and Metal Ions Active ingredient is not expected to be stable at elevated temperatures.

830.6317 Storage Stability Study is in progress. As a term of the Phyllom SDS

- 502 MP registration, the EPA is requiring that the results of a one

year storage stability study be submitted by June 6, 2014

. 487168

- 01 Rose (2013b)

830.6319 Miscibility

N/A Soluble when mixed in water or oil but settles out over time. 487168

- 01

830.6320 Corrosion Characteristics Study is in progress. As a term of the Phyllom SDS

- 502 MP registration, the EPA is requiring that the results of a

one - year storage corrosion characteristics be submitted by June 6, 2014

. 487168

- 01 Rose (2013b)

830.7000 pH As a term of the Phyllom SDS

- 502 MP registration, the EPA is requiring the results of a pH study by December 6, 2013

. N/A

487168

- 01

830.7100 Viscosity N/A Not required because Phyllom SDS

- 502 MP is not a liquid form of microbial pesticide (refer to test note #5 of 40 C.F.R. § 158.2120(d)).

830.7300 Density/Relative Density/Bulk Density (Specific Gravity) 25

– 35 lb/ft 3 N/ABacillus thuringiensis subspecies galleriae strain SDS

- 502 Page 35 of 42 Biopesticides Registration Action Document

TABLE 4. Physical and Chemical Characteristics for the End

- Use Pesticide Product (EP s ), Phyllom grubGONE! G and Phyllom beetleGONE! TM biological insecticide (40 C

. F

. R

. § 158.2120)

Harmonized

Guideline Number Data Requirement Results MRID No. or Other Refe rence Phyllom grubGONE! G Phyllom beetleGONE! TM biological insecticide 830.6302 3 Color Tan or light brown 487168

- 02 1
489401

- 06 2 830.6303 3 Physical State Dry granules, ~ 1 mm in diameter Wettable powder, less than 100 mesh 830.6304 3 Odor Mild, earthy

830.6313 Stability to Normal and Elevated Temperatures, Metals, and Metal Ions N/A N/A

830.6317 Storage Stability Studies are in progress. As a term of the Phyllom grubGONE! G and Phyllom beetleGONE! TM biological insecticide registrations, the EPA is requiring that the results of one

- year storag e stability studies be submitted by June 6, 2014 (Phyllom grubGONE! G) or by July 11, 2014 (Phyllom beetleGONE! TM biological insecticide)

. 487168

- 02 1
489401

- 06 2 Rose (2013b)

830.6319 Miscibility When mixed with water, the EPs form fine

- particulate suspensions that settle out over time and easily re

distribute in the water column when agitated. 487168

- 02 1
489401

- 06 2

830.6320 Corrosion Characteristics Studies are in progress. As a term of the Phyllom grubGONE! G and Phyllom beetleGONE! TM biological insecticide registrations, the EPA is requiring that the results of one

- year corrosion characteristics studies be submitted by June 6, 2014 (Phyllom grubGONE! G) or by July 11, 2014 (Phyllom beetleGONE! TM biological insecticide)

. 487168

- 02 1
489401

- 06 2 Rose (2013b)

830.7000 pH N/A N/A

830.7100 Viscosity Not required because the EPs are not liquid forms of microbial pesticides (refer to test note #5 of 40 C.F.R. § 158.2120(d)). 487168

- 02 1
489401

- 06 2
830.7300 3 Density/Relative Density/Bulk Density

(Specific Gravity) 45 lb/ft 3 33.9 lb/ft 3

1 Study supporting Phyllom grubGONE! G (EPA Reg. No. 88347

- 2).
2 Study supporting Phyllom beetleGONE! TM biological insecticide ( EPA Reg. No. 88347

- 3).
3 According to 40 C.F.R. § 158.2120, these data are only required for the technical grade of the active ingredient. Since Phyllom, LLC

, included this information with its application for Phyllom grubGONE! G and/or Phyllom beetleGONE! TM biological insecticide , it is summarized appropriately in this table.Bacillus thuringiensis subspecies galleriae strain SDS

- 502 Page 36 of 42 Biopesticides Registration Action Document

T ABLE 5

. Toxicology Data Requirements for the Technical Grade of the Active Ingredient (TGAI), Bacillus thuringiensis subspecies galleriae strain SDS

- 502, and Its Manufacturing

- Use Pesticide Product (MP), Phyllom SDS

- 502 MP (40 C

. F

. R

. § 158.2140) Harmonized

Guideline N umber Data Requirement Results MRID N o. TGAI MP
Tier I

885.3050 Acute Oral Toxicity/Pathogenicity Not toxic, infective, and/or pathogenic to rats when administered by oral gavage in a single dose of 2.4 x 10 8 cfu per rat. Classification: Acceptable

N/A 485532

- 08 485532

- 11 489401

- 08

885.3150 Acute Pulmonary Toxicity/Pathogenicity No conclusions can be made regarding infectivity, pathogenicity or toxicity based on the study results. Attributing lethal effects to dosing was not followed up with alternate dosing methods. Replaced with MRID No. 489731

- 01 ( acute inhalation toxicity stud y conducted with the MP, see below)

. Classification: Un a cceptable 485532

- 09 485532

- 11 489401

- 08

885.3200 Acute Injection Toxicity/Pathogenicity Not toxic, infective, and/or pathogenic to rats when administered intraperitoneally in a single dose of 2.02 x 10 7 cfu per rat. Classification: Acceptable 485532

- 10 485532

- 11 489401

- 08

885.3400 Hypersensitivity Incidents The applicant reported that n o hypersensitivity incidents, including immediate

- type or delayed

- type reactions of humans and domestic animals

, occurred during research, development, or testing of the TGAI or MP . Any future hypersensitivity incidents must be reported to the EPA (refer to test note #3 of 40 C

. F

. R

. § 158.2140(d))

. 485532

- 12

885.3500 Cell Culture Not required because Bacillus thuringiensis subspecies galleriae strain SDS

- 502 is not a virus (refer to test note #4 of 40 C

. F

. R

. § 158.2140(d)). N/A 485532

- 13

870.1100 Acute Oral Toxicity N/A Testing waived based on the applicant’s scientific rationale, which cited to the results of MRID No. 485532

- 08 (acute oral toxicity/pathogenicity study conducted with the TGAI, see above in this table), and because this formulation contains inert ingredients that are not expected to be of toxicological concern through the oral route of exposure (refer to test note #5 of 40 C.F.R. § 158.2140(d)). Classification: Acceptable TOXICITY CATEGORY III 485532

- 14Bacillus thuringiensis subspecies galleriae strain SDS

- 502 Page 37 of 42 Biopesticides Registration Action Document

T ABLE 5

. Toxicology Data Requirements for the Technical Grade of the Active Ingredient (TGAI), Bacillus thuringiensis subspecies galleriae strain SDS

- 502, and Its Manufacturing

- Use Pesticide Product (MP), Phyllom SDS

- 502 MP (40 C

. F

. R

. § 158.2140) Harmonized

Guideline N umber Data Requirement Results MRID N o. TGAI MP

870.1200 Acute Dermal Toxicity N/A Testing waived based on the results of MRID No. 485532

- 18 (acute dermal toxicity and pathology study conducted with this MP, see below in this table), and because this formulation contains inert ingredients that are not expected to be of toxicological concern through the dermal route of exposure (refer to test note #5 of 40 C.F.R. § 158.2140(d)). Classification: Acceptable TOXICITY CATEGORY III 485532

- 15

870.1300 Acute Inhalation Toxicity Inhalation LC 50 > 4.30 mg/L Classification: Acceptable TOXICITY CATEGORY IV 489731

- 01

870.2400 Acute Eye Irritation

N/A Phyllom SDS

- 502 MP was mildly

- irritating to the eyes of rabbits. Classification: Acceptable TOXICITY CATEGORY I II 485532

- 17

870.2500 Primary Dermal Irritation Phyllom SDS

- 502 MP was essentially non

irritating to the skin of rabbits. All animals were free of very slight erythema or other skin

- related findings by day 7

. Classification: Acceptable TOXICITY CATEGORY I II 485532

- 18

Tiers II and III Not required for Bacillus thuringiensis subspecies galleriae strain SDS

- 502 based on the lack of acute toxicity/pathogenicity attributed to the bacterium in the Tier I studies.Bacillus thuringiensis subspecies galleriae strain SDS

- 502 Page 38 of 42 Biopesticides Registration Action Document

TABLE 6 . Toxicology Data Requirements for the End

- Use Pesticide Product s (EP s ) , Phyllom grubGONE! G and Phyllom beetleGONE! TM biological insecticide (40 C

. F

. R

. § 158.2140)

Harmonized

Guideline Number Data Requirement Results MRID No. or Other Reference Phyllom grubGONE! G Phyllom beetleGONE! TM biological insecticide

885.3050 Acute Oral Toxicity/Pathogenicity

N/A N/A 885.3150 Acute Pulmonary Toxicity/Pathogenicity

885.3200 Acute Injection Toxicity/Pathogenicity

885.3400 Hypersensitivity Incidents The applicant reported than n o hypersensitivity incidents, including immediate

type or delayed

- type reactions of humans and domestic animals

, occurred during research, development, or testing of the EPs

. Any future hypersensitivity incidents must be reported to the EPA (refer to test note #3 of 40 C

. F

. R

. § 158.2140(d)). 485698

- 08 1
489731

- 02 2

885.3500 Cell Culture N/A N/A

870.1100 Acute Oral Toxicity Testing waived based on the applicant’s scientific rationale, which cited to the results of MRID No. 485532

- 08 (acute oral toxicity/pathogenicity study conducted with the TGAI, see Table 5), and because this formulation contains inert ingredients that are not expected to be of toxicological concern through the oral route of exposure (refer to test note #5 of 40 C.F.R. § 158.2140(d)). Classification: Acceptable TOXICITY CATEGORY III 485698

- 09 1
489731

- 03 2
490940

- 01 1,2 Rose (2013b) 2

870.1200 Acute Dermal Toxicity Testing waived based on the results of MRID No. 485698

- 13 (acute dermal toxicity and pathology study conducted with this EP, see below in this table )

. Because the EPA classified the product

- specific acute eye irritation study done with this EP as Toxicity Category II, a Toxicity Category of no higher than III is to be used for labeling of dermal hazards. Classification: Acceptable TOXICITY CATEGORY III Testing waived based on the applicant’s scientific rationale, which cited to the results of MRID No. 485532

- 18 (acute dermal toxicity and pathology study conducted with the MP, see Table 5), and because this formulation contains inert ingredients that are not expected to be of toxicological concern through the dermal route of exposure (refer to test note #5 of 40 C.F.R. § 158.2140(d)). Classification: Acceptable TOXICITY CATEGORY I II 485698

- 10 1
489731

- 04 2
490940

- 01 1,2 Rose (2013b) 2

870.1300 Acute Inhalation Toxicity Testing waived based on the applicant’s scientific rationale, which cited to the results of MRID No. 489731

- 01 (acute inhalation toxicity study conducted with the MP, see Table 5). Because the EPA classified the product

- specific acute e ye irritation study done with this EP as Toxicity Category II, a Toxicity Category of no higher than III is to be used for labeling of inhalation hazards. Classification: Acceptable TOXICITY CATEGORY III Testing waived based on the applicant’s scientific rationale, which cited to the results of MRID No. 489731

- 01 (acute inhalation toxicity study conducted with the MP, see Table 5), and because this formulation contains inert ingredients that are not expected to be of toxicological concern through the inhal ation route of exposure (refer to test note #5 of 40 C.F.R. § 158.2140(d)). Classification: Acceptable TOXICITY CATEGORY IV 489731

- 05 2
489731

- 08 1
490940

- 01 1,2 Rose (2013b) 2Bacillus thuringiensis subspecies galleriae strain SDS

- 502 Page 39 of 42 Biopesticides Registration Action Document

TABLE 6 . Toxicology Data Requirements for the End

- Use Pesticide Product s (EP s ) , Phyllom grubGONE! G and Phyllom beetleGONE! TM biological insecticide (40 C

. F

. R

. § 158.2140)

Harmonized

Guideline Number Data Requirement Results MRID No. or Other Reference Phyllom grubGONE! G Phyllom beetleGONE! TM biological insecticide

870.2400 Acute Eye Irritation Phyllom grubGONE! G was severely irritating to the eyes of rabbits.

Classification: Acceptable TOXICITY CATEGORY II Testing waived based on the applicant’s scientific rationale, which cited to the results of MRID No. 485532

- 17 (acute eye irritation study conducted with the MP, see Table 5), and because this formulation contains inert ingredients that are not expected to be of toxicological concern through this route of exposure (refer to test note #5 of 40 C.F.R. § 158.2140(d)). Classification: Acceptable TOXICITY CATEGORY III 485698

- 12 1
489731

- 06 2
490940

- 01 2 Rose (2013b) 2

870.2500 Primary Dermal Irritation Phyllom grubGONE! G was essentially

non

- irritating to the skin of rabbits. All

animals were free of very slight to moderate erythema or other skin

related findings by day 7. Classification: Acceptable TOXICITY CATEGORY III Testing waived based on the applicant’s scientific rationale, which cited to the results of MRID No. 485532

- 18 (acute dermal toxicity and pathology study conducted with the MP, see Table 5), and because this formulation contains inert ingredients that are not expected to be of toxicological concern through this route of exposure (refer to test note #5 of 40 C.F.R. § 158.2140(d)). Classification: Acceptable TOXICITY CATEGORY I II 485698

- 13 1
489731

- 07 2
490940

- 01 2 Rose (2013b) 2

1 Study supporting Phyllom grubGONE! G (EPA Reg. No. 88347

- 2).
2 Study supporting Phyllom beetleGONE! TM biological insecticide ( EPA Reg. No. 8 8 347

- 3).Bacillus thuringiensis subspecies galleriae strain SDS

- 502 Page 40 of 42 Biopesticides Registration Action Document

TABLE 7 . Nontarget Organism Toxicity and Environmental Fate Data Requirements for the Technical Grade of the Active Ingredient (TGAI), Bacillus thuringiensis subspecies galleriae strain SDS

- 502 (40 C

. F

. R

. § 158.2150) Harmonized Guideline Number Data Requirement Results MRID No. or Other Reference Tier I

885.4050 Avian Oral Toxicity The maximum daily dose for Bacillus thuringiensis subspecies galleriae strain SDS

- 502 was determined as 3.0 x 10 11 cfu/kg body weight. Classification: Acceptable 485532

- 19 489401

- 07 Rose (2013b)

885.4100 Avian Inhalation Toxicity/Pathogenicity Scientific rationale was sufficient to satisfy the data requirement. Bacillus thuringiensis subspecies galleriae strain SDS

- 502 is not considered toxic or pathogenic to birds. Classification: Acceptable 485532

- 20

885.4150 Wild Mammal Toxicity/Pathogenicity Tests required by 40 C.F.R. § 158.2140 are adequate and appropriate for assessment of hazards to wild mammals. The oral LD 50 is greater than 2.4 x 10 8 cfu Bacillus thuringiensis subspecies galleriae strain SDS

- 502 per rat. Bacillus thuringiensis subspecies galleriae strain SDS

- 502 was not infective, pathogenic or toxic to laboratory rats. Classification: Acceptable 485532

- 08 485532

- 11 485532

- 21 489401

- 08

885.4200 Freshwater Fish Toxicity/Pathogenicity Scientific rationale was sufficient to satisfy the data requirement. Bacillus thuringiensis subspecies galleriae strain SDS

- 502 is not considered toxic or pathogenic to fish. Classification: Acceptable 485532

- 22

885.4240 Freshwater Invertebrate Toxicity/Pathogenicity A 21

- day study shows that the EC 50 for survival, reproduction, and growth of Daphnia magna is greater than 100 mg/L. The NOEC for survival is 100 mg/L (2.5 x 10 7 cfu/mL), and the NOEC for reproduction and growth is 50 mg/L ( 8.9 x 10 6 cfu/mL). Classification: Acceptable 485532

- 04 485532

- 23 489401

- 07 Rose (2013b)

885.4280 Estuarine/Marine Fish and Invertebrate Testing Scientific rationale was sufficient to satisfy the data requirement. Bacillus thuringiensis subspecies galleriae strain SDS

- 502 is not considered toxic or pathogenic to estuarine or marine organisms. Classification: Acceptable 485532

- 24

885.4300 Nontarget Plant Testing Scientific rationale was sufficient to determine that adverse effects are not expected in plants (terrestrial and aquatic) as a result of exposures to Bacillus thuringiensis subspecies galleriae strain SDS

- 502. Classification: Acceptable 485532

- 25

885.4340 Nontarget Insect Testing The 9

- day LC 50 and NOEC was greater than 1,700 ppm for adult ladybird beetles ( Hippodamia convergens ) exposed to artificial diets amended with Bacillus thuringiensis subspecies galleriae strain SDS

- 502 Cry8Da protein. Maximum hazard doses were confirmed. Classification: Acceptable 485532

- 04 485532

- 26 489401

- 07 Rose (2013b)Bacillus thuringiensis subspecies galleriae strain SDS

- 502 Page 41 of 42 Biopesticides Registration Action Document

TABLE 7 . Nontarget Organism Toxicity and Environmental Fate Data Requirements for the Technical Grade of the Active Ingredient (TGAI), Bacillus thuringiensis subspecies galleriae strain SDS

- 502 (40 C

. F

. R

. § 158.2150) Harmonized Guideline Number Data Requirement Results MRID No. or Other Reference

885.4340 Nontarget Insect Testing The 96

- hour LC 50 for Bacillus thuringiensis subspecies galleriae strain SDS

- 502 was greater than 500 μg Cry8Da protein/mL diet for first instars of Colorado p otato b eetle ( Leptinotarsa decemlineata ). The maximum hazard dose could not be confirmed. Classification: Supplemental 485532

- 28 489401

- 07 Rose (2013b)

885.4340 Nontarget Insect Testing No mortalities were observed in four species of adult parasitic wasps ( Tetrastichus planipennisi

, Oobius agrili

, Spathius agrili

, Atanycolus cappaerti , Order Hymenoptera), whereas 52% mortality was noted for the adult e merald a sh borer ( Agri lus planipennis ) when exposed to 0.4 μg Cry8Da/μL honey diet. Maximum hazard doses could not be confirmed. Classification: Supplemental 487168

- 03 489401

- 07 Rose (2013b)

885.4340 Nontarget Insect Testing Data and information provided on three lepidopteran insect species ( Bombyx mori

, Plutella xylostella , and Spodoptera litura ) are useful for demonstrating potential spectra of insecticidal activity. The test duration (2 to 5 days) may have been of insufficient length for insect species that showed no mortality ( Bombyx mori and Spodoptera litura ). Maximum hazard doses could not be confirmed. Classification: Supplemental 485532

- 27 489401

- 07 Rose (2013b)

885.4340 Nontarget Insect Testing No adverse effects were observed when larvae of Southern corn rootworm ( Diabrotica undecimpunctata howardi ) were exposed to reported concentrations of purified Cry8Da protein in artificial diets. There were no quantitative data or information to confirm the concentrations of the test material and reported resul ts. Maximum hazard doses could not be confirmed. Classification: Unacceptable 485532

- 29 489401

- 07 Rose (2013b)

885.4380 Honey Bee Testing The 20

- day NOEC for Bacillus thuringiensis subspecies galleriae strain SDS

- 502 was 500 ng Cry8Da protein/honeybee larval cell. There is uncertainty in the amount of Cry8Da protein that might remain in the environment after applications of the microbial pesticide. The study provides useful information for a honeybee la rval toxicity profile for the Cry8Da protein at the screening level. The maximum hazard dose could not be confirmed. Classification: Supplemental 485532

- 30 489401

- 07 Rose (2013b)

Tiers II, III, and IV Not required for Bacillus thuringiensis subspecies galleriae strain SDS

- 502 based on the acceptability of the scientific rationale and studies provided for Tier I

.Bacillus thuringiensis subspecies galleriae strain SDS

- 502 Page 42 of 42 Biopesticides Registration Action Document

APPENDIX B. PESTICIDE PRODUCTS

EPA Registration

Number Registration Name Percentage Active Ingredient Formulation Type Use Site(s) Method(s) of Application Application Rate (s) Target Pest

88347

- 1 Phyllom SDS

502 MP 90.0% Technical N/A

88347

- 2 Phyllom grubGONE! G 9.0% End Use

– Granule Turf and

o rnamentals Calibrated
g ranular

e quipment 42

– 55 ounces of product per 1,000 square feet of turf or

ornamental plants White grubs of certain beetles (e.g., Japanese beetle)

88347

- 3 Phyllom beetleGONE! TM

biological insecticide 76.5% End Use

– Water Dispersible

Powder Ornamentals,

s elect plants/ c rops for f ood and a nimal f eed u ses, and

p oultry
p remises /houses Ground or a erial

a pplication

e quipment Variable (see stamped, accepted label in docket 1 for or the Pesticide Product

Label System for specifics) Certain beetle pests, particularly those in the Families Buprestidae

,

Scarabaeidae , and

Curculionidae

1 Go to www.regulations.gov . Search for Docket Identification Number “EPA

- HQ

- OPP

- 2011

- 0862. ”