### BIOPESTICIDES REGISTRATION ACTION DOCUMENT



## Bacillus thuringiensis subspecies galleriae strain SDS-502

Pesticide Chemical (PC) Code: 006399

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

DRAFT – May 15, 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 OVERVIEW .............................................................................................. ....... 6

III. REGULATORY BACKGROUND .................................................................................................. .......... 7

A. Applications for Pesticide Product Registration ......................................................................... .. 7

B. Food Tolerance Exemption ................................................................................................ ............. 7

IV. RISK ASSESSMENT ......................................................................................................... ......................... 8

A. Product Analysis Assess ment (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 JUSTICE .................................................................................................... ........... 18

VI. RISK MANAGEMENT DECISION ................................................................................................ ....... 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 Produc t Registrations ....................................................................... ............. 23

B. Environmental Protection Agen cy Risk Assessment Memoranda ............................................ 29

C. Other References ........................................................................................................ .................... 29

APPENDIX A. MICROBIAL PESTIC IDES DATA REQUIREMENTS ........................................................ 32

APPENDIX B. PESTICIDE 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 a pplications for a manufacturing-use pesticide product, Phyllom SDS-502 MP (EPA File Symbo l: 88347-R), and an end-use pesticide product, Phyllom grubGONE! G (EPA File Symbol: 88347-E), to the U.S. Environmental Protection Agency (EPA) under section 3 of the Fede ral Insecticide, Fungicide, and Rodenticide Act (FIFRA). In November 2012, Phyllom, LLC, submitted another a pplication for an end-use pesticide product, Phyllom beetleGONE! TM biological insecticide (E PA 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 commonl y found in soil. It forms parasporal crystalline bodies, which are toxic fo r certain invertebrates, especially species of inse ct larvae belonging to the insect orders Coleoptera (beetles), Diptera (flies), and Lepidoptera (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 during his st udy of a bacterial disease of silkworms. Later, in 1915, a German scie ntist named Berliner found a similar bacillus killing the larvae of flour moths; he published a descrip tion of the bacterium and its properties, naming it Bacillus thuringiensis

. 1 In the 1930’s, French researchers recogni zed its potential for use in agriculture and developed 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, th ere 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 ag ainst 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 co leopteran insects, Phyllom, LLC, proposed to register a manufacturing-use pesticid e product and two end-use pesticide products that are intended to be applied to turf, ornamentals, and food crops, as well as in poultry pr emises, to control white grubs of

beetles and adult beetles.

The EPA’s scientists 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, an d 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 The applicant fulfilled most of the product analysis data requirements for Bacillus thuringiensis subspecies galleriae strain SDS-502, including product chemistr y and composition, analysis of samples, and physical and chemical characteristics, by acceptabl e guideline studies. As term s of registration, the EPA will require the applicant to submit storage st ability, corrosion characteristics, and pH studies.

Toxicology The applicant submitted adequate mammalian toxicology data and information to support the Bacillus thuringiensis subspecies galleriae strain SDS-502 pesticide produc ts. Acute oral and injection toxicity/pathogenicity studies and an acute inhalation toxicity study showed that, at a single high dose, Bacillus thuringiensis subspecies galleriae strain SDS-502 is not toxic, in fective, and/or pathogenic via these routes of exposure. Moreover, the applican t reported that no hypersensitivity incidents occurred during research, development, or testing of Bacillus thuringiensis subspecies galleriae strain SDS-502. In light of the results of the acu te toxicity/pathogenicity data a nd 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 diet ary risks are expected from use of Bacillus thuringiensis subspecies galleriae strain SDS-502 as an active i ngredient in pesticide products. Although exposure to Bacillus thuringiensis subspecies galleriae strain SDS-502 through consumed food and drinking water may be possible, the EPA antic ipates these types of exposures not to be of concern because of suppor ting toxicological data. Bacillus thuringiensis subspecies galleriae strain SDS-502 currently meets the criteria of 40 C.F.R. § 180.1011.

Occupational Exposure Despite the low toxicological profile of Bacillus thuringiensis subspecies galleriae strain SDS-502, the EPA is requiring baseline persona l protective equipment (PPE) fo r handlers who, due to their occupation, may be exposed to th e active ingredient fo r 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, waterproof gloves, protective eyewear and/or a dust/mist filt ering 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., ch emical-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 a nd for risk assessment purposes. Further testing of nontarget organisms at higher tier leve ls (i.e., Tiers II, III, and IV) is not required. The EPA performed an environmental risk assessment, based on informa tion provided by the applicant, and determined that no effects are anticipated for mo st nontarget species exposed to Bacillus thuringiensis subspecies galleriae strain SDS-502 as a result of th e proposed labeled applications.Bacillus thuringiensis subspecies galleriae strain SDS-502 Page 6 of 42 Biopesticides Registration Action Document

Public Participation On October 1, 2009, the EPA announced a policy to pr ovide a more meaningful opportunity for the public to participate in major regist ration decisions before they occur. According to this policy, the EPA provides a public comment period prior to making a registration decision fo r the following types of applications: new active ingredients; first food uses; fi rst outdoor uses; first resi dential uses; or any other registration actions for which the EPA believe s there may be signifi cant public interest.

Consistent with the policy of making registration actions more tr ansparent, the proposed pesticide products containing Bacillus thuringiensis subspecies galleriae strain SDS-502, a new active ingredient, will be subject to a comment period. In addition to c ontaining a new active ingred ient, the registration of these pesticide products would result in the first food, outdoor, and residential uses for Bacillus thuringiensis subspecies galleriae strain SDS-502. The docket identification number, which is associated with these registration ac tions and can be accessed through either http://www.regulations.gov

or http://www.epa.gov/pesticides/regu lating/registration-status.html , is EPA-HQ-OPP-2011-0862. The following documents are available for co mment in EPA-HQ-OPP-2011-0862: (1) a draft Bacillus thuringiensis subspecies galleriae strain SDS-502 Biopesticides Registration Action Document (BRAD); (2) an environmental risk assessment for Bacillus thuringiensis subspecies galleriae strain SDS-502; (3) a draft label for the manufacturing-use pesticide product, Phyllom SDS-502 MP (EPA File Symbol: 88347-R); (4) a draft label for the end-use pesticide product, Phyllom grubGONE! G (EPA File Symbol: 88347-E); and (5) a draft label for the end-use pesticide product, Phyllom beetleGONE! TM
biological insecticide (EPA File Symbol: 88347-G). While a final decisi on on registration is contingent upon review and consideration of public comments, th e EPA presently believes that, based upon the risk assessment and information submitted in support of Phyllom SDS-502 MP, Phyllom grubGONE! G, and Phyllom beetleGONE! TM biological insecticide, it is appropriate to issue these registrations. The basis for this preliminary decision can be found in the risk assessment for Bacillus thuringiensis subspecies galleriae strain SDS-502, which is characte rized throughout this 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: 006399

Type of Pesticide: Microbial Pesticide – Insecticide

See Appendix B for specific information (e.g., use sites, application rates, methods of application, formulation types, and ta rget pests) regarding the proposed pesticide pr oducts containing this active ingredient.Bacillus thuringiensis subspecies galleriae strain SDS-502 Page 7 of 42 Biopesticides Registration Action Document

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 applica tions to register a manufacturi ng-use pesticide product, Phyllom SDS-502 MP (EPA File Symbol: 88347-R), and an e nd-use pesticide product, Phyllom grubGONE! G (EPA File Symbol: 88347-E), under FIFR A section 3. On February 29, 2012 ( 77 FR 12295 ), the EPA announced receipt of these applications to register pesticide products containing a new active ingredient and opened a 30-day public comment period pursuant to the provisions of FIFR A section 3(c)(4). No comments were received following this publication.

In November 2012, Phyllom, LLC, submitted an applic ation to register an end-use pesticide product, Phyllom beetleGONE! TM (EPA File Symbol 88347-G) under FIFR A section 3. On April 19, 2013, the EPA announced receipt of this application to re gister a pesticide produc t containing a new active ingredient ( 78 FR 23558 ) and opened a 30-day public comment pe riod pursuant to the provisions of FIFRA section 3(c)(4). No comments were received fo llowing this publication.

B. Food Tolerance Exemption

An exemption from the requirement of a tolerance is established for viable spor es of the microorganism Bacillus thuringiensis Berliner in or on honey and honeycom b 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, pr ovided the specifications set fo rth 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 conforming to the morphological and biochemi cal characteristics of Bacillus thuringiensis as described in Bergey's

Manual of Determinative B acteriology, Eighth Edition.

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

(3) Each lot of spore prepara tion, prior to the addition of other materi als, 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 infecti on or injury in the test animals when observed for 7 days following injection.

(4) Spore preparations shall be free of the Bacillus thuringiensis β -exotoxin when tested with the fly larvae toxicity test (“Microbial Contro l of Insects and Mites,” R.P.M. Bond et al ., p. 280 ff., 1971). This specification can be satisf ied either by 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 pe riodically determining that β -exotoxin synthesized during spore production is eliminated by the subse quent spore-harvesting procedure.Bacillus thuringiensis subspecies galleriae strain SDS-502 Page 8 of 42 Biopesticides Registration Action Document

The EPA determined that Bacillus thuringiensis subspecies galleriae strain SDS-502 currently meets the criteria of 40 C.F.R. § 180.1011. To ensure that pr oducers 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 exem ption criteria, the EPA will also be integrating these quality cont rol measures into the text of the Phyllom SDS-502 MP registration

notice as terms.

IV. RISK ASSESSMENT

In the Federal Registers of October 26, 2007 ( 72 FR 60988 ) and August 30, 2012 ( 77 FR 52610 ), the EPA issued Final Rules on the data requirements to support registration of mi crobial pesticides and updated the definition for microbial pesticides. The da ta and information evaluated for this BRAD were considered in light of these requirements.

The classifications that are found for each data subm ission are assigned by the EPA’s science reviewers

and are an indication of the usef ulness of the information contai ned in the documents for risk assessment. A rating of “acceptable” indicates the study is scientifica lly sound and is useful for risk assessment. A “supplemental” rating i ndicates the data provide some info rmation that can be useful for risk assessment. The studies may have certain asp ects determined not to be scientifically acceptable (“supplemental: upgradeable”). If a study is rate d as “supplemental: upgradeable,” the EPA always provides an indication of what is l acking or what can be provided to change the rating to “acceptable.” If there is simply a “supplemental” rating, the reviewer will often state that the study is not required by 40 C.F.R. Part 158. Both “acceptable” and “supplemental” studies may be used in the risk assessment process as appropriate. An “unacceptable” ratin g indicates that new data must be submitted.

For the acute toxicity data requirements, T oxicity Categories are assigned based on the hazard(s) identified from studies and/or other inform ation submitted to the EPA in support of a pesticide registration. The active ingredient or particular product is clas sified into Toxicity Category I, II, III, or IV, where Toxicity Category I indi cates the highest toxici ty and Toxicity Cate gory IV indicates the lowest toxicity (see section II of U.S. EPA (2012b)).

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, the applicant fulfilled all product analysis data requirements for Bacillus thuringiensis subspecies galleriae strain SDS-502. Refer to Tables 1, 2, 3, and 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 all toxicology data requirements for Bacillus thuringiensis subspecies galleriae

strain SDS-502. Acceptable Tier I mammalian toxicology data and information support registration of

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.Bacillus thuringiensis subspecies galleriae strain SDS-502 Page 9 of 42 Biopesticides Registration Action Document

For a summary of the generic toxicology data requ irements described 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 (H armonized Guideline 885.3050; Master Record Identification Numbers (MRID Nos.) 485532-08, 485532-11, and 489401-08) : In an acute oral toxicity and pathogenicity study, groups of fast ed, 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 p hosphate 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 sacrifices on days 3, 7, and 14. Three ma les and three females we re treated with heatinactivated Bacillus thuringiensis subspecies galleriae strain SDS-502 as inactivat ed controls, two males and two females were untreated shelf controls, and two males and two 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 deat hs, clinical signs, necropsy findi ngs, 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 organi sm was recovered from the cecum contents or any organ or blood from treated an imals sacrificed on day 21. The EPA rated this study acceptable.

Acute Pulmonary Toxicity/Pathogenicity – Rat (Harmonized Guideline 885.3150; MRID Nos. 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 phosphate 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 initially mis-dosed at approximately 0.2 x 10 8 cfu per animal, and three male and three female activetreated animals were killed (on schedule) before the mis-dosing was recognized. The following day,

three male inactive controls were treated w ith a dose approximately equivalent to 2 x 10 8 cfu per animal with immediate mortality. The remaining animals sc heduled 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 inac tive-treated 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 formulation of the inactive control, 2 of 3 males and 3 of 5 females treated with the further diluted and homogenized inactive suspensi on were found dead on days 0–1, and an additional female in this group was sacrificed in extremis on day 0. One female treate d with the active suspension was found dead on day 1 after exhibiting lethargy, 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 clini cal signs among the surviving active-treated animals included inappetence, ocular and/or nasal discharge, lethargy, noisy breathing, and staining of the fur and skin. The study author attributed these signs to the ph ysical properties of the dosing formulations. Body weight losses were noted for the active-treated animals of both sexes killed on day 3, and the activetreated males had decreased body weight gain dur ing days 0–7. Abnormal gross necropsy findings among animals killed on schedule were limited to ta n, mottled discoloration of the lungs in the three males and three females that were killed on day 7. Av erage cfu counts decreased over time in all of the evaluated tissues or samples, but the test organisms did remain pres ent through day 21 with the highestBacillus thuringiensis subspecies galleriae strain SDS-502 Page 10 of 42 Biopesticides Registration Action Document

counts seen in lung tissue (126 cfu per gram). No conclusions can be made regarding infectivity, pathogenicity, or toxicity based on these study results. Attributing lethal effects to dosing was not followed up with alternate dosing methods. The EPA ra ted this study unacceptabl e and concluded that a replacement pulmonary toxicity/pat hogenicity study or an inhalati on toxicity study was required to address pulmonary effects. (See review su mmary for MRID No. 489731-01, directly below.)

Acute Inhalation Toxicity – Rat (Harm onized Guideline 870.1300; MRID No. 489731-01) : All animals survived and gained weight with exception of 1 ma le that lost 7 grams after day 7. Symptoms noted between 4.5 and 6 hours after exposure included red na sal discharge, red crust around eyes, decreased activity and piloerection noted as slight 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. Th e inhalation median lethal concentration (LC 50 ) was greater than 4.30 mg/L, and the EP A rated this study acceptable.

Acute Injection Toxicity/Pathogeni city (Intraperitoneal) – Rat (H armonized Guideline 885.3200; MRID Nos. 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 per animal and observed for up to 21 days with interim s acrifices of 3 animals per sex done on days 0, 3, 7, and 14. Two additional groups of two males and two female s were either left untre ated or treated with a suspension of the heat-in activated test material. Bacillus thuringiensis subspecies galleriae strain SDS502 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 ga in was not significantly affected. The test organism was recovered from cecum contents, organ and blood samples from ac tive-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 (Harmoni zed Guideline 885.3400; MRID No. 485532-12) : The applicant reported that no hypersensitivity incidents, including immediate-type or delayed-type reactions of humans and domestic animals, occurred dur ing 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 viru s (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/Pathogeni city 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/pat hogenicity in the Tier I st udies attributed to the bacterium. Any uncertainties about the to xicity, pathogenicity, or infectivity of Bacillus thuringiensis subspecies galleriae strain SDS-502 raised in the acute pu lmonary toxicity/pathogenicity study (MRID No. 485532-09) were addressed by a subsequent acute inhalation toxi city study (MRID No. 489731-01 ) and information to support lack of infectivity of Bacillus thuringiensis subspecies galleriae strain SDS502 (MRID No. 489401-08).Bacillus thuringiensis subspecies galleriae strain SDS-502 Page 11 of 42 Biopesticides Registration Action Document

c. Endocrine Disruptors

As required by the Administrator under FFDCA sec tion 408(p), the EPA has developed the Endocrine Disruptor 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 wildlife si milar to an effect produced by a “naturally occurring estrogen, or ot her such endocrine effects as th e 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 occurr ing estrogenic substance.

Between October 2009 and February 2010, the EPA issued test orders/data call-ins for the first group of 67 chemicals, which contains 58 pesticide act ive ingredients and 9 inert ingredients. Bacillus thuringiensis subspecies galleriae strain SDS-502 is not among the group of 58 pesticide active ingredients on the initi al 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 would not produce any effect in humans similar to an effect produced by a naturally occurring estrog enic substance. As su ch, pursuant to Section 408(p)(4), the EPA will determine in the future whether it can exempt Bacillus thuringiensis subspecies galleriae strain 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, th e policies and procedures, th e list of 67 chemicals, the test guidelines and the Tier 1 screen ing battery, please visit our website 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 pest icide chemical residue in or on a food) only if the EPA determines that the exemption is “safe.” Sectio n 408(c)(2)(A)(ii) of FFDCA defines “safe” to mean that “there is a reasonable certainty that no harm will result from a ggregate exposure to the pesticide chemical residue, including all anticipated dietar y exposures and all other exposur es for which there is reliable information.” This includes exposure through drinki ng water and in residential settings but does not include occupational exposure. Pursuant to FFDCA sec tion 408(b)(2)(B), in establishing or maintaining in effect an exemption from the requirement of a tole rance, 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 chem ical residue in establis hing 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 re sidue....” Additionally, FFDCA section 408(b)(2)(D) requires that 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.”Bacillus thuringiensis subspecies galleriae strain SDS-502 Page 12 of 42 Biopesticides Registration Action Document

The EPA performs a number of analyses to determin e 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 ava ilable 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 th is information to human risk. The

EPA also considered available information concerni ng the variability of the sensitivities of major identifiable subgroups of consumers, including infants and children.

Based on the acute toxicity/pathogeni city data and information discu ssed previously and presented in

Tables 5 and 6 in Appendix A , the 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 di rects the EPA to consider available information concerning exposures from the pest icide residue in food and all other non-occupational exposures, including drinking water from ground water or su rface water and exposure through pesticide use in gardens, lawns, or buildings (re sidential and othe r indoor uses).

Food Exposure and Risk Characterization : Exposure to residues of Bacillus thuringiensis subspecies galleriae strain SDS-502 in food may occur through the consumption of food commodities that have been treated with microbial pestic ides containing this bacterium. Th e 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 individu als are already exposed in the natural environment— Bacillus thuringiensis is commonly found in soil and on plants (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 certai n food commodities, processing procedures (e.g., peeling and washing) may reduce such resi dues (U.S. EPA 1996). Regardless 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 in fectivity is likely to occur with any food exposure resulting from the use of Bacillus thuringiensis subspecies galleriae strain SDS-502 as a pesticide when manufactured to meet the spec ifications of 40 C.F.R. § 180.1011 and applied in

accordance with label directions a nd good agricultural practices (see se ction IV(B)(1)(a) and Tables 5

and 6 in Appendix A ).

Drinking Water Exposure and Risk Characterization : Exposure to residues of Bacillus thuringiensis subspecies galleriae strain SDS-502, a naturally occurring bacterium (Damgaard 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 different 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 wastewat er treatment systems and drinking f acilities (e.g., pH adjustments and chlorination). According to the World Health Organization, Bacillus species, such as thuringiensis , are often detected in drinking water even after going through acceptable wa ter treatment processes, largelyBacillus thuringiensis subspecies galleriae strain SDS-502 Page 13 of 42 Biopesticides Registration Action Document

because the spores are resistant to these disinf ection 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 strain SDS-502 as a pesticide when manufactured to meet the speci fications of 40 C.F.R. § 180.1011 and applied in accordance with label di rections and good agricu ltural practices (see s ection IV(B)(1)(a) and

Tables 5 and 6 in Appendix A ).

Non-occupational, Residentia l 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 occu rring. Additional exposure to Bacillus thuringiensis subspecies galleriae strain SDS-502 may occur because the end-us e pesticide products are labeled for application to sites like athletic fields, residential lawn s, and playgrounds. Even if additio nal non-occupational exposures were to occur, no adverse e ffects attributed to Bacillus thuringiensis subspecies galleriae strain SDS-502 are anticipated, particularly in light of testing results that indicated this ba cterium is not toxic, pathogenic, or infective and is essentially non-irr itating to skin at high doses (see se ction 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 re quires that, when considering wh ether to establish, modify, or revoke a tolerance, the EPA consider “available info rmation concerning the cumulative effects of [a particular pesticide’s] . . . resi dues 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, and Bacillus thuringiensis subspecies galleriae strain SDS-502 does not appear to produce a toxic metabolit e produced by other substances. For the purposes of this tolerance action, therefore, the EPA has assumed that Bacillus thuringiensis subspecies galleriae

strain SDS-502 does not have a comm on mechanism of toxicity with other substances. Following from this, therefore, the EPA concludes that ther e are no cumulative effects associated with Bacillus thuringiensis subspecies galleriae strain SDS-502 that need to be considered. For information regarding

the EPA’s efforts to determine chemicals that have a common mechanism of toxic ity 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 Un ited States Population, Infants and Children

FFDCA section 408(b)(2)(C) pr ovides that, in considering the estab lishment of a tolera nce or tolerance exemption for a pesticide chemical residue, the EPA shall assess the available information about consumption patterns among infants and children, speci al susceptibility of infants and children to pesticide chemical residues, and the cumulative effect s on infants and children of the residues and other

substances with a common mechanism of toxicit y. In addition, FFDCA sect ion 408(b)(2)(C) provides that the EPA shall apply an additiona l tenfold (10X) margin of safety for infants and children in the case of threshold effects to account for prenatal and post natal toxicity and the comp leteness of the database on toxicity and exposure, unless the EPA determines that a different margin of safety will be safe for infants and children. This additional margin of safe ty is commonly referred to as the Food Quality Protection Act Safety Factor. In a pplying this provision, the EPA either retains the default value of 10X,Bacillus thuringiensis subspecies galleriae strain SDS-502 Page 14 of 42 Biopesticides Registration Action Document

or 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 pa thogenicity data/information disc ussed in section IV(B)(1)(a) and

Tables 5 and 6 in 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 EP A concludes that no additional margin of exposure (safety) is necessary.

Moreover, based on the same data and analysis as pres ented directly above, the EPA is able to conclude 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 acco rding to the criteria set forth in 40 C.F.R. § 180.1011 and used as labeled as an insecticide. Such exposure include s all anticipated dietary exposures and all other exposures for which there is reliable informati on. 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, pat hogenic, 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 requiri ng appropriate personal protective equipment and precautionary statements on pesticide product labels to mitigate any potential risk s 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, waterpr oof 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 th e standard described above, on a product-specific basis.

4. Human Health Risk Characterization

The EPA considered human exposure to Bacillus thuringiensis subspecies galleriae strain SDS-502 in light of the standard for regi stration in FIFRA and the releva nt safety factors in FFDCA. A

determination has been made that no unreasonable adve rse effects to the U.S. population in general, and to infants and children in particular, will result from incidental exposures when Bacillus thuringiensis subspecies galleriae strain SDS-502 pesticide products are applie d to turf, to ornamentals, to various crops, and in poultry premises in accord ance 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 a nd for risk assessment purposes. Further testing of nontarget organisms at higher tier leve ls (i.e., Tiers II, III, and IV) is not required. The EPA performed an environmental risk assessment, based on informa tion provided by the applicant, and determined that no effects are anticipated for mo st nontarget species exposed to Bacillus thuringiensis subspecies galleriae strain SDS-502 as a result of th e proposed labeled applications.

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

.

1. Ecological Exposure and Ri sk Characterization

The end-use pesticide product (EP), Phyllom beetleGONE! TM biological insecticide, is proposed for application to many types of grain, vegetable an d 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 incl ude landscapes, interiors capes, nurseries, fields and greenhouses. Since the proposed applications are expected to result in contac t of the active ingredient (a.i.) with foliar surfaces and soil, and spray drift is expected to occur, exposure to terrestrial and aquatic nontarget orga nisms is possible.

Based on current information concer ning the amount of active ingredient 2 in the EP, Phyllom

beetleGONE! TM biological insecticide, the EPA estimat es terrestrial maximum environmental concentrations (EC) of Bacillus thuringiensis subspecies galleriae strain SDS-502 as approximately 6,078 g/A. An aquatic maximum EC was estimat ed 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 th e 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 w ith laboratory 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 Nos. 485532-11 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 with respect to effects on plants and is not taxonomically related to known plant pathogens. Th erefore, adverse effects on nontarget plants (terrestrial and aquatic) are not expected to result from the proposed applications of Bacillus thuringiensis subspecies galleriae

strain SDS-502.

c. Nontarget Insects (Harmonized Gu ideline 885.4340) and Honey Bees (Harmonized Guideline 885.4380) (MRID Nos. 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) provide d limited information regarding pot ential dietary to xicity of the Cry8Da protein, extracte d and purified from Bacillus thuringiensis subspecies galleriae strain SDS-502 (Table 7 in Appendix A ). These studies may have been of insu fficient duration to note potential activity by an insecticide that must be ingested by susceptibl e insect larvae for toxic effects. Insect bioassays may be influenced by relative proportions of the differe nt proteins within the crystal, thereby obscuring the role an individual toxin may pl ay in insect toxicity. Variability in the composition and potency of crystals within the same strain ma y be caused by factors such as mob ile genetic elements and nutritional requirements (Masson et al . 1998). At least seven different de lta-endotoxins were identified in Bacillus thuringiensis subspecies galleriae (Chestukhina et al . 1994).

It is not possible to determine whether thes e 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 me thods and did not report the per centage of the Cry8Da protein.

The EPA determined that the itemized studies below (studies b through f) are supplemental; however, all

studies are useful 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 effects ar e anticipated for populations of Hippodamia 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 pota to beetle).

e. No Cry8Da toxicity to 4 species of adult parasi tic Hymenoptera.

f. High toxicity of purified Cry8Da from Bacillus thuringiensis subspecies galleriae strain SDS502 to the Emerald Ash Borer ( Agrilus planipennis or Agrilus marcopoli ) and to Scarabaeidae beetle “white grubs” ( Anomala cuprea ) was demonstrated in another submitted study.Bacillus thuringiensis subspecies galleriae strain SDS-502 Page 17 of 42 Biopesticides Registration Action Document

Asano et al . (2003) noted that other species of the Scarabaeidae (e.g., cupreous chafer, Anomala cuprea ; Japanese beetle, Popillia japonica Newman) are affected by othe r strains or subspecies of Bacillus thuringiensis (e.g., Bacillus thuringiensis subspecies galleriae NRRL B-4027 and Bacillus thuringiensis subspecies japonensis ) and other Cry8 genes were active against Cotinis spp. (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 spp.) in honeybee hives (Me lin and Cozzi 1990).

Based on these studies and the information presented a bove, effects to some insect species in the orders Coleoptera and Lepidopt era as a result of Bacillus thuringiensis subspecies galleriae strain SDS-502 applications cannot be precluded. More refined estimates of the potenti al risk are not possible because (1) the majority of the studies were not conducted at ma ximum hazard concentrations and (2) there are

uncertainties associated with the bi oassays because of variation in the protein extraction methods used in those studies. These bioassays 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. Ba sed on this study, the no observed effect concentration (NOEC) was 500 nanograms Bacillus thuringiensis subspecies galleriae strain SDS-502 Cry8Da protein/honeybee larval cell; howeve r, the study was supplemental because of the uncertainty in dosing. However, a commercial formulation containing Bacillus thuringiensis subspecies galleriae (strain not identified) with Le pidopteran activity was discussed in a review paper by Melin and Cozzi (1990). The formulation (no potency informa tion provided) was devel oped 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 provided) in a 40% sucrose solution showed no adverse effect on adult workers or colony life as determined by egg laying, brood producti on, 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 Gu ideline 885.4200), Freshwater Invertebrates (Harmonized Guideline 885.4240), and Estuarin e/Marine Fish and Invertebrates (Harmonized Guideline 885.4280) (MRID Nos. 485532-04, 485532-22, 485532-23, 485532-24, and 489401-07; Rose 2013b)

The applicant submitted scientific rationales 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 fi sh species (U.S. EPA 1998) and a limited host range to certa in insect orders.

The applicant also submitted a study with Daphnia magna in which daphnids were exposed to water containing a range of nominal concentrations of the technical grade of the tec hnical grade of the active ingredient (TGAI) for 21 days. Based on the percentage of the active ingredient 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 re production and growth (i.e., 50 mg/L) and for survival (i.e., 100 mg/L). Although high daphnid mortalit y and low neonate produc tion were noted in the filtered treatment, the adverse effects did not affect the overall conclusions of the study and wereBacillus thuringiensis subspecies galleriae strain SDS-502 Page 18 of 42 Biopesticides Registration Action Document

attributed to low dissolved oxygen content, and anom alous values for conductiv ity and water hardness. Since exposure resulting from the proposed applicatio ns is well below the defi ned toxicity endpoints,

adverse effects to freshwater i nvertebrates are not expected.

2. Environmental Fate Data

Microbial pesticides containing Bacillus thuringiensis readily break down in the environment, degrade

within 48 hrs under ultraviole t light exposures (Myasnik et al . 2001), and have a half-life of 3.8 hours under normal sunlit conditions (USDA 1994). The microor ganism can persist for several months in soil (Melin and Cozzi 1990), and certain Cry proteins ma y 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 anti cipated for most nontarget species exposed to Bacillus thuringiensis subspecies galleriae strain SDS-502 as a result of th e proposed labeled applications, including mammals, birds, terrestrial and aquatic plan ts, and freshwater and ma rine/estuarine animals. Therefore, a “No Effect” determination is made for direct and indirect eff ects to federally listed endangered and threatened (“listed”) mammals, birds, terrestrial and aquatic plants, and freshwater and

marine/estuarine animals and for th eir designated critical habitat.

Sustained population-level effects to nontarget ins ects are not anticipated; however, it is known that Bacillus thuringiensis subspecies galleriae can affect insects in the orde rs Coleoptera and Lepidoptera.

Direct effects to individuals of lis ted insect species (USFWS 2013) ca nnot be precluded, nor can indirect effects to listed species that depe nd specifically on a listed insect. Addi tional analysis of the potential effects to these is necessary to make a determination for these species.

V. ENVIRONMENTAL JUSTICE

The EPA seeks to achieve environmen tal justice—the fair treatment a nd meaningful involvement of all people regardless of race, color, national origi n, or income—with respect to the development, implementation, and enforcement of environmental laws , regulations, and policies. Fair treatment means that no group of people, incl uding racial, ethnic, or soci oeconomic groups, should bear a disproportionate share of the negative environmental co nsequences 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 affect ed community residents have an appropriate opportunity to particip ate 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 pa rticipants involved will be considered in the decision-making process; and (4)

the decision-makers seek out and f acilitate the involvement of thos e potentially affected. The EPA has this goal for all communities and pe rsons across the United States.

To help address potential environm ental justice issues, the EPA seeks information on any groups or segments of the population who, as a result their loca tion, cultural practices, or other factors, may have atypical, unusually high exposure to Bacillus thuringiensis subspecies galleriae strain SDS-502,Bacillus thuringiensis subspecies galleriae strain SDS-502 Page 19 of 42 Biopesticides Registration Action Document

compared to the general population. Please comment if you are aware of any subpopulations that may have atypical, unusually high exposure compared to the general population.

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

.

VI. RISK MANAGEMENT DECISION

Section 3(c)(5) of FIFRA permits for the registrati on 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 functi on without unreasonable adverse effects on the environment; AND (4) When used in accordance with wi despread 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 subspecies galleriae strain SDS502 is well-known to have insectic idal 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 su ch pesticide products are not expect ed to cause unreasonable adverse effects on the environment when used according to their respective label instructions. Criterion 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 en vironment, and Phyllom grubGONE! G and Phyllom

beetleGONE! TM biological insecticide (end-us e pesticide products), in part icular, are likely to control certain beetle pests as claimed, satisfying crite rion 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 subspecies galleriae strain SDS-502 as a ne w active ingredient, are eligible for registration under FIFRA s ection 3(c)(5) for the labeled uses.

VII. ACTIONS REQUIRED OF THE REGISTRANT

A. Final Printed Labeling

Before releasing pestic ide products containing Bacillus thuringiensis subspecies galleriae strain SDS502 for shipment, the registrant is required to provi de 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 P hyllom SDS-502 MP (EPA File Symbol: 88347-R) registration:

(1) Storage Stability (Har monized Guideline 830.6317) : Within one year of registration, 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) : Within one year of registration, Phyllom, LLC, must provide the EPA with the resu lts of a one-year corrosion characteristics study on Phyllom SDS-502 MP.

(3) pH (Harmonized Guideline 830.7000) ; Within six months of regi stration, Phyllom, LLC, must provide the EPA with the results of a pH study on Phyllom SDS-502 MP.

(4) Producers of Phyllom SDS-502 MP will conduct the fo llowing production quality control measures

to detect contamination or ch anges 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 contro l measures during production to detect any changes fr om 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 s pores into each of 5 laboratory mice weighing 17 grams to 23 grams. Such test shall show no eviden ce of infection or injury 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 Contro l of Insects and Mites,” R.P.M. Bond et al ., p. 280 ff., 1971). This specification can be satis fied 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 determining that β -exotoxin synthesized during spore production is eliminated by the subsequent spore-harvesting procedure.

(5) Producers of Phyllom SDS-502 MP will maintain r ecords of the quality c ontrol 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 items will be terms of the Phyllom grubGONE! G (EPA File Symbol: 88347-E) registration:

(1) Storage Stability (Har monized Guideline 830.6317) : Within one year of registration, Phyllom, LLC, must provide the EPA with the results of a one-y ear storage stability study on Phyllom grubGONE!

G.

(2) Corrosion Characteristics (Harmonized Guideline 830.6320) : Within one year of registration, Phyllom, LLC, must provide the EPA with the resu lts of a one-year corrosion 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 (Har monized Guideline 830.6317) : Within one year of registration, 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) : Within one year of registration, Phyllom, LLC, must provide the EPA with the resu lts 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 previ ous sections, it should be clearly understood 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 advers e effects to the environment must be submitted to the EPA under the provisions stated in FIFRA section 6(a)(2). Additionally, a ll incidents of hypersensi tivity (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 Environmental Protection Agency FFDCA Federal Food, Dr ug, and Cosmetic Act FIFRA Federal Insecticide, F ungicide, 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 de rived concentration of a substance that can be expected to cause death in 50% of test animals. It is usually expressed as the weight of substance per weight or volume of water, air, or feed (e.g., mg/L, mg/kg, or ppm).

LD 50 median lethal dose. A statistically derived si ngle dose that can be expected to cause death

in 50% of the test animals when administer ed by the route indicated (oral, dermal, or inhalation). It is expressed as a weight of subs tance per unit weight of animal (e.g., mg/kg).

μ g/ μ L micrograms per microliter

μ g/mL micrograms per milliliter mg/cm 2 milligrams per square centimeter mg/kg milligrams 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 O ccupational 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 SDS-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 prepared 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, Di scussion of Formation of Unintentional Ingredients. Unpublished study prep ared 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, Ce rtification 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 Ra ts. 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 Inje ction Toxicity/Pathogenicity St udy in Rats. Project Number: DUPONT/20581 16248 1015. Unpublished study prepar ed 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 In cidents. 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, Ac ute 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 pr epared by Stillmeadow, Inc., 16 pages.

485532-18 Kuhn J. 2006. SDS-502 TGAI - Acute Dermal Toxi city Study in Rabbits with a Microbial Pest Control Agent (MPCA) (Final Repor t). 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 Ge nesis Laboratories, Inc., 58 pages.

485532-20 Libs J, Rose R. 2010. Phyllom SDS-502 MP, Avian Inhalation Toxicity/Pathogenicity (Waiver Request). Unpublished st udy 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 (Wai ver Request). Unpublished study prepared by Phyllom, LLC, 3 pages.

485532-25 Libs J, Rose R. 2011. Phyllom SDS-502 MP, N ontarget Plant Testi ng (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 Toxic ity 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 th e Colorado Potato Beetle ( Leptinotarsa decemlineata ) (Coleoptera: Chrysomelidae), Acute Nontarget Insect Testing, Tier I. Project Number: PHY/SDS502/GRUB. Unpublished study prepar ed by the University of Maryland, 5

pages.

485532-29 Libs J. 2010. Phyllom SDS-502 MP, Bacillus thuringiensis subsp. galleriae , Strain SDS502 Toxicity to the Spotted Cucumber Beetle/Southern Corn Rootworm ( Diabrotica undecimpunctata howardi ) (Coleoptera: Chrysomelidae), Acute Nontarget Insect Testing, Tier I. Unpublished study prepar ed by Phyllom, LLC, 4 pages.

485532-30 Richards R. 2006. Evaluation of Dietary E ffects of SDS-502 Cry8D a on Larval Honeybee Development ( Apis mellifera L.). Project Number: CAR/152/05. Unpublished study prepared by California Agricultu ral 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 prep ared 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 Irr itation Study in Rabbits (Final Report). Project Number: 9643/06. Unpublished study pr epared 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 Char acteristics for Phyllom GrubGONE Granular (G). Unpublished study prepared by Phyllom, LLC, 15 pages.

487168-03 Bauer L, Londono D. 2012. Acute Nontarget Insect Testing, Ti er I: Parasitic Hymenoptera Wasps: Tetrastichus planipennisi

, Oobius agrili

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

489401-01 Libs J. 2012. Phyllom boreGONE! EP, Pr oduct Identity and Characterization. Unpublished study prepared by Phyllom, LLC, 3 pages.

489401-02 Libs J. 2012. Phyllom boreGONE! Biologica l Insecticide, Manufacturing Process. Unpublished study prepared by Phyllom, LLC, 4 pages.

489401-03 Libs J. 2012. Phyllom boreGONE! Biological In secticide, Discussion of Formation of Unintentional Ingredients. Unpublished st udy 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, Cer tification of Lim its. Unpublished study prepared by Phyllom, LLC, 2 pages.

489401-06 Libs J. 2012. Physical an d 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 th e End-Use Pesticide Product, Phyllom grubGONE! Granular (G), and the Manufactur ing-Use Pesticide Product, Phyllom SDS502 MP. Unpublished study prepared by Robert I. Rose, 8 pages.

489401-08 Libs J. 2012. Phyllom Bacillus thuringiensis subsp./var. galleriae , Strain SDS-502, Primary Powder TGAI/MUP/MP – Rationale/Clear ance Demonstration of Lack of, or No Infectivity in Studies Conducted for Regi stration 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 Inhalation 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 In secticide, Hypersensi tivity Incidents. Unpublished study prepared by Phyllom, LLC, 3 pages.

489731-03 Libs J. 2012. Phyllom boreGONE! Biological In secticide End-Use Product, Acute Oral Toxicity. Unpublished study prepar ed by Phyllom, LLC, 3 pages.

489731-04 Libs J. 2012. Phyllom boreGONE! Biological In secticide End-Use Product, Acute Dermal Toxicity. Unpublished study prepar ed by Phyllom, LLC, 4 pages.

489731-05 Libs J. 2012. Phyllom boreGONE! Biologica l 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 prepared 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 prepar ed 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: Musc idae. Project Nu mber: 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 prepar ed 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. U npublished study prepared by San Diego State University, 12 pages.

490940-05 Matthews T. 2013. Phyllom Bacillus thuringiensis subsp. galleriae , Strain SDS-502, grubGONE! Granular (G) Colony Forming Un its (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. 2013a. 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 pe sticide, 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 assessm ent 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. th rough S. Borges to J. Kausch, dated May 14, 2013.

U.S. EPA. 2012a. 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, Yama naka 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 (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. 1998. 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 89(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 Lepi dopteran 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.

Mendelsohn 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 an d 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 Letter 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 Preliminary Technica l Screening Letter from the EPA).

USDA. 1994. Extension Service, National Agricu ltural Pesticide Impact Assessment Program.Bacillus thuringiensis subspecies galleriae strain SDS-502 Page 31 of 42 Biopesticides Registration Action Document

U.S. EPA. 2012b. Chapter 7 of the Label Review Ma nual (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 Pesticid e Test Guidelines – Background for Residue Analysis of Microbial Pest Control Agents (OPPT S 885.2000). Available from http://www.epa.gov/ocspp/pubs/frs/publica tions/Test_Guide lines/series885.htm

.

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

.

World Health Organization. 2011. Guidelines for Dri nking-water Quality (Fourth Edition). Available

from http://whqlibdoc.who.int/ publications/2011/9789241548151_eng.pdf

.Bacillus thuringiensis subspecies galleriae strain SDS-502 Page 32 of 42 Biopesticides Registration Action Document

APPENDIX 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 Pr oduct (MP), Phyllom SDS-502 MP (40 C.F.R. § 158.2120)

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 90.0% by weight Bacillus thuringiensis subspecies galleriae strain SDS-502 fermentation solids, spores, and insecticidal toxins (contains a

minimum of 1 x 10 10 cfu per gram). 485532-01 Rose (2013a) Rose (2013b)

885.1200 Manufacturing Process Submitted data fulfill the requireme nt for manufacturing process. As a term of the Phyllom SDS-502 MP registrat ion, all producers of Phyllom SDS502 MP will conduct c ertain production qualit y control measures to detect contamination or changes to the parent strain and will maintain records of these quality co ntrol tests for a period of at least five years

from the date each batch is released. 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 BP7667. 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 Phy llom SDS-502 MP will conduct certain production quality control measures to detect contamination or changes to the parent strain and will maintain records of thes e 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 fu lfill 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 Requ irements for the End-Use Pesticide Prod ucts (EPs), Phyllo m 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 fermentation 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 fu lfill 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 Lim its listed on the confidential statemen t of formula are adequate/acceptable. 485698-05 1

489401-05 2
Rose (2013b) 1,2

1 Study supporting Phyllom grubGONE! G, EPA File Symbol 88347-E

2 Study supporting Phyllom beetleGONE! TM biological insecticide, EPA File Symbol 88347-GBacillus 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 Pr oduct (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 yeastlike

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 requiri ng that the results of a oneyear storage stability study be submitted within one year of registration. 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 within one year of registration. 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. 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 Characteri stics for the End-Use Pestic ide Product (EPs), Phyllo m grubGONE! G and Phyllom

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

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 storage stability studies be submitted within one year of registration. 487168-02 1

489401-06 2
Rose (2013b)

830.6319 Miscibility When mixed with water, the EPs form fine-particulatesuspensions that settle out over time and easily redistribute 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 corrosio n characteristics studies be submitted within one year of registration. 487168-02 1

489401-06 2
Rose (2013b)

830.7000 pH N/A N/A

830.7100 Viscosity Not required because the EP s 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 File Symbol 88347-E

2 Study supporting Phyllom beetleGONE! TM biological insecticide, EPA File Symbol 88347-G

3 According to 40 C.F.R. § 158.2120, these data are only required for the technical grade of the active ingredient. Since Phyllo m, LLC, included this information with its application fo r 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

TABLE 5. Toxicology Data Requireme nts 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

Number Data Requirement Results MRID No. 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 study conducted with the MP, see below).

Classification : Unacceptable 485532-09

485532-11

489401-08

885.3200 Acute Injection Toxicity/Pathogenicity Not toxic, infective, and/or pathogenic to rats when

administered in traperitoneally 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 no 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 iner t 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

TABLE 5. Toxicology Data Requireme nts 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

Number Data Requirement Results MRID No. 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 iner t 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 wa s mildly-irritating to the eyes of rabbits. Classification: Acceptable

TOXICITY CATEGORY III 485532-17

870.2500 Primary Dermal

Irritation Phyllom SDS-502 MP was essentially nonirritating 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 III 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 Products (E Ps), Phyllom grubGO NE! 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 no hypersensitivity incidents, including immediatetype 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 acu te 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 III 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 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 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 inhalation 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 Products (E Ps), Phyllom grubGO NE! 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 tomoderate erythema or other skinrelated 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 III 485698-13 1

489731-07 2

490940-01 2
Rose (2013b) 2

1 Study supporting Phyllom grubGONE! G, EPA File Symbol 88347-E

2 Study supporting Phyllom beetleGONE! TM biological insecticide, EPA File Symbol 88347-GBacillus thuringiensis subspecies galleriae strain SDS-502 Page 40 of 42 Biopesticides Registration Action Document

TABLE 7. Nontarget Organism Toxicity a nd 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 a nd 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 potato beetle ( 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 emerald ash borer ( Agrilus 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 concentratio ns of purified Cry8Da protein in artificial diets. There were no quantitative data or information to confirm the concentrations of the test material and reported

results. 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 larval 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 File Symbol Registration

Name Percentage

Active Ingredient Formulation Type Use Site(s) Method(s) of Application Application Rate(s) Target Pest

88347-R Phyllom SDS502 MP 90.0% Technical N/A

88347-E Phyllom grubGONE! G 9.0% End Use –

Granule Turf and

ornamentals Calibrated granular equipment 115–150 pounds per

acre White grubs of certain

beetles (e.g., Japanese beetle)

88347-G Phyllom

beetleGONE!

biological

insecticide 76.5% End Use –

Water Dispersible

Powder Ornamentals, select crops for

food and animal feed uses, and poultry premises Ground or aerial application equipment Variable (see draft label in docket for specifics) Certain beetle pests, particularly those in the

Families Buprestidae

,

Scarabaeidae , and

Curculionidae