BIOPESTICIDES REGISTRATION ACTION DOCUMENT



## Trichoderma virens strain G

- 41

Pesticide Chemical (PC) Code: 176604

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

DRAFT

- December 8 , 2011Trichoderma virens strain G

- 41 Page 2 of 34 Biopes ticides Registration Action Document

TABLE OF CONTENTS

I. EXECUTIVE SUMMARY

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

II. ACTIVE INGREDIENT OV ERVIEW

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

III. REGULATORY BACKGROUN D

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

A. Applications for Pesticide Product Registration

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

B. North American Free Trade Agreement Joint Review

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

C. Food Tolerance Exemption

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

IV. RISK ASSESSMENT

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

A. Product Analysis Assessment (40 CFR § 158.2120)

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

B. Human Health Assessment (40 CFR § 158.2140)

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

C. Environmental Assessment (40 CFR § 158.2150)

................................ ................................ ........ 14

V. ENVIRONMENTAL JUSTIC E

................................ ................................ ................................ ................. 16

VI. RISK MANAGEMENT DECI SION

................................ ................................ ................................ .......... 17

VII. ACTIONS REQUIRED OF THE REGISTRANT

................................ ................................ .................... 17

VIII. GLOSSARY OF ACRONYMS AND ABBREVIATIONS

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

IX. BIBLIOGRAPHY

................................ ................................ ................................ ................................ ....... 20

A. Studies Submitted or Cited to Support the Trichoderma virens strain G

- 41 Pest icide Product Registrations

................................ ................................ ................................ ..................... 20

B. Environmental Protection Agency Risk Assessme nt Memoranda

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

C. Other References

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

APPENDIX A. MICROBI AL PESTICIDES DATA R EQUIREMENTS

................................ ............................ 24

APPENDIX B. PESTICI DE PRODUCT S

................................ ................................ ................................ .............. 33Trichoderma virens strain G

- 41 Page 3 of 34 Biopes ticides Registration Action Document

BIOPESTICIDES REGISTRATION ACTION DOCUMENT TEAM

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

Science Reviews

Ibrahim S. Barsoum , Ph.D. Product Analysis, Human Health United States Environmental Pr otection Agency

John L. Kough, Ph.D. Product Analysis, Human Health Shannon Borges Environmental Effects

Leila Heikkila Hu man Health Health Canada Pest Management Regulatory Agency

James Elwin Product Analysis, Environmental Effects

Regulations

Sheryl K. Reilly, Ph.D. Chief, Microbial Pesticides Branch United States Environmental Protection Agency

Jeannine Kausch Regulatory Action LeaderTrichoderma virens strain G

- 41 Page 4 of 34 Biopes ticides Registration Action Document

I. EXECUTIVE S UMMARY

In September 2009, BioWorks, Inc. submitted application s for a manufacturing

- use pesticide product, G

- 41 Technical (EPA File Symbol 68539

- I), and an end

- use pesticide product, BW240 WP Biological Fungicide (EPA File Symbol 68539

- O), to b oth the United States Environmental Protection Agency (EPA) and the Health Canada Pest Management Regulatory Agency (PMRA) and requested a North American Free Trade Agreement (NAFTA) Joint Review of these application s

. Background

1 Concurrently with these applications , BioWorks, Inc. submitted an application for another end

- use pesticide product, BW240 G Biological Fungicide (EPA File Symbol 68539

- RN), and filed a petition for a tolerance exemption for residues of Trichoderma virens strain G

41, the new active ingredie nt contained in all three proposed pesticide products, with EPA only.

Trichoderma virens , such as strain G

- 41

, is a naturally occ urring fungus that is native to the United States and is widely distributed throughout the world, inhabiting forest, agricult ural and orchard soils, as well as plant litter (Samuels 1996). As such, Trichoderma virens strain G

- 41 was isolated from soil samples taken from Aphanamyces

- suppressive fields in Livingston County, New York. According to BioWorks, Inc., the new active ing redient, Trichoderma virens strain G

- 41

, inhibits or kills plant

- pathogenic fungi (e.g., Rhizoctonia spp. and Fusarium spp. ) through several mechanisms: (1) competition for food and space, (2) mycoparasitism, and (3) induction of plant defenses. Given the fungicidal capabilities of Trichoderma virens strain G

- 41, BioWorks

, Inc. has proposed to register these pestici de products to control soilborne plant pathogens and plant root diseases that adversely affect agricultural and greenhouse crops

, and plants in residential settings (e.g., vegetables, fruit, and ornamentals).

EPA and /or PMRA scientists reviewed product analysis, toxicology, and nontarget organism data and information (40 Code of Federal Regulations (CFR) §§ 158.2120, 158.2140, and 158.21 50, respectively) submitted to support the registration of the t hree proposed Trichoderma virens strain G

- 41 pesticide products. Overall, such data and information are adequate for risk assessment purposes , fulfill the current microbial pesticide data requ irements

, and allow for registration under Section 3(c)(5) of the Federal Insecticide, Fungicide, and Rodenticide Act (FIFRA)

.

Most of the p roduct analysis data requirements for Trichoderma virens strain G

- 41 , including product chemistry and composition, analysis of samples, and physical and chemical characteri stics, were fulfilled by acceptable guideline studies. Although a one

- year storage stability and corrosion characteristics study is outstanding, EPA has reviewed a n interim report of

a storage stability and corrosion characteristics study and , as a term of registration, is requiring the final report of this study be submitted approximately one year from the date of registration

. Product Analysis

Adequate mamm alian toxicology data and infor mation were submitted to support the Trichoderma virens strain G

- 41 pesticide products. An acute injection toxicity/pathogenicity study showed t hat, at a single high dose, Trichoderma virens strain G

- 41 is not toxic , infective, and/o r pathogenic via the in traperitoneal route of exposure , while an acute oral Toxicology

1 See http://www.epa.gov/oppfead1/international/naftatwg/guidance/jointreview

- biope.pdf

.Trichoderma virens strain G

- 41 Page 5 of 34 Biopes ticides Registration Action Document

toxicity/pathogenicity study showed that, at a single high dose, Trichoderma virens strain G

- 41 is not toxic via the oral route of exposure. A cute p ulmonary toxicity/patho genicity test ing was waived afte r considering the resul ts of the intraperitoneal test
, a BW240 WP Biological Fungicide

- specific acute inhalation toxicity test, and a pulmonary toxicity/pathogenicity test for functionally similar Trichoderma virens strain GL

- 21 ( formerly known as Glioclad ium virens strain GL

- 21)

. Moreover, no hypersensitivity incidents, occurring during rese arch, development, or testing of Trichoderma virens strain G

- 41 , were reported by the applicant . In light of the results of the intra peritoneal and oral toxicity/pathog enicity data , the acceptable rationale provided to waive the acute pulmonary toxicity/pathogenicity test , and the absence of hypersensitivity incidents, testing at higher tiers (i.e., Tiers II and III) was not required.

EPA concludes t hat there is a reasonable certainty that no harm will result to the United States population, including infants and children, from aggregate exposure to residues of Trichoderma
virens strain G

- 41 . No dietary risks are expected from use of Trichoderma viren s strain G

- 41 as an active ingredient in pesticide products. Significant exposure to Trichoderma virens strain G

41

— through food and drinking water and exceeding already present background levels

— is not anticipated due to : (1) the proposed application meth ods of the end

- use pesticide products (soil directed and soil incorporated ; no aquatic applications ) ; (2) the potential for this fungus to degrade due to competition with other biological organisms and exposure to environmental factors (e.g., varying tempe ratures ) in the soil ; (3) the filtering effect of many particulate soil types; and (4) the conditions (e.g., filtration and pH adjustments) water is subjected to in wastewater treatment systems and drinking water facilities

. Should Trichoderma virens strai n G

41 be present on food or in drinking water, the acute oral toxicity an d pathogenicity data available for Trichoderma virens strain G

- 41 and functionally similar Trichoderma virens strain GL

- 21 demonstrated no toxicity, infectivity, and/or pathogenicity is likely to oc cur

. Tolerance Exemption

Despite the low toxicological profile of Trichoderma virens strain G

- 41 , baseline personal protective equipment (PPE) is required for handlers that may be exposed to the active ingredient, due to their occupation , for prolonged periods or numerous times . Handlers working with Trichoderma virens strain G

- 41 in agricultural settings are directed to wear a long

- sleeved shirt, long pants, socks, shoes, and a dust/mist filtering respirator meeting National Institute fo r Occupational Safety and Health (NIOSH) standards of at least N

- 95, R

- 95, or P

- 95. EPA may require additional PPE (e.g., protective eyewear) , other than the standard described above, on a product

- specific basis. Occupational Exposure

Data and other i nfor mation (e.g., scientific literature) submitted by the applicant to support requests to waive nontarget organism testing for Trichoderma virens strain G

- 41 are sufficient to fulfill the relevant microbial pesticide data requirements and for risk assessment purposes. Further testing of nontarget organisms at higher tier levels (i.e., Tiers II, III, and IV) is not required. EPA performed an environment al risk assessment based on data and other information provided by the ap plicant

, and has determined that adve rse effects to nontarget organisms are not anticipated from the proposed pesticidal uses of Trichoderma virens strain G

- 41. Moreover, EPA has made a “No Effect” determination for direct and indirect effects to listed species and their designated critical h abitats resulting from these same proposed pesticidal uses. Nontarget OrganismsTrichoderma virens strain G

- 41 Page 6 of 34 Biopes ticides Registration Action Document

On October 1, 2009, EPA announced a new policy to provide a more meaningful opportunity for the public to participate i n major registration decisions before they occur. According to this policy, EPA intends to provide a public comment period prior to making a registration decision for, at minimum, the following types of appli cations: new active ingredients; first food uses; first outdoor uses; first residential use s; or a ny other registration actions for which EPA believes there may be significant public interest

. Public Participation

Consistent with the policy of making registration actions more transparent, the proposed

pe sticide products containing Trichoderma virens strain G

- 41

, a new a ctive ingredient, are subject to a 30

- day comment period. In addition to containing a new active ingredient, the registration of these pesticide products would result in the first outdoor, food, and residential uses for Trichoderma virens strain G

- 41. The docket identification number, associated with these registration actions and accessed through either http://www.regulations.gov or http://www.epa.gov/pesticides/regulating/registration

- status.html , is EPA

- HQ

- OPP

- 2010

- 0057. The following documents are available for comment in EPA

- HQ

- OPP

- 2010

- 0057: (1) draft Trichoderma virens strain G

- 41 Biopesticides Registration Action Document (BRA D); (2) environmental risk assessment for Trichoderma virens strain G

- 41; (3) draft label for the manufacturing

- use pesticide product, G

- 41 Technical (EPA File Symbol 68539

- I); (4) draft label for the end

- use pesticide product, BW240 WP Biological Fungicid e (EPA File Symbol 68539

- O); and (5) draft label for the end

- use pesticide product, BW240 G Biological Fungicide (EPA File Symbol 68539

- RN). While a final decision on registration is contingent upon review and consideration of public comments, EPA presentl y believes that, based upon the risk assessment and information submitted in support of G

- 41 Technical, BW240 WP Biological Fungicide, and BW240 G Biological Fungicide, it is in the best interest of the public and the environment to issue these registratio ns. The basis for this preliminary decision can be found in the risk assessment for Trichoderma virens strain G

- 41, which is characterized throughout this BRAD.

II. ACTIVE INGREDIENT OVERVIEW

Biological Name: Trichoderma virens strain G

- 41

Cultur e Deposit: American Type Culture Collection in Manassas

, Virginia under Accession N umber ATCC 20906

OPP Chemical Code: 176604

Type of Pesticide: Microbial Pesticide

– Fungicide

See Appendix B for spec ific information (e.g., use sites, application rate s , methods of application, formulation type s , and target pests) regarding the registered pesticide products containing this active ingredient.Trichoderma virens strain G

- 41 Page 7 of 34 Biopes ticides Registration Action Document

III. REGULATORY BACKGROUND

A. Applications for Pestici de Product Registration

On September 18, 2009

, T echnology Sciences Group, Inc. (address: 1150 18 th Street NW, Suite 1000; Washington

, District of Columbia 20036 ) , on behalf of BioW orks, Inc. (address: 100 Rawson Road, Suite 205; Victor, New York 14564), s ubmitted application s to register a manufacturing

- use pesticide product, G

- 41 Technical (EPA File Symbol 68539

- I )

, and two end

use pesticide product s , BW240 WP Biological Fungicide (EPA File Symbol 68539

- O ) and BW240 G Biological Fungicide (EPA File Symbol 68539

- RN), under FIFRA section 3. On
M arch 10, 2010

, EPA announced receipt of these application s to register pesticide product s containing a new active ingredient ( 75 Federal Reg ister (FR) 11175 ) and opened a 3 0

- day public comment period pursuant to the provisions of FIFRA section 3(c)(4). No comments were received following this publication.

B. North American Free Trade Agreement Joint Review

Trichoderma virens strain G

- 41 was the subject of a NAFTA Joint Review conducted by EPA and PMRA.

C . Food Tolerance Exemption

Concurrent with its registration application s and under Federal Food, Drug, and Cosmetic Act (FFDCA) section 408(d), Technology Sciences Group, Inc. , on beha lf of BioWorks, Inc.

, submitted a petition to establish an exemption from the requirement of a tolerance for Trichoderma virens strain G

- 41 (Pesticide Petition (PP) 9F7 618 )

. In the Federal Register of March 10, 2010 ( 75 FR 11171 ), EPA announced that Technology Sciences Group, Inc., on behalf of BioWorks, Inc., proposed to establish an exemption from the requirement of a tolerance for residues of the fungicide

, Trichoderma virens s train G

- 41

, in or on all food commodities and opened a 30

- day comment period. No comments were received following this publication.

IV. RISK ASSESSMENT

In the Federal Register of October 26, 2007, EPA issued a Final Rule on the data requirements to

s upport registration of microbial pesticides and updated the definition for microbial pesticides ( 72 FR 61002 )

. The rule became effective on December 26, 2007. The data and informa tion evaluated for this BRAD were considered in light of these requirements.

The classifications that are found for each data submission are assigned by EPA science reviewers and are an indication of the usefulness of the information contained in the docu ments for risk assessment. A rating of “ acceptable ” indicates the study is scientifically sound and is useful for risk assessment. A “ supplemental ” rating indicates the data provide some information that can be useful for risk assessment. The studies may h ave certain aspects determined not to be scientifically acceptable (“ supplemental : upgradeable ”). If a study is rated as “supplemental: upgradeable ,” EPA always provides an indication of what is lacking or what can be provided to change the rating to “ acce ptable .” If there is simply a “ supplemental ” rating, the reviewer willTrichoderma virens strain G

- 41 Page 8 of 34 Biopes ticides Registration Action Document

often state that t he study is not required by 40 C FR P art 158. Both “ acceptable” and “supplemental ” studies may be used in the risk assessment process as appropriate. An “unacceptable ” rating indicates that new data must be submitted.

For the acu te toxicity data requirements, Toxicity C ategories are assigned based on the hazard(s) identified from studies and/or other infor mation submitted to EPA in support of a pesticide registration. The active ingredient or particular product is classified into Toxicity Category I, II, III, or IV, where Toxicity Category I indicates the highest toxicity and Toxicity Category IV indicates the lowest toxicity (see 40 CFR § 156.62 )

.
A. Product Analysis Assessment ( 40 CFR § 158.2120 )

With the exception of a one

- year storage stability and corrosion characteristic s study, which EPA will require as a term of regis tration, a ll product analysis data requirements for Trichoderma
virens strain G

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

- s pecific information.

B. Human Health Assessment ( 40 CFR § 158.2140 )

1. Toxicity

All toxico logy data requirements for Trichoderma virens strain G

- 41 have been fulfilled

. Acceptable Tier I mammalian toxicology data and information support registration of the Trichoderma virens strain G

- 41 pesticide products

. Furthermore, Tier II and Tier III stud ies were not required for Trichoderma virens strain G

- 41

, based on the lack of acute toxicity/pathogenicity in the Tier I studies.

For a summary of the generic toxicology data requirements described in sections IV(B)(1)(a) and IV(B)(1)(b), as well as addi tional product

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

.
a. Acute Toxicity/Pathogenicity

– Tier I

Acute Oral Toxicity/Pathogenicity

– Rat ( Harmonized Guideline 885.3050; Master Record Identification Number (MRID No.) 4 83438

- 01 ) : In an acute oral toxicity study, groups of young Sprague

- Dawley albino rats (12 per sex) were given a single oral dose of G

- 41 technical grade of the active ingredient (TGAI) (1.5 x 10 9 c olony

- forming units per milliliter (cfu/mL)) in Phosphate Buffered Saline at doses of 1.5 x 10 8 colony

- forming units (cfu) per animal. The animals were then observed for a period of up to 21 days with interim scheduled sacrifices on Days 3, 7, and 14. A gr oup of animals (5 per sex) were also dosed with inactivated test substance. A group of control (untreated) animals (5 per sex) and a group of shelf contro l animals (2 per sex) were concurrently conducted. There were no treatment

- related clinical signs, and no necropsy findings other than an empty digestive system in one animal from the test substance group. There were no changes in body weight, other than in one animal from the test substance group who lost weight on Day 3. The test substance had cleared fr om all tissu es and organs by Day 7; however, the sensitivity of detection indicated low recovery of the microbial pest control agent (MPCA) fr om tissues and fluids (0.5

– 9%). C learance of the MPCA by Day 7 is uncertain

. Trichoderma virens strain G

- 41 is not toxic to rats when administered by oral gavage in a singleTrichoderma virens strain G

- 41 Page 9 of 34 Biopes ticides Registration Action Document

dose of 1.5 x 10 8 cfu per animal. This study was rated acceptable.

Acute Pulmonary Toxicity/Pathogenicity ( Harmonized Gui deline 885.3150; MRID No s

. 478708

- 01 and 482526

- 01 ) : Upon consideration o f results of other definitive toxicological data and/or information submitted or cited by the applicant, EPA waived acute pulmonary toxicity/pathogenicity testing for Trichoderma virens strain G

- 41

. First, i n an acute inhalation toxicity study (MRID No. 4 7 8650

- 04 ), rats were exposed to undiluted BW240 WP, containing both Trichoderma virens strain G

- 41 and Trichoderma harzianum Rifai strain T

- 22, at 5.14 milligrams per liter (mg/L) for 4 hours. There were no mortalities and only minor signs of toxicity (acti vity decrease; piloerection), which resolved by Day 2

. The gross necropsy conducted at study termination rev ealed no signs of pathogenicity; c learance, however, was not directly followed. Second, in an acute injection toxicity/pathogenicity study (MRID Nos

. 478651

- 02 and 482368

- 01)

, Trichoderma virens strain G

- 41 was not toxic to rats when administered intraperitoneally in a single dose of 10 7 cfu per animal. While clearance was not directly assessed in this study, the lack of clinical findings upon necrops y, in combination with the lack of mortality and signs of toxicity in the animals, strongly suggests that Trichoderma virens strain G

41 is also not pathogenic by intraperitoneal injection. Third, based on the similarity of Trichoderma virens strain G

- 41 a nd Trichoderma virens strain GL

- 21 (Kenerley 2010), it is reasonable to expect that toxicological data on Trichoderma virens strain GL

- 21 , such as an acute pulmonary toxicity/pathogenicity study (MRID Nos. 407198

- 05 and 408640

- 02) that demonstrated Trichod erma virens strain GL

- 21 was not toxic to, infective in, or pathogenic for rats, would be representative of the toxicological nature of Trichoderma virens strain G

- 41. Finally, the growth of Trichoderma virens strain G

- 41 is weak, at best, at the temperatu re of the human body (37°C), which further supports the lack of pathogenicity of Trichoderma virens strain G

- 41. EPA believes these data and information , when taken together, clearly indicate that Trichoderma virens strain G

- 41 would not be toxic, infectiv e, and/or pathogenic through the inhalation route of exposure

, and that further testing is not necessary.

Acute Injection Toxicity/Pathogenicity (Intra peritoneal)

– Rat (Harmonized Guideline 885.3200;
MRID No s

. 478651

- 02 and 482368

- 01 ) : In an acute intrap eritoneal infectivity study, young adult Wistar rats were injected with approximately 10 7 cfu of the G

- 41 TGAI in distilled water by intraperitoneal route (3 per sex; 1 milliliter per animal). A vehicle control group, which received sterile, distilled wate r, was also included. The animals were observed once daily for 21 days. The observation included changes in the skin and fur, eyes and mucous membrane, respiratory system, circulatory system, autonomic and central nervous system, somatomotor activity, and behavior pattern. Particular attention was directed to observations of tremors, convulsions, diarrhea, lethargy, salivation, sleep and coma; all observed toxic signs were recorded. Clearance, however, was not unequivocally assessed as it is not specified a s a requirement for infectivity testing by intraperitoneal injection. There were no mortalities, no toxic signs in any of the animals, no abnormal findings upon necropsy, and body weight gain was not affected. While clearance was not directly assessed in t his study, the lack of clinical findings upon necropsy, in combination with the lack of mortality and signs of toxicity in the animals, strongly suggests that Trichoderma virens strain G

- 41 is also not pathogenic by intraperitoneal injection. Trichoderma

v irens strain G

- 41 is not toxic , infective, and/or pathogenic to rats when administered intraperitoneally in a single dose of 10 7 cfu per animal. This study was rated acceptable.Trichoderma virens strain G

- 41 Page 10 of 34 Biopes ticides Registration Action Document

Hypersensitivity Incidents ( Harmonized Guideline 885.3400; MRID No. 4825 26

- 01 ) : No hypersensitivity incidents, including immediate

- type or delayed

- type reactions of humans and domestic animals that occurred during research, development, or testing of Trichoderma virens strain G

- 41

, were reported by the applicant . Any future hy persensitivity incidents must be reported to EPA (refer to test note #3 of 40 CFR § 158.2140(d))

.
Cell Culture ( Harmonized Guideline 885.3500) : This study is not required because Trichoderma
virens strain G

- 41 is not a virus (refer to test note #4 of 40 C FR § 158.2140(d)).

b. Acute Toxicology and Subchronic Toxicity/Pathogenicity

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

– Tier III

Tier II and Tier III studies were not r equired for Trichoderma virens strain G

- 41

, based on the lack of acute toxicity/pathogenicity in the Tier I studies.

c. Endocrine Disruptors

As required under FFDCA section 408(p), EPA has developed the Endo crine Disruptor Screening Program (EDSP) to determine whether certain substances (including pesticide active and other ingredients) may have an effect in humans or wildlife similar to an effect produced by a “naturally occurring estrogen, or other such end ocrine effects as the Administrator may designate.” The EDSP employs a two

- tiered approach to making the statutorily required determinations. Tier 1 consists of a battery of 11 screening assays to identify the potential of a chemical substance to interact with the estrogen, androgen, or thyroid (E, A, or T) hormonal systems. Chemicals that go through Tier 1 screening and are found to have the potential to interact with E, A, or T hormonal systems will proceed to the next stage of the EDSP where EPA will det ermine which, if any, of the Tier 2 tests are necessary based on the available data. Tier 2 testing is designed to identify any adverse endocrine

- related effects caused by the substance, and establish a quantitative relationship between the dose and the E, A, or T effect.

Between October 2009 and February 2010, EPA issued test orders/data call

- ins for the first group of 67 chemicals, which contains 58 pesticide active ingredients and 9 inert ingredients. This list of chemicals was selected based on the pot ential for human exposure through pathways such as food and water, residential activity, and certain post

- application agricultural scenarios. This list should not be construed as a list of known or likely endocrine disruptors.

Trichoderma virens strain G

- 41 is not among the group of 58 pesticide active ingredients on the initial list to be screened under the EDSP. Under FFDCA section 408(p), EPA must screen all pesticide chemicals. Accordingly, EPA anticipates issuing future EDSP orders/data call

- ins for

a ll pesticide active ingredients.

For further information on the status of the EDSP, the policies and procedures, the list of 67 chemicals, the test guidelines and the Tier 1 screening battery, please visit our website: http://www.epa.gov/endo/

.Trichoderma virens strain G

- 41 Page 11 of 34 Biopes ticides Registration Action Document

2

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

Section 408(c)(2)(A)(i) of FFDCA allows EPA to establish an exemption from the requirement for a tolerance (the legal limit for a pesticide chemical residu e in or on a food) only if EPA determines that the exemption is “safe.” Section 408(c)(2)(A)(ii) of FFDCA defines “safe” to mean that “there is a reasonable certainty that no harm will result from aggregate exposure to the pesticide chemical residue, inclu ding all anticipated dietary exposures and all other exposures for which there is reliable information.” This includes exposure through drinking water and in resi dential settings

, but does not include occupational exposure. Pursuant to section 408(c)(2)(B) of FFDCA, in establishing or maintaining in effect an exemption from the requirement of a tolerance, EPA must take into account the factors set forth in section 408(b)(2)(C) of FFDCA, which require EPA to give special consideration to exposure of infants and children to the pesticide chemical residue in establishing a tolerance exemption

, and to “ensure that there is a reasonable certainty that no harm will result to infants and children from aggregate exposure to the pesticide chemical residue....” Additi onally, section 408(b)(2)(D) of FFDCA requires that EPA consider “available information concerning the cumulative effects of [a particular pesticide's]

. . . residues and other substances that have a common mechanism of toxicity.”

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

Consistent with section 408(b)(2)(D) of FFDCA, EPA has reviewed the available scientific data and other relevant information, and considered its validity, completeness, and reliability and the relationship of this information to human risk. EPA has also considered available information concerning the variability of the sensitivities of major identifiable subgroups of consumers, including infants and children.

Based on the acute toxicity/pathogenicity data and infor mation discussed previously and presented in Table s 5 and 6 in Appendix A , t he data required for a FFDCA risk assessment for Trichoderma virens strain G

- 41 have been fulfilled

.
a. Aggregate Exposure

In examining aggregate exposure, section 408 of FFDCA directs EPA to consider available information concerning exposures from the pesticide residue in food and all other nonoccupational exposures, including drinking water from ground water or surface water and exposure through pesticide use in gardens, lawns, or buildings (residential and other indoor uses).

Food Exposure and Risk Characterization : A ll p roposed Trichoderma virens strain G

- 41 applications are soi l directed or soil incorporated because of the targeted soilborne pests (e.g., Rhizoctonia spp. and Fusarium spp. ). These applications are not expected to significantly increase the populations of this fungus above natural levels in the soil . No reports were available in the literature describing natural concentrations o f Trichoderma virens ; however, Trichoderma species have been reported in various types of soils at concentrations of 10 4 to 10 6 cfu/g (Leandro et al . 2007, Liu et al . 2008). Based on the maximum application rate of the proposed end

- use pesticide products c ontaining Trichoderma virens strain G

- 41 and its reported minimum viabilityTrichoderma virens strain G

- 41 Page 12 of 34 Biopes ticides Registration Action Document

in these pesticide products, the estimated amount of Trichoderma virens applied to the soil surface is approximately 6.7 x 10 3 colony

- forming units per square centimeter ( cfu/cm 2 ). Assuming a bulk density of 1 to 2 grams per cubic centimeter ( g/cm 3 ), the maximum application rate will not result in soil concentrations that are substantially greater than concentrations of Trichoderma virens naturally found in the soil, and increased e xposure to Trichoderma virens in the terrestrial environment , including through inadvertent contact with aboveground plant parts, is not expected. Should this microbial pesticide be present on food, the acute oral toxicity and pathogenicity data available for Trichoderma virens strain G

- 41 and functionally similar Trichoderma virens strain GL

- 21 demonstrated no toxicity, infectivity, and/or pathogenicity is likely to occur with any exposure level of Trichoderma virens strain G

- 41 resulting from application in accordance with good agricultural practices (see section IV(B)(1)(a) and Table 5 in Appendix A ).

Drinking Water Exposure and Risk Characterization : Exposure to residues of Trichoderma
virens strain G

- 41 in consumed drinki ng water is unlikely. The proposed use patterns for Trichoderma virens strain G

- 41 are soil directed and soil incorporated , thereby limiting contact with surface water by drift and runoff

. Furthermore, ground water is not expected to have significant expos ure to Trichoderma virens strain G

- 41 since , like other Trichoderma species

, this fungus would likely be filtered out by the particulate nature of many soil types and be concentrated in upper soil horizons (Longa et al . 2009, Sariah et al . 2005) near plant roots (USPTO 2010)

. I f Trichoderma virens strain G

- 41 were to be transferred to surface or ground

water s that are intended for eventual human consumption (e.g., through spray drift or runoff) and directed to wastewater treatment systems or drinking water facilities

, it likely w ould not survive the cond itions water is subjected to in such systems or facilities , including high temperatures, chlorination, pH adjustments, and/or filtration (Center s for Disease Control and Prevention 2009; U.S. EPA 2004)

. For i nstance, Trichoderma virens strain G

- 41 does not grow well at 37°C (Kenerley 2010, Lumsden et al . 1996) and test data submitted b y the petitioner has shown it to be unstable at elevated temperatures; there fore, any heat treatment applied to water containin g Trichoderma virens strain G

- 41 would probably render the fungus inviable. In the remote likelihood that this microbial pesticide is present in drinking water (e.g., water not subject to treatment systems or facilities), the acute oral toxicity and pathog enicity data available for Trichoderma virens strain G

- 41 and functionally similar Trichoderma virens strain GL

- 21 demonstrated no toxicity, infectivity, and/or pathogenicity is likely to occur with any exposure level of Trichoderma virens strain G

- 41 resu lting from application in accordance with good agricultural practice s (see section IV(B)(1)(a) and Table 5 in Appendix A )

.

Non

- occupational, Residential Risk Characterization : Given Trichoderma virens ’ natural occurrence in soil, non

- occupational exposure to the fungus is likely already occurring

. E ven with the proposed pesticide applications of Trichoderma virens strain G

- 41, it is not likely that there will be a significant increase in these exposures due to the expected ma intenance of background levels in the soil (see calculations in the food exposure section above). I f significant

non

- occupational exposures were to occur, such exposures would not exceed EPA’s level of concern given test results that indicated Trichoderma virens strain G

- 41 is not toxic (acute inhalation and dermal toxicity), is non

- irritating (primary dermal irritation), and is not pathogenic or infective (acute injection toxicity/pathogenicity) (see section IV(B)(1)(a) and Table s 5 and 6 in Appendix A ).Trichoderma virens strain G

- 41 Page 13 of 34 Biopes ticides Registration Action Document

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

Section 408(b)(2)(D)(v) of FFDCA requires that, when considering whether to establish, modify, or revoke a tolerance exemption, EPA consider “avail able information concerning the cumulative effects of [a particular pesticide’s]

. . . residues and other substances that have a common mechanism of toxicity.”

There are several isolates of Trichoderma species used as active ingredient s in registered pesti cide products. While these isolates may produce similar metabolites, the likelihood of adverse cumulative effects via a common mechanism of toxicity is likely to be minimal

, based on the lack of toxicity/pathogenicity potential of these active ingredients (U.S. EPA 2008)

. For information regarding EPA ’s efforts to determine which chemicals have a common mechanism of toxicity and to evaluate the cumulative effects o f such chemicals, see EPA ’s website at http://www.epa.gov/pesticides/cumulative

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

FFDCA section 408(b)(2)(C) provides that EPA shall assess the available information about consumption patterns among infants and children, special susceptibility of infants and children to pesticide chemical residues, and the cumulative effects on infants and children of the residues and other substances with a common mechanism of toxicity. In addition, FFDCA section 408(b)(2)( C) provides that EPA shall apply an additional tenfold (10X) margin of safety for infants and children in the case of threshold effects to account for prenatal and postnatal toxicity and the completeness of the database on toxicity and exposure unless EPA determines that a different margin of safety will be safe for infants and children. This additional margin of safety is commonly referred to as the Food Quality Protection Act Safety Factor. In applying this provision, EPA either retains the default value of 10X or uses a different additional safety factor when reliable data available to EPA support the choice of a different factor.

Based on the acute toxicity and pathogenicity data / information summarized in section IV(B)(1)(a) and Table s 5 and 6 in Appendix A

, EPA concludes that there is a reasonable certainty that no harm will result to the United States population, including infants and children, from aggregate exposure to the residues of Trichoderma virens strain G

- 41

. Such exposure includes all anticipated dietary exposures and all other exposures for which there is reliable information. EPA has arrived at this conclusion because , considered collectively, the data and information available on Trichoderma virens strain G

- 41, as well as data available on functionally similar Trichoderma virens strain GL

- 21

, do not demonstrate toxic, pathogenic, and/ o r infective potential to sensitive populations from exposure to this microbial pest control agent . Thus, there are no threshold e ffects of concern and, as a result, the provision requiring an additional margin of safety is not necessary

.
3. Occupational Exposure and Risk Characterization

Handler exposure to Trichoderma virens strain G

- 41 is not expected to pose any undue risk.
R egardless, appropriate personal protective equipment and precautionary statements are required on pesticide product labels to mitigate any potential risks to pesticide handlers due to prolonged or numerous exposures. Handlers applying Trichoderma virens st rain G

- 41 end

- use pesticide products in agricultural settings must wear a long

- sleeved shirt, long pants, socks, shoes, and aTrichoderma virens strain G

- 41 Page 14 of 34 Biopes ticides Registration Action Document

dust/mist filtering respirator meeting NIOSH standards of at least N

- 95, R

- 95, or P

- 95. Additional PPE (e.g., protective eyewear)

, other than the standard described above, may be required on a product

- specific basis.

4. Human Health Risk Characterization

EPA considered human exposure to Trichoderma virens strain G

- 41 in light of the standard for registration in FIFRA and the releva nt safety factors in FFDCA. A determination has been made that no unreason able adverse effects to the United States population in general, and to infants and children in particu lar, will result when Trichoderma virens strain G

- 41 pesticide products are

use d in accordance with EPA

- approved labeling.

C. Environmental Assessment ( 40 CFR § 158.2150 )

D ata and other i nformation (e.g., scientific literature) submitted by the applicant to support requests to waive nontarget organism testing for Trichoderma virens strain G

- 41 are sufficient to fulfill the relevant microbial pesticide data requirements and f or risk assessment purposes. Further testing of nontarget organisms at higher tier levels (i.e., Tiers I I, III, and IV) is not required . EPA performed an environmental risk assessment based on data and other information provided by the ap plicant

, and has d etermined that adverse effects to nontarget organisms are not anticipated from the proposed pesticidal uses of Trichoderma virens strain G

- 41. Moreover, EPA has made a “No Effect” determination for direct and indirect effects to listed species and their de signated critical habitats resulting from these same proposed pesticidal uses.

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

.

1. Ecological Exposure and Ri sk Characterization

a. Terrestrial Animals and Plants

Birds ( Harmonized Guideline 885.4050) and Wild Mammals (Harmonized Guideline 885.4150) (MRID Nos. 478651

- 03 and 482526

- 01) : Data and other i nformation submitted by BioWorks, Inc. to support its w aiver requests showed that Trichoderma virens strain G

- 41 is not expected to be toxic or pathogenic to birds or mammals because ( 1) birds and mammals are commonly exposed to this fungus; ( 2) Trichoderma virens strain G

- 41 does not grow at mammalian or avia n body temperatures; and ( 3) Trichoderma virens strain G

- 41 is similar to Trichoderma virens strain GL

- 21, which was previously determined to present minimal risk to birds and mammals

. Trichoderma virens is a ubiquitous fung us in terrestrial environments, inhabiting soil of all types and plant litter (Samuels 1996). Therefore, wild mammals and birds are commonly exposed to this fungus. Trichoderma virens strain G

- 41 also do es not grow at 3 9 oC or above (Kenerley 2010 ). Kenerley (2010) also provides data to s how that Trichoderma virens strain G

- 41 is biologically similar to Trichoderma virens strain GL

- 21 and is expected to have a similar toxicity/pathogenicity profile. Trichoderma virens strain G

- 41 is not expected to adverse ly affect wild mammals, since no t oxicity or pathogenicity was shown in laboratory rats exposed to G

- 41 Technical by injection at 10 7 units of MPCA per animal (submitted in MRID Nos. 478651

- 02 and 482 368

- 01). Based on this information, adverse effects to wild mammals and birds are not anti cipated to result from the proposed labeled uses of Trichoderma virens strain G

- 41.Trichoderma virens strain G

- 41 Page 15 of 34 Biopes ticides Registration Action Document

Nontarget Insects ( Harmonized Guideline 885.4340) and Honey Bees (Harmonized Guideline 885.4380) (MRID Nos.478651

- 03 and 482526

- 01) : BioWorks, Inc. presented s imilar data and information to support waiver requests and a conclusion of minimal exposure and risk to nontarget insects and honey bees. Trichoderma virens strain G

- 41 is not expected to be toxic or pathogenic to honey bees and other insects

, because it is a natural ly occurring fungus that is not an insect pathogen, and it also has not been known to produce insect epizootics (i.e., widespread outbreaks of disease that spread quickly) . It is also native to the United States and is widely distributed throughout the wor ld, inhabiting forest, agricultural and orchard soils as well as plant litter (Samuels 1996). The end

- use pesticide products are proposed for use in greenhouses, nurseries, and soil treatments, which will reduce exposure to honey bees and many other insect s. Since Trichoderma virens is naturally found in soils and plant litter, soil

- and ground

- dwelling insects are commonly exposed to this fungus. As discussed below (see “nontarget plants” summary) , the proposed applications of T richoderma virens strain G

- 4 1 are not expected to increase the concentrations of this fungus above naturally occurring levels. Based on these lines of evidence, the proposed pesticidal uses of Trichoderma virens strain G

- 41 are not expected to result in adverse effects to honey bees and other nontarget insects.

Nontarget Plants ( Harmonized Guideline 885.4300 ; MRID Nos. 478651

- 03 and 482526

- 01 ) : BioWorks, Inc. presented data and other information to support waiver requests and show that Trichoderma virens strain G

- 41 will not cause a dverse effects in nontarget plants

. Trichoderma
virens is a naturally occurring fungus that is native to the United States and is widely distributed throughout the world, inhabiting forest, agricultural and orchard soils

, as well as plant litter (Samue ls 1 996). Therefore, nontarget terrestrial plants are commonly exposed to Trichoderma
virens . The proposed applications are also not expected to significantly increase the populations of this fungus in soil above natural levels. No reports were available in th e literature describing natural concentrations of T richoderma virens ; however, Trichoderma spp. have been reported in various types of soils at concentrations of 10 4 to 10 6 cfu/g (Leandro et al . 2007, Liu et al . 2008). Based on the maxi mum application rate of the end

- use pesticide products containing
T

richoderma virens strain G

- 41

, and its report ed minimum viability in the end

- use pesticide product s (5 x 10 6 cfu/g), the estimated amount of Trichoderma virens applied to the soil surface is approximately 6.7 x 10 3 cfu/cm 2

. Assuming a soil bulk density of 1 to 2 g/cm 3

, the maximum application rate will not result in soil concentrations that are substantially greater than concentrations of Trichoderma virens naturally found in the soil , and increased exposure to Trichoderma virens in the terrestria l environment is not expected. Trichoderma virens is also not related to known plant pathogens. The refore, based on the data and information presented, adverse effects to terrestrial nontarget plants resulting from the proposed pesticidal applications of Trichoderma virens strain G

- 41 are not expected.

b. Aquatic Animals and Plants

Freshwater Fish ( Harmonized Guideline 885.4200) , Freshwater Invertebrates ( Harmonized Guideline 885.4240)

, Estuarine/Marine Fish and Inverte brates (Harmonized Guideline 885.4280) , and Nontarget Plants (Harmonized Guideline 885.4300) (MRID Nos. 478651

- 03 and 482526

- 01) : BioWorks, Inc. presented s imilar data and information to support waiver requests and show that adverse effects to fres hwater and marine/estuarine fish and invertebrates will not occur as a result of the proposed applications of Trichoderma virens strain G

- 41. Similar data and information was als o provided for aquatic plants. Trichoderma virens is native to the UnitedTrichoderma virens strain G

- 41 Page 16 of 34 Biopes ticides Registration Action Document

Stat es and is widely distributed

, inhabiting forest, agricultural and orchard soils

, as well a s plant litter (Samuels 1996). Given the ubiquitous distribution of Trichoderma virens in soil, this microbe is very likely washed into freshwater and marine/estuarin e systems in nature, and fish, invertebrates, and aquatic plants are commonly exposed to this fungus

. As discussed above (see “nontarget plants” summary) , the maximum proposed application rate is not expected to raise the concentration of Trichoderma viren s in the soil above naturally occurring levels, so runoff from treated areas is expected to contain similar amounts of Trichoderma virens as would be expected in runoff from natural areas. Trichoderma virens is also not known to proliferate in water, so
Tr ichoderma virens deposited in water from runoff would not become established in aquatic environments. Based on this data and information, the proposed pesticidal applications of Trichoderma virens strain G

- 41 are not expected to result in significant expos ure of nontarget aquatic organisms to levels above those that occur naturally, and adverse effects to freshwater and marine/estuarine fish, invertebrates, and plants are not expected.

2. Environmental Fate Data

As t he data and information provided are sufficient to fulfill the Tier I nontarget organism data requirements and allow for nontarget organism risk asse ssment for Trichoderma virens strain G

41 , f urther testing at higher tier levels (i.e., Tiers II, III, and IV) is not required.

3

. Threatene d and Endangered Species Assessment

Since EPA has determined that no effects are ant icipated for any non target species exposed to Trichoderma virens strain G

- 41 as a result of the proposed applications, effects to threatened and endangered species and the ir designated critical ha bitats are also not expected. Therefore, a “No Effect” determination is made for direct and indirect effects to listed species and their designated critical habitats resulting from the proposed pesticidal uses of Trichoderma virens strain G

- 41 , as labeled.

V. ENVIRONMENTAL JUSTICE

EPA seeks to achieve environmental justice

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

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

- making process; and (4) the decision

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

To help address potential environmental justice issues

, EPA seeks information on any groups or segments of the population who, as a result their location, cultural practices, or other factors, mayTrichoderma virens strain G

- 41 Page 17 of 34 Biopes ticides Registration Action Document

have atypical, unusually high exposure to Trichoderma virens strain G

- 41 , compared to the general population. Ple ase 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 EPA’s web site at http://www.epa.gov/compliance/environmentaljustice/index.html

.

V I

. RISK MANAGEMENT DECISION

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

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

To satisfy criterion 1, the Trichoderma virens strain G

- 41 pesticide products have well

- known properties. EPA has no knowledge that would contradict the claims made on the G

- 41 Technical, BW240 WP Biological Fungicide, and BW240 G Biological Fungicide label s, and such pesticide produ cts are not expected to cause unreasonable adverse effects on the environment when used according to the ir respective label instructions. Criterion 2 is satisfied by the current product label s, as well as the data and information presented in this document . It is believed that the Trichoderma virens strain G

- 41 pesticide products will not cause any unreasonable adverse effects on the environment, and BW240 WP Biological Fungicide and BW240 G Biological Fungicide (end

- use pesticide product s ) , in particular, are likel y to provide protection against fungal pests as claimed, satisfying criterion 3. Criterion 4 is satisfied in that the Trichoderma
virens strain G

- 41 pesticide products are not expected to cause unreasonable adverse effects when used according to l abel instructions. Therefore, G

- 41 Technical, BW240 WP Biological Fungicide, and BW240 G Biological Fungicide

, containing Trichoderma virens strain G

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

u ses.

V I I

. ACTIONS REQUIRED OF THE REGISTRANT

A. Final Printed Labeling

Before releasing pesticide products containing Trichoderma virens strain G

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

.Trichoderma virens strain G

- 41 Page 18 of 34 Biopes ticides Registration Action Document

B. Terms of Registration

As terms of the G

- 41 Technical, BW240 WP Biological Fungicide, and BW240 G Biological Fungicide registrations, BioWorks, Inc. must submit the results of a one

- year s torage s tability (Harmonized Guideline 830.6317) and c orrosion c ha racteristics (Harmonized Guideline 830.6320) study within one year of registration

.
Additionally, as a term of the BW240 G Biological Fungicide registration, BioWorks, Inc. must submit additional information (confirmatory) on the d iscussion of fo rmation o f u nintentional i ngredients (Harmonized Guideline 885.1300) and a nalysis of s amples (Harmonized Guideline 885.1400) data requirements within six months of registration.

C. Reporting of Adverse Effects and Hypersensitivity Incidents

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

Reports of all incidents of adverse effects to the environment must be submitted to EPA under the provisions stated in FIFRA section 6(a)(2). Additionally, all incidents of hypersensitivity (including both suspected and confirmed incidents) must be reported to EPA under the provisions of 40 CFR § 158.2140(d).Trichoderma virens strain G

- 41 Page 19 of 34 Biopes ticides Registration Action Document

VIII. GLOSSARY OF ACRONYMS AND ABBREVIATIONS

BRAD Biopesticides Registration Action Document CFR Code of Federal Regulations

c fu colony

- forming unit (s) cfu/cm 2 colon y

- forming units per square centimeter cfu/g colony

- forming units per gram cfu/mL colony

- forming unit s per milliliter EDSP Endocrine Disruptor Screening Program
EP end

- use pesticide product EPA United States Environmental Protection Agency (the “Agency”)
FF DCA Federal Food, Drug, and Cosmetic Act FIFRA Federal Insecticide, Fungicide, and Rodenticide Act
FR Federal Register

g gram g/cm 3 grams per cubic centimeter LC 50 median lethal concentration. A statistically derived concentration of a substance that can be expected to cause death in 50% of test animals. It is usually expressed as the weight of substance per weight or volume of water, air, or feed (e.g., mg/L, mg/kg, or ppm).
LD 50 median lethal dose. A statistically derived single dose that can be exp ected to cause death in 50% of the test animals when administered by the route indicated (oral, dermal, or inhalation). It is expressed as a weight of substance per unit weight of animal (e.g., mg/kg). mg/kg milligram s per kilogram mg/L milligram s pe r liter
MP manufacturing

- use pesticide product MPCA microbial pest control agent MRID No. Master Record Identification Number NAFTA North American Free Trade Agreement NIOSH National Institute for Occupational Safety and Health OPP Office of Pesticide Pro grams PC Code Pesticide Chemical Code PMRA Health Canada Pest Management Regulatory Agency
PP Pesticide Petition
PPE personal protective equipment

ppm parts per million TGAI technical grade of the active ingredient U.S. United States USPTO United States Pa tent and Trademark OfficeTrichoderma virens strain G

- 41 Page 20 of 34 Biopes ticides Registration Action Document

I X. B IBLIOGRAPHY

A. Studies Submitted or Cited to Support the Trichoderma virens strain G

- 41 Pesticide Product Registrations

MRID No. Study Information

407198

- 04 David R. 1988. Acute Oral Toxicity/Pathogen icity Study of Alginate Prill of Strain GL21 in Rats

– Final Report. Study Number: G

- 7131.222. Unpublished study prepared by Microbiological Associates, Inc., 22 pages.

407198

- 05 David R. 1988. Acute Pulmonary Toxicity/Pathogenicity Study of Spores of Str ain GL21 in Rats

– Interim Report. Study Number: G

- 7131.225. Unpublished study prepared by Microbiological Associates, Inc., 20 pages.

407198

- 06 David R. 1981. Acute Intravenous Toxicity/Pathogenicity Study of Mycelia of Strain GL21 in Rats

– Interim Repo rt. Study Number: G

- 7131.224. Unpublished study prepared by Microbiological Associates, Inc., 21 pages.

408640

- 01 David R. 1988. Acute Intravenous Toxicity/Pathogenicity Study of Mycelia of Strain GL21 in Rats

– Final Report. MBA Study Number: G

- 7131.224. Unpublished study prepared by Microbiological Associates, Inc., 23 pages.

408640

- 02 David R. 1988. Acute Pulmonary Toxicity/Pathogenicity Study of Spores of Strain GL21 in Rats

– Final Report. MBA Study Number: G

- 7131.225. Unpublished study prepared by M icrobiological Associates, Inc., 23 pages.

478649

- 01 Brookman D. 2009. Product Chemistry of BW240 Granules. Project Number: BW240G/001. Unpublished study prepared by Technology Sciences Group, Inc., 57 pages.

478650

- 01 Brookman D. 2009. Product Chemistry of BW240 WP. Project Number: BW240/001. Unpublished study prepared by Technology Sciences Group, Inc., 79

pages.

478650

- 02 Kuhn J. 2009. BW240 WP: Acute Oral Toxicity Study (UDP) in Rats

– Final Report. Project Number: 12982/09. Unpublished study prepare d by Stillmeadow, Inc., 11 pages.

478650

- 03 Kuhn J. 2009. BW240 WP: Acute Dermal Toxicity Study in Rats

– Final Report. Project Number: 12983/09. Unpublished study prepared by Stillmeadow, Inc., 12

pages.

478650

- 04 Younger C. 2009. BW240 WP: Acute Inhala tion Toxicity Study in Rats

– Final Report. Project Number: 12984/09. Unpublished study prepared by Stillmeadow, Inc., 18 pages.Trichoderma virens strain G

- 41 Page 21 of 34 Biopes ticides Registration Action Document

MRID No. Study Information

478650

- 05 Kuhn J. 2009. BW240 WP: Acute Eye Irritati on Study in Rabbits

– Final Report. Project Number: 12985/09. Unpublished study prepared by Stillmeadow, Inc., 17

pages.

478650

- 06 Kuhn J. 2009. BW240 WP: Acute Dermal Irritation Study in Rabbits

– Final Report. Project Number: 12986/09. Unpublished study prepared by Stillmeadow, Inc., 12 pages.

478651

- 01 Brookman D. 2009. Product Chemistry of G

- 41 Technical. Project Number: BWG41/001, G6690/PHYCHEM, G6690. Unpublished study prepared by Technology Sciences Group, Inc., 95 pages.

478651

- 02 Prakash P. 200 9. G41 Technical Grade Active Ingredient: Acute Injection Toxicity/Pathogenicity Study in Wistar Rats. Project Number: G6692/AP/R, G6692. Unpublished study prepared by Advinus Therapeutics Private Ltd., 28

pages.

478651

- 03 Mileson B. 2009. Response to Tie r I Microbial Pesticide Data Requirements for G

- 41 Technical. Unpublished study prepared by Technology Sciences Group, Inc., 47 pages.

478708

- 01 Even M. 2009. Response to Tier I Microbial Pesticide Data Requirements for G

- 41 Technical. Unpublished study prepared by Technology Sciences Group. Inc., 32 pages.

481964

- 02 Even M. 2010. Response to Tier I Microbial Pesticide Data Requirements for BW240 G. Unpublished study prepared by Technology Sciences Group, Inc., 4

pages.

482104

- 01 Brookman D. 2010. Suppl emental Product Chemistry of BW240 WP. Project Number: BW240WP/002. Unpublished study prepared by Technology Sciences Group, Inc., 22 pages.

482104

- 02 Even M. 2010. Response to Tier I Microbial Pesticide Data Requirements for BW240 WP. Unpublished study p repared by Technology Sciences Group, Inc., 4

pages.

482120

- 01 Brookman D, Bjornson H. 2010. Supplemental Product Chemistry of G

- 41 Technical. Project Number: BWG41/002. Unpublished study prepared by Technology Sciences Group, Inc., 57 pages.

482368

- 01 P rakash P. 2009. G41 Technical Grade Active Ingredient: Acute Injection Toxicity/Pathogenicity Study in Wistar Rats

– Final Report. Project Number: G6692. Unpublished study prepared by Advinus Therapeutics Private Ltd., 7

pages.Trichoderma virens strain G

- 41 Page 22 of 34 Biopes ticides Registration Action Document

MRID No. Study Information

482526

- 01 Mileson B, Even M. 2010. Response to Tier I Microbial Pesticide Data Requirements for G

- 41 Technical. Unpublished study prepared by BioWorks, Inc., 220 pages.

483438

- 01 Monds K. 2010. G41 TGAI: Acut e Oral Toxicity/Pathogenicity Study in Rats

– Final Report. Project Number: 14392/10. Unpublished study prepared by Stillmeadow, Inc., 28 pages.

B. Environmental Protection Agency Risk Assessment Memoranda

U.S. EPA. 2011a. Review of Product Ch emistry, Manufacturing Process, Discussion of Formation of Unintentional Ingredients, Analysis of Samples, Certification of Limits, and Physical and Chemical Characteristics for Section 3 Registration of an End Use Product BW240 G (EPA Reg. No. 68539

- RN). Memorandum from I.S. Barsoum, Ph.D. and J.L. Kough, Ph.D. to J. Kausch dated July 7, 2011.

U.S. EPA. 20 11 b

. Environmental Risk Assessment of the FIFRA Section 3 registration of Trichoderma virens strain G

- 41 (PC Code 176604 ; EPA File Symbols 68539

- I,

- O,

- RN). Memorandum from S. Borges to J. Kausch dated November 14, 2011.

U.S. EPA. 2011c. Review of Product Chemistry, Manufacturing Process, Discussion of Formation of Unintentional Ingredients, Analysis of Samples, Certification of Limits, Physical and Chemical Characteristics, and Acute Toxicity Studies for Section 3 Registration of an MUP G

- 41 Technical (EPA Reg. No. 68539

- I) and an EP BW240 WP (EPA Reg. No. 68539

- O). Memorandum from I.S. Barsoum, Ph.D. and J.L. Kough, Ph.D. to J. Kausch dated Decembe r 2011.

C. Other References

Bjornsen HR. 2009. Cover Letter Accompanying Application for Pesticide Registration (re: Application to Register BW240 G). Letter from H.R. Bjornsen to S. Reilly dated September 18, 2009.

Brookman D. 2010. Supplement Product Chemistry of BW240 Granules. Unpublished study prepared by Technology Sciences Group, Inc., 16 pages (No MRID No.).

Centers for Disease Control and Prevention. 2009. Drinking Water

- Water Treatment. Available from http://www.cdc.gov/healthywater/drinking/public/water_treatment.html

.
Kenerley CM. 2010. Report on studies comparing two strains of Trichoderma virens ; GL21 and G41. Texas A&M University, Depar tment of Plant Pathology and Microbiology. Unpublished report.

Leandro LFS, Guzman T, Ferguson LM, Fernandez GE, Louws FJ. 2007. Population dynamics of Trichoderma in fumigated and compost

- amended soil and on strawberry roots. Appl

. Soil Ecol . 35 :237

– 246.Trichoderma virens strain G

- 41 Page 23 of 34 Biopes ticides Registration Action Document

Liu B, Glenn D, Buckley K. 2008. Trichoderma communities in soils from organic, sustainable, and conventional farms, and their relation with Southern blight of tomato. Soil Biol

. Biochem . 40:1124

– 1136.

Longa CMO, Savazzini F, Tosi S, Elad Y, Pertot I. 2009. Evaluating the survival and environmental fate of the Biocontrol agent Trichoderma atroviride SC1 in vineyards in northern Italy. J. Appl

. Microbiol . 106:1549

– 1557.

Lumsden RD, Walter JF, Baker CP. 1996. Development of Gliocladiu m virens for damping

- off disease control. Can

. J

. Plant Pathol . 18:463

– 468.

Samuels GJ. 1996. Trichoderma : A review of biology and systematics of the genus. Mycol

. Res

. 100:923

– 935.

Sariah M, Choo CW, Zakaria H, Norihan MS. 2005. Quantification an d characterization of Trichoderma spp. from different ecosystems. Mycopathologia 159:113

– 117.

U.S. EPA. 2008. Trichoderma Species Final Registration Review Decision (signed September 11, 2008). Available from http://www.regulations.gov/#!documentDetail;D=EPA

- HQ

OPP

- 2006

- 0245

- 0008;oldLink=false

.
U.S. EPA. 2004. Primer for Municipal Wastewater Treatment Systems. EPA 832

- R

- 04

- 001. Available f rom http://www.epa.gov/npdes/pubs/primer.pdf

.
USPTO. 2010. Control of Plant Diseases and Enhancing Plant Growth Using a Combination of a Trichoderma virens Species and a Rhizosphere Competent Tr ichoderma harzianum Species (United States Patent Application

– US 2010/0028303 A1). Available from http://www.freepatentsonline.com/20100028303.pdf

.Trichoderma virens strain G

- 41 Page 24 of 34 Biopes ticides Registration Action Document

A PPENDIX A. MI CROBIAL PESTICIDES DATA REQUIREMENTS (40 CFR PART 158

– SUBPART V )

TABLE 1. Product Analysis Data Requirements for the Technical Grade of the Active Ingredient (TGAI) , Trichoderma virens strain G

- 41, and the Manufacturing

- Use Pesticide Product ( MP ) , G

- 4 1 Technical (40 CFR § 158.2 120 )
Harmonized

G uideline N umber Data Requirement Results MRID No. Trichoderma virens strain G

- 41 G

- 41 Technical

885.1100 Product Identity N/A Submitt ed data fulfill the requirement for product identity. G

- 41 Technical

co ntains 12.10% by weight Trichoderma virens strain G

- 41.

478651

- 01 482120

- 01 885.1200 Manufacturing Process Submitted data fulfill the requiremen t for manufacturing process

.

Not applicable Deposition of a Sample in a Nationally Recognized Culture Coll ection Trichoderma virens strain G

- 41 is on deposit with the American Type Culture Collection in Manassas, Virginia under Accession Number ATCC 20906. N/A

885.1300 Discussion of Formation of Unintentional

Ingredients Submitted data fulfill the requiremen t for discussion of formation of unintentional ingredients

.

885.1400 Analysis of S amples Submitted data fulfill the requirement for analysis of samples. 478651

- 01

8 85 .1 50 0 Certification of
L imits N/A Limits listed o n the confidential statement of formul a are adequate/a cceptable. 478651

- 01 482120

- 01Trichoderma virens strain G

- 41 Page 25 of 34 Biopes ticides Registration Action Document

TABLE 2. Product Analysis Data Requirements for the End

- Use Pesticide Product ( E P s ), BW240 WP Biological Fungicide and BW240 G Biological Fungicide (40 CFR § 158.2120)
Harmonized

Gui deline Number Data Requirement Results MRID No.

BW240 WP* BW240 G**

885.1100 Product Identity Submitted data fulfill the requirement for product identity. BW240 WP contains 1.15% by weight Trichoderma harzianum Rifai strain T

- 22 and 0.61% by weight Tr ichoderma
virens strain G

- 41. Submitted data fulfill the requirement for product identity. BW240 G contains 1.15% by weight Trichoderma harzianum Rifai strain T

- 22 and 0.61% by weight Trichoderma
virens strain G

- 41. 478649

- 01* * Brookman (2010)* * 478650

- 01* 482104

- 01*

885.1200 Manufacturing Process Submitted data fulfill the requirement for manufacturing

process. 478649

- 01** 478650

- 01* 482104

- 01*

N/A Deposition of a Sample in a Nationally Recognized

Culture Collection N/A N/A

885.1300 Discussion of Format ion of Unintentional Ingredients Submitted data fulfill the requirement for discussion of

formation of unintentional ingredients. Submitted data fulfill the requirement for discussion of formation of unintentional ingredients for purposes of FIFRA section 3(c)(5) registration. As a term of the BW240 G registration, EPA is requiring additional information (confirmatory) on this data requirement. 478649

- 01** 478650

- 01*

885.1400 Analysis of Samples Submitted date fulfill the requirement for analysis of sample s

. Submitted data fulfill the requirement for discussion of formation of unintentional ingredients for purposes of FIFRA section 3(c)(5) registration. As a term of the BW240 G registration, EPA is requiring additional information (confirmatory) on this dat a requirement. 478649

- 01** 478650

- 01*

885.1500 Certification of Limits Limits listed on the confidential statement of formula are adequate/acceptable. 478649

- 01** Brookman (2010)** 478650

- 01* 482104

- 01*Trichoderma virens strain G

- 41 Page 26 of 34 Biopes ticides Registration Action Document

TABLE 3

. Physical and Chemical Characteristics fo r the Technical Grade of the Active Ingredient (TGAI), Trichoderma virens strain G

- 41, and the Manufacturing

- Use Pesticide Product (MP), G

- 41 Technical (40 CFR § 158.2120)
Harmonized G uideline N umber Data Requirement Results MRID No.

Trichoderma viren s strain G

- 41 G

- 41 Technical

830.6302 1 Color Gray

- green

478651

- 01 830.6303 1 Physical State Solid 830.6304 1 Odor Slight musty odor

830.6313 1 Stability to Normal and
E levated Temperatures, Metals, and Metal I ons Unstable at elevated temperatures; s table to metals or metal ions (iron metal, aluminum metal, iron acetate, and aluminum acetate).

830.6317 Storage Stability Stable at 4°C and 21°C for 6 months. As a term of the G

- 41 Technical registration, EPA is requiring the results of a

one - year stor age stability and corrosion characteristics study be submitted within one year of registration

.

830.6319 Miscibility N/A Not required because G

- 41 Technical is not an

e mulsifiable liquid form of a microbial pesticide (refer to test note #2 of 40 CFR § 15 8.2120(d)). N/A

830.6320 Corrosion Characteristics Not corrosive to polyethylene bag in a six

- month study. As a term of the G

- 41 Technical registration, EPA is requiring the results of a one

- year storage stability and corrosion characteristics study be s ubmitted within one year of registration

. 478651

- 01

830.7000 1 pH 5.6 (1% solution)

830.7100 Viscosity N/A Not required because G

- 41 Technical is not a liquid form of a microbial pesticide (refer to test note #4 of 40 CFR § 158.2120(d)). N/A

830.7300 1 Density/Relative Density/Bulk Density

(Specific Gravity) 0.178 g/cm 3 478651

- 01

1 According to 40 CFR § 158.2120, these data are only required for the technical grade of the active ingredient. Since BioWorks

,

Inc. included this information with its appl ication for G

- 41, it is summarized appropriately in this table.Trichoderma virens strain G

- 41 Page 27 of 34 Biopes ticides Registration Action Document

TABLE 4. Physical and Chemical Characteristics for the End

- Use Pesticide Product s (EP s ), BW240 WP Biological Fungicide and BW240 G Biological Fungicide (40 CFR § 158.2120)
Ha rmonized Guideline Number Data Requirement Results MRID No. BW240 WP* BW240 G** 830.6302 Color N/A N/A 830.6303 Physical State N/A N/A 830.6304 Odor N/A N/A

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

830 .6317 Storage Stability Stable at 4°C and 21°C for 6 months

. As terms of the BW240 WP and BW240 G registration s , EPA is requiring the results of a one - year storage stability and corrosion characteristics study be submitted within one year of registration

. 478649

- 01** 478650

- 01*

830.6319 Miscibility Not required because the end

- use pesticide products, BW240 WP and BW240 G, are not emulsifiable liquid forms of microbial pesticides (refer to test note #2 of 40 CFR § 158.2120(d)). N/A

830.6320 Corrosion Chara cteristics Not corrosive to polyethylene bag in a six

- month study

. As term s of the BW240 WP and BW240 G registration s , EPA is requiring the results of a one year storage stability and corrosion characteristics study be submitted within one year of registra tion

. 478649

- 01** 478650

- 01*

830.7000 1 pH 4.35 (1% solution) N/A 482104

- 01*

830.7100 Viscosity Not required because the end

- use pesticide products, BW240 WP and BW240 G, are not liquid forms of microbial pesticides (refer to test note #4 of 40 CFR § 158. 2120(d)). N/A

830.7300 1 Density/Relative Density/Bulk Density

(Specific Gravity) 0.147 g/cm 3 N/A 2

1 According to 40 CFR § 158.2120, these data are only required for the technical grade of the active ingredient. Since BioWorks,

Inc. included this info rmation with its application for BW240 WP and/or BW240 G

, it is summarized appropriately in this table

. 2 Obtained from the confidential statements of formula for the end

- use pesticide products.Trichoderma virens strain G

- 41 Page 28 of 34 Biopes ticides Registration Action Document

T ABLE 5

. Toxicology Data Requirements for the Technical Gra de of the Active Ingredient (TGAI) , Trichoderma virens strain G

- 41, and the Manufacturing

- Use Pesticide Product (MP) , G

- 41 Technical (40 CFR § 158.2140) Harmonized

Guideline N umber Data Requirement Results MRID N o.

Trichoderma virens strain G

- 41 G

- 41 T echnical

Tier I

885.3050 Acute Oral Toxicity/Pathogenicity Not toxic to rats when administered by oral gavage in a single dose of 1.5 x 10 8 cfu per animal. Although a pattern of pathogenicity was established, pathogenicity was not unequivocally assessed given a sensitivity of detection that demonstrated low recovery of the MPCA from tissues and fluid (0.5

– 9%). Classification: Acceptable TOXICITY CATEGORY IV N/A 483438

- 01

885.3150 Acute Pulmonary Toxicity/Pathogenicity Waived based on the results of MRI D Nos. 478651

- 02 and 482368

- 01 (acute injection toxicity/pathogenicity study ) ; MRID No. 478650

- 04 ( acute inhalation toxicity study ); and MRID Nos. 407198

- 05 and 408640

- 02 (acute pulmo nary toxicity/pathogenicity study ). Classification: Acceptable N/A 47 8708

- 01 482526

- 01

885.3200 Acute Injection Toxicity/Pathogenicity (Intra peritoneal ) N ot toxic, infective, and/or pathogenic

to rats when administered intraperitoneally in a single dose of 10 7 cfu per animal

. Classification: Acceptable N/A 478651

- 02 48236 8

- 01

885.3400 Hypersensitivity Incidents No hypersensitivity incidents, including immediate

- type or delayed

- type reactions of humans and domestic animals that occurred durin g research, development, or testing of the TGAI or MP , were reported by the applic ant. Any future hypersensitivity incidents must be reported to EPA (refer to test note #3 of 40 CFR § 158.2140(d))

. 482526

- 01

885.3500 Cell Culture Not required because Trichoderma
virens strain G

- 41 is not a virus (refer to test note #4 of 40 CFR § 158. 2140(d)). N/A N/A

870.1100 Acute Oral Toxicity N/A Waived based on the results of MRID No. 4 83438

- 01 and because this formulation contains inert ingredients that are not expected to be of toxicological

conce rn. Classification: Acceptable TOXICITY CATEGORY III 478651

- 03

870.1200 Acute Dermal Toxicity N/A Waived because this formulation contains inert ingredients that are not expected to be of toxicological conce rn. Classification: Acceptable TOXICITY CATEGORY III 478651

- 03Trichoderma virens strain G

- 41 Page 29 of 34 Biopes ticides Registration Action Document

T ABLE 5

. Toxicology Data Requirements for the Technical Gra de of the Active Ingredient (TGAI) , Trichoderma virens strain G

- 41, and the Manufacturing

- Use Pesticide Product (MP) , G

- 41 Technical (40 CFR § 158.2140) Harmonized

Guideline N umber Data Requirement Results MRID N o.

Trichoderma virens strain G

- 41 G

- 41 T echnical

870.1300 Acute Inhalation Toxici ty N/A Waived based on the results of MRID No s

. 478651

- 02, 482368

- 01, 478650

- 04, 407198

- 05, and 408640

- 02 and because this formulation contains inert ingredients that are not expected to be of toxicological conce rn. Classification: Acceptable TOXICITY CATE GORY III 478651

- 03

870.2400 Acute Eye Irritation N/A Waived because this formulation contains inert ingredients that are not expected to be of toxicological conce rn. Classification: Acceptable TOXICITY CATEGORY III 478651

- 03

870.2500 Primary Dermal Irrit ation N/A Waived because this formulation contains inert ingredients that are not expected to be of toxicological conce rn. Classification: Acceptable TOXICITY CATEGORY III 478651

- 03

Tiers II and III Not required for Trichoderma virens strain G

- 41 based o n the lack of acute toxicity/pathogenicity in the Tier I studies.

Additional Studies

885.3050 Acute Oral Toxicity/Pathogenicity Not toxic to, infective in, or pathogenic for rats when given a single oral dose of 10 8 cfu/animal ( Trichoderma virens

stra in GL

- 21). Classification: Acceptable but supplemental in this situation (test organism is not the MPCA proposed for registration). N/A 407198

- 04

885.3150 Acute Pulmonary Toxicity/Pathogenicity Not toxic to, infective in, or pathogenic for rats when given a single intratracheal dose of 10 8 cfu/animal ( Trichoderma virens strain GL

- 21). Classification: Acceptable but supplemental in this situation (test organism is not the MPCA proposed for registration). N/A 407198

- 05 408640

- 02Trichoderma virens strain G

- 41 Page 30 of 34 Biopes ticides Registration Action Document

T ABLE 5

. Toxicology Data Requirements for the Technical Gra de of the Active Ingredient (TGAI) , Trichoderma virens strain G

- 41, and the Manufacturing

- Use Pesticide Product (MP) , G

- 41 Technical (40 CFR § 158.2140) Harmonized

Guideline N umber Data Requirement Results MRID N o.

Trichoderma virens strain G

- 41 G

- 41 T echnical

885.3200 Acute Injection Tox icity/Pathogenicity (Intravenous) Not infective in or pathogenic for rats when given a single intravenous dose of 0.1 gram (g)/animal ( Trichoderma
virens strain GL

- 21). The data supports the conclusion, however, that the test material is acutely toxic and lethal for test animals via mechanical clogging of capillaries. Classification: Acceptable but supplemental in this situation (test organism is not the MPCA proposed for registration). N/A 407198

- 06 408640

- 01

TABLE 6

. Toxicology Data Requirements for t he End

- Use Pesticide Product s (EPs) , BW240 WP Biological Fungicide and BW240 G Biological Fungicide (40 CFR § 158.2140)
Harmonized

Guideline Number Data Requirement Results MRID No. BW240 WP * BW240 G **

885.3050 Acute Oral Toxicity/Pathogenicity N/A N /A

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

885.3200 Acute Injection Toxicity/Pathogenicity N/A N/A

885.3400 Hypersensitivity Incidents No hypersensitivity incidents, including immediate

- type or delayed

- type reactions of humans and domest ic animals that occurred during research, development, or testing of the EP s , were reported by the applicant. Any future hypersensitivity incidents must be reported to EPA (refer to test note #3 of 40 CFR § 158.2140(d)). 482104

- 02* 481964

- 02 **

885.3500 Ce ll Culture N/A N/A

870.1100 Acute Oral Toxicity Oral LD 50 female rats > 5,000 mg/kg Classification: Acceptable TOXICITY CATEGORY I V Waived based on the results of MRID No. 478650

- 02, which evaluated a substance with identical active ingredients (1.15%
Tri choderma harzianum Rifai strain
T

- 22 and 0.61% Trichoderma virens strain G

- 41) and nearly identical inert ingredients to BW240 G. Classification: Acceptable TOXICITY CATEGORY I V 478650

- 02 * Bjornsen (2009)**

870.1200 Acute Dermal Toxicity Dermal LD 50 combi ned (male and female) rats > 5,050 mg/kg Classification: Acceptable TOXICITY CATEGORY I V Waived based on the results of MRID No. 478650

- 03, which evaluated a substance with identical active ingredients (1.15% Trichoderma harzianum Rifai strain
T

- 22 and 0.6 1% Trichoderma virens strain G

- 41) and nearly identical inert ingredients to BW240 G. Classification: Acceptable TOXICITY CATEGORY I V 478650

- 03* Bjornsen (2009)**Trichoderma virens strain G

- 41 Page 31 of 34 Biopes ticides Registration Action Document

TABLE 6

. Toxicology Data Requirements for t he End

- Use Pesticide Product s (EPs) , BW240 WP Biological Fungicide and BW240 G Biological Fungicide (40 CFR § 158.2140)
Harmonized

Guideline Number Data Requirement Results MRID No. BW240 WP * BW240 G **

870.1300 Acute Inhalation Toxicity 4

- Hour Inhalation LC 50 combined (male and female) rats > 5.14 mg/L Classification: Acceptable TOXICITY CATEGORY I V Waived based on the results of MRID No. 478650

- 04, which evaluated a substance with identical active ingredients (1.15% Trichoderma harzianum Rifai strain
T

- 22 and 0.61% Trichoderma virens strain G

- 41) and nearly identical inert ingredients to BW240 G. Classification: Acceptable TOXICITY CATEGORY I V 478650

- 04* Bjornsen (2009)**

870.2400 Acute Eye Irritation BW240 WP was minimally irritating to the eye of rabbits based on the maximum irritation score of 18 obtained 24 hours after

treatment. P ositive effects cleared by Day 4

. Classification: Acceptable TOXICITY CATEGORY I II Waived based on the results of MRID No. 478650

- 0 5 , which evaluated a substance with identical active ingredients (1.15% Trichoderm a harzianum Rifai strain
T

- 22 and 0.61% Trichoderma virens strain G

- 41) and nearly identical inert ingredients to BW240 G. Classification: Acceptable TOXICITY CATEGORY I II 478650

- 05* Bjornsen (2009)**

870.2500 Primary Dermal Irritation BW240 WP was non

- ir ritating to the skin of rabbits. Classification: Acceptable TOXICITY CATEGORY I V Waived based on the results of MRID No. 478650

- 0 6 , which evaluated a substance with identical active ingredients (1.15% Trichoderma harzianum Rifai strain
T

- 22 and 0.61% Trich oderma virens strain G

- 41) and nearly identical inert ingredients to BW240 G. Classification: Acceptable TOXICITY CATEGORY IV 478650

- 06* Bjornsen (2009)**Trichoderma virens strain G

- 41 Page 32 of 34 Biopes ticides Registration Action Document

TABLE 7

. Nontarget Organism Toxicity and Environmental Fate Data Requirements for the Technical Grade of the Active Ingredient (TGAI), Trichoderma virens strain G

- 41 (40 CFR § 158.2150)
Harmonized Guideline Number Data Requirement Results MRID No.

Tier I

885.4050 Avian Oral Toxicity Data and other information provide sufficient inf ormation to determine that toxicity/pathogenicity to avian wildlife is not expected as a result of the proposed pesticidal

uses. Classification: Acceptable 478651

- 03 482526

- 01

885.4100 Avian Inhalation Toxicity/Pathogenicity Not required as the nature of the microbial pesticide does not indicate potential pathogenicity to birds or relatedness to any known bird pathogens (refer to test note #3 of 40 CFR § 158.2150(e)). N/A

885.4150 Wild Mammal Toxicity/Pathogenicity

Data and other information provide suff icient information to determine that toxicity/pathogenicity to wild mammals, freshwater and marine/estuarine fish and invertebrates, honey bees, nontarget insects, and nontarget plants is not expected as a result of the proposed pesticidal uses. Classifica tion: Acceptable 478651

- 03 482526

- 01 885.4200 Freshwater Fish Toxicity/Pathogenicity

885.4240 Freshwater Invertebrate Toxicity/Pathogenicity

885.4280 Estuarine/Marine Fish and Invertebrate Testing

885.4300 Nontarget Plant Testing

885.4340 Nont arget Insect Testing

885.4380 Honey Bee Testing
Tiers II, III, and IV Not required for Trichoderma virens strain G

- 41 based on the acceptability of the data and other information provided for Tier I

.Trichoderma virens strain G

- 41 Page 33 of 34 Biopes ticides Registration Action Document

APPENDIX B. PESTICIDE PROD UCTS

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

68539

- I G

- 41 Technical 12.10% 3 Technical N/A N/A N/A N/A

68539

- O BW240 WP Biological Fungicide 1.15 % 2
0.61% 3 End Use

– Wettable Powder Various agricultural and greenhouse crops and residential plants

(e.g., vegetables, ornamentals, and fruit) Chemigation (agricultural and

commercial applications) 1

– 32 ounces BW240WP per 100 gallons of water

Various fun gal pests Cutting, bulb, or bare

root transplant dip (agricultural and

commercial applications) (1) 0.25

– 5 pounds of BW240WP per 20 gallons of

water

(2) Direct dip into BW240 WP powder

Soil drench (agricultural and

commercial applications) (1) Shallow beds or pots (up to 4

- inch depth)

– 50

– 100 gallons of prepared mixture 1 per 800 square feet of beds or pots

(2) Deep beds or pots (greater than 4

- inch depth)

– 100 gallons of prepared mixture 1 per 400 square feet of beds or pots or 1/2

– 1 cup o f prepared mixture 1 per 3

– 6 inch diameter pot

In

- furrow spray or transplant starter solution (agricultural and

commercial applications) 1

– 32 ounces of BW240 WP per acre of soil

Bulb, tuber, or cut potato seed piece

treatment (agricultural an d

commercial applications) (1) 0.03

– 3 ounces of BW240 WP per 100 pounds of bulbs, tubers, or cut potato seed pieces

(2) Dip into suspension of 0.25

– 5 pounds of BW240 WP per 20 gallons of water

Seed drench (residential applications) Prepared mixtur e (1

– 3 tablespoons of BW240 WP per gallon of water) to 25 feet of planting furrow

Transplants or potted

plants (residential

applications) 1/2

– 1 cup of prepared mixture 1 per transplant or per 4

– 8 inch diameter pot
New and established plant be ds (residential applications) 1 gallon of prepared mixture 1 per 25 square feet of plant bedTrichoderma virens strain G

- 41 Page 34 of 34 Biopes ticides Registration Action Document

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

68539

- RN BW240 G Biological Fungicide 1.15% 2
0.61% 3 End Use

– Granular Various agricultural and greenhouse crops

(e.g., vegetables, ornamentals, and fruit) In

- fur row soil

treatment 5

– 12 pounds of BW240 G per acre of soil Various fungal pests Planting mix amendment (greenhouse and nursery applications) 1

– 3 pounds of BW240 G per cubic yard of planting mix

Bed incorporation (nursery applications) 0.5

– 7 .5 pounds of BW240 G per 1,000 square feet of bed Transplant 0.2

– 8 ounces of BW240 G per planting hole

Broadcast (turf only) (1) New turf seedlings

– 1

– 4 pounds of BW240 G per square feet of turf or 40

– 120 pounds of BW240 G per acre of turf

(2) Established turf

– 1

– 3 pounds of BW240 G per 1,000 square feet or turf or 40

– 120 pounds of BW240 G per acre of turf

1 Prepared mixture = BW240 WP + water

2 Trichoderma harzianum Rifai strain T

- 22

3 Trichoderma virens strain G

- 41