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

February 6, 2012
Trichoderma virens strain G-41 Page 2 of 34

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. 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 JUSTICE.................................................................................................................17

VI. RISK MANAGEMENT DECISION..........................................................................................................17

VII. ACTIONS REQUIRED OF THE REGISTRANT....................................................................................18

A. Final Printed Labeling ...................................................................................................................18

B. Terms of Registration.....................................................................................................................18

C. Reporting of Adverse Effects and Hypersensitivity Incidents....................................................18

VIII. GLOSSARY OF ACRONYMS AND ABBREVIATIONS ......................................................................19

IX. BIBLIOGRAPHY .......................................................................................................................................20

A. Studies Submitted or Cited to Support the Trichoderma virens strain G-41 Pesticide 

 Product Registrations.....................................................................................................................20

B. Environmental Protection Agency Risk Assessment Memoranda .............................................22

C. Other References............................................................................................................................22

APPENDIX A. MICROBIAL PESTICIDES DATA REQUIREMENTS............................................................24

APPENDIX B. PESTICIDE PRODUCTS..............................................................................................................33
Trichoderma virens strain G-41 Page 3 of 34

Biopesticides 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 Protection Agency

John L. Kough, Ph.D. Product Analysis, Human Health

Shannon Borges Environmental Effects

Leila Heikkila Human 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 Leader
Trichoderma virens strain G-41 Page 4 of 34

Biopesticides Registration Action Document

I. EXECUTIVE SUMMARY

In September 2009, BioWorks, Inc. submitted applications 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 both 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

applications.

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 G41, the new active ingredient contained in all three proposed pesticide products, with EPA only. 

Trichoderma virens, including strain G-41, 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 (Samuels 1996). Trichoderma virens strain G-41 was isolated 

from soil samples taken from Aphanomyces-suppressive fields in Livingston County, New York. 

Much like other Trichoderma species, Trichoderma virens strain G-41 inhibits or kills certain 

plant-pathogenic fungi (e.g., Rhizoctonia species and Fusarium species) through several 

mechanisms: (1) competition for food and space, (2) mycoparasitism, (3) antibiosis, and (4)

induction of plant defense responses (Kenerley 2010, Samuels 1996). Given the fungicidal 

capabilities of Trichoderma virens strain G-41, BioWorks, Inc. proposed to register these

pesticide products to control soilborne plant pathogens and plant root diseases that adversely 

affect agricultural, greenhouse, and nursery crops, as well as 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 158.2150, 

respectively) submitted to support the registration of the three 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 requirements, and allow for 

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

(FIFRA).

Most of the product analysis data requirements for Trichoderma virens strain G-41, including 

product chemistry and composition, analysis of samples, and physical and chemical

characteristics, were fulfilled by acceptable guideline studies. Although a one-year storage 

stability and corrosion characteristics study is outstanding, EPA reviewed an 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 mammalian toxicology data were submitted or cited to support the Trichoderma virens

strain G-41 pesticide products. An acute injection toxicity/pathogenicity study showed that, at a 

single high dose, Trichoderma virens strain G-41 is not toxic, infective, and/or pathogenic via the 

Toxicology

 1 See http://www.epa.gov/oppfead1/international/naftatwg/guidance/jointreview-biope.pdf.
Trichoderma virens strain G-41 Page 5 of 34

Biopesticides Registration Action Document

intraperitoneal route of exposure, while two acute oral toxicity/pathogenicity studies (one 

conducted with Trichoderma virens strain G-41 and the other conducted with functionally similar 

Trichoderma virens strain GL-21 (formerly known as Gliocladium virens strain GL-21)) indicate

that, at a single high dose, Trichoderma virens strain G-41 is not toxic and/or pathogenic via the 

oral route of exposure.2 Acute pulmonary toxicity/pathogenicity testing demonstrated that 

Trichoderma virens strain GL-21, a functionally similar strain to Trichoderma virens strain G-41, 

was not toxic to, infective in, or pathogenic for rats when given a single intratracheal dose. Given 

the functional similarity of these two strains, EPA concluded that Trichoderma virens strain G-41 

is also not likely to be toxic, infective, and/or pathogenic through the inhalation route. Moreover, 

no hypersensitivity incidents, occurring during research, development, or testing of Trichoderma 

virens strain G-41, were reported by the applicant. In light of the results of the injection, 

pulmonary, and oral toxicity/pathogenicity data, as well as the absence of hypersensitivity 

incidents, testing at higher tiers (i.e., Tiers II and III) was not required.

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 Trichoderma virens strain 

G-41. No dietary risks are expected from use of Trichoderma virens 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 methods of the end-use pesticide products (soil 

directed and soil incorporated; no aquatic applications); (2) the filtering effect of many 

particulate soil types; and (3) the conditions (e.g., filtration and pH adjustments) water is 

subjected to in wastewater treatment systems and drinking water facilities. Should this microbial 

pesticide be present on food or in drinking water, the acute oral toxicity and pathogenicity data

available for Trichoderma virens strain G-41 and functionally similar Trichoderma virens strain 

GL-21 demonstrated that 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.

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, protective eyewear, and a dust/mist filtering respirator meeting National 

Institute for Occupational Safety and Health (NIOSH) standards of at least N-95, R-95, or P-95. 

EPA may require different PPE, other than the standard described above, on a product-specific 

basis. 

Occupational Exposure

Data and other information (e.g., scientific literature) submitted by the applicant to support 

Nontarget Organisms

 2 With its pesticide product applications, BioWorks, Inc. cited to toxicological data done with a similar, previously 

registered strain of Trichoderma virens, GL-21 (Kenerley 2010). Although GL-21 and G-41 are not identical, the 

two strains share many characteristics typical of Trichoderma virens (e.g., particular morphological features and 

weak growth at the temperature of the human body (37°C)), and thus are considered to be functionally similar. Based 

on these similarities, EPA concluded that data on Trichoderma virens strain GL-21 would be representative of the 

toxicological nature of Trichoderma virens strain G-41. 
Trichoderma virens strain G-41 Page 6 of 34

Biopesticides Registration Action Document

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 environmental risk assessment based on data and other information 

provided by the applicant, and determined 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 

designated critical habitats resulting from these same proposed pesticidal uses. 

On October 1, 2009, EPA announced a new policy to provide a more meaningful opportunity for 

the public to participate in 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 applications: new active ingredients; first food uses; first 

outdoor uses; first residential uses; or any 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 

pesticide products containing Trichoderma virens strain G-41, a new active ingredient, and 

allowing for this active ingredient’s first outdoor, food, and residential uses were subject to a 30-

day comment period. During this comment period, no comments were received on EPA’s 

preliminary decision to register the Trichoderma virens strain G-41 pesticide products, G-41 

Technical, BW240 WP Biological Fungicide, and BW240 G Biological Fungicide. Therefore, 

EPA maintained that, based upon the risk assessment and information submitted in support of 

registration of such pesticide products, it was appropriate to issue the G-41 Technical, BW240 

WP Biological Fungicide, and BW240 G Biological Fungicide registrations under FIFRA section 

3(c)(5). The basis for this decision can be found in the risk assessment for Trichoderma virens 

strain G-41, which is characterized throughout this Biopesticides Registration Action Document 

(BRAD). 

II. ACTIVE INGREDIENT OVERVIEW

Biological Name: Trichoderma virens strain G-41

Culture Deposit: American Type Culture Collection in Manassas, Virginia 

under Accession Number ATCC 20906

OPP Chemical Code: 176604

Type of Pesticide: Microbial Pesticide – Fungicide

See Appendix B for specific information (e.g.,

use sites, application rates, methods of application, 

formulation types, and target pests) regarding

the registered pesticide products containing this 

active ingredient.
Trichoderma virens strain G-41 Page 7 of 34

Biopesticides Registration Action Document

III. REGULATORY BACKGROUND

A. Applications for Pesticide Product Registration

On September 18, 2009, Technology Sciences Group, Inc. (address: 1150 18th Street NW, Suite 

1000; Washington, District of Columbia 20036), on behalf of BioWorks, Inc. (address: 100 

Rawson Road, Suite 205; Victor, New York 14564), submitted applications to register a

manufacturing-use pesticide product, G-41 Technical (EPA File Symbol 68539-I), and two enduse pesticide products, BW240 WP Biological Fungicide (EPA File Symbol 68539-O) and 

BW240 G Biological Fungicide (EPA File Symbol 68539-RN), under FIFRA section 3. On 

March 10, 2010, EPA announced receipt of these applications to register pesticide products

containing a new active ingredient (75 Federal Register (FR) 11175) and opened a 30-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 applications and under Federal Food, Drug, and Cosmetic Act 

(FFDCA) section 408(d), Technology Sciences Group, Inc., on behalf 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) 9F7618). 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 strain G-41, in or on all food commodities and 

opened a 30-day comment period. No comments were received following this publication. 

On February 1, 2012, EPA established an exemption from the requirement of a tolerance for 

residues of Trichoderma virens strain G-41, in or on all food commodities, when applied as a 

fungicide and used in accordance with good agricultural practices (40 CFR § 180.1310; 77 FR 

4903).

IV. RISK ASSESSMENT

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

support 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 information 

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 documents 

for risk assessment. A rating of “acceptable” indicates the study is scientifically sound and is 

useful for risk assessment. A “supplemental” rating indicates the data provide some information 

that can be useful for risk assessment. The studies may have certain aspects determined not to be 
Trichoderma virens strain G-41 Page 8 of 34

Biopesticides Registration Action Document

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 “acceptable.” If there is simply a “supplemental” rating, the reviewer will 

often state that the study is not required by 40 CFR Part 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 acute toxicity data requirements, Toxicity Categories are assigned based on the 

hazard(s) identified from studies and/or other information 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 characteristics study, which EPA 

will require as a term of registration, all 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-specific information. 

B. Human Health Assessment (40 CFR § 158.2140)

1. Toxicity

All toxicology 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 studies 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 additional product-specific data submitted to support the individual 

registrations, refer to Tables 5 and 6 in Appendix A.

a. Acute Toxicity/Pathogenicity – Tier I

Acute Oral Toxicity/Pathogenicity – Rat (Harmonized Guideline 885.3050; Master Record 

Identification Number (MRID No.) 483438-01): 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 109 colony-forming units per milliliter (cfu/mL)) in 

Phosphate Buffered Saline at doses of 1.5 x 108 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 group 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 control 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 from all tissues and organs by 
Trichoderma virens strain G-41 Page 9 of 34

Biopesticides Registration Action Document

Day 7; however, the sensitivity of detection indicated low recovery of the microbial pest control 

agent (MPCA) from tissues and fluids (0.5–9%). Clearance 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 single 

dose of 1.5 x 108 cfu per animal. This study was rated acceptable.

Acute Oral Toxicity/Pathogenicity – Rat (Harmonized Guideline 885.3050; MRID No. 407198-

04): In a 14-day acute oral toxicity/pathogenicity study in Sprague-Dawley rats (13 per sex) with 

Trichoderma virens strain GL-21, there were no mortalities, and no signs of 

infectivity/pathogenicity were observed following oral administration at 108 cfu in 2.0% 

carboxymethyl cellulose per animal in a maximum dosing volume of 5 milliliters. General signs 

of toxicity (lethargy, rapid breathing) were observed 1 hour post treatment, but all signs had 

resolved by Day 1. There were no observable abnormalities upon necropsy, and all animals 

gained weight throughout the study. Interim sacrifices (Days 1, 7 and 14) were carried out to 

analyze for the presence of the microbe in kidney, lungs, liver, brain, spleen, mesenteric lymph 

nodes, and blood as a clearance assessment of the test organism. The test organism was detected 

in the feces on Day 1 and in low levels in the lungs (one animal) on Day 7 only; the test organism 

had cleared from all organs/tissue by Day 14. Trichoderma virens strain GL-21 is not toxic to, 

infective in, or pathogenic for rats when administered in a single oral dose at 108 cfu per animal. 

This study was originally rated acceptable but is supplemental in this situation (test organism is 

not the active ingredient proposed for registration).

Acute Pulmonary Toxicity/Pathogenicity (Harmonized Guideline 885.3150; MRID Nos. 407198-

04 and 408640-02): In a 21-day acute pulmonary toxicity/pathogenicity study in rats with 

Trichoderma virens strain GL-21, no mortality, toxicity, or pathogenicity was observed following 

intratracheal administration at 108 cfu per animal. General signs of toxicity (poor weight gain; 

lethargy; rapid breathing; rough hair coat) were observed after dosing, but all signs had resolved 

by Day 4. Interim sacrifices (Days 1, 7 and 14) were carried out to analyze for the presence of the 

microbe in kidney, lungs, liver, brain, spleen, mesenteric lymph nodes, and blood as a clearance 

assessment of the test organism. The test organism was steadily cleared from the lungs of test 

animals after 2–3 weeks. For other organs/tissues, the test organism was detected on Day 1 in 

low numbers in the liver, blood, mesenteric lymph nodes, spleen, kidney, and brain. The test 

organism had cleared in all organs/tissues by Day 7, except for one animal that had low levels of 

the organism in the spleen on Day 14. Upon necropsy, treated animals appeared normal except 

for the appearance of grey spots in the lungs at each interim sacrifice. The peritracheal lymph 

nodes of dosed animals often appeared enlarged and pale. This can most likely be attributed to 

the administration of the test substance and probably represents the test material itself.

Trichoderma virens strain GL-21 is not toxic to, infective in, or pathogenic for rats when 

administered in a single intratracheal dose at 108 cfu per animal. This study was originally rated 

acceptable but is supplemental in this situation (test organism is not the active ingredient 

proposed for registration).

Acute Injection Toxicity/Pathogenicity (Intraperitoneal) – Rat (Harmonized Guideline 885.3200; 

MRID Nos. 478651-02 and 482368-01): In an acute intraperitoneal infectivity study, young adult 

Wistar rats were injected with approximately 107 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 water, 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 
Trichoderma virens strain G-41 Page 10 of 34

Biopesticides Registration Action Document

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 as 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 this 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 

virens strain G-41 is not toxic, infective, and/or pathogenic to rats when administered 

intraperitoneally in a single dose of 107 cfu per animal. This study was rated acceptable.

 

Hypersensitivity Incidents (Harmonized Guideline 885.3400; MRID No. 482526-01): 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 hypersensitivity 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 CFR § 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 required 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 Endocrine 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 endocrine 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 determine 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 potential 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 Page 11 of 34

Biopesticides Registration Action Document

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 

all 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/.

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 residue in or on a food) only if EPA 

determines that the exemption is “safe.” Section 408(c)(2)(A)(ii) of FFDCA defines “safe” to 

mean that “there is a reasonable certainty that no harm will result from aggregate exposure to the

pesticide chemical residue, including all anticipated dietary exposures and all other exposures for 

which there is reliable information.” This includes exposure through drinking water and in 

residential settings, but does not include occupational exposure. Pursuant to 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....” Additionally, 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 information discussed previously and 

presented in Tables 5 and 6 in Appendix A, the 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).
Trichoderma virens strain G-41 Page 12 of 34

Biopesticides Registration Action Document

Food Exposure and Risk Characterization: All proposed Trichoderma virens strain G-41 

applications are soil directed or soil incorporated because of the targeted soilborne pests (e.g., 

Rhizoctonia spp. and Fusarium spp.). Based on calculations made in EPA’s environmental risk 

assessment, 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 of Trichoderma virens; however, Trichoderma species have been reported 

in various types of soils at concentrations of 104 to 106 colony-forming units per gram (cfu/g) 

(Leandro et al. 2007, Liu et al. 2008). Based on the maximum application rate of the proposed 

end-use pesticide products containing Trichoderma virens strain G-41, the estimated amount of 

Trichoderma virens applied to the soil surface is approximately 6.7 x 103 colony-forming units 

per square centimeter (cfu/cm2). Assuming a bulk density of 1 to 2 grams per cubic centimeter 

(g/cm3), 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 overall 

increased exposure to Trichoderma virens in the terrestrial environment, including on aboveground plant parts such as food commodities, is not expected. Work by Jackson et al. (1991) 

supports this conclusion given that, after Trichoderma virens and three other Trichoderma

isolates were incorporated into soil, fungal numbers either transiently increased, remained stable, 

or declined. 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 drinking 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 exposure 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). If Trichoderma virens strain G-41 were to be transferred to surface or ground 

waters (e.g., through spray drift or runoff) that are intended for eventual human consumption and 

directed to wastewater treatment systems or drinking water facilities, it likely would not survive 

the conditions water is subjected to in such systems or facilities, including chlorination, pH 

adjustments, filtration, and occasionally high temperatures (Centers for Disease Control and 

Prevention 2009, U.S. EPA 2004). For instance, Trichoderma virens strain G-41 does not grow 

well at 37°C (Kenerley 2010, Lumsden et al. 1996), and test data has shown it to be unstable at 

elevated temperatures; therefore, any heat treatment applied to water containing Trichoderma 

virens strain G-41 would probably render the fungus non-viable. 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 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).
Trichoderma virens strain G-41 Page 13 of 34

Biopesticides Registration Action Document

Non-occupational, Residential Risk Characterization: Given Trichoderma virens’ natural 

occurrence in soil (Samuels 1996), non-occupational exposure to the fungus is likely already

occurring. Even 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 relative stability of 

typical background levels in the soil (see calculations and information presented in the food 

exposure section above). If significant non-occupational exposures were to occur, such exposures 

would not exceed EPA’s level of concern in light of test results that indicated Trichoderma 

virens strain G-41 is not toxic (acute pulmonary toxicity/pathogenicity, acute inhalation toxicity, 

and acute dermal toxicity), is non-irritating (primary dermal irritation), and is not pathogenic or 

infective (acute pulmonary toxicity/pathogenicity) (see section IV(B)(1)(a) and Tables 5 and 6 in 

Appendix A).

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

Section 408(b)(2)(D)(v) of FFDCA requires that, when considering whether to establish, modify, 

or revoke a tolerance exemption, EPA consider “available information concerning the cumulative 

effects of [a particular pesticide’s] . . . residues and other substances that have a common 

mechanism of toxicity.”

There are several Trichoderma species used as active ingredients in registered pesticide products. 

While these different microbial pest control agents may produce similar metabolites, the 

likelihood of adverse cumulative effects via a common mechanism of toxicity is not anticipated,

based on the lack of toxicity/pathogenicity potential of the active ingredients used on food and/or 

labeled for residential uses (U.S. EPA 2008, 2010a, 2010b, and 2010c). For information 

regarding EPA’s efforts to determine which chemicals have a common mechanism of toxicity 

and to evaluate the cumulative effects of 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, in considering the establishment of a tolerance or 

tolerance exemption for a pesticide chemical residue, 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 Tables 5 and 6 in Appendix A, as well as use of Trichoderma pesticide products 

since 1989 without reported adverse effects to humans, EPA concludes that there are no 

threshold effects of concern to infants, children, or adults when Trichoderma virens strain G-41 

is used as labeled in accordance with good agricultural practices. As a result, EPA concludes that 
Trichoderma virens strain G-41 Page 14 of 34

Biopesticides Registration Action Document

no additional margin of exposure (safety) is necessary to protect infants and children and that not 

adding any additional margin of exposure (safety) will be safe for infants and children.

Moreover, based on the same data and EPA analysis as presented directly above, the Agency 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

Trichoderma virens strain G-41 when it is used as labeled and in accordance with good 

agricultural practices as a fungicide. 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/or infective potential to mammals, including infants and 

children. 

3. Occupational Exposure and Risk Characterization

Handler exposure to Trichoderma virens strain G-41 is not expected to pose any undue risk. 

Regardless, 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 strain G-41 end-use pesticide

products in agricultural settings must wear a long-sleeved shirt, long pants, socks, shoes,

protective eyewear, and a dust/mist filtering respirator meeting NIOSH standards of at least N95, R-95, or P-95. EPA may require different PPE, other than the standard described above, 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 relevant safety factors in FFDCA. A determination has been made 

that no unreasonable adverse effects to the United States population in general, and to infants and 

children in particular, will result when Trichoderma virens strain G-41 pesticide products are 

used in accordance with EPA-approved labeling.

C. Environmental Assessment (40 CFR § 158.2150)

Data and other information (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 environmental risk assessment based on data and other information

provided by the applicant, and has determined 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 

designated 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.
Trichoderma virens strain G-41 Page 15 of 34

Biopesticides Registration Action Document

1. Ecological Exposure and Risk 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 information submitted by BioWorks, 

Inc. to support its waiver 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 avian 

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 fungus 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 does not grow at 39oC or above (Kenerley 

2010). Kenerley (2010) also provides data to show 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 adversely affect 

wild mammals, since no toxicity or pathogenicity was shown in laboratory rats exposed to G-41 

Technical by injection at 107 units of MPCA per animal (submitted in MRID Nos. 478651-02 

and 482368-01). Based on this information, adverse effects to wild mammals and birds are not 

anticipated to result from the proposed labeled uses of Trichoderma virens strain G-41.

Nontarget Insects (Harmonized Guideline 885.4340) and Honey Bees (Harmonized Guideline 

885.4380) (MRID Nos.478651-03 and 482526-01): BioWorks, Inc. presented similar 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 naturally 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 world, 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 insects. 

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 Trichoderma virens strain G-41 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 adverse 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 

(Samuels 1996). 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 the literature describing 

natural concentrations of Trichoderma virens; however, Trichoderma spp. have been reported in 
Trichoderma virens strain G-41 Page 16 of 34

Biopesticides Registration Action Document

various types of soils at concentrations of 104 to 106 cfu/g (Leandro et al. 2007, Liu et al. 2008). 

Based on the maximum application rate of the end-use pesticide products containing 

Trichoderma virens strain G-41, and its reported minimum viability in the end-use pesticide 

products (5 x 106 cfu/g), the estimated amount of Trichoderma virens applied to the soil surface 

is approximately 6.7 x 103 cfu/cm2

. Assuming a soil bulk density of 1 to 2 g/cm3, 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 terrestrial environment is not expected. Trichoderma virens is also not

related to known plant pathogens. Therefore, 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 Invertebrates (Harmonized Guideline 

885.4280), and Nontarget Plants (Harmonized Guideline 885.4300) (MRID Nos. 478651-03 and 

482526-01): BioWorks, Inc. presented similar data and information to support waiver requests 

and show that adverse effects to freshwater 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 also provided for aquatic plants. Trichoderma virens is native to the United 

States and is widely distributed, inhabiting forest, agricultural and orchard soils, as well as plant 

litter (Samuels 1996). Given the ubiquitous distribution of Trichoderma virens in soil, this 

microbe is very likely washed into freshwater and marine/estuarine 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 virens 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 

Trichoderma 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 exposure 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 the data and information provided are sufficient to fulfill the Tier I nontarget organism data 

requirements and allow for nontarget organism risk assessment for Trichoderma virens strain G41, further testing at higher tier levels (i.e., Tiers II, III, and IV) is not required. 

3. Threatened and Endangered Species Assessment

Since EPA has determined that no effects are anticipated for any nontarget species exposed to 

Trichoderma virens strain G-41 as a result of the proposed applications, effects to threatened and 

endangered species and their designated critical habitats 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 
Trichoderma virens strain G-41 Page 17 of 34

Biopesticides Registration Action Document

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, 

implementation, and enforcement of environmental laws, regulations, and policies. Fair treatment 

means that no group of people, including racial, ethnic, or socioeconomic groups, should bear a 

disproportionate share of the negative environmental consequences resulting from industrial, 

municipal, and commercial operations or the execution of federal, state, local, and tribal 

environmental programs and policies. Meaningful involvement means that (1) potentially 

affected community residents have an appropriate opportunity to participate in decisions about a 

proposed activity that will affect their environment and/or health; (2) the public’s contribution 

can influence the regulatory agency’s decision; (3) the concerns of all participants involved will 

be considered in the decision-making process; and (4) the decision-makers 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, during the 30-day public participation 

comment period, EPA sought information on any groups or segments of the population who, as a 

result their location, cultural practices, or other factors, may have atypical, unusually high 

exposure to Trichoderma virens strain G-41, compared to the general population. No public 

comments were received on this particular matter.

For additional information regarding environmental justice issues, please visit EPA’s web site at 

http://www.epa.gov/compliance/environmentaljustice/index.html.

VI. RISK MANAGEMENT DECISION

Section 3(c)(5) of FIFRA permits for the registration of a pesticide provided that all the 

following determinations are made:

(1) Its composition is such as to warrant the proposed claims for it;

(2) Its labeling and other material required to be submitted comply with the

 requirements of FIFRA;

(3) It will perform its intended function without unreasonable adverse effects on 

 the 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 labels, and such pesticide 

products are not expected to cause unreasonable adverse effects on the environment when used 

according to their respective label instructions. Criterion 2 is satisfied by the current product
Trichoderma virens strain G-41 Page 18 of 34

Biopesticides Registration Action Document

labels, 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 products), in particular, are likely 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 label 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 

uses. 

VII. 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.

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 storage stability 

(Harmonized Guideline 830.6317) and corrosion characteristics (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 discussion of formation of unintentional 

ingredients (Harmonized Guideline 885.1300) and analysis of samples (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

Biopesticides Registration Action Document

VIII. GLOSSARY OF ACRONYMS AND ABBREVIATIONS

BRAD Biopesticides Registration Action Document

CFR Code of Federal Regulations

cfu colony-forming unit(s)

cfu/cm2 colony-forming units per square centimeter

cfu/g colony-forming units per gram

cfu/mL colony-forming units per milliliter

EDSP Endocrine Disruptor Screening Program

EP end-use pesticide product

EPA United States Environmental Protection Agency (the “Agency”)

FFDCA Federal Food, Drug, and Cosmetic Act

FIFRA Federal Insecticide, Fungicide, and Rodenticide Act

FR Federal Register

g gram

g/cm3 grams per cubic centimeter

LC50 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).

LD50 median lethal dose. A statistically derived single dose that can be expected 

to cause death in 50% of the test animals when administered by the route 

indicated (oral, dermal, or inhalation). It is expressed as a weight of 

substance per unit weight of animal (e.g., mg/kg).

mg/kg milligrams per kilogram

mg/L milligrams per 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 Programs

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 Patent and Trademark Office
Trichoderma virens strain G-41 Page 20 of 34

Biopesticides Registration Action Document

IX. BIBLIOGRAPHY

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/Pathogenicity 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 

Strain 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 Report. 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 Microbiological 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 prepared 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 Inhalation 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

Biopesticides Registration Action Document

MRID No. Study Information

478650-05 Kuhn J. 2009. BW240 WP: Acute Eye Irritation 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. 2009. 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 Tier 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. Supplemental 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 prepared 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 Prakash 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

Biopesticides 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: Acute 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 Chemistry, 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. 2011b. 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 December 20, 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.

Jackson AM, Whipps JM, Lynch JM. 1991. Production, delivery systems, and survival in soil of 

four fungi with biocontrol potential. Enzyme Microb. Tech. 13:636–642.

Kenerley CM. 2010. Report on studies comparing two strains of Trichoderma virens; GL21 and 

G41. Texas A&M University, Department of Plant Pathology and Microbiology. 

Unpublished report.
Trichoderma virens strain G-41 Page 23 of 34

Biopesticides Registration Action Document

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.

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 Gliocladium 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 and characterization of 

Trichoderma spp. from different ecosystems. Mycopathologia 159:113–117.

U.S. EPA. 2010a. Trichoderma asperellum strain ICC 012 Biopesticides Registration Action 

Document dated March 4, 2010 (available as “Supporting & Related Material” within 

docket ID number EPA-HQ-OPP-2009-1004 at http://www.regulations.gov).

U.S. EPA. 2010b. Trichoderma gamsii strain ICC 080 Biopesticides Registration Action 

Document dated March 4, 2010 (available as “Supporting & Related Material” within 

docket ID number EPA-HQ-OPP-2009-1003 at http://www.regulations.gov).

U.S. EPA. 2010c. Trichoderma hamatum isolate 382 Biopesticides Registration Action 

Document dated July 13, 2010 (available as “Supporting & Related Material” within 

docket ID number EPA-HQ-OPP-2010-0489 at http://www.regulations.gov).

U.S. EPA. 2008. Trichoderma Species Final Registration Review Decision (signed September 

11, 2008). Available from http://www.regulations.gov/#!documentDetail;D=EPA-HQOPP-2006-0245-0008;oldLink=false.

U.S. EPA. 2004. Primer for Municipal Wastewater Treatment Systems. EPA 832-R-04-001. 

Available from 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 Trichoderma 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

Biopesticides Registration Action Document

APPENDIX A. MICROBIAL 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-41 Technical (40 CFR § 158.2120)

Harmonized 

Guideline 

Number 

Data Requirement

Results

MRID No.

Trichoderma virens strain G-41 G-41 Technical

885.1100 Product Identity N/A

Submitted data fulfill the requirement

for product identity. G-41 Technical 

contains 12.10% by weight 

Trichoderma virens strain G-41. 

478651-01

482120-01

885.1200 Manufacturing 

Process Submitted data fulfill the requirement for manufacturing process.

Not applicable

Deposition of a 

Sample in a 

Nationally 

Recognized Culture 

Collection

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 requirement for discussion of formation of 

unintentional ingredients.

885.1400 Analysis of Samples Submitted data fulfill the requirement for analysis of samples. 478651-01

885.1500 Certification of 

Limits N/A

Limits listed on the confidential 

statement of formula are 

adequate/acceptable.

478651-01

482120-01
Trichoderma virens strain G-41 Page 25 of 34

Biopesticides Registration Action Document

TABLE 2. Product Analysis Data Requirements for the End-Use Pesticide Product (EPs), BW240 WP Biological Fungicide and 

BW240 G Biological Fungicide (40 CFR § 158.2120)

Harmonized

Guideline 

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 Trichoderma 

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 Formation 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 

samples.

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*

885.1500 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

Biopesticides Registration Action Document

TABLE 3. Physical and Chemical Characteristics for 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 

Guideline Number Data Requirement

Results

MRID No. Trichoderma virens

strain G-41 G-41 Technical

830.63021 Color Gray-green

478651-01

830.63031 Physical State Solid

830.63041 Odor Slight musty odor

830.63131

Stability to Normal and 

Elevated

Temperatures, Metals,

and Metal Ions

Unstable at elevated temperatures; stable 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 storage 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 

emulsifiable liquid form of a 

microbial pesticide (refer to 

test note #2 of 40 CFR § 

158.2120(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 submitted within one year of 

registration.

478651-01

830.70001 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

830.73001

Density/Relative 

Density/Bulk Density 

(Specific Gravity)

0.178 g/cm3 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 application for G-41 Technical, it is summarized appropriately in this table.
Trichoderma virens strain G-41 Page 27 of 34

Biopesticides Registration Action Document

TABLE 4. Physical and Chemical Characteristics for the End-Use Pesticide Products (EPs), BW240 WP Biological Fungicide and 

BW240 G Biological Fungicide (40 CFR § 158.2120)

Harmonized

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 registrations, 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 Characteristics

Not corrosive to polyethylene bag in a six-month study. 

As terms of the BW240 WP and BW240 G 

registrations, EPA is requiring the results of a oneyear storage stability and corrosion characteristics 

study be submitted within one year of registration.

478649-01**

478650-01*

830.70001 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

830.73001

Density/Relative 

Density/Bulk Density 

(Specific Gravity)

0.147 g/cm3 N/A2

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 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

Biopesticides Registration Action Document

TABLE 5. Toxicology Data Requirements for 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 § 158.2140)

Harmonized

Guideline 

Number

Data Requirement

Results

MRID No.

Trichoderma virens strain G-41 G-41 Technical

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 

108 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

Fulfilled based on the results of MRID 

Nos. 407198-05 and 408640-02 (acute 

pulmonary toxicity/pathogenicity 

study) and MRID No. 478650-04 

(acute inhalation toxicity study). 

N/A 478708-01

482526-01

885.3200

Acute Injection 

Toxicity/Pathogenicity 

(Intraperitoneal)

Not toxic, infective, and/or pathogenic 

to rats when administered 

intraperitoneally in a single dose of 107

cfu per animal.

Classification: Acceptable

N/A 478651-02

482368-01

885.3400 Hypersensitivity

Incidents

No hypersensitivity incidents, including immediate-type or delayed-type 

reactions of humans and domestic animals that occurred during research, 

development, or testing of the TGAI or MP, 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)). 

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. 483438-01 and 

because this formulation contains 

inert ingredients that are not 

expected to be of toxicological 

concern.

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 concern.

Classification: Acceptable

TOXICITY CATEGORY III

478651-03
Trichoderma virens strain G-41 Page 29 of 34

Biopesticides Registration Action Document

TABLE 5. Toxicology Data Requirements for 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 § 158.2140)

Harmonized

Guideline 

Number

Data Requirement

Results

MRID No.

Trichoderma virens strain G-41 G-41 Technical

870.1300 Acute Inhalation 

Toxicity N/A

Waived based on the results of 

MRID Nos. 478650-04, 407198-

05, and 408640-02 and because 

this formulation contains inert 

ingredients that are not expected 

to be of toxicological concern.

Classification: Acceptable

TOXICITY CATEGORY 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 concern.

Classification: Acceptable

TOXICITY CATEGORY III

478651-03

870.2500 Primary Dermal 

Irritation N/A

Waived because this formulation 

contains inert ingredients that are 

not expected to be of 

toxicological concern.

Classification: Acceptable

TOXICITY CATEGORY III

478651-03

Tiers II and III

Not required for Trichoderma virens strain G-41 based on 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 108 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 407198-04

885.3150 Acute Pulmonary 

Toxicity/Pathogenicity

Not toxic to, infective in, or pathogenic 

for rats when given a single 

intratracheal dose of 108 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

408640-02
Trichoderma virens strain G-41 Page 30 of 34

Biopesticides Registration Action Document

TABLE 5. Toxicology Data Requirements for 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 § 158.2140)

Harmonized

Guideline 

Number

Data Requirement

Results

MRID No.

Trichoderma virens strain G-41 G-41 Technical

885.3200

Acute Injection 

Toxicity/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 the End-Use Pesticide Products (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 domestic animals that occurred during research, 

development, or testing of the EPs, 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 Cell Culture N/A N/A

870.1100 Acute Oral Toxicity

Oral LD50 female rats > 5,000 

mg/kg

Classification: Acceptable

TOXICITY CATEGORY IV

Waived based on the results of 

MRID No. 478650-02, 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 IV

478650-02*

Bjornsen (2009)**

870.1200 Acute Dermal 

Toxicity

Dermal LD50 combined (male and 

female) rats > 5,050 mg/kg

Classification: Acceptable

TOXICITY CATEGORY IV

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 0.61% Trichoderma virens

strain G-41) and nearly identical inert 

ingredients to BW240 G.

Classification: Acceptable

TOXICITY CATEGORY IV

478650-03*

Bjornsen (2009)**
Trichoderma virens strain G-41 Page 31 of 34

Biopesticides Registration Action Document

TABLE 6. Toxicology Data Requirements for the End-Use Pesticide Products (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 LC50 combined 

(male and female) rats > 5.14 mg/L

Classification: Acceptable

TOXICITY CATEGORY IV

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 IV

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. Positive effects cleared 

by Day 4.

Classification: Acceptable

TOXICITY CATEGORY III

Waived based on the results of 

MRID No. 478650-05, 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 III

478650-05*

Bjornsen (2009)**

870.2500 Primary Dermal 

Irritation

BW240 WP was non-irritating to 

the skin of rabbits.

Classification: Acceptable

TOXICITY CATEGORY IV

Waived based on the results of 

MRID No. 478650-06, 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 IV

478650-06*

Bjornsen (2009)**
Trichoderma virens strain G-41 Page 32 of 34

Biopesticides 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 

information 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 sufficient 

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.

Classification: 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 Nontarget 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

Biopesticides Registration Action Document

APPENDIX B. PESTICIDE PRODUCTS

EPA Reg.

Number

Registration 

Name

Percentage 

Active 

Ingredient

Formulation 

Type

Use Site(s) Method(s) of 

Application

Application Rate(s) Target 

Pest

68539-8 G-41 

Technical 12.10%3 Technical N/A N/A N/A N/A

68539-9

BW240 WP 

Biological 

Fungicide

1.15%2

0.61%3

End Use –

Wettable 

Powder

Various agricultural, 

greenhouse, and 

nursery crops, as well 

as residential plants 

(e.g., vegetables, 

ornamentals, and fruit)

Chemigation 

(agricultural and 

commercial 

applications)

1–32 ounces BW240 WP per 

100 gallons of water

Various 

fungal 

pests

Cutting or bare-rooted

transplant dip 

(agricultural and 

commercial 

applications)

(1) 0.25–5 pounds of BW240

WP 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 mixture1

per 800 square feet of beds or 

pots

(2) Deep beds or pots (greater 

than 4-inch depth) – 100 

gallons of prepared mixture1

per 400 square feet of beds or

pots, 1⁄2 cup of prepared 

mixture1 per 3-inch diameter 

pot, or 1 cup of prepared 

mixture1 per 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 and 

commercial 

applications)

(1) 0.03–3 pounds of BW240 

WP per 100 pounds of bulbs 

or cut potato seed pieces

(2) Dip bulbs, tubers, or cut 

potato seed pieces into 

suspension of 0.25–5 pounds 

of BW240 WP per 20 gallons 

of water 

Seed drench 

(residential 

applications)

Prepared mixture (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 

mixture1 per transplant or per 

4–8 inch diameter pot

New and established 

plant beds (residential 

applications)

1 gallon of prepared mixture1

per 25 square feet of plant bed
Trichoderma virens strain G-41 Page 34 of 34

Biopesticides Registration Action Document

EPA Reg.

Number

Registration 

Name

Percentage 

Active 

Ingredient

Formulation 

Type

Use Site(s) Method(s) of 

Application

Application Rate(s) Target 

Pest

68539-10

BW240 G 

Biological 

Fungicide

1.15%2

0.61%3

End Use –

Granular

Various agricultural, 

greenhouse, and 

nursery crops (e.g., 

vegetables, 

ornamentals, and fruit)

In-furrow soil 

treatment 

2.5–6 pounds of BW240 G 

per 1⁄2 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 seedings – 1–4 

pounds of BW240 G per

1,000 square feet of turf 

(2) Established turf (early 

spring) – 1–4 pounds of 

BW240 G per 1,000 square 

feet of turf 

(3) Established turf (late 

summer/early fall) - 1–2 

pounds of BW240 G per 

1,000 square feet of turf

1 Prepared mixture = BW240 WP + water

2 Trichoderma harzianum Rifai strain T-22

3 Trichoderma virens strain G-41