Federal Food, Drug, and Cosmetic Act (FFDCA) Considerations for Trichoderma asperelloides strain 

JM41R

Docket ID Number: EPA-HQ-OPP-2012-0963

Date: April 20, 2015

Section 408(c)(2)(A)(i) of FFDCA allows the U.S. Environmental Protection Agency (EPA) to establish an 

exemption from the requirement for a tolerance (the legal limit for a pesticide chemical residue in or on a 

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

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

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

there is reliable information.” This includes exposure through drinking water and in residential settings but 

does not include occupational exposure. Pursuant to FFDCA section 408(c)(2)(B), in establishing or 

maintaining in effect an exemption from the requirement of a tolerance, the EPA must take into account the 

factors set forth in FFDCA section 408(b)(2)(C), which require the EPA to give special consideration to 

exposure of infants and children to the pesticide chemical residue in establishing a tolerance or tolerance 

exemption, and to “ensure that there is a reasonable certainty that no harm will result to infants and children 

from aggregate exposure to the pesticide chemical residue . . . .” Additionally, FFDCA section

408(b)(2)(D) requires that the EPA consider “available information concerning the cumulative effects of [a 

particular pesticide’s] . . . residues and other substances that have a common mechanism of toxicity.”

The EPA performs a number of analyses to determine the risks from aggregate exposure to pesticide 

residues. First, the EPA determines the toxicity of the pesticide. Second, the EPA examines exposure to the 

pesticide through food, drinking water, and other exposures that occur as a result of pesticide use in 

residential settings.

I. Summary of Petitioned-for Tolerance Exemption

In the Federal Register of February 21, 2014 (79 FR 9870), the EPA issued a notice pursuant to FFDCA 

section 408(d)(3), 21 U.S.C. 346a(d)(3), announcing the filing of a pesticide tolerance petition (PP 2F8102) 

by BASF Corporation, 26 Davis Dr., Research Triangle Park, NC 27709. The petition requested that 40 

CFR part 180 be amended by establishing an exemption from the requirement of a tolerance for residues of 

Trichoderma fertile strain JM41R in or on all food commodities. That document referenced a summary of 

the petition prepared by the petitioner BASF Corporation, which is available in Docket ID Number EPAHQ-OPP-2012-0963 via http://www.regulations.gov.

Subsequently, the petitioner provided additional data (i.e., DNA sequence work based on newly published 

taxonomies) on the identity of the pesticide to the EPA. After reviewing these data, the EPA concluded that 

the correct identity of the pesticide was Trichoderma asperelloides strain JM41R and not Trichoderma 

fertile strain JM41R. In order to give the public an opportunity to comment on this new information, the 

EPA republished its receipt of this tolerance exemption petition filing with an updated and accurate 

description in the Federal Register of January 28, 2015 (80 FR 4525) and placed a revised petition from 

BASF Corporation into the docket.

II. Toxicological Profile

Consistent with FFDCA section 408(b)(2)(D), the EPA reviewed the available scientific data and other 

relevant information on Trichoderma asperelloides strain JM41R, and considered its validity, 

completeness, and reliability, as well as the relationship of this information to human risk. The EPA also 

considered available information concerning the variability of the sensitivities of major identifiable 

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subgroups of consumers, including infants and children.

A. Overview of Trichoderma asperelloides strain JM41R

Fungi of the genus Trichoderma are abundant in nature, especially in soil and close to plant roots (Refs. 1, 

2, 3, and 4). Since 1989, several Trichoderma species (e.g., Trichoderma polysporum and Trichoderma

viride) have been used in pesticide products—notably without reported incidents—to control various fungal 

plant pathogens such as Pythium species, Phytophthora species, Heterobasidion annosum, and 

Chondrostereum purpureum. In conjunction with the registration of some of these pesticide products, the

EPA established the following tolerance exemptions:

(1) Gliocladium virens isolate GL-21 (now recognized as Trichoderma virens isolate GL-21) (40 CFR 

180.1100) – see the Federal Register of September 20, 1995 (60 FR 48657) and October 5, 1995 (60 

FR 52248).

(2) Trichoderma harzianum Rifai strain T-22 (40 CFR 180.1102) – see the Federal Register of April 7,

1999 (64 FR 16856).

(3) Trichoderma harzianum strain T-39 (40 CFR 180.1201) – see the Federal Register of June 22, 2000 

(65 FR 38753).

(4) Trichoderma gamsii strain ICC 080 (40 CFR 180.1293) – see the Federal Register of February 25, 

2010 (75 FR 8504).

(5) Trichoderma asperellum strain ICC 012 (40 CFR 180.1294) – see the Federal Register of March 3, 

2010 (75 FR 9527).

(6) Trichoderma hamatum isolate 382 (40 CFR 180.1298) – see the Federal Register of July 23, 2010 

(75 FR 43072). 

(7) Trichoderma virens strain G-41 (40 CFR 180.1310) – see the Federal Register of February 1, 2012 

(77 FR 4903).

In 2003, a University of Pretoria student isolated Trichoderma asperelloides strain JM41R in South Africa. 

This particular fungus colonizes root surfaces close to inoculated plants, parasitizing and competitively 

displacing invasive fungi (e.g., Rhizoctonia species and Fusarium species) and mineralizing insoluble 

phosphate to create a form of this substance that is available to plants. In an October 2012 submission to 

the EPA, the petitioner proposed to register an end-use pesticide product that contains Trichoderma fertile 

(now Trichoderma asperelloides) strain JM41R as an active ingredient and that is to be applied to soil and 

growing media in commercial greenhouses to manage soilborne diseases caused by organisms such as 

Rhizoctonia species and Fusarium species.

B. Microbial Pesticide Toxicology Data Requirements

All applicable mammalian toxicology data requirements supporting the request for an exemption from the 

requirement of a tolerance for residues of Trichoderma asperelloides strain JM41R in or on all food 

commodities have been fulfilled with data submitted by the petitioner or data waiver requests that have 

been granted by the EPA.

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The toxicity tests (acute oral, dermal, and inhalation toxicity) and irritation tests (acute eye and primary 

dermal irritation) that address potential routes of exposure to the active ingredient are all classified in 

Toxicity Category III or IV (see section II.B. of Ref. 5) and reveal little to no toxicity or irritation attributed 

to Trichoderma asperelloides strain JM41R. Moreover, during a typical toxicity/pathogenicity test done 

with microbial pesticides, Trichoderma asperelloides strain JM41R did not show any signs of toxicity, 

pathogenicity, or infectivity via the pulmonary route of exposure. Finally, the petitioner reported that, to 

date, no hypersensitivity incidents have occurred during research, development, and testing of this fungus. 

The conclusions/classifications from all toxicological information submitted by the petitioner are briefly 

described below. In-depth synopses of the study results can be found in either the associated toxicological 

risk assessment or the draft Biopesticides Registration Action Document provided as references in Section 

VII of this document (Refs. 2 and 6).

1. Acute oral toxicity – rat (Harmonized Guideline 870.1100; Master Record Identification Number (MRID 

No.) 489476-03). An acceptable acute oral toxicity study demonstrated that Trichoderma asperelloides

strain JM41R is not toxic to female rats when dosed at 5,000 milligrams per kilogram (mg/kg) of body 

weight. The oral median lethal dose (LD50), which is a statistically derived single dose that can be expected 

to cause death in 50% of test animals, is greater than 5,000 mg/kg for female rats (Toxicity Category IV)

(Refs. 2 and 6).

2. Acute oral toxicity/pathogenicity (Harmonized Guideline 885.3050; MRID No. 489476-09). Upon 

consideration of the petitioner’s scientific rationale that cited to results of other toxicological data and 

public literature, the EPA waived acute oral toxicity/pathogenicity testing for Trichoderma asperelloides

strain JM41R. An acute oral toxicity study conducted on female rats (MRID No. 489476-03) demonstrated 

that Trichoderma asperelloides strain JM41R is not toxic, while an acute pulmonary toxicity/pathogenicity 

study conducted on rats (MRID No. 489476-02) showed that Trichoderma asperelloides strain JM41R is

not toxic, pathogenic, or infective. Further, Trichoderma asperelloides strain JM41R does not grow at or 

above 35°C (normal human body temperature is 37°C). Finally, although a literature search yielded two 

reports of infections in humans from Trichoderma species, the fungi involved were Trichoderma 

longibranchiatum or Trichoderma harzianum and not Trichoderma asperelloides, and the infections were 

likely due to immune system dysfunction, invasive procedures involved with therapy for the overlying 

disease, and/or immunosuppressive drugs. The EPA believes these data and information, when taken 

together, indicate that this fungus would not be toxic, pathogenic, or infective through the oral route of 

exposure and that further testing is not necessary (Refs. 2 and 6). 

3. Acute inhalation toxicity – rat (Harmonized Guideline 870.1300; MRID No. 489476-05). A supplemental 

acute inhalation toxicity study demonstrated that Trichoderma asperelloides strain JM41R is not toxic to 

male and female rats when exposed to 2.34 milligrams per liter (mg/L). The inhalation median lethal 

concentration (LC50), which is a statistically derived concentration that can be expected to cause death in 

50% of test animals, is greater than 2.34 mg/L for both male and female rats combined (Toxicity Category 

IV). The EPA rated this study as supplemental because the 6.5-micrometer (μm) particle size used for 

testing was outside the acceptable guideline range of 1–4 μm needed for adequate lung penetration. Even 

with this deficiency, however, the EPA still considered these data useful for risk assessment purposes

(Refs. 2 and 6). 

4. Acute pulmonary toxicity/pathogenicity – rat (Harmonized Guideline 885.3150; MRID No. 489476-02).

A supplemental acute pulmonary toxicity/pathogenicity study demonstrated that Trichoderma asperelloides

strain JM41R is not toxic, pathogenic, or infective when administered by intratracheal instillation in a 

single dose of 4.7 x 107 colony-forming units (CFU) per animal. The EPA rated this study as supplemental 

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because clearance was not definitively established and the 4.7 x 107 CFU per animal dose used was below 

the acceptable guideline dose of 108 CFU per animal. Even with these deficiencies, however, the EPA still 

concluded that Trichoderma asperelloides strain JM41R showed a general pattern of clearance from lung 

tissues and considered these data useful for risk assessment purposes (Refs. 2 and 6). 

5. Acute injection toxicity/pathogenicity (Harmonized Guideline 885.3200; MRID No. 489476-09). Upon 

consideration of the petitioner’s scientific rationale that cited to results of other toxicological data and 

public literature, the EPA waived acute injection toxicity/pathogenicity testing for Trichoderma 

asperelloides strain JM41R. An acute oral toxicity study conducted on female rats (MRID No. 489476-03) 

and an acute inhalation toxicity study conducted on male and female rats (MRID No. 489476-05) 

demonstrated that Trichoderma asperelloides strain JM41R is not toxic, while an acute pulmonary 

toxicity/pathogenicity study conducted on rats (MRID No. 489476-02) showed that Trichoderma 

asperelloides strain JM41R is not toxic, pathogenic, or infective. Further, Trichoderma asperelloides strain 

JM41R does not grow at or above 35°C (normal human body temperature is 37°C). Finally, although a 

literature search yielded two reports of infections in humans from Trichoderma species, the fungi involved 

were Trichoderma longibranchiatum or Trichoderma harzianum and not Trichoderma asperelloides, and 

the infections were likely due to immune system dysfunction, invasive procedures involved with therapy 

for the overlying disease, and/or immunosuppressive drugs. The EPA believes these data and information, 

when taken together, indicate that this fungus would not be toxic, pathogenic, or infective through the 

injection route of exposure (when the skin is bypassed as a barrier) and that further testing is not necessary

(Refs. 2 and 6).

6. Hypersensitivity incidents (Harmonized Guideline 885.3400; MRID No. 489476-09). The petitioner 

reported that, to date, no hypersensitivity incidents, including immediate-type or delayed-type reactions of 

humans and domestic animals, have occurred during research, development, or testing of Trichoderma 

asperelloides strain JM41R (Ref. 6).

7. Cell culture (Harmonized Guideline 885.3500; MRID No. 489476-09). Not required because 

Trichoderma asperelloides strain JM41R is not a virus (refer to test note #4 of 40 CFR § 158.2140(d))

(Ref. 6).

8. Acute dermal toxicity – rat (Harmonized Guideline 870.1200; MRID No. 489476-04). An acceptable 

acute dermal toxicity study demonstrated that Trichoderma asperelloides strain JM41R is not toxic to male 

and female rats when dosed at 5,050 mg/kg of body weight for 24 hours. The dermal LD50 is greater than 

5,050 mg/kg for both male and female rats combined (Toxicity Category IV) (Refs. 2 and 6). 

9. Primary dermal irritation – rabbit (Harmonized Guideline 870.2500; MRID No. 489476-07). An 

acceptable primary dermal irritation study demonstrated that Trichoderma asperelloides strain JM41R is 

mildly irritating to the skin of rabbits (all animals free of slight dermal erythema without edema by 48 

hours; Toxicity Category IV) (Refs. 2 and 6). 

10. Acute eye irritation – rabbit (Harmonized Guideline 870.2400; MRID No. 489476-06). An acceptable 

acute eye irritation study demonstrated that Trichoderma asperelloides strain JM41R is mildly irritating to 

the eyes of rabbits (all animals free of conjunctival irritation (e.g., redness) by 72 hours; Toxicity Category 

III) (Refs. 2 and 6).

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III. Aggregate Exposure

In examining aggregate exposure, FFDCA section 408 directs the EPA to consider available information

concerning exposures from the pesticide residue in food and all other non-occupational exposures, 

including drinking water from ground water or surface water and exposure through pesticide use in 

gardens, lawns, or buildings (residential and other indoor uses).

Food Exposure: Exposure to residues of Trichoderma species, fungi commonly found in agricultural 

settings (i.e., soils and plant roots) and on fresh produce (Refs. 1, 2, 3, and 4), in consumed food is possible.

For optimal management of the target pests (i.e., soilborne diseases caused by organisms such as 

Rhizoctonia species and Fusarium species), Trichoderma asperelloides strain JM41R needs to be applied to 

soil and growing media in commercial greenhouses in a manner that facilitates the fungus’ movement into 

the root zones of plants. This requires that end users take certain actions, depending on the treatment type, 

that would inevitably minimize the amount of Trichoderma asperelloides strain JM41R residues on aboveground commodities. That is, although Trichoderma asperelloides strain JM41R can be applied to soil, 

pesticide applications made to the soil include enough water or are followed immediately by irrigation to 

properly incorporate Trichoderma asperelloides strain JM41R into the soil. As Trichoderma asperelloides 

strain JM41R colonizes the soil or growing media in close proximity to plant roots or the root surfaces of 

plants, it is possible that food commodities that develop underground (e.g., root and tuber vegetables) or 

adjacent to the soil/ground (e.g., cucumber) could be directly exposed to Trichoderma asperelloides strain 

JM41R. Although residues from use of pesticides containing Trichoderma asperelloides strain JM41R may 

be present on certain food commodities, processing procedures (e.g., peeling and washing) may reduce 

such residues (Ref. 7). Finally, since Trichoderma species can be endophytes (i.e., organisms that live 

within plants) (Ref. 8), it is conceivable that Trichoderma asperelloides strain JM41R could be present 

within food commodities. Should Trichoderma asperelloides strain JM41R be present in or on food, 

supporting toxicological data and information indicate that no toxicity, pathogenicity, or infectivity is likely 

to occur with this type of exposure resulting from the use of this microbial pesticide when applied in 

accordance with label directions and good agricultural practices (see additional discussion in Section II of 

this document). 

Drinking Water Exposure: Trichoderma asperelloides strain JM41R is to be applied to soil or growing 

media in commercial greenhouses and is not intended for use in drinking water. Once applied, the risk of 

Trichoderma asperelloides strain JM41R passing through the soil to ground water is minimal to unlikely 

because the growing organism forms a cottony web that is stationary, and the spores are not the type (i.e., 

chlamydospores) that are able to survive unfavorable conditions such as water (Ref. 9). If Trichoderma 

asperelloides strain JM41R were to be transferred to surface or ground waters, it likely would not survive 

the conditions water is subjected to in wastewater treatment systems and drinking facilities (e.g., pH 

adjustments and chlorination) (Refs. 10 and 11). Should Trichoderma asperelloides strain JM41R be

present in drinking water, supporting toxicological data and information indicate that no toxicity, 

pathogenicity, or infectivity is likely to occur with this type of exposure resulting from the use of this 

microbial pesticide when applied in accordance with label directions and good agricultural practices (see 

additional discussion in Section II of this document).

Other Non-occupational Exposure: Dermal or inhalation exposure to the general population due to use of 

Trichoderma asperelloides strain JM41R is not likely to occur, based on product labeling that limits the use 

site to commercial greenhouses. Even if additional non-occupational exposures were to occur (e.g., through 

eventual expansion of use sites), no adverse effects attributed to Trichoderma asperelloides strain JM41R

are anticipated, particularly in light of testing that indicates that this fungus is not toxic (acute inhalation 

toxicity, acute dermal toxicity, and acute pulmonary toxicity/pathogenicity tests), is only mildly irritating 

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(primary dermal irritation test), and is not pathogenic or infective (acute pulmonary toxicity/pathogenicity 

test) at high doses (see additional discussion in Section II of this document).

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

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

a tolerance, the EPA consider “available information concerning the cumulative effects of [a particular 

pesticide’s] . . . residues and other substances that have a common mechanism of toxicity.”

Trichoderma asperelloides strain JM41R is not toxic via the oral, dermal, or inhalation routes of exposure.

Thus, for the purposes of this tolerance action, the EPA has concluded that Trichoderma asperelloides

strain JM41R does not have a common mechanism of toxicity with other substances. Following from this, 

therefore, the EPA concludes that there are no cumulative effects associated with Trichoderma 

asperelloides strain JM41R that need to be considered. 

For information regarding the EPA’s efforts to determine chemicals that have a common mechanism of 

toxicity and to evaluate the cumulative effects of such chemicals, see the EPA’s website at

http://www.epa.gov/pesticides/cumulative.

V. Determination of Safety for the U.S. Population, Infants and Children

A. U.S. Population

For all of the reasons discussed previously, there is reasonable certainty that no harm will result to the U.S. 

population, including infants and children, from aggregate exposure to residues of Trichoderma 

asperelloides strain JM41R. This includes all anticipated dietary exposures and all other exposures for 

which there is reliable information.

B. Infants and Children

FFDCA section 408(b)(2)(C) provides that the 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 the EPA determines based on reliable 

data 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, the 

EPA either retains the default value of 10X or uses a different additional safety factor when reliable data 

available to the EPA support the choice of a different factor. Humans, including infants and children, have

likely been exposed and continue to be exposed to Trichoderma asperelloides through soil and fresh 

produce, where it is commonly found, with no known or reported adverse effects. As discussed above, the 

EPA has concluded that Trichoderma asperelloides strain JM41R is not toxic, pathogenic, or infective to 

mammals, including infants and children. Because there are no threshold levels of concern to infants, 

children, and adults when Trichoderma asperelloides strain JM41R is used according to label directions 

and good agricultural practices, the EPA concludes that no additional margin of safety is necessary to 

protect infants and children.

VI. Conclusions

The EPA concludes that there is a reasonable certainty that no harm will result to the U.S. population, 

including infants and children, from aggregate exposure to residues of Trichoderma asperelloides strain 

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JM41R. Therefore, an exemption from the requirement of a tolerance is established for residues of 

Trichoderma asperelloides strain JM41R in or on all food commodities when used in accordance with label 

directions and good agricultural practices.

VII. References

1. Druzhinina IS, Kopchinskiy AG, Kubicek CP. 2006. The first 100 Trichoderma species characterized by 

molecular data. Mycoscience 47:55–64. Available from http://isth.info/library/pdf/61/isth_library_61.pdf. 

2. U.S. EPA. 2015a. Trichoderma asperelloides strain JM41R. Memorandum from J.V. Gagliardi, Ph.D. 

through J.L. Kough, Ph.D. to J. Kausch, dated January 21, 2015.

3. U.S. EPA. 2015b. Environmental risk assessment of Trichoderma asperelloides strain JM41R for a 

FIFRA Section 3 registration for use in greenhouses. Memorandum from S. Borges to J. Kausch, dated 

February 18, 2015.

4. Harman GE, Howell CR, Viterbo A, Chet I, Lorito M. 2004. Trichoderma species – opportunistic, 

avirulent plant symbionts. Nature Reviews Microbiology 2:43–56. Available from 

http://naldc.nal.usda.gov/download/25508/PDF. 

5. U.S. EPA. 2014. Chapter 7 of the Label Review Manual (Precautionary Statements) (Revised July 2014). 

Available from http://www2.epa.gov/sites/production/files/2014-

07/documents/chapter7_revised_final_0714.pdf. 

6. U.S. EPA. 2015c. Draft Trichoderma asperelloides strain JM41R Biopesticides Registration Action 

Document (Dated April 8, 2015) (available as a “Supporting Document” within docket ID number EPAHQ-OPP-2012-0962 at http://www.regulations.gov).

7. U.S. EPA. 1996. Microbial Pesticide Test Guidelines – Background for Residue Analysis of Microbial 

Pest Control Agents (OPPTS 885.2000) (Dated February 1996). Available from 

http://www.epa.gov/ocspp/pubs/frs/publications/Test_Guidelines/series885.htm. 

8. Chaverri P, Gazis RO, Samuels GJ. 2011. Trichoderma amazonicum, a new endophytic species on 

Hevea brasiliensis and H. guianensis from the Amazon basin. Mycologia 103(1):139–151. Available from 

http://www.mycologia.org/content/103/1/139.full. 

9. Chaverri P, Castlebury LA, Overton BE, Samuels GJ. 2003. Hypocrea/Trichoderma: species with 

conidiophore elongations and green conidia. Mycologia 95(6):1100–1140. Available from 

http://www.mycologia.org/content/95/6/1100.full.

10. Centers for Disease Control and Prevention. 2015. Drinking Water - Water Treatment. Available from 

http://www.cdc.gov/healthywater/drinking/public/water_treatment.html.

11. 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. 

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