Federal Food, Drug, and Cosmetic Act (FFDCA) Considerations for Trichoderma asperelloides strain
JM41R

Docket ID Number: EPA

- HQ

- OPP

- 201 2

- 0 963
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 drin king 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 reasonabl e certainty that no harm will result to infants and children from aggregate exposure to th e 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 fo od, 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 . Th at document referenced a summary of the petition prepared by the petitioner BASF Corporation , which is available in D ocket ID Number EPA

HQ

- OPP

- 201 2

- 0963 via http://w ww.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

1subgroups of consumers, including infants and children.

A. Overview of T richoderma asperelloides strain JM41R

Fungi of the genus Trichoderma a re 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 s ome 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 Re gister 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 )

– s ee 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 pho sphate to create a form of this substance that is available to plants

. In a n October 20 12 submission to the EPA, the petitioner proposed to register an end

- use pe sticide 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 a sperelloides 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.

2The toxicity tests (acute oral, dermal, and inhalation toxicity) and irritation te sts (acute eye and primary dermal irritation) that address potential routes of exposure to the active ingredient are all cla ssified 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 toxic ity, pathogenic ity, or infectivity via the pulmonary route of exposure. Finally, the petitioner reported that , to date, no hypersensitivity incidents have occurred during research, developmen t, and testing of this fungus

.

The conclusions /classifications from all toxicological information submitted by the petitio ner are br iefly
described below. I n

- depth synopses of the study results can be found in either the associated toxicological
risk assessment or the draft Biopesticides Registratio n Action Document provided as reference s in Section

VII of this document (Ref s . 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 (LD 50 ), which is a statistically derived single dose that can be expected
to cause d eath in 50% of test animals, is greater than 5,000 mg/kg for female rats (Toxicity Category IV) (R efs. 2 and 6 ).

2

. Acute oral toxicity/pathogenicity (Harmonized Guidel ine 885.3050; MRID No

. 489476

- 09 )

. Upon consideration of the petitioner’s scientific rationale that cited to results of other toxicologic al 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 strai n JM41R is no t toxic , while an acute pulmonary toxicity/pathogenicity study conducted on r ats (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 Trichod erma harzianum and not Trichoderma asperelloides

, and the infect i ons were likely due t o immune system dysfunction, invasive procedures involved with therapy for the overlying disease, and/or immunosuppressive drugs. The EPA believes these data and informat ion, when taken together, indicate that this fungus would not be toxic, pathogenic, or infective through the oral route of

ex posure 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 le thal concentration (LC 50 ), which is a statistically derived concentration that can be expected to cause death in 50% of test animals, is greater tha n 2.34 mg/L for both male and female rats combined (Toxicity Category IV) . The EPA rated this study as suppl emental 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 assess ment purposes (Re fs. 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 10 7 colony

- forming units (CFU) per animal. The EPA rated this study as supplemental

3because clearance was not definitively establ ished and the 4.7 x 10 7 CFU per animal dose used was below the acceptable guideline dose of 10 8 CFU per animal. Even with these d eficiencies, 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 tox icity 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 tempera ture 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 w ere 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 . 4 89476

- 09 ) . The p etitioner 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 c ulture (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 LD 50 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 (Ha rmonized 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 b y 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; Toxic ity Category III) (Refs. 2 and 6 ).

4III. Aggregate Exposure

In examining aggregate exposure, FFDCA section 408 di rects 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 targ et pests (i.e.

, soilborne disea ses 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 gree nhouses in a manne r 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 above

ground 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 immediatel y by irrigation to properly incorpor ate 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, i t is possible that food commodities that develop unde rground (e.g., root and tuber v egetables) or adjacent to the soil/gr ound (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 E xposure : Trichoderma asperelloides strain JM41R is to be applied to soil or growing
media in commercial greenhouse s 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 a s water ( Ref. 9 )

. If Trichoderma asperelloides strain JM41R were to be transf erred 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 a nd 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 expo sure 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 : D ermal or inhalation exposure to the general popula tion due to use of Trichoderma asperelloides strain JM41R is no t 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 toxic ity, and acute pulmonary toxicity/pathogenicity tests), is only mildly irritating

5(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 t his 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, f or 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 cumu lative 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 c hemicals, 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 ant icipated 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 childre n. 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
availa ble 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 r eported 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,

c hildren, 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

6JM41R . Therefore, an exemption from the requirement of a to lerance is established for residues of

T

richoderma 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 Biopesticide s Registration Action Document (D ated April 8 , 2015 ) (available as a “Supporting Document” within docket ID number EPA

HQ

- 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, Overto n 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

.

1 1 . 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

.

7