Document ID: EPA-HQ-OPP-2009-1003-0005
Agency: epa
Document Type: Supporting & Related Material
Title: 
Posted Date: 2009-12-30T05:00Z

BIOPESTICIDES REGISTRATION ACTION DOCUMENT 

 

Trichoderma gamsii strain ICC 080 

PC Code: 119207

U.S. Environmental Protection Agency

Office of Pesticide Programs

Biopesticides and Pollution Prevention Division

Last Updated – December 23, 2009



Table of Contents

  TOC \o "1-3" \h \z \u    HYPERLINK \l "_Toc249333118"  I.  EXECUTIVE
SUMMARY	  PAGEREF _Toc249333118 \h  4  

  HYPERLINK \l "_Toc249333119"  II.  ACTIVE INGREDIENT OVERVIEW	 
PAGEREF _Toc249333119 \h  6  

  HYPERLINK \l "_Toc249333120"  III.  REGULATORY BACKGROUND	  PAGEREF
_Toc249333120 \h  6  

  HYPERLINK \l "_Toc249333121"  IV.  RISK ASSESSMENT	  PAGEREF
_Toc249333121 \h  8  

  HYPERLINK \l "_Toc249333122"  A.  Product Analysis Assessment	 
PAGEREF _Toc249333122 \h  8  

  HYPERLINK \l "_Toc249333123"  1.  Product Chemistry and Composition	 
PAGEREF _Toc249333123 \h  8  

  HYPERLINK \l "_Toc249333124"  2.  Analysis and Certified Limits	 
PAGEREF _Toc249333124 \h  9  

  HYPERLINK \l "_Toc249333125"  3.  Physical and Chemical
Characteristics	  PAGEREF _Toc249333125 \h  9  

  HYPERLINK \l "_Toc249333126"  B.  Human Health Assessment	  PAGEREF
_Toc249333126 \h  9  

  HYPERLINK \l "_Toc249333127"  1.  Toxicology	  PAGEREF _Toc249333127
\h  9  

  HYPERLINK \l "_Toc249333128"  2.  Dietary Exposure and Risk
Characterization	  PAGEREF _Toc249333128 \h  13  

  HYPERLINK \l "_Toc249333129"  3.   Drinking Water Exposure and Risk
Characterization	  PAGEREF _Toc249333129 \h  14  

  HYPERLINK \l "_Toc249333130"  4.  Acute and Chronic Dietary Exposure
and Risks for Sensitive	  PAGEREF _Toc249333130 \h  14  

  HYPERLINK \l "_Toc249333131"  Subpopulations, Particularly Infants and
Children	  PAGEREF _Toc249333131 \h  14  

  HYPERLINK \l "_Toc249333132"  5.  Occupational, Residential, School,
and Daycare Exposure and Risk	  PAGEREF _Toc249333132 \h  15  

  HYPERLINK \l "_Toc249333133"  Characterization	  PAGEREF _Toc249333133
\h  15  

  HYPERLINK \l "_Toc249333134"  6.  Aggregate Exposure from Multiple
Routes Including Dermal, Oral, and	  PAGEREF _Toc249333134 \h  15  

  HYPERLINK \l "_Toc249333135"  Inhalation	  PAGEREF _Toc249333135 \h 
15  

  HYPERLINK \l "_Toc249333136"  7.  Cumulative Effects	  PAGEREF
_Toc249333136 \h  15  

  HYPERLINK \l "_Toc249333137"  8.  Risk Characterization	  PAGEREF
_Toc249333137 \h  16  

  HYPERLINK \l "_Toc249333138"  C.  Environmental Assessment	  PAGEREF
_Toc249333138 \h  16  

  HYPERLINK \l "_Toc249333139"  1.  Summary of Non-target Organism
Testing and Waiver Rationales	  PAGEREF _Toc249333139 \h  16  

  HYPERLINK \l "_Toc249333140"  2.  Environmental Effects Conclusions	 
PAGEREF _Toc249333140 \h  20  

  HYPERLINK \l "_Toc249333141"  3. Threatened and Endangered Species
Assessment	  PAGEREF _Toc249333141 \h  22  

  HYPERLINK \l "_Toc249333142"  V.  ENVIRONMENTAL JUSTICE	  PAGEREF
_Toc249333142 \h  22  

  HYPERLINK \l "_Toc249333143"  VI.   RISK MANAGEMENT AND REGISTRATION
DECISIONS	  PAGEREF _Toc249333143 \h  22  

  HYPERLINK \l "_Toc249333144"  A.  Determination of Eligibility	 
PAGEREF _Toc249333144 \h  22  

  HYPERLINK \l "_Toc249333145"  B.  Regulatory Decision	  PAGEREF
_Toc249333145 \h  23  

  HYPERLINK \l "_Toc249333146"  C.  Labeling	  PAGEREF _Toc249333146 \h 
23  

  HYPERLINK \l "_Toc249333147"  VII.   ACTIONS REQUIRED BY THE APPLICANT
  PAGEREF _Toc249333147 \h  24  

  HYPERLINK \l "_Toc249333148"  A.  Reporting of Adverse Effects and
Hypersensitivity Incidents	  PAGEREF _Toc249333148 \h  24  

  HYPERLINK \l "_Toc249333149"  VIII.  GLOSSARY OF ACRONYMS AND
ABBREVIATIONS	  PAGEREF _Toc249333149 \h  25  

  HYPERLINK \l "_Toc249333150"  IX.  BIBLIOGRAPHY	  PAGEREF
_Toc249333150 \h  26  

  HYPERLINK \l "_Toc249333151"  A.  Studies Submitted in Support of
Trichoderma gamsii Strain ICC 080	  PAGEREF _Toc249333151 \h  26  

  HYPERLINK \l "_Toc249333152"  B.  Environmental Protection Agency Risk
Assessment Memorandums	  PAGEREF _Toc249333152 \h  29  

  HYPERLINK \l "_Toc249333153"  C.  Other References Cited	  PAGEREF
_Toc249333153 \h  30  

  HYPERLINK \l "_Toc249333154"  APPENDIX A – MICROBIAL PESTICIDE DATA
REQUIREMENTS	  PAGEREF _Toc249333154 \h  31  

  HYPERLINK \l "_Toc249333155"  APPENDIX B – Trichoderma gamsii strain
ICC 080 Products	  PAGEREF _Toc249333155 \h  38  

 

		

		



BIOPESTICIDES REGISTRATION ACTION DOCUMENT TEAM

Office of Pesticide Programs

Biopesticides and Pollution Prevention Division

Microbial Pesticides Branch

Science Reviews

Ibrahim Barsoum, Ph.D				   Product Analysis, Human Health

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

Shannon Borges				  	   Environmental Effects

Zigfridas Vaituzis, Ph.D.                                              
Environmental Effects

Regulations

Sheryl Reilly, Ph.D.				               Chief, Microbial Pesticides
Branch

Susanne Cerrelli                     				   Regulatory Action Leader

I.  EXECUTIVE SUMMARY

The Biopesticides and Pollution Prevention Division (BPPD) received
applications to register, under Section 3(c)(5) of the Federal
Insecticide, Fungicide and Rodenticide Act (FIFRA), two products
containing the active ingredient, Trichoderma gamsii strain ICC 080, as
a microbial fungicide.  The manufacturing-use product (MP; Trichoderma
gamsii strain ICC 080 Technical, EPA File Symbol 80289RN) is intended to
be used to formulate end-use pesticide products with Trichoderma gamsii
strain ICC 080.  The end-use product (EP; Tenet WP, Bioten WP, Remedier
WP; Tenet  T&O; EPA File Symbol 80289-O)   contains 2% Trichoderma
gamsii strain ICC 080, as well as 2% of another similar microbial active
ingredient, Trichoderma asperellum strain ICC 012.  The EP is intended
to control pathogenic fungi on several food and non-food crops,
including ornamentals, fruiting vegetables, leafy vegetables, cole
crops, legumes, aromatic herbs, cucurbits, berries and small fruits, and
turf.  Depending on the use, the EP may be applied as a cutting and bare
root application, substrate mix, greenhouse and nursery drench,
broadcast application, or through chemigation.  

The active ingredient, Trichoderma gamsii strain ICC 080 is a strain of
asexual fungi that was originally classified as Trichoderma viride, and
subsequently recharacterized. Trichoderma species are asexual fungi that
are isolated primarily from soil and decomposing matter. 

Trichoderma gamsii strain ICC 080 was isolated from a soil in Sardinia,
Italy that was found to suppress plant disease.  Trichoderma gamsii
strain ICC 080 is intended to be used for control of many soil-borne
fungal plant pathogens [i.e., Pythium species (spp.), Phytophthora spp.,
Sclerotinia spp., Sclerotium spp., Thielaviopsis basicola, Rhizoctonia
spp., Verticillum spp.]   Trichoderma gamsii strain ICC 080 acts as a
pathogen antagonist, colonizing in soil and around plant roots to
compete with plant pathogenic fungi for living space and nutrients. 
Moreover, Trichoderma gamsii strain ICC 080 also uses enzymes to attack
the cell walls of pathogenic fungi. 

Adequate mammalian toxicology data, data waivers and other information
were submitted to fulfill the Tier I acute toxicity data requirements
for the registration of 

Trichoderma gamsii strain ICC 080 as a microbial fungicide. 

The toxicological data demonstrated that Trichoderma gamsii strain ICC
080 is not toxic, infective or pathogenic to mammals.  No acute,
sub-chronic, chronic, immune, endocrine, or non-dietary exposure issues
were identified.  The intended uses of this microbial fungicide do not
pose a dietary risk to the U.S. population in general, including infants
and children.  Dietary exposure via drinking water is also not expected
to pose harm to populations, because the microbial fungicide is not
known to grow or thrive in aquatic environments, nor would be expected
to survive municipal treatment of drinking water. Further, in the
unlikely event that humans are exposed to Trichoderma gamsii strain ICC
080 from drinking water, the toxicological data base indicates no
adverse effects would be expected.

The potential for aggregate, non-occupational exposure from agricultural
applications of Trichoderma gamsii strain ICC 080 is unlikely, because
the intended use sites are not expected to be in close proximity to
residential areas.  However, the product is also intended to be used in
a residential setting, and if inadvertent residential exposures occur
from agricultural applications, the low toxicity of the active
ingredient would not be expected to cause adverse effects to humans.

A final rule establishing an exemption from the requirement of a
tolerance was signed (publication date will be entered here), and
published in the Federal Register (publication citation will be entered
here).

Exposure of non-target organisms to Trichoderma gamsii strain ICC 080 is
possible from the intended uses of this organism as a microbial
fungicide.  Isagro, S.p.A. submitted studies and data waiver rationales
to satisfy data requirements for non-target organism risk assessment
with the active ingredient (TGAI).  The honey bee study that was
submitted was of too short a duration to discern the potential hazards
to these organisms, and a non-target insect study on one species was
submitted.  BPPD has evaluated the information available and has
determined that additional studies are not necessary for Trichoderma
gamsii strain ICC 080.  Some strains of Trichoderma harzianum are
reported to affect bark beetles, however, an extensive literature search
did not produce reports of Trichoderma gamsii or Trichoderma viride
having adverse effects on bees.  Additionally, Gouli et al. 2008 report
a lack of adverse effects of a strain of Trichoderma viride in insects
(thrips).  Therefore, BPPD has determined that adverse effects of the
proposed uses of Trichoderma gamsii strain ICC 080 to honey bees and
other insects are unlikely.  

Adverse effects are not anticipated to non-target species, including
threatened or endangered species, exposed to the microbial fungicide
Trichoderma gamsii strain ICC 080, when used in accordance with the
label directions.  Therefore, the Agency has made a “No Effect”
determination for threatened or endangered species from the fungicidal
use of Trichoderma gamsii strain ICC 080. 

The information provided is sufficient to satisfy the Tier I non-target
organism data requirements for the active ingredient, Trichoderma gamsii
strain ICC 080, contained in the proposed MP and EP products.  Further
testing of non-target organisms at higher tier levels is not required.

Consistent with the new policy of making registration actions more
transparent, Trichoderma asperellum strain ICC 080 is subject to a 30
day comment period as a “new active ingredient” whose registration
would result in a “first outdoor use” and a “first residential
use.”  While a final decision on registration is contingent upon
review and consideration of public comments, EPA presently believes that
based on the risk assessment and information submitted in support of the
registration of Trichoderma gamsii strain ICC 080 that it is in the best
interests of the public and the environment to issue the registration
for Trichoderma gamsii strain ICC 012. The basis for this preliminary
decision can be found in the risk assessment for Trichoderma gamsii
strain ICC 080, which is characterized in this BRAD.  Acute toxicity and
pathogenicity data demonstrate that it has low toxicity (category III
and IV).  EPA has no concerns for non-target organisms, included
threatened or endangered species, exposed to Trichoderma gamsii strain
ICC 012 when it is used in accordance with approved label directions. 
EPA has not identified any toxic endpoints for non-target mammals,
birds, plants, aquatic, or soil organisms. Thus, EPA concludes that use
of the microbial fungicide Trichoderma gamsii strain ICC 080 in
accordance with the proposed label directions will not cause any
unreasonable adverse effects on the environment.  

The Environmental Protection Agency (EPA) considered information
submitted in support of an application for a registration under section
Section 3(c)(5) of the Federal Insecticide, Fungicide and Rodenticide
Act (FIFRA) for the microbial fungicide Trichoderma gamsii strain ICC
080 and determined that these data and information adequately satisfy
current data requirements (refer to 40 CFR Subpart U § 158.2000).  If
the Agency receives comments during the 30 day public comment period
that inform EPA’s initial decision, EPA will address such new
information and take appropriate action.  

II.  ACTIVE INGREDIENT OVERVIEW

	Biological Name:		Trichoderma gamsii strain ICC 080 

Culture Deposit:	CABI Bioscience International Mycological Institute-
Egham, UK under IMI CC No. 392151

	Trade/Other Names:  	Bioten WP; Tenet WP; Remedier WP

Office of Pesticide

Programs (OPP)

Chemical Code:		119207

	Type of Pesticide:		Microbial Pesticide, fungicide 

					See   HYPERLINK  \l "Table1_AppB"  Appendix B  for specific
information (i.e., 

use sites, application rate, method of application, 

formulation type, and target pests) 

III.  REGULATORY BACKGROUND  

On February 8, 2008, Mel Graben of Isagro, USA (address: 430 Davis
Drive, Suite 240, Morrisville, NC 27560), acting as the United States
authorized agent for Isagro S.p.A.,  (address: Centro Uffici San-Edifico
D-ala 3, Via Caldera, 21-20153 Milan, Italy) submitted an application to
register Bioten WP (EPA File Symbol 80289-O)  and  Trichoderma gamsii
strain ICC 080 (EPA File Symbol 80289- RN) under Section 3 of the
Federal Insecticide, Fungicide, and Rodenticide Act. On October 29,
2008, the EPA announced receipt of this application to register a
pesticide product containing a new 

active ingredient [ HYPERLINK
"http://www.epa.gov/fedrgstr/EPA-PEST/2008/March/Day-28/p6266.htm" 73
Federal Register (FR)  64325] and opened a 60-day public comment period
on the receipt of this application. 

In response to this notice of receipt, EPA did not receive any
substantive comments relevant to the subject Trichoderma pesticide
products. 

Concurrent with their registration application, and under the Federal
Food, Drug, and Cosmetic Act (FFDCA), as amended by the Food Quality
Protection Act of 1996, Isagro S.p.A. submitted a petition to establish
an exemption from the requirement of a tolerance for Trichoderma gamsii
strain ICC 080 [Pesticide Petition (PP) 8F 7327]. In the Federal
Register dated November 12, 2008 (73 FR  66897), the EPA announced that
Isagro, S.p.A., proposed to establish an exemption from the requirement
of a tolerance for residues of the microbial pesticide, Trichoderma
gamsii strain ICC 080.   One comment from an anonymous American citizen
was received following the publication of a notice of filing of a
petition under the FFDCA.   

EPA is proposing to promulgate a final rule establishing the exemption
from tolerance for Trichoderma gamsii strain ICC 080.  EPA has placed
the draft final rule that would establish such a tolerance exemption in
the docket along with this proposed FIFRA decision to permit comment on
both actions the Agency is considering. 

IV.  RISK ASSESSMENT

On October 26, 2007, the Agency issued a Final Rule in the Federal
Register on the data requirements to support registration of biochemical
and microbial pesticides, and updated the definitions for biochemical
and microbial pesticides (  HYPERLINK
"http://www.epa.gov/fedrgstr/EPA-TOX/2007/October/Day-26/t20828.htm"  72
FR 61002 ). The rule became effective on December 26, 2007. The data and
information evaluated for this Biopesticides Registration Action
Document (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 scientifically
acceptable (“SUPPLEMENTAL: UPGRADABLE”). If a study is rated as
“SUPPLEMENTAL: UPGRADABLE,” the Environmental Protection Agency
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 the current 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 need to be submitted.

For product-specific acute toxicity data requirements, toxicity
categories are assigned based on the hazard(s) identified from studies
and/or information submitted to the Agency. The 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.  

A.  Product Analysis Assessment 

All product analysis data requirements (  HYPERLINK
"http://ecfr.gpoaccess.gov/cgi/t/text/text-idx?c=ecfr&sid=f25d5426f66f63
ffeb9c986db92bf243&rgn=div8&view=text&node=40:23.0.1.1.9.16.1.3&idno=40"
 40 CFR § 158.2120 ) for Section 3(c)(5) registration of Trichoderma
gamsii strain ICC 080 Technical and Tenet WP, containing Trichoderma
gamsii strain ICC 080 as an active ingredient, have been satisfied by
either acceptable guideline studies or waiver rationales. For a
comprehensive guideline-by-guideline summary of the product analysis
data requirements described in sections IV(A)(1), IV(A)(2), and
IV(A)(3), refer to   HYPERLINK  \l "Table1"  Table 1  in Appendix A. 

	1.  Product Chemistry and Composition

Trichoderma gamsii strain ICC 080 Technical is an MPCA soil fungicide
for manufacturing use only.  The product contains 99.9% w/w Trichoderma
gamsii strain ICC 080 as active ingredient with a minimum and nominal
content of 1 x 109 and 2.5 x 109 conida cfu/g dry weight, respectively. 
The label indicates that the product also contains 0.10% other
ingredients.  The strain ICC 080, originally characterized as
Trichoderma viride, has been reclassified as Trichoderma gamsii, and has
been characterized by molecular biology techniques.  Trichoderma gamsii
strain ICC 080 was isolated from a soil in Sardinia, Italy that was
found to suppress plant disease.  The strain has been deposited in the
CABI Bioscience International Mycological Institute-Egham, UK IMI CC No.
392151. No relationships are known between the Trichoderma genus and any
pathogen of man, animals, or plants.  The extensive Moniliaceae family,
which includes phytopathogenic genera Botrytis and Verticillium, and the
human pathogenic genus Blastomyces, includes Trichoderma, but no
Trichoderma or closely related genera are known to be pathogenic to
mammals.

Submitted data satisfied the requirements for manufacturing process, and
discussion of formation of unintentional ingredients. [(OPPTS)
Guidelines 885.1100, 885.1200, and 885.1300] 

	2.  Analysis and Certified Limits 

Results of a five-batch preliminary analysis were provided, and the
requirement for analysis of samples has been satisfied.  The requirement
for certified limits has also been satisfied.

3.  Physical and Chemical Characteristics

Submitted data and waiver rationales satisfied the requirements for the
physical and chemical characteristics, including color, physical state,
odor, stability to normal and elevated temperatures, metals, and metal
ions, storage stability, miscibility, corrosion characteristics, pH,
viscosity, and density/relative density/bulk density (specific gravity).

B.  Human Health Assessment

	1.  Toxicology 

Acceptable Tier I mammalian toxicology data and data waivers information
support the registration of the manufacturing product, Trichoderma
gamsii strain ICC 080 Technical, and the end-use product, Bioten™ WP,
which contains Trichoderma gamsii strain ICC 080. Tier II and Tier III
studies were not required for Trichoderma gamsii strain ICC 080, based
upon the lack of acute toxicity/pathogenicity in the Tier I studies. 

For a comprehensive guideline-by-guideline summary of the toxicology
data requirements and additional studies described in sections
IV(B)(1)(a), IV(B)(1)(b), and IV(B)(1)(c), refer to   HYPERLINK  \l
"Table2"  Table 2  in Appendix A. 

	a.  Acute Toxicity/Pathogenicity – Tier I (  HYPERLINK
"http://ecfr.gpoaccess.gov/cgi/t/text/text-idx?c=ecfr&sid=76805ba87ba7a2
c69b53feed9af452bb&rgn=div8&view=text&node=40:23.0.1.1.9.16.1.5&idno=40"
 40 CFR § 158.2140 )

Acute Oral Toxicity – Rat [OPPTS Guideline 885. 3550; (MRID# 47345801)

An acute oral toxicity study was performed on rats given a single oral
dose of Trichoderma gamsii (formerly known as Trichoderma viride) strain
ICC 080 of (7.5 x 108 CFU/g) in 0.9% NaCl solution at a dose of 2000
mg/kg of body weight in a limit test.  The animals were observed for a
period of up to 14 days. The Oral LD50 for males, females, and the
combined test animals were: males > 2000 mg/kg of body weight, females >
2000 mg/kg of body weight, combined > 2000 mg/kg of body weight.  No
mortalities occurred during the study. Based on the results of this
study, Trichoderma gamsii strain ICC 080 was found to be of low acute
oral toxicity. Microbial enumeration was not done so it is unknown if
Trichoderma gamsii is infective or pathogenic in the rat.  There were no
treatment related clinical signs, changes in body weight or pathological
findings at necropsy. This study was rated “ACCEPTABLE” for risk
assessment purposes.

Acute Intraperitoneal Injection Toxicity– Rat (OPPTS Guideline
885.3200;  MRID# 47345802)  An acute intraperitoneal injection toxicity
study was submitted, in which groups of fasted, 41-48 days old rats
(3/sex) were injected with Trichoderma gamsii strain ICC 080 (at 7.5 x
108 CFU/g) in 0.9% NaCl solution at a dose of 1 x 107 CFU/g.  Animals
were then observed for up to 21 days. Control animals (2/sex) were
injected with 0.9% NaCl solution only. Trichoderma gamsii strain ICC 080
is not toxic based on the results of this study.  There were no
treatment-related necropsy findings or changes in body weight. 
Microbial enumeration was not done so it is unknown if Trichoderma
gamsii is infective or pathogenic in the rat.  All of the animals
treated with the test material experienced slightly reduced mobility,
slight ataxia, slightly reduced muscle tone, slight dyspnea, mydriasis,
and writhing, observed 60 minutes after administration.  All of these
clinical signs were completely resolved within 24 hours.   This study
was rated “ACCEPTABLE” for risk assessment purposes.

Acute Pulmonary Toxicity/Pathogenicity Studies– Rats (OPPTS Guideline
885.3150; MRID# 47345803, 47345804)  Acute pulmonary
toxicity/pathogenicity studies were submitted, in which groups of fasted
43-56 days old rats (31/sex) were exposed by the intratracheal route to
Trichoderma gamsii strain ICC 080 at a dose of 2.5 x 106 CFU/animal. 
Animals were observed for up to 22 days. Rats in the control group were
administered the vehicle, [0.1% solution of Tween 20 in aqua ad
iniectabilia (water for injection)] only.  Rats in the reference groups
were administered inactivated test item.  Samples of feces, lungs, lymph
nodes, kidneys, brain, liver, spleen, and blood were taken for microbial
enumeration in those tissues.  None of the administered Trichoderma
gamsii conidia from lung tissue of the animals appeared in other organ
tissue.  Conidia could not be detected in blood samples at any time
during the study.  Conidia were detected in the feces up to 21 days post
administration.  Conidia density in the lung tissue decreased to 0
within 21 days post administration.  This shows a pattern of clearance
and lack of infectivity of  Trichoderma gamsii strain ICC 080.  The
recorded Pulmonary LD50 was greater than 2.5 x 106 CFU/animal in males,
females and in the combined group of test animals.  No mortality
occurred.  Based upon these results, Trichoderma gamsii strain ICC 080
is of low toxicity, and Trichoderma gamsii was not infective or
pathogenic in the rat.  There were no treatment related clinical signs,
changes in body weight, or pathological changes observed at necropsy.
This study was rated “ACCEPTABLE” for risk assessment purposes.

Acute Oral Toxicity/Pathogenicity– Rats   (OPPTS Guideline 870.1100;
MRID# 47346301).  In an acute oral toxicity study (MRID 47346201),
groups of fasted, 6-7 week old CD/CRL:CD rats (5/sex) were given a
single oral dose of Bioten WP (originally referred to as Remedier WP in
the acute studies below)(Trichoderma conidia (1.2 x 108 CFU/g);
Trichoderma asperellum strain ICC 012  (originally classified as T.
harzianum)  (7.8 x 107 CFU/g); Trichoderma gamsii strain ICC 080 (4.2 x
107 CFU/g)) in 0.9% NaCl solution at a dose of 2000 mg/kg bw in a limit
test.  The animals were then observed for a period of up to 14 days.
Oral LD50 were as follows: Males > 2000 mg/kg bw, Females > 2000 mg/kg
bw , Combined > 2000 mg/kg bw. Limit test: no mortality occurred during
the study. Based on the results of this study BiotenWP (Trichoderma
asperellum 7.8 x 107 CFU/g; Trichoderma gamsii  4.2 x 107 CFU/g) is not
toxic and has an LD50 greater than 2000 mg/kg bw.(EPA Toxicity Category
III.) Microbial enumeration was not performed, therefore, the
infectivity and pathogenicity of Trichoderma conidia, Trichoderma
asperellum strain ICC 012, and Trichoderma gamsii strain ICC 080, are
unknown. There were no treatment related clinical signs, necropsy
findings or changes in body weight. This study was rated
“ACCEPTABLE” for risk assessment purposes.

Acute Dermal Toxicity (OPPTS Guideline 870.1200; MRID# 47346302)

In an acute dermal toxicity study (MRID 43746302), groups of fasted, 6-7
week old CD/CRL:CD rats (5/sex) were given a single dermal dose of
BiotenWP (Trichoderma conidia (1.2 x 108 CFU/g); Trichoderma asperellum 
strain ICC 012 (7.8 x 107 CFU/g); Trichoderma gamsii  strain ICC 080
(4.2 x 107 CFU/g)) in 0.9% NaCl at a dose of 2000 mg/kg bw for 24 hours
to an area of approximately 5 x 6 cm (~10% of body surface).  Following
exposure, the animals were observed for a period of 14 days. Dermal LD50
were as follows: Males > 2000 mg/kg bw,Females > 2000 mg/kg bw, Combined
> 2000 mg/kg bw. Limit test; no mortality occurred during the study.
Based on the results of this study Remedier WP (Trichoderma asperellum 
7.8 x 107 CFU/g; Trichoderma gamsii  4.2 x 107 CFU/g) is of MODERATE
Toxicity, EPA Toxicity Category III. This study was rated
“ACCEPTABLE” for risk assessment purposes.

Acute Inhalation Toxicity (OPPTS Guideline 870.1300; MRID# 47346303)

In an acute inhalation toxicity study (MRID 47346303), groups of fasted,
44-55 day old CD/CRL:CD rats (5/sex) were exposed by the inhalation
route to Bioten WP (Trichoderma conidia (1.2 x 108 CFU/g); Trichoderma
asperellum strain ICC 012 (7.8 x 107 CFU/g); Trichoderma gamsii strain
ICC 080 (4.2 x 107 CFU/g)) as supplied at a concentration of 5.20 mg/L. 
Animals then were observed for 14 days. The MMAD was 2.573 µm and the
GDS was 11.375. Inhalation LC50were as follows: Males > 5.20 mg/L, 

Females > 5.20 mg/ L, Combined > 5.20 mg/L. No mortality occurred during
the study. Bioten WP (Trichoderma asperellum 7.8 x 107 CFU/g;
Trichoderma gamsii  4.2 x 107 CFU/g) is of LOW Toxicity, EPA Toxicity
Category IV, based on lack of mortality in male and female rats. There
were no treatment related clinical signs, necropsy findings or changes
in body weight. This study was rated “ACCEPTABLE” for risk
assessment purposes. Toxicity Category IV

Acute Eye Irritation – Rabbit (OPPTS Guideline 870.2400; MRID#
473463034)

In a primary eye irritation study (MRID 47346304), 100 mg of Bioten WP
(Trichoderma conidia (1.2 x 108 CFU/g); Trichoderma asperellum ICC 012
(7.8 x 107 CFU/g); Trichoderma gamsii strain ICC 080 (4.2 x 107 CFU/g)),
as supplied, was instilled into the right conjunctival sac of 3 male
fasted young adult Himalayan rabbits  for 24 hours. Animals then were
observed for 72 hours. Irritation was scored by the method that was not
named but similar to the method of Draize (1959). Conjunctival redness
was observed in all animals 1 hour after instillation, and resolved by
24 hours.  The cornea and the iris were not affected by instillation of
the test item. In this study, Bioten WP (Trichoderma asperellum 7.8 x
107 CFU/g; Trichoderma gamsii 4.2 x 107 CFU/g) was not an eye irritant
and is in EPA Toxicity Category IV. This study was rated
“ACCEPTABLE” for risk assessment purposes.

Acute Dermal Irritation Toxicity- Rabbit (OPPTS 870.2500; MRID #
47346305)

In a primary dermal irritation study (MRID 47346305), fasted young adult
Himalayan rabbits (3 males) were dermally exposed to 500 mg of Bioten WP
(Trichoderma conidia (1.2 x 108 CFU/g); Trichoderma asperellum strain
ICC 012 (7.8 x 107 CFU/g); Trichoderma gamsii strain ICC 080 (4.2 x 107
CFU/g)) in aqua ad iniectabilia (water for injection) for 4 hours on a
shaved area of 6 cm2.  Animals then were observed for 6 days. 
Irritation was scored by the method that was not named but similar to
the method of Draize (1944). Very slight erythema was observed on all
three animals 24 hours through 5 days after patch removal.  Very slight
edema was observed on one animal 72 hours through 4 days after patch
removal.  The Primary Irritation Index (PII) was 0.8. In this study,
Remedier WP (Trichoderma asperellum 7.8 x 107 CFU/g; Trichoderma gamsii
4.2 x 107 CFU/g) is slightly irritating and is in EPA Toxicity Category
IV. This study was rated “ACCEPTABLE” for risk assessment purposes.

Skin Sensitization Study-Guinea Pigs	OPPTS 870.2600; MRID # 47346306)

   SEQ CHAPTER \h \r 1 In a skin sensitization study (MRID 47346306)
with Bioten WP (Trichoderma conidia (1.2 x 108 CFU/g); Trichoderma
asperellum strain ICC 012 (7.8 x 107 CFU/g); Trichoderma gamsii strain
ICC 080 (4.2 x 107 CFU/g)) in 0.9% NaCl, young adult Dunkin-Hartley
guinea pigs (15 males) were tested using the method of Magnusson and
Kligman (Maximization Test). The test substance was administered as a
suspension for intradermal application and applied as a suspension for
topical application.  Benzocaine was used as a positive control.
Discrete or patchy erythema was observed on all test animals exposed to
the 10% suspension of Bioten WP (Trichoderma asperellum  and Trichoderma
gamsii  wettable powder) in 0.9% NaCl solution 25 and 48 hours after the
induction phase.  Some animals exposed to a 50% suspension of Bioten WP
(Trichoderma asperellum and Trichoderma gamsii wettable powder) in 0.9%
NaCl solution had moderate, confluent erythema 49 and 72 hours after
application.  There were no skin reactions in the test animals 24, 48,
or 72 hours after the final challenge using a 1% suspension of BiotenWP
(Trichoderma asperellum and Trichoderma gamsii  wettable powder) in 0.9%
NaCl solution.  In this study, (Trichoderma asperellum  7.8 x 107 CFU/g;
Trichoderma gamsii  4.2 x 107 CFU/g) was not a dermal sensitizer. This
study was rated “ACCEPTABLE” for risk assessment purposes.

	b.	Acute Toxicology and Subchronic Toxicity/Pathogenicity – Tier II;

	    	Reproductive Fertility Effects, Carcinogenicity, Immunotoxicity,
and 

	   	Infectivity/Pathogenicity Analysis – Tier III (  HYPERLINK
"http://ecfr.gpoaccess.gov/cgi/t/text/text-idx?c=ecfr&sid=9061faa75a746f
eda99a9bbe8072f5d3&rgn=div8&view=text&node=40:23.0.1.1.9.16.1.5&idno=40"
 40 CFR § 158.2140 )

Tier II and Tier III studies were not required for Trichoderma gamsii
strain ICC 080 based on the lack of acute toxicity/pathogenicity in the
Tier I studies.

	

	c.  Effects on the Endocrine System

Section 408(p) of the Federal Food, Drug, and Cosmetic Act (FFDCA)
requires EPA to develop a screening program to determine whether certain
substances (including all pesticide active and other ingredients) “may
have an effect in humans that is similar to an effect produced by a
naturally occurring estrogen, or such other endocrine effect as the
Administrator may designate.” Following the recommendations of its
Endocrine Disruptor Screening and Testing Advisory Committee (EDSTAC),
EPA determined that there were scientific bases for including, as part
of its program, androgen and thyroid hormone systems, in addition to the
estrogen hormone system. The Environmental Protection Agency also
adopted EDSTAC’s recommendation that the Program include evaluations
of potential effects on wildlife.

The Agency has no information to suggest that Trichoderma gamsii strain
ICC 080 has an effect on the endocrine system.  The submitted acute
pulmonary toxicity/pathogenicity study in rodents indicated that
following pulmonary exposure, the immune system is still intact and able
to process and clear the active ingredient.  Trichoderma gamsii strain
ICC 080 is a ubiquitous organism in the environment and there have been
no reports of the organism affecting endocrine systems.  Therefore, it
is unlikely that this organism would have estrogenic or endocrine
effects and it is practically non-toxic to mammals. 

Additional data, specifically on the endocrine effects of this microbial
pesticide, are not required at this time. When the appropriate screening
and/or testing protocols being considered under the Agency’s Endocrine
Disruptor Screening Program (EDSP) have been developed and vetted,
Trichoderma gamsii strain ICC 080 may be subjected to additional
screening and/or testing to better characterize effects related to
endocrine disruption.

	2.  Dietary Exposure and Risk Characterization

Dietary exposure to the microbial pesticide is likely to occur, but the
lack of acute oral toxicity, infectivity, and pathogenicity support the
establishment of an exemption from the requirement of a tolerance for
Trichoderma gamsii strain ICC 080.

Dietary exposure to the microbial active ingredient is expected to be
minimal. The product is typically applied to soil, but sometimes may be
applied when the crops are growing in the field, resulting in residues
on the crops.  The Agency expects residues on food to be minimal because
of the typical method used to apply the microbial fungicide to soils.  
Moreover, Trichoderma lives in soils and is unlikely to live on the
plants because any spores that do end up on the plant due to application
would decrease over time due to weathering, desiccation and ultraviolet
radiation, which can kill even quiescent forms of the fungus.  In the
remote likelihood that the applied fungus grew on the edible portions of
a treated crop, the results of the oral toxicity testing demonstrated
that no toxicity or pathogenicity in treated animals occurred, even when
dosed with the fungus at high levels by the oral route of exposure. (See
  HYPERLINK  \l "Acute_Toxicity_Pathogenicity_TierI"  section
IV(B)(1)(a) .) 

 

3.   Drinking Water Exposure and Risk Characterization 

Drinking water exposure is expected to be negligible because the
microbial fungicide will not be applied to water.  Further, Trichoderma
gamsii is a soil microorganism, and would not proliferate in aquatic
environments.  Moreover, the Agency believes that Trichoderma within the
soil will not likely percolate into water due to the large size of the
fungal spores and the fact that they adhere to soil particles.  Even in
the unlikely event that dietary exposure occurs through drinking water,
the Agency concludes that there is a reasonable certainty that no harm
will result because of the lack of acute oral toxicity/pathogenicity to
mammals as previously described. 

	4.  Acute and Chronic Dietary Exposure and Risks for Sensitive 

 	     Subpopulations, Particularly Infants and Children

Section 408(b)(2)(C) of the FFDCA provides that EPA shall assess the
available information about consumption patterns among infants and
children, the special susceptibility of infants and children to
pesticide chemical residues, and the cumulative effects on infants and
children from the residues and other substances with a common mechanism
of toxicity. In addition, FFDCA section 408(b)(2)(C) also provides that
EPA shall apply an additional tenfold 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 unless EPA
determines that a different margin of safety will be safe for infants
and children. Margins of exposure (safety), which are often referred to
as uncertainty factors, are incorporated into EPA risk assessment either
directly or through the use of a margin of exposure analysis, or by
using uncertainty (safety) factors in calculating a dose level that
poses no appreciable risk.

Based on the acute toxicity and pathogenicity data discussed in  
HYPERLINK  \l "Acute_Toxicity_Pathogenicity_TierI"  section IV(B)(1)(a)
, the Environmental Protection Agency concludes that there is a
reasonable certainty that no harm to sensitive subpopulations, including
infants, children, and adults, will result from the use of Trichoderma
gamsii strain ICC 080. This includes all anticipated dietary exposures
and all other exposures for which there is reliable information. The
Agency has arrived at this conclusion because the data available on
Trichoderma gamsii strain ICC 080 do not demonstrate toxic, pathogenic,
or infective potential to mammals. Thus, there are no threshold effects
of concern and, as a result, the provision requiring an additional
margin of safety does not apply.  Further, the considerations of
consumption patterns, special susceptibility, and cumulative effects do
not apply to pesticides without a demonstrated significant adverse
effect.

	5.  Occupational, Residential, School, and Daycare Exposure and Risk 

	     Characterization

Occupational Exposure and Risk Characterization

In light of the Tier I acute toxicity/pathogenicity studies, which did
not show any toxicity via the oral, pulmonary, or intraperitoneal
injection routes of exposure (  HYPERLINK  \l
"Acute_Toxicity_Pathogenicity_TierI"  see section IV(B)(1)(a) ), or
pathogenic effects to rats via the pulmonary route of exposure, handler
exposure to Trichoderma gamsii strain ICC 080 is not expected to pose
any undue risk. Regardless, requirements for the use of appropriate
personal protective equipment, and precautionary statements are required
on the product label to mitigate any potential risks to pesticide
handlers due to prolonged exposure.  Handlers working with Trichoderma
gamsii strain ICC 080 must wear a long-sleeved shirt, long pants, socks,
shoes, waterproof gloves, and a dust/mist filtering respirator meeting
NIOSH standards of at least N-95, R-95, or P-95 when mixing, loading, or
applying the product.

Residential, School, and Daycare Exposure and Risk Characterization

Trichoderma gamsii strain ICC 080 is a naturally occurring microorganism
and is ubiquitous in the environment.  Trichoderma gamsii strain ICC 080
will be applied to substrate mixes, ornamental plants, agricultural
fields, turf, and various plants grown in greenhouses. Although some
applications to turf or ornamental plants may be in residential areas,
non-dietary exposure would be expected to be below the Agency’s level
of concern because of its low toxicity classification, and because the
lab results indicate Trichoderma gamsii strain ICC 080  is not
pathogenic to mammals.  

	6.  Aggregate Exposure from Multiple Routes Including Dermal, Oral, and

                 Inhalation

In examining aggregate exposure, Section 408 of the FFDCA directs 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). 

Trichoderma gamsii strain ICC 080 will be applied to substrate mixes,
ornamental plants, agricultural fields, turf, and various plants grown
in greenhouses. Although some applications to turf or ornamental plants
may be in residential areas, non-dietary exposure would be expected to
be below the Agency’s level of concern because of its low
toxicity/pathogenicity to mammals. 

7.  Cumulative Effects

Section 408(b)(2)(D)(v) of the FFDCA requires the Agency to consider the
cumulative effect of exposure to Trichoderma gamsii strain ICC 080 and
to other substances that have a common mechanism of toxicity. These
considerations include the possible cumulative effects of such residues
on infants and children.  Based on tests in mammalian systems,
Trichoderma gamsii strain ICC 080 does not appear to be toxic to humans
via dietary and pulmonary exposure.  Therefore, the requirement to
consider cumulative effects does not apply.

	8.  Risk Characterization

The Agency considered human exposure to Trichoderma gamsii strain ICC
080 in light of the standard for registration and safety factors in
FIFRA and FFDCA, as amended by the FQPA. 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 from the
use of Trichoderma gamsii strain ICC 080 when used in accordance with
EPA-approved labeling.

	

C.  Environmental Assessment

Exposure of non-target organisms to Trichoderma gamsii strain ICC 080 is
possible from the proposed uses of microbial pesticides containing this
active ingredient.  Isagro, S.p.A. submitted studies and data waiver
rationales to satisfy data requirements for non-target organism risk
assessment with the active ingredient (TGAI).  The following is a
summary of the information submitted to support registration and an
assessment of environmental risk.     

		1.  Summary of Non-target Organism Testing and Waiver Rationales

The data and other information provided to the Agency in support of the
applcations for registration for the MP and EP are sufficient to satisfy
the Tier I non-target organism data requirements for the active
ingredient, Trichoderma gamsii strain ICC 080, contained in the
technical product and the EP. Summaries of each study are presented
below.  Table 3 in Appendix A provides the status of the data
requirements for the TGAI.  

.a.  Avian Oral Toxicity/Pathogenicity (OPPTS Guideline 885.4050); Avian
Inhalation Toxicity/Pathogenicity (OPPTS Guideline 885.4100)

A data waiver was requested in lieu of data requirements for avian
testing.  Justification for these waivers was largely based on the
temperatures at which Trichoderma species can grow compared with higher
avian body temperatures. The supporting rationale cited data that showed
that Trichoderma species grow at temperatures below 37°C, whereas
normal bird body temperatures are typically above 38°C, with active
body temperatures above 40°C (Prinzinger et al. 1991).  As the
registrant states, EPA has acknowledged the validity of this rationale
in the past (USEPA 2008).  The registrant also argued that no toxic or
pathogenic effects to birds should be expected, since none were observed
in the rat acute oral toxicity/pathogenicity tests with the active
ingredient or the EP.  However, the comparability of the effects between
these two taxa for Trichoderma gamsii strain ICC 080 is not known. 
Nonetheless, the growth temperature rationale presented is sufficient to
conclude that adverse effects to birds are not expected due to exposure
to T. gamsii strain ICC 080.  This rationale is sufficient to fulfill
the data requirement. 

b.  Freshwater Fish Toxicity/Pathogenicity (OPPTS 885. 4200) and 

(MRID No.: 47345805)

 cfu/L).  Likewise, the NOEC was ≥ 100 mg/L ( ≥ 1.4 x 1010 cfu/L,
measured).  The study is scientifically sound and acceptable for
assessment of risk to freshwater fish from exposure to Trichoderma
gamsii strain ICC 080.  Based on these results, adverse effects to
freshwater fish resulting from proposed label applications of
Trichoderma gamsii strain ICC 080 are not expected. This study was
classified as “ACCEPTABLE.”

c.  Freshwater Invertebrate Toxicity/Pathogenicity (OPPTS 885.4240)

and (MRID Nos.: 47345806 and 47346307)

In a 21-day toxicity/pathogenicity study, (MRID No.: 47345806) neonate
Daphnia magna received an aqueous exposure to nominal concentrations of
6.25, 12.5, 25, 50, and 100 mg Trichoderma gamsii strain ICC 080
Technical/L (corresponding to measured concentrations of 1.6 x 109, 2.9
x 109, 4.4 x 109, 1.0 x 1010, and 2.1 x 1010 cfu/L, respectively) under
semi-static conditions.  A dilution water control and an attenuated
control were also included.  Daphnia survival was 95%, 90%, 85%, 70%,
and 50% in the 6.25, 12.5, 25, 50 and 100 mg/L Trichoderma gamsii strain
ICC 080 test concentrations, respectively, and was 100% and 80% in the
negative and attenuated controls, respectively.  Thus, the survival rate
was considered to be significantly reduced only in the highest test
concentration (100 mg/L).  Statistical calculation of the
mortality/immobility EC50 could not be performed, since no test
concentration produced >50% mortality.   Results of statistical analysis
of the reproduction rates indicate reproduction rate was reduced only in
Daphnia exposed to the 100 mg/L test concentration (38% reduction). 
Based on the mortality/immobility and the reproduction rates of
surviving Daphnia, the 21-day NOEC was determined to be 50 mg/L highest
(1.0 x1010 cfu/L, measured).  Based on these results, adverse effects to
freshwater invertebrates resulting from proposed label applications of
Trichoderma gamsii strain ICC 080 are not expected. This study was
classified as “ACCEPTABLE.”

This 48-hour Immobilization test (MRID No.: 47346307) was performed with
the end-use product.  In a 48-hour immobilization study, Daphnia magna
received an aqueous exposure to the nominal concentration of 100 mg/L of
Remedier WP under semi-static conditions (test and control water was
renewed at 24 hours).  The active ingredients in Remedier WP are
Trichoderma asperellum strain ICC 012 and Trichoderma gamsii strain ICC
080 (corresponding to 3.2 x 108 cfu/L total, measured) and are each
contained in the product at 2% by weight.  A dilution water control was
also included.  No adverse effects were observed in daphnids exposed to
the 100 mg/L limit test concentration which indicates a 48-hour EC50 of
>100 mg/L (3.2 x 108 cfu/L).  This study is classified as unacceptable
as a Tier I pathogenicity study for freshwater aquatic invertebrate with
a microbial pesticide due to the 48-hour duration.  The study was
classified as “unacceptable”. It cannot be upgraded and cannot be
used to fulfill the pathogenicity data requirement for either TGAI. 

 

Non-target Insect Testing (OPPTS 885.4340) (MRID No.: 47346308)

This dose response study is not a data requirement for microbial
fungicides.  However, the registrant submitted a 14-day contact
toxicity/pathogenicity study using the predatory mite, Typhlodromus
pyri.  The test insects were exposed to nominal concentrations of 0.062,
0.185, 0.556, 1.670, and 5.0 kg Remedier WP/ha (corresponding to 0.002,
0.007, 0.022, 0.067, and 200 g total active ingredient/ha).  Mite
mortality was monitored for the first 7-days and the effects of Remedier
WP on reproduction potential were monitored in adult mites from day 7
thru 14.  The active ingredients in Remedier WP are Trichoderma
asperellum strain ICC 012 and Trichoderma gamsii strain ICC 080
(corresponding to 1.2 x 108 cfu/L, total).  A dilution water control and
positive control were included in the study.  Statistically significant
mortality was not observed in mites exposed to any of the Remedier WP
treatment concentrations.  Thus the LD50 was determined to be > 5 kg/ha,
which is higher than the highest broadcast application rate. 
Confirmation of infection by the Trichoderma active ingredients in dead
mites was not performed.  Reproduction was affected in the 0.062 kg,
0.185 kg and 5.0 kg Remedier WP/ha treatment groups, but no
statistically significant effects on reproduction occurred in the 0.556
kg or 1.670 kg Remedier WP/ha treatment groups.  The study authors
concluded that effects observed on reproduction were not of biological
significance and not due to exposure to the test item.  However, these
results are inconclusive and it appears that the study design is not
sensitive enough to detect biologically significant reductions in
reproduction in this species as a result of exposure to the test
material.  Because mortality is the primary endpoint of concern, this
study is classified as acceptable.

Honey Bee Testing (OPPTS 885.4380) (MRID No.: 47345807)

Adult honey bees (Apis mellifera L.) were exposed to 50 µg per bee
Trichoderma gamsii strain ICC 080 Technical (0.85 x 105 cfu/bee) in a
contact toxicity test, and 112.1 µg/bee Trichoderma gamsii strain ICC
080 Technical (1.9 x 105 cfu/bee) in an oral toxicity test.  The
duration of both tests was 48 hours.  A negative control (for contact
toxicity this was water plus Adhasit, an adhesive; for oral toxicity,
this was water plus sugar) and positive control (dimeothate) were also
included.  Two percent mortality was observed in the contact toxicity
test, and 4% mortality was observed in the acute oral toxicity test. 
There were no signs of abnormal behavior in the surviving honey bees
throughout the tests.  The 24 and 48 hour contact LD50 of Trichoderma
gamsii strain ICC 080 was >50 µg product (0.85 x 105 cfu/bee).  The 24
and 48 hour oral LD50 of Trichoderma gamsii strain ICC 080 was >112.1
µg product (1.9 x 105 cfu/bee).  Because the study was not long enough
to allow for observation of pathogenic effects, it is classified as
unacceptable/not upgradeable, and cannot be used to satisfy the
requirement for honey bee data.

Non-guideline Studies

i) Acute Toxicity (14 Days) of Trichoderma gamsii to the Earthworm
Eisenia fetida in Artificial Soil   (MRID No.: 47345810)

Adult earthworms (Eisenia fetida) were exposed to 198, 296, 444, 667,
and 1000 mg Trichoderma gamsii strain ICC 080 Technical/kg soil dry
weight in artificial soil corresponding to 3.37 x 108, 5.03 x108, 7.55 x
108, 1.13 x 109, and 1.7 x 109 cfu/kg soil dry weight.  A negative
control (untreated soil) was also included.  The test duration was 14
days.  No mortality was observed in any of the soil test groups with
Trichoderma gamsii and there were no signs of abnormal behavior in the
surviving worms throughout the test.  Body weight gain in the treatment
groups was not significantly different from the negative control at test
termination.  The 14-day contact LD50 and LOEC of Trichoderma gamsii was
>1000 mg/kg soil (1.7 x 109 cfu/kg soil) corresponding to the highest
concentration tested.  The NOEC was 1000 mg/kg soil (1.7 x 109 cfu/kg
soil).  This non-guideline study is scientifically sound and acceptable
for earthworm toxicity testing.

ii)  Effects of Trichoderma gamsii to the Activity of Soil Microflora in
the Laboratory (MRID No.: 47345809)

The effects of Trichoderma gamsii strain ICC 080 on the activity of soil
microflora as determined by carbon and soil nitrogen transformation was
assessed in a 28-day laboratory study.  Test concentrations of
Trichoderma gamsii strain ICC 080 were 0.07 and 0.67 mg/kg soil dry
weight were applied to loamy, sandy soils collected from a fallow field.
 On Day 28 after application, the respiration rates of Trichoderma
gamsii strain ICC 080 treated soils, as determined by CO2 production,
were 8.86% and 3.40% higher than in the untreated control for the test
concentrations of 0.07 mg and 0.67 mg Trichoderma gamsii strain ICC
080/kg soil dry weight, respectively.  These values were not
statistically different from the crontrol and did not exceed the 25%
inhibition threshold according to OECD, EPPO and SETAC guidelines,
indicating no long-term effects of the test substance on soil
respiration.  Nitrate content was 18.7% and 10.4% lower in the soils
treated at 0.07 mg and 0.67 mg Trichoderma gamsii strain ICC 080/kg soil
dry weight, respectively, compared to the negative control, which was
statistically significant for the lower test concentration.  A minor
difference (-5.5% from the control) was observed for nitrate formation
rate.  The differences for the soil mineral nitrogen content were -18.6%
and -10.0%, respectively, which was statistically significant for the
0.07 mg/kg soil test level.  However, the soil nitrate content, nitrate
rate of formation and soil mineral nitrogen content were all below the
25% trigger values indicating no long-term effects to soil microflora. 
This non-guideline study is scientifically sound and classified as
acceptable for assessing hazard of the effects of Trichoderma gamsii
strain ICC 080 on soil microflora. This study was classified as
“ACCEPTABLE.”

2.  Environmental Effects Conclusions

a. Terrestrial Animals and Plants 

Exposure is expected to terrestrial animals and plants as a result of
some, but not all, proposed use patterns for the EP containing
Trichoderma gamsii strain ICC 080.  Greenhouse applications will limit
environmental release of the active ingredient, and exposure to
non-target organisms is not expected from these uses.  Applications made
to cuttings and bare roots, in substrate mixes, and through soil
drenches would limit the presence of the active ingredient to the soil
immediately around treated plants, which will significantly reduce the
potential for non-target exposure to only those organisms that utilize
those areas.  This exposure will be limited even further if the treated
plants are in containers (e.g., pots, flats, etc.).  A greater potential
for non-target exposure exists with broadcast applications and
chemigation that result in residues of the active ingredient on foliar
surfaces in addition to the soil.  Exposure to terrestrial non-target
organisms is anticipated as a result of these applications.  

The supporting rationale to waive the non-target data requirement for
avian wildlife study was sufficient to conclude that adverse effects are
not expected in avian wildlife as a result of exposure to Trichoderma
gamsii strain ICC 080, due to the growth temperatures required for the
microbial active ingredient compared with the higher body temperature of
birds.  

The data submitted for toxicity/pathogenicity to honey bees (Apis
mellifera) were unacceptable due to the short duration (48 hours) of the
oral and contact tests that were performed in the study that was
submitted.  Honeybee studies should be long enough (e.g., 30 days) to
allow for the observation of pathogenic effects.  While BPPD recognizes
that some strains of Trichoderma harzianum are known to affect
non-target insects (e.g., Glinski and Buczek 2003, Santamarina et al.
2002), there are no reports in the scientific literature that
Trichoderma gamsii or Trichoderma viride causes adverse effects on bees.
 Therefore, BPPD has concluded that adverse effects of the proposed uses
of Trichoderma gamsii strain ICC 080 to honey bees are unlikely.

Although not a data requirement for microbial fungicides, an insect
toxicity/pathogenicity study with the predatory mite (Typhlodromus pyri,
MRID 47346308) was submitted and reviewed.  This study showed that
adverse effects on these and related insects are not expected at field
application rates  Therefore, BPPD concluded that the adverse effects to
non-target insects as a result of the proposed fungicidal uses of
Trichoderma gamsii strain ICC 080 are not expected.  

Non-target plant testing is not required because Trichoderma gamsii
strain ICC 080 is not related to any known plant pathogen.  Adverse
effects on plants are not expected to result from proposed fungicidal
uses of Trichoderma gamsii strain ICC 080.  

Two non-guideline studies on earthworm toxicity/pathogenicity and
effects on soil microflora activity were also submitted.  While these
studies are not required for registration of microbial pesticides, they
provided information to allow BPPD to conclude that the proposed
fungicidal uses of Trichoderma gamsii strain ICC 080 are not expected to
have adverse effects in soil.

Based on the data and other information submitted by the applicant,
adverse effects are not expected to occur to terrestrial animals or
plants as a result of proposed labeled applications of Trichoderma
gamsii strain ICC 080.  

b. Aquatic Animals and Plants 

Exposure of non-target organisms to Trichoderma gamsii strain ICC 080 in
the aquatic environment is not expected to be significant.  For example,
an application made directly to a pond that is only 6 inches deep (a
standard size used by OPP to estimate aquatic exposure) at the highest
broadcast application rate (5 lbs/acre) would yield a concentration of
184 cfu/mL, so the amount reaching nearby aquatic environments due to
runoff or drift would be expected to be less.  This is far below the
maximum hazard concentrations at which the aquatic testing was
conducted.  Furthermore, if Trichoderma gamsii strain ICC 080 enters the
aquatic environment it is not expected to proliferate in the aquatic
environment, since it is a soil-borne fungus.

Studies with the TGAI were submitted for the freshwater fish and
freshwater invertebrate toxicity/pathogenicity testing data
requirements.  No significant adverse effects were observed in the
subjects of either study.  Both of their endpoint values were above the
maximum hazard dose for aquatic testing with microbial pesticides,
indicating a lack of effects above environmental concentrations that may
occur in aquatic environments as a result of the proposed applications
of Trichoderma gamsii strain ICC 080.  A 48-hour Daphnia test with the
EP was submitted but was determined to be unacceptable due to the
inadequate study duration; however, the available data on the TGAI are
adequate to assess the potential risk of Trichoderma gamsii strain ICC
080.  Trichoderma species are ubiquitous in the environment, and do not
proliferate in aquatic ecosystems, so significant exposure in freshwater
environments is not expected from the proposed applications of
Trichoderma gamsii strain ICC 080.  Therefore, adverse effects to
freshwater aquatic animals are not anticipated.

Trichoderma species are not related to any known plant pathogens, and as
determined above, Trichoderma gamsii strain ICC 080 is not expected to
occur in freshwater environments in significant amounts.  Therefore,
data from aquatic plant testing are not required, and no effects are
anticipated to this taxon as a result of the proposed applications.  A
study with the aquatic vascular plant, Lemna gibba, was submitted and
reviewed, and results show that the 7-day growth inhibition NOEC for the
aquatic vascular plant Lemna gibba is 2.1x107 cfu/mL.  These results
confirm the conclusion of no anticipated adverse effects to aquatic
plants.

As with freshwater environments, significant amounts of Trichoderma
gamsii strain ICC 080 are not expected to reach marine/estuarine
environments.  Therefore data are not required for these taxa, and
adverse effects in these environments are not anticipated.

Based on the data submitted and what is known about the nature of
Trichoderma species in aquatic environments, adverse effects to
freshwater and marine/estuarine fish, invertebrates, and plants are not
expected as a result of the proposed label applications of Trichoderma
gamsii strain ICC 080.  

3. Threatened and Endangered Species Assessment

There are no listed endangered or threatened species related to the
target pests.  Since it is concluded that effects are not anticipated
for non-target species exposed to Trichoderma gamsii strain ICC 080 as a
result of proposed labeled applications, effects to listed species are
also not expected.  Therefore, a “No Effect” determination is made
for use of Trichoderma gamsii strain ICC 080 as labeled. 

For a comprehensive guideline-by-guideline summary of the non-target
toxicity data requirements, refer to   HYPERLINK  \l "Table4"  Table 3 
in Appendix A.

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, in the development, implementation, and
enforcement of environmental laws, regulations, and policies. To help
address potential environmental justice issues, the Agency seeks
information on any groups or segments of the population who, as a result
of their location, cultural practices, or other factors, may have
atypical, unusually high exposure to Trichoderma gamsii strain ICC 080,
compared to the general population.  Please comment if you are aware of
any sub-populations that may have atypical, unusually high exposure
compared to the general population.

VI.   RISK MANAGEMENT AND REGISTRATION DECISIONS tc "V.   RISK
MANAGEMENT AND REGISTRATION DECISION" 

	A.  Determination of Eligibility 

 tc "A.  Determination of Eligibility " \l 2 

Section 3(c)(5) of FIFRA provides for the registration of new active
ingredients if it is determined that (A) its composition is such as to
warrant the proposed claims for it; (B) its labeling and other materials
required to be submitted comply with the requirements of FIFRA; (C) it
will perform its intended function without unreasonable adverse effects
on the environment; and (D) when used in accordance with widespread and
commonly recognized practice it will not generally cause unreasonable
adverse effects on the environment. 

The four criteria of the Eligibility Determination for Pesticidal Active
Ingredients are satisfied by the science assessments supporting products
containing Trichoderma gamsii strain ICC 080.  Such products are not
expected to cause unreasonable adverse effects, and are likely to
provide protection as claimed when used according to label instructions.
Therefore, Trichoderma gamsii strain ICC 080 is eligible for
registration as a microbial fungicide.  

B.  Regulatory Decision

On October 1, 2009, EPA announced a new policy to provide a more
meaningful opportunity for the public to participate on major
registration decisions before they occur.  According to this new 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 use; first outdoor
use; and first residential use.  Accordingly, this pesticide is subject
to a 30-day comment period as a new active ingredient with both outdoor
uses and residential uses.  Any regulatory decision will be informed by
the comments EPA receives.

At this time, EPA believes, the data submitted fulfill the requirements
of registration for products containing Trichoderma gamsii strain ICC
080 as a microbial fungicide.  Acute toxicity data for Trichoderma
gamsii strain ICC 080 demonstrate that it has low toxicity (category III
and IV).  EPA has no concerns for any non-target organisms, including
threatened or endangered species, exposed to Trichoderma gamsii strain
ICC 080 when it is used in accordance with approved label directions. 
EPA has not identified any toxic endpoints for non-target mammals,
birds, plants, aquatic, or soil organisms.  EPA supports its
registration under Section 3(c) (5) of the Federal Insecticide,
Fungicide, and Rodenticide Act (FIFRA).  Refer to Appendix B for
product-specific information.

		1.  Conditional/Unconditional Registration

The Environmental Protection Agency (EPA) considered information
submitted for granting registration under Section 3(c)(5) of the Federal
Insecticide, Fungicide and Rodenticide Act (FIFRA) and determined that
the data and information submitted in support of Trichoderma gamsii
strain ICC 012 adequately satisfy current data requirements (refer to 40
CFR Subpart U § 158.2000).  If the Agency receives comments during the
30 day public comment period that inform EPA’s initial decision, EPA
will address such new information and take appropriate action.   

		C.  Labeling

Before releasing pesticide products containing Trichoderma gamsii strain
ICC 080 for shipment, the applicant is required to provide appropriate
labels.

VII.   ACTIONS REQUIRED BY THE APPLICANT

The Agency evaluated the data submitted in connection with the initial
registration of Trichoderma gamsii strain ICC 080 and determined that
these data fulfill current registration requirements.  No additional
data are required to be submitted to the Agency at this time. 
Additional data may be required for new uses and/or changes to existing
uses.  

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

	A.  Reporting of Adverse Effects and Hypersensitivity Incidents

Reports of all incidents of adverse effects to the environment must be
submitted to the Agency 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 the Agency under
the provisions of 40 CFR Part 158.2140 (Guideline reference number OPPTS
885.3400).

		

VIII.  GLOSSARY OF ACRONYMS AND ABBREVIATIONS

BCCM	Belgian Coordinated Collections of Microorganisms

BPPD	Biopesticides and Pollution Prevention Division

BRAD	Biopesticides Registration Action Document

CFR	Code of Federal Regulations

cfu 	colony-forming unit

cm2 	square centimeter

°C 	degrees Celsius

EDSP	Endocrine Disruptor Screening Program

EDSTAC	Endocrine Disruptor Screening and Testing Advisory Committee

EP	end-use product

EPA	Environmental Protection Agency (the “Agency”)

FFDCA	Federal Food, Drug, and Cosmetic Act

FIFRA	Federal Insecticide, Fungicide, and Rodenticide Act

FQPA 	Food Quality Protection Act

FR	Federal Register

g 	gram

kg	kilogram

L 	Liter

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).

LLC 	limited liability company

LOEC	lowest observable effect concentration

MRID No.	Master Record Identification Number

μm 	micrometer

mg	milligram

mL	milliliter

mm	millimeter

MUCL 	Mycothèque de l'Université Catholique de Louvain

NIOSH 	National Institute for Occupational Safety and Health

NOEC	no observable effect concentration

OPP	Office of Pesticide Programs

OPPTS 	Office of Prevention, Pesticides, and Toxic Substances 

PII 	Primary Irritation Index

P.O. Box	Post Office Box

PP 	Pesticide Petition

PPE 	personal protective equipment

TGAI 	technical grade of the active ingredient

IX.  BIBLIOGRAPHY

A.  Studies Submitted in Support of Trichoderma gamsii Strain ICC 080

MRID	Citation	Receipt Date

47345800	Isagro U.S.A. (2008) Submission of Product Chemistry and
Toxicity Data in Support of the Application for Registration of
Trichoderma ICC 080 gamsii. Transmittal of 13 Studies.	08-Feb-2008

47345801	Leuschner, P. (2004) Acute Toxicity Study of Trichoderma viride
Strain ICC 080 By Oral Administration to Rats. Project Number: 17290/03.
Unpublished study prepared by Laboratory of Pharmacology & Toxicology .
30 p.	08-Feb-2008

47345802	Leuschner, P. (2004) Acute Toxicity Study of Trichoderma viride
by Intraperitoneal Injection to Rats. Project Number: 17472/03.
Unpublished study prepared by Laboratory of Pharmacology & Toxicology .
32 p.	08-Feb-2008

47345803	Leuschner, P. (2004) Acute Pulmonary Toxicity / Pathogenicity
of Trichoderma viride Strain ICC 080 by Intratracheal Administration to
Rats. Project Number: 17291/03. Unpublished study prepared by Laboratory
of Pharmacology & Toxicology . 48 p.	08-Feb-2008

47345804	Dengler, D. (2004) Analysis of the Occurrence of Test Substance
Trichoderma viride Conidia in Animal Tissue. Project Number:
20031372/01/AMAT. Unpublished study prepared by GAB Biotechnologie Gmbh.
25 p.	08-Feb-2008

47345805	Pawlowski, S. (2004) Toxicity of Trichoderma viride Strain ICC
080 to Rainbow Trout (Oncorhynchus mykiss) in a Prolonged Toxicity Test.
Project Number: 17113231. Unpublished study prepared by Institut fuer
Biologische Analytik und Consulting IBACON. 67 p.	08-Feb-2008

47345806	Pawlowski, S. (2004) Influence of Trichoderma viride Strain ICC
080 to Daphnia magna in a Reproduction Test. Project Number: 17112221.
Unpublished study prepared by Institut fuer Biologische Analytik und
Consulting IBACON. 68 p.	08-Feb-2008

47345807	Schmitzer, S. (2004) Effects of Trichoderma viride (Acute
Contact and Oral) on Honey Bees (Apis mellifera L.) in the Laboratory.
Project Number: 17115035. Unpublished study prepared by Institut fuer
Biologische Analytik und Consulting IBACON. 30 p.	08-Feb-2008

47345808	Spatz, B.; Pawlowski, S. (2004) Toxicity of Trichoderma viride
Strain ICC 080 to the Aquatic Plant Lemna gibba in a Growth Inhibition
Test. Project Number: 17114240. Unpublished study prepared by Institut
fuer Biologische Analytik und Consulting IBACON. 69 p.	08-Feb-2008

47345809	Reis, K. (2004) Effects of Trichoderma viride on the Activity
of the Soil Microflora in the Laboratory. Project Number: 17119080.
Unpublished study prepared by Institut fuer Biologische Analytik und
Consulting IBACON. 47 p.	08-Feb-2008

47345810	Gossmann, A.; Luhrs, U. (2004) Acute Toxicity (14 Days) of
Trichoderma viride to the Earthworm Eisenia fetida in Artificial Soil.
Project Number: 17118021. Unpublished study prepared by Institut fuer
Biologische Analytik und Consulting IBACON. 28 p.	08-Feb-2008

47345811	Winkler, V. (2007) Trichoderma gamsii Strain ICC 080 (Formerly
Classified as Trichoderma viride): Group A: Product Analysis. Project
Number: 110207. Unpublished study prepared by Isagro S.P.A. 74 p.
08-Feb-2008

47345812	Winkler, V. (2007) Trichoderma gamsii Strain ICC 080 (Formerly
Trichoderma viride): Group B: Physical and Chemical Properties (Self
Certification). Project Number: 112507. Unpublished study prepared by
Chemservice S.P.A. 7 p.	08-Feb-2008

47345813	Garofani, S. (2007) Trichoderma viride: Complete Analysis of
Five Batch Samples. Project Number: CH/317/2007. Unpublished study
prepared by Chemservice S.r.l. 28 p.	08-Feb-2008

47346300	Isagro S.P.A. (2008) Submission of Product Chemistry and
Toxicity Data in Support of the Application for Registration of Bioten
WP (Remedier WP). Transmittal of 10 Studies.	08-Feb-2008

47346301	Leuschner, P. (2004) Acute Toxicity Study of Remedier WP
(Trichoderma harzianum and Trichoderma viride Wettable Powder) by Oral
Administration to Rats. Project Number: 17795/04. Unpublished study
prepared by Laboratory of Pharmacology & Toxicology . 30 p.	08-Feb-2008

47346302	Leuschner, P. (2004) Acute Toxicity Study of Remedier WP
(Trichoderma harzianum and Trichoderma viride Wettable Powder) in Rats
by Dermal Administration. Project Number: 17796/04, 140185. Unpublished
study prepared by Laboratory of Pharmacology & Toxicology . 33 p.
08-Feb-2008

47346303	Leuschner, P. (2004) Acute Inhalation Toxicity Study of
Remedier WP (Trichoderma harzianum and Trichoderma viride Wettable
Powder) by Oral in Rats. Project Number: 17797/04, 140185. Unpublished
study prepared by Laboratory of Pharmacology & Toxicology . 35 p.
08-Feb-2008

47346304	Leuschner, P. (2004) Acute Eye Irritation Study of Remedier WP
(Trichoderma harzianum and Trichoderma viride Wettable Powder) by
Instillation into the Conjunctival Sac of Rabbits. Project Number:
17799/04, 140185. Unpublished study prepared by Laboratory of
Pharmacology & Toxicology. 27 p.	08-Feb-2008

47346305	Leuschner, P. (2004) Acute Skin Irritation Test (Patch Test) of
Remedier WP (Trichoderma harzianum and Trichoderma viride Wettable
Powder) in Rabbits. Project Number: 17798/04, 140185. Unpublished study
prepared by Laboratory of Pharmacology & Toxicology . 27 p.	08-Feb-2008

47346306	Leuschner, P. (2004) Examination of Remedier WP (Trichoderma
harzianum and Trichoderma viride Wettable Powder) in a Skin
Sensitisation Test in Guinea Pigs According to Magnusson and Kligman
(Maximisation Test). Project Number: 17800/04, 140185. Unpublished study
prepared by Laboratory of Pharmacology & Toxicology . 40 p.	08-Feb-2008

47346307	Pawlowski, S. (2004) Acute Toxicity of Remedier WP to Daphnia
magna in a 48-hour Immobilization Test. Project Number: 19561220.
Unpublished study prepared by Institut fuer Biologische Analytik und
Consulting IBACON. 46 p.	08-Feb-2008

47346308	Rosenkranz, B. (2004) Effects of Remedier WP on the Predatory
Mite Typhlodromus pyri in the Laboratory - Dose Response Test. Project
Number: 19562063. Unpublished study prepared by Institut fuer
Biologische Analytik und Consulting IBACON. 40 p.	08-Feb-2008

47346309	Winkler, V. (2007) Bioten WP: Group A: Product Analysis.
Project Number: 111907. Unpublished study prepared by Isagro S.P.A. 50
p.	08-Feb-2008

47346310	Winkler, V. (2007) Bioten WP: Group B: Physical and Chemical
Properties (Self Certification). Project Number: 112607. Unpublished
study prepared by Chemservice S.P.A. 7 p.	08-Feb-2008

47460900	Isagro USA, Inc. (2008) Submission of Product Chemistry Data in
Support of the Applications for Registration of Trichoderma gamsii
strain ICC 080 Technical, Trichoderma asperellum ICC 012 Technical, and
Bioten WP. Transmittal of 1 Study.	30-Jun-2008

47460901	Winkler, V (2008) Response to EPA April 24, 2008 Letter
"Deficiencies Noted During Preliminary Screen of Submissions for
Trichoderma gamsii strain ICC 080 Technical: Trichoderma asperellum ICC
012 Technical: and Bioten WP" . Project Number: 062308. Unpublished
study prepared by Chemservice S.P.A., DSMZ-Deutsche Sammlung von
Mikroorganismen und Zellkulturen GmbH, and Isagro S.p.A. Unita prodotti
Biologic. 88 p.	30-Jun-2008

47643900	Isagro S.P.A. (2009) Submission of Product Chemistry Data in
Support of the Application for Registrations of Bioten WP, Trichoderma
Gamsii strain ICC 080 Technical and Trichoderma asperellum ICC 012
Technical. Transmittal of 1 Study.	14-Jan-2009

47643901	Lodini, S. (2008) Mesophilic Contaminants in Active Ingredients
Trichoderma gamsii Strain ICC 080 and Trichoderma asperellum ICC 012:
Final Report. Project Number: REM/80122. Unpublished study prepared by
Isagro S.P.A. 30 p.	14-Jan-2009

47802700	ISAGRO S.P.A. (2009) Submission of Product Chemistry Data in
Support of the Applications for Registration of Trichoderma gamsii
strain ICC 080 Technical, Trichoderma asperellum ICC 012 Technical, and
Tenet WP. Transmittal of 1 Study. 	15-Jun-2009

47802701	Graben, M. (2009) Response to EPA on Phone Conference of June
4, 2009 and April 2, 2009 Letter: Deficiencies Noted During Review of
Submissions for Trichoderma gamsii strain ICC 080 Technical: Trichoderma
asperellum ICC 012 Technical: and Bioten WP (EPA File Symbols: 80289-RN:
and 8028-O [PRIA Category: B590]. Including Two Updated End Use Labels
and Refined CSF. Project Number: 061209, 901, 800/02REV. Unpublished
study prepared by ISAGRO S.P.A. 88 p.	15-Jun-2009

B.  Environmental Protection Agency Risk Assessment Memorandums

U.S. EPA.  1998.  Non-target organism data requirements for Makhtashim
Chemical Works Ltd.’s Trichodex containing the microbial fungicide
Trichoderma harzianum T-39.  Memorandum from M. Mendelsohn to S. Bacchus
and P. Hutton.

U.S. EPA.  2000.  Trichoderma harzianum Rifai strain T-39 (119200)
technical document.  URL:   HYPERLINK
"http://epa.gov/oppbppd1/biopesticides/ingredients/tech_docs/tech_119200
.htm" 
http://epa.gov/oppbppd1/biopesticides/ingredients/tech_docs/tech_119200.
htm . 

U.S. EPA 2008.  Trichoderma species final Reregistration Review
decision.  Case 6050.  Docket No. EPA-HQ-OPP-2006-0245.

C.  Other References Cited

Glinski, Z. and K. Buczek.  2003.  Response of the Apoidea to fungal
infections.  Apiacta 38:183-189.

Gouli, S., V. Gouli, M. Skinner, B. Parker, J. Marcelino, and M.
Shternshis.  2008.  Mortality of Western flower thrips (Frankliniella
occidentalis) under influence of single and mixed fungal inoculations. 
Journal of Agricultural Technology 4(2):37-47. 

Howell, C. R.  2003.  Mechanisms employed by Trichoderma species in the
biological control of plant diseases: the history and evolution of
current concepts.  Plant Disease 87(1):4-10.

Prinzinger, R., A. Pressmar, and E. Schleucher.  1991.  Body temperature
in birds.  Comparative Biochemistry and Physiology 99A(4):499-506.

Santaramarina, M. P., J. Rosello, R. Llacer, and V. Sanchis.  2002. 
Antagonistic activity of Penicillium oxalicum Corrie and Thom,
Penicillum decumbens Thom, and Trichoderma harzianum Rifai isolates
against fungi, bacteria and insects in vitro.  Revista Iberoamericana de
Micologia 19:99-103.

Jijakli MH, Lepoivre P, Tossut P, Thornard P. 1993. Biological control
of Botrytis

		cinerea and Penicillium sp. on post-harvest apples by two antagonistic
yeasts. 

		Med. Fac. Landbouww. Univ. Gent 53(3b):1349–1358.

  

Yemma JJ, Berk MP. 1994. Chemical and physiological effects of Candida
albicans 

		toxin on tissues. Cytobios 77(310):147–158.

APPENDIX A – MICROBIAL PESTICIDE DATA REQUIREMENTS

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

                  Trichoderma gamsii strain ICC 080, and Its Associated
End-Use Product (EP), Tenet WP (40 CFR § 158.2120)

Data Requirement

 (OPPTS Guideline)	Results	MRID Number

	TGAI	EP

	Product Chemistry and Composition

Product Identity

 (885.1100)	Trichoderma gamsii strain ICC 080, originally characterized
as Trichoderma viride, has been recharacterized as Trichoderma gamsii
and also has been characterized by molecular biology techniques.
Trichoderma gamsii strain ICC 080 was isolated from a suppressive soil
in Sardinia, Italy.  Submitted data satisfy the requirements 

Classification: Acceptable 	Submitted data satisfy the requirements of
Product Identity for the EP.

 Classification: Acceptable	47345811

47346309

47460901

47643901

Manufacturing Process (885.1200)	Submitted data satisfy the requirements
of manufacturing process for both the TGAI and EP. 

Classification: Acceptable	47345811

47346309

47460901

Deposition of a Sample in a Nationally Recognized Culture Collection

(Not applicable)	Trichoderma gamsii strain ICC 080 has been deposited in
the CABI Bioscience International Mycological Institute-Egham, UK with
the Accession Number IMI CC No. 392151.

	Discussion of Formation of Unintentional Ingredients (885.1300)
Submitted data satisfy the requirements of discussion of formation of
unintentional ingredients for both the TGAI and EP.

Classification: Acceptable	47345811

47346309

47460901

47802701

 

Analysis and Certified Limits

Analysis of Samples (885.1400)	Submitted data satisfy the requirements
of analysis of samples for both the TGAI and EP.

Classification: Acceptable	47345811

47345813 

47346309

47460901

 47802701

Certification of Limits (885.1500)	The certified limits for the Active
ingredient were within the specified range.

Classification: Acceptable 	 The certified limits for the

 active ingredients exceed the

 OPPTS Guideline 830.1750

 specified ranges, but an

 acceptable explanation was

 provided.

 Classification: Acceptable	47345811

47346309

47460901

47802701

Physical and Chemical Characteristics

Color

(830.6302)	Gray green 	Not applicable	47345812

47346310

Physical State

(830.6303)	Solid powder	Not applicable	47345812

47346310

Odor

(830.6304)	Slight odor	Not applicable	47345812

47346310

Stability to Normal and Elevated Temperatures, Metals, and Metal Ions 

(830.6313)	Not required for MP 

	Not applicable	47345812

47346310

Storage Stability

(830.6317)	Stable for 6 months at 20ºC, not stable at 54ºC	Active
ingredient stable for 15 months at 20-25ºC 

	47345812

47346310

Miscibility

(830.6319)	Product is not a suspension.	Product is not a suspension.

	47345812

Corrosion Characteristics

(830.6320)	Product is not corrosive.	Product is not corrosive.	47345812.

pH

(830.7000)	 6.6 (1% aqueous solution)	Not applicable	47345812.

Viscosity

(830.7100)	Product is not a liquid. 	Product is not a liquid. 	47345812

47346310

Density/Relative Density/Bulk Density

(830.7300)	0.2 g/mL.	Not applicable	47345812

47346310



TABLE 2. Toxicology Data Requirements for the Technical Grade of the
Active Ingredient (TGAI), Trichoderma gamsii Strain ICC 080, and Its
Associated End-Use Product (EP), Bioten WP(40 CFR § 158.2140)

Data Requirement

(OPPTS Guideline)	Results	MRID Number

	TGAI	EP

	Tier I

Acute Oral Toxicity/Pathogenicity

(885.3050)	Trichoderma gamsii strain ICC 080 (7.5 x 108 CFU/g) in 0.9%
NaCl solution was not toxic at a dose of 2000 mg/kg bw.  

Trichoderma gamsii  strain ICC 080 was of Low Toxicity.  Microbial
enumeration was not done so it is unknown if Trichoderma gamsii  is
infective or pathogenic in the rat. This study is classified
SUPPLEMENTAL, but is upgraded by the Pulmonary Toxicity study which has
the clearance data.

Classification: Acceptable	Not applicable

	47345801

Acute Pulmonary Toxicity/Pathogenicity

(885.3150)	Not toxic, infective, and/or pathogenic to rats by pulmonary
dose of 2.5 x 106 cfu/animal.

Classification: Acceptable	Not applicable	

47345803 47345804

Acute Intraperitoneal Injection Toxicity/Pathogenicity

(885.3200)	Not toxic to rats by injection at dose of 1 x 107 cfu/animal.

This study is classified SUPPLEMENTAL, but is upgraded by the Pulmonary
Toxicity study which has the clearance data.

Classification: Acceptable	Not applicable	47345802

Hypersensitivity Incidents 

(885.3400)	Any hypersensitivity incidents must be reported per OPPTS
Guideline 885.3400. 

	Cell Culture

(885.3500)	Not required because Trichoderma gamsii Strain ICC 080 is not
a virus (refer to test note #4 of 40 CFR § 158.2140(d)).	Not applicable
Not applicable

Acute Oral Toxicity

(870.1100)

	Not applicable	Bioten WP/ Remedier WP (Trichoderma asperellum  7.8 x
107 CFU/g; Trichoderma gamsii 4.2 x 107 CFU/g) is not toxic 5.3.9) in
the rat.

Oral LD50  was 

> 2000 mg/kg bw 

Limit test; no mortality occurred during the study

Classification: Acceptable

TOXICITY CATEGORY III 	47346301

Acute Dermal Toxicity

(870.1200)	Not applicable	When dosed with Bioten WP/ Remedier WP in 0.9%
NaCl at a dose of 2000 mg/kg bw is of MODERATE Toxicity.  There were no
treatment related clinical signs, necropsy findings or changes in body
weight.

Classification: Acceptable

TOXICITY CATEGORY III 	43746302

Acute Inhalation Toxicity

(870.1300)	Not applicable	 Bioten WP/ Remedier WP Inhalation LC50were as
follows: Males > 5.20 mg/L, 

Females > 5.20 mg/ L, Combined > 5.20 mg/L. 

Bioten WP/ Remedier WP is of LOW Toxicity, based on lack of mortality in
male and female rats. There were no treatment related clinical signs,
necropsy findings or changes in body weight. 

Classification: Acceptable

TOXICITY CATEGORY IV 	47346303

Acute Eye Irritation

(870.2400)	Not applicable	Bioten WP was not an eye irritant.

Classification: Acceptable

TOXICITY CATEGORY IV 	47346304

Primary Dermal Irritation

(870.2500)	Not applicable	When dosed with 500 mg of Bioten WP/ Remedier
WP (Trichoderma conidia (1.2 x 108 CFU/g); Trichoderma asperellum ICC
012 (7.8 x 107 CFU/g); Trichoderma gamsii strain ICC 080 (4.2 x 107
CFU/g)), the product is slightly irritating.

 

Classification: Acceptable

TOXICITY CATEGORY IV	47346305

Skin Sensitization

(870.2600)	Not applicable	This study is not required

Bioten WP was not a dermal sensitizer.

Classification: Acceptable	47346306

Tiers II and III

Not required for Trichoderma gamsii Strain ICC 080  based on the lack of
acute toxicity/pathogenicity in the Tier I studies.

TABLE 3.   Summary of data submitted to comply with non-target organism
data requirements published in 40 CFR § 158.2150 for support of the
registration of Trichoderma gamsii strain ICC 080.

Data Requirement	OPPTS Guideline No.	Results Summary and Classification
MRID No.

Avian oral toxicity	885.4050	Data waiver rationale provides sufficient
information to determine that toxicity/pathogenicity to avian wildlife
is not expected. 

Classification:  Acceptable	Contained in tolerance petition

Avian inhalation toxicity/pathogenicity	885.4100	Data are not required. 
Nature of microbial pesticide does not indicate potential pathogenicity
to birds or relatedness to any known bird pathogens.	N/A

Wild mammal toxicity/pathogenicity	885.4150	Tests required by 40 CFR §
158.2140 are adequate and appropriate for assessment of hazards to wild
mammals.  Testing indicates no adverse effects to laboratory rats at 7.7
x 107 spores/mL (2000 mg TGAI/kg bw) when dosed orally.  Testing with
the EP indicates no adverse effects to laboratory rats dosed orally at
2.3 x 107 spores/mL (2000 mg EP/kg bw).

Classification:  Acceptable for wild mammal risk assessment	473458-01

473463-01

Freshwater fish toxicity/pathogenicity	885.4200	The 30-day LC50 for
rainbow trout (Oncorhynchus mykiss) is >4.9 x 1010 cfu/L. 

Classification:  Acceptable	47345805

Freshwater invertebrate toxicity/pathogenicity	885.4240	The 21-day NOEC
for Daphnia based on mortality/immobility and reproduction was ≥ 1x107
cfu/mL.  A 48-hour Daphnia test with the EP (containing T. gamsii strain
ICC 080 and T. asperellum ICC 012) was also submitted but was determined
to be unacceptable due to the inadequate study duration.

Classification:  Acceptable	47345806

47346307

Estuarine/marine fish and invertebrate testing	885.4280	Data are not
required.  T. gamsii strain ICC 080 will not be applied directly to
water and is not expected to enter marine/estuarine environments in
amounts that are significantly higher than naturally-occurring
concentrations.	N/A

Non-target plant testing	885.4300	Data are not required, since T. gamsii
strain ICC 080 is not related to known plant pathogens, and is not
expected to have adverse effects on plants.  Additionally, a 7-day
growth inhibition NOEC for the aquatic vascular plant Lemna gibba was
2.1x107 cfu/mL.

Classification:  Acceptable	473458-08

Non-target insect testing	885.4340	LD50 >5 kg/ha for EP (containing T.
gamsii strain ICC 080 and T. asperellum ICC 012) for predatory mite
(Typhlodromus pyri) and no significant effects on reproduction observed.
 

Classification:  Acceptable	47346308

Honey bee testing	885.4380	48-hour contact and oral LD50 were > 0.85 x
105 cfu/bee and >1.9 x 105 cfu/bee, respectively.  The study was of too
short a duration.  

Classification:  Uncceptable

	47345807

Non-guideline testing

Earthworm (Eisenia fetida) testing	N/A	The 14-day earthworm (Eisenia
fetida) contact LD50 was determined to be >1.7 x 109 cfu/kg soil.

Classification: Acceptable 	47345810

Non-guideline testing

Effects on Soil Microflora

Activity	N/A	Soil respiration, nitrate content, nitrogen formation rate,
and mineral nitrogen content were below the 25 % threshold value,
indicating no significant perturbation of the soil microflora community 

Classification: Acceptable	47345809

APPENDIX B – Trichoderma gamsii strain ICC 080 Products

EPA Registration Number	Registration Name	Percentage Active Ingredient
Formulation Type	Use Site	Method of Application	Application Rate	Target
Pests

80289-9	Bioten  WP

 (or TenetWP)	2.0% T. gamsii

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 asperellum	Wettable Powder	Alfalfa

Berries

Cereal Grains

Citrus:

Clover 

Cole Crops

Corn:

Cotton

Cucurbits 

Grass, Forage, Fodder, and Hay 

Fruiting Vegetables

Ginseng

Herbs (fresh, dried and for oil)

Leafy Vegetables (except Brassica) Legume Vegetables 

Olive 

Onions

Dry bulb

Pineapple 

Peanut

Pomegranate Root , Tuber, and Corm Vegetables Sunflower

Tobacco

Tree Fruits 

Tree Nuts

Tropical Fruit

At Greenhouse, nursery and field sites

Vines:

(table grapes, wine grapes, muscadines, hops, kiwi)	In-furrow and banded
application. 

Broadcast application

Cutting and bare root application

Drip, drench and chemigation.

Substrate mix.	Rates vary by crop and application method	Armillaria sp.,
Fusarium spp., Phytophthora spp., Pythium spp., Rhizoctonia spp.,
Rosellinia sp., Sclerotinia spp. Sclerotium rolfsii, Thielaviopsis
basicola, Verticillium spp 

80289-10	Trichoderma gamsii strain ICC 080	99.9%

Formulating use only	N/A	N/A

	

Trichoderma gamsii strain ICC 080 

Biopesticides Registration Action Document (BRAD)	December 2009

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 PAGE   37