**UNITED STATES ENVIRONMENTAL PROTECTION AGENCY**
**WASHINGTON, D.C. 20460**

OFFICE OF CHEMICAL SAFETY

AND POLLUTION PREVENTION

**September 22, 2010**

SUBJECT: Environmental Risk Assessment for a FIFRA Section 3 Registration for Trichoderma asperellum , strain T34 for Control of Fungal Root Disease in

Ornamental and Carnation Plants in Greenho uses [EPA File No. 87301

- R ] ; Decision # 425700, DP Barcode # 373656, Submission # 865549 MEMORANDUM

FROM: Shannon Borges, Biologist / signed /

Microbial Pesticides Branch Biopesticides and Pollution Prevention Division, 7511P

THROUGH: Zigfridas Vaituz is, Senior Scientist / signed /

Microbial Pesticides Branch Biopesticides and Pollution Prevention Division, 7511P

TO: Anna Gross, Regulatory Action Leader

Microbial Pesticides Branch Biopesticides and Pollution Prevention Division, 7511P

Bio control Technologies, S.L. has applied for a FIFRA Section 3 registration for Trichoderma asperellum strain T34 use on ornamental and carnation plants in greenhouses for control of fungal root diseases. This memorandum contains the ecological risk assessm ent for this proposed registration.

I. Introduction

Trichoderma asperellum strain T34 is an active ingredient proposed for a FIFRA Section 3 registration for use on ornamental and carnation plants in greenhouses

. T. asperellum strain T34

is intended to control fungal diseases within the rhizosphere of these plants resulting from pathogenic microorganisms , primarily Fusarium oxysporum . The primary mode of fungicidal

action of T. asperellum strain T34 is competition with pathogenic fungi for space and nu trients through colonization of plant root zones and construction of a physical barrier to infection

, parasitization of pathogenic fungi, and induction of plant resistance responses. The proposed

end

- use product containing T. asperellum strain T34 is appl ied via treatment of potting media before planting, treatment of the potting media surface at or after planting either through a spray

or through an irrigation system

, or root dip. If needed, reapplication is suggested every 2

- 3

months or prior to a situa tion that might cause increase d risk of disease

.II. Environmental Exposure

The proposed label directs the user to apply T. asperellum strain T34 at a rate of 10g/m 3 of potting media prior to planting , at a rate of 0.5 g per m 2 surface area immediately after planting,

or at a r ate of 5 g per 1000 1

- L pots containing potting medium

. These maximum application

rates result in concentrations of 6.6 x 10 3

– 1.0 x 10 4 cfu T. asperellum strain T34 /mL potting

medium. Applications via root dip are expected to r esult in much lower concentrations within the potting media, since the T. asperellum strain T34 will be localized at the roots. Trichoderma

spp. concentrations in soil have been reported at 10 3

– 10 6 cfu/g (Harman et al. 2004 , Leandro et al. 2007, Liu et al. 2008 ) , and T. asperellum strain T34 was found to persist at 10 4 cfu/mL potting medium at four months after application

, based on data submitted by the registrant in

MRID 48067407. Therefore, the population size of T. asperellum strain T34 is n ot expec ted to significantly increase beyond that resulting from the proposed application rates and will not be larger than that of Trichoderma spp. naturally found in soil . In the event that treated potting media were disposed of outdoors, nontarget organisms wo uld not be exposed to Trichoderma concentrations above naturally occurring levels. Nonetheless, exposure in outdoor environments is expected to be very limited, since the product is intended for use indoors in greenhouses. Therefore, exposure to nontarge t organisms is expected to be negligible, if it occurs at all.

II I . Environmental Effects Data

Table 1 provides the status of the data requirements for the technical grade active ingredient (TGAI). T he data waiver rationale s used for this assessment ar e summarized within the

environmental risk conclusions in the section below , and a Data Evaluation Record of the

rationale submitted and its supporting documentation is attached. The information provided is sufficient to satisfy the Tier I nontarget organ ism data requirements and for nontarget organism

risk assessment for the TGAI . Further testing of nontarget organisms at higher tier levels is not required.

Table 1

. Status of data required to comply with nontarget organism data requirements published in 40 CFR § 158.2150 for support of the registration of Trichoderma asperellum Strain T34

.

Data Requirement OPPTS

Guideline No. Results Summary and

Classification MRID No.

• Avian oral toxicity/pathogenicity

• Freshwater fish toxicity/pathogenicity

• Freshwater

invertebrate toxicity/pathogenicity

• Estuarine/marine fish and invertebrate testing • Nontarget insect testing • Honey bee testing 885.4050

885.4200

885.4240

885.4280

885.4340

885.4380 Data waiver rationale provides

sufficient informati on to determine that toxicity/pathogenicity to avian wildlife, wild mammals,

freshwater and marine/estuarine fish and invertebrates, honey bees, and nontarget insects is not expected as a result of the proposed label uses. Classification: Acceptable 4706 8102Data Requirement OPPTS

Guideline No. Results Summary and

Classification MRID No.

Wild mammal toxicity/pathogenicity 885.4150 Studies required by 40 CFR § 158.2140 are adequate and appropriate for assessment of

hazards to wild mammals. Acute oral toxicity/pathogenicity testing

indicated no significant adverse

effects to laborat ory rats dosed at

1.1 x 10 8
Classification: Acceptable for

wild mammal risk assessment cfu/animal after 21. 4 8067401

• Avian inhalation toxicity/pathogenicity • Nontarget plant testing 885.4100

885.4300 Not required

– active ingredient is

not related t o known avian or plant pathogens N/A

IV. Environmental Risk Conclusions

A. Terrestrial animals and plants

The proposed registration limits use to indoor environments within greenhouses and the proposed maximum application use rates result in concen trations that are not above measured

concentrations of Trichoderma spp. naturally occurring in soil. Therefore, exposure in the terrestrial environment is expected to be negligible, if it occurs at all.

The registrant submitted data waiver rationale to fulfill the requirement of acute oral and avian inhalation toxicity/pathogenicity studies. Data from an optimum growth temperature study was also submitted, and showed that T. asperellum strain T34 does not grow at normal avian body temperatures. Based on this information, exposure of birds to T. asperellum strain T34 is not expected to result in adverse effects on birds and risk to birds resulting from the proposed use of T. asperellum strain T34 is not anticipated.

Acute oral toxicity/pathogenicity te sting indicated no significant adverse effects to laboratory

rats dosed at 1.1 x 10 8 cfu/animal after 21 days (MRID 48067401)

. There is no reason that tests on laboratory mammals are not representative of the potential hazard to wild mammals in this

case; therefore, based on this information, adverse effects on wild mammals are not expected from exposure to T. asperellum strain T34. Since the proposed use is also expected to minimize

exposure
, risk to wild mammals resulting from the proposed registration o f T. asperellum strain T34 is not anticipated.

The registrant submitted information on the toxicity/pathoge nicity data for several insect species exposed to T. harzianum ( the former classification of some strains of T. asperellum ) and T. gamsii . While th ese data cannot be used directly to determine the potential hazard of this particular strain of T. asperellum , they do provide some evidence of the lack of effects ofTrichoderma spp. on insects. The Agency has previously expressed concern over the potent ial

hazard of T. harzianum strain T39 (USEPA 2000)

, and several papers have shown adverse

effects of this related fungus on nont arget insects, particularly coleopteran species (Cardoza et al. 2006, Jassim et al. 1990, Shakeri and Foster 2006). Nontarget i nsect testing has not been performed for T. asperellum strain T34; however, because exposure is expected to be minimal risk to nontarget insects is not anticipated. F uture expansion of the uses of T. asperellum strain T34 may require additional nontarget insect toxicity/pathogenicity testing.

As with other nontarget insects, t he Agency had previously expressed concerns for potential effects on honey bees for a related Trichoderma spp.
, T. harzianum strain T39 (USEPA 2000); however, data were subsequently submitted to show that adverse effects are not expected from exposure to that strain. Trichoderma spp. are widely understood to present no potential adverse

effects to honey bees, which is supported by data submitted by the registrant showing lack of

effe cts of honey bees and bumble bees ( Bombus terrestris ) following exposure to several

Trichoderma spp. Therefore, BPPD concludes that T. asperellum strain T34 will no t pose risk to honey bees as a result of the proposed labeled uses.

Studies on the toxicit y/pathogenicity of T. asperellum strain T34 to terrestrial plants were not required because T. asperellum , and other Trichoderma spp., are not taxonomically related to any known plant pathogens. Therefore, adverse effects to plants are not expected as a r esult of the proposed uses of T. asperellum strain T34, and risk is not anticipated.

B . Freshwater and marine/estuarine fish and invertebrates

The registrant presented extensive rationale to show that exposure of T. asperellum strain T34 in aquatic env ironments would be negligible, if it occurred at all, and would have no effects on fish or aquatic invertebrates if it w ere to reach surface waters. Data from a study was submitted to show that at four months after the last application of T. asperellum st rain T34 to potted plants, leachate from the pots contains 10 2 cfu/mL of T

. asperellum strain T34, which is less than the

amount of Trichoderma spp. found naturally in soil. Therefore, spillage onto greenhouse soil and outdoor disposal of treated potting material would not increase the amount of Trichoderma spp. normally found in soil or in any runoff. Incidents such as these are expected to have negligible contribution to exposure. The registrant presented additional data from public

literature to show that if T. asperellum strain T34 were to reach surface waters, it would be unable to proliferate and would be unlikely to survive. Based on the information provided,

exposure to aquatic environments is expected to be negligible and adverse effects in thes e environments are not expected. Therefore, BPPD concludes that the proposed use of T. asperellum strain T34 will not result in risk of adverse effects to fish and aquatic invertebrates in

freshwater and marine environments.

Studies on the toxicity/path ogenicity of T. asperellum strain T34 to aquatic plants were not required because T. asperellum , and other Trichoderma spp., are not taxonomically related to any known plant pathogens and exposure in these environments is not expected. Therefore,

adverse effects to aquatic plants are not expected as a result of the proposed uses of T. asperellum strain T34, and risk is not anticipated.C . Risk to Federally Listed Threatened and Endangered Species

Since EPA has determined that no effects are anticipated for any non

- target species exposed to T. asperellum strain T34 as a result of the proposed labeled applications, effects to threatened and

e ndangered species and their designated critical habitats are also not expected. There fore, a “No

Effect” determina tion is made for direct and indirect effects to listed species and their designated critical habitats resulting from the registered uses of T. asperellum strain T34 , as labeled.

V. Environmental Assessment Summary

The Agency has performed an environment al risk assessment based on data waiver rationale and open literature studies

, and has determined that the proposed uses of Trichoderma asperellum

strain T34 do not pose risk to nontarget organisms

. T . asperellum strain T34 is a naturally occurring strain of T . asperellum . This and other Trichoderma species are ubiquitous and occur widely in soils in concentrations ranging up to10 6 cfu/g dry soil. Based on the concentration of the active ingredient in the end use product and the application instructions on the label, concentrations in treated potting media result in concentrations of 6.6 x 10 3

– 1.0 x 10 4 cfu T. asperellum strain T34/mL. T . asperellum strain T34 is intended for inoculation of potting media

and root treatment of carnation and other ornament al plants, and is restricted to application in greenhouses only. Due to the nature of the application method, exposure to terrestrial nontarg et organisms is expected to be very l ow , if not nonexistent . The use of T . asperellum strain T34 as proposed is n ot expected to significantly increase the amount of Trichoderma spp. beyond levels that are already present in soils.

The registrant has submitted data waiver rationale to satisfy data requirements for nontarget organism risk assessment with the TGAI. Submitted data, data waiver rationale , and a search by BPPD of several literature databases provides sufficient information to determin e that toxicity/pathogenicity to wild mammals, avian wildlife, freshwater and marine/estuarine

vertebrates and inverteb rates, honey bees, nontarget insects, and terrestrial and aquatic plants is not expected from the proposed label uses. As a result, a “No Effect” determination is also made for direct and indirect effects to listed species and their designated critical hab itats resulting from the registered uses of T . asperellum strain T34 , as labeled.

The information provided is sufficient to satisfy the Tier I nontarget organism data requirements

and for ecological risk assessment for the active ingredient. Further te sting of nontarget organisms at higher tier levels is not required.

VI. References

Cardoza, Y.J., K.D. Klepzig, and K.F. Raffa. 2006. Bacteria in oral secretions of an endophytic insect inhibit antagonistic fungi. Ecol. Ent. 31:636

- 645.

Harman, G.E., C.R. Howell, A. Viterbo, I. Chet, and M. Lorito. 2004. Trichoderma species

– opportunistic, avirulent plant symbionts. Nature Reviews 2:43

- 56. MRID 48067407.Jassim, H.K., H.A. Foster, and C.P. Fairhurst. 1990. Biological control of Dutch e lm disease: larvicidal activity of Trichoderma harzianum

, T. polystrum and Scydalidium lignicola

, Scolytus scolytus and S. multistriatus reared in artificial culture. Ann. Appl. Biol. 117:187

- 196.

Leandro, L. F. S., T. Guzman, L. M. Ferguson, G. E. F ernandez, and F. J. Louws. 2007. Population dynamics of Trichoderma in fumigated and compost

- amended soil and on strawberry roots. Applied Soil Ecology 35:237

- 246.

Liu, B., D. Glenn, and K. Buckley. 2008. Trichoderma communities in soils from organic

, sustainable, and conventional farms, and their relation with Southern blight of tomato. Soil Biology and Biochemistry 40:1124

- 1136.

Shakeri, J. and H.A. Foster. 2006. Proteolytic activity and antibiotic production by Trichoderma
harzianum in relati on to pathogenicity to insects. Enzyme and Micro. Tech. 40:961

- 968.

U.S. Environmental Protection Agency (USEPA). 2000. Trichoderma harzianum Rifai strain T

39 (119200) Fact Sheet. Available at: http://www.epa.gov/pesticides/biopesticides/ingredients/tech_docs/tech_119200.htm

.