BIOPESTICIDES REGISTRATION ACTION DOCUMENT

Clonostachys rosea strain 321U

previously known as Gliocladium roseum strain 321U

Pesticide Chemical (PC) Code: 021000

U.S. Environmental Protection Agency

Office of Pesticide Programs

Biopesticides and Pollution Prevention Division

May 22, 2014
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TABLE OF CONTENTS

I. EXECUTIVE SUMMARY.............................................................................................................................................4

II. ACTIVE INGREDIENT OVERVIEW.........................................................................................................................5

III. REGULATORY BACKGROUND................................................................................................................................6

A. Applications for Pesticide Product Registration ...................................................................................................6

B. Food Tolerance Exemption ......................................................................................6Error! Bookmark not defined.

IV. RISK ASSESSMENT .....................................................................................................................................................7

A. Product Analysis Assessment (40 C.F.R. § 158.2120) ...........................................................................................7

B. Human Health Assessment (40 C.F.R. § 158.2140)...............................................................................................8

C. Environmental Assessment (40 C.F.R. § 158.2150)...............................................................................................9

V. ENVIRONMENTAL JUSTICE ..................................................................................................................................10

VI. RISK MANAGEMENT DECISION ...........................................................................................................................10

VII. ACTIONS REQUIRED OF THE REGISTRANT.....................................................................................................11

A. Final Printed Labeling...........................................................................................................................................11

B. Terms of Registration............................................................................................................................................11

C. Reporting of Adverse Effects and Hypersensitivity Incidents ...........................................................................12

VIII. GLOSSARY OF ACRONYMS AND ABBREVIATIONS........................................................................................12

IX. BIBLIOGRAPHY .........................................................................................................................................................13

A. Studies Submitted to Support the Clonostachys rosea strain 321U Pesticide Product Registrations.13

B. Environmental Protection Agency Risk Assessment Memoranda ....................................................................15

C. Other References....................................................................................................................................................15

APPENDIX A. MICROBIAL PESTICIDES DATA REQUIREMENTS .........................................................................17

APPENDIX B. PESTICIDE PRODUCTS............................................................................................................................29
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BIOPESTICIDES REGISTRATION ACTION DOCUMENT TEAM

Office of Pesticide Programs (OPP)

Biopesticides and Pollution Prevention Division

Microbial Pesticides Branch

Science Review

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

Shannon Borges Ecological Effects

Regulatory Review

Kimberly Nesci Chief, Microbial Pesticides Branch

Shanaz Bacchus Regulatory Action Leader

Shannon Borges Microbial Pesticides Team Leader

U.S. EPA-PMRA Coordinator

Gail Tomimatsu, Ph.D. USEPA PMRA Coordinator
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I. EXECUTIVE SUMMARY

Background

On March 25, 2012, W.F. Stoneman Company, LLC, on behalf of AEF Global, Inc., 201, Mgr-Bourget 

Levis Quebec, G6V 6Z3, Canada, submitted applications for two pesticide products to the U.S. 

Environmental Protection Agency (EPA or the Agency) under section 3 of the Federal Insecticide, 

Fungicide, and Rodenticide Act (FIFRA). These pesticide products, FPSolano®Technical (EPA File 

Symbol: 89046-E), and an end-use product (EP), FPSolano®(EPA File Symbol: 89046-R), contain a 

new active ingredient (a.i.), Clonostachys rosea strain 321U previously known as Gliocladium roseum

strain 321U. The application was reviewed jointly with Canada’s Pesticide Management Regulatory

Agency (PMRA). Both PMRA and the EPA scientists reviewed product analysis, acute mammalian 

toxicology, and nontarget organism data and information (40 C.F.R. §§ 158.2120, 158.2140, and 

158.2150, respectively) submitted to support the registration of the two proposed Clonostachys rosea

strain 31U pesticide products. Summaries of these reviews are available in Appendix A in this 

Biopesticides Registration Action Document (BRAD). 

Clonostachys rosea strain 321U is a Microbial Pest Control Agent (MPCA) that is intended as an 

antisapstain product for control of blue stain and mold on freshly cut lumber and logs in hardwoods and 

softwoods. FPSolano®Technical, which is provided as a solution, is proposed for formulation into the 

wettable powder EP FPSolano®. When applied, the pesticide displaces wild molds and staining fungi 

from taking up their natural ecological niches on sawn wood products and logs, thus allowing the 

MPCA to become established before the pest species. The MPCA metabolizes simple, non-structural 

nutrients present in woody plant materials, thereby minimizing nutrients for staining fungi.

Clonostachys rosea strain 321U, a fungus in the family Bionectriaceae, was isolated in Ottawa. It 

colonizes living plants as an endophyte, digests material in soil as a saprophyte and is also known as a 

parasite of other fungi and of nematodes. Its identity was confirmed by Dr. K.A. Seifert at the Centre of 

Land and Biological Resources Research (Agriculture Canada). Clonostachy rosea has a cosmopolitan 

distribution. It is common in a wide range of habitats in tropical, temperate, sub-arctic and desert 

regions of the world. It has been reported from cultivated grassland and woodland, forest, heathland, 

freshwater and coastal soils. This fungus has been frequently associated with cysts of Heterodera sp., 

Globodera sp. and other nematodes in soil and with sclerotia of Sclerotnia sclerotorium, 

Phymatotrichum omnivorum, Rhizoctonia solani, Botryis sp., Verticillium sp. and other soil fungi and 

plant materials.

For product identity, the applicant submitted adequate product chemistry data except for metabolite 

characterization and storage stability. Three studies were conducted to address the acute dermal, acute 

intraperitoneal injection and primary dermal irritation data requirements for the technical. Requests to 

waive testing for other Tier 1 acute human health toxicology data requirements were based on the results 

of these studies and published information. The EPA accepted the rationales for acute oral, acute 

pulmonary toxicity and infectivity and pathogenicity testing for the technical product as well as the 

acute oral, primary eye irritation. Similar rationales were submitted to waive testing for the acute 

toxicology data requirements for the EP and considered adequate to support its proposed registration. A 

cell culture test was not required for this fungal active ingredient. The applicant reported that no 

hypersensitivity incidents occurred during research, development, or testing of Clonostachys rosea

strain 321U. In light of the results of the acute toxicity/ pathogenicity data and information and the 

absence of hypersensitivity incidents, the EPA is not requiring testing at Tiers II and III (Appendix A).
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The proposed use pattern is expected to limit exposure to nontarget organisms. The a.i. will be confined 

to dip tanks used for treatment, and is not expected to result in exposure under labeled use. Birds, wild 

mammals, and terrestrial plants are not expected to be exposed to Clonostachys rosea strain 321U in dip 

tanks, and would be expected to have very limited exposure to the a.i. that is present on freshly treated 

wood stored at the treatment facility. Nontarget insects and honey bees may be incidentally exposed to 

freshly treated lumber, though this exposure is also expected to be very limited. Aquatic organisms may 

be exposed if the treatment solution is released into the environment, such as with splash during 

treatment or wash-off of the treatment during rain events. Language is present on the label to limit 

exposure in aquatic environments (Appendix A).

Occupational Exposure

Despite the low toxicological profile of Clonostachys rosea strain 321U, the EPA is requiring baseline 

personal protective equipment (PPE) for handlers who, due to their occupation, may be exposed to the 

active ingredient for prolonged periods and/or numerous times. Handlers working with Clonostachys 

rosea strain 321U in pesticide applications are directed to wear a long-sleeved shirt, long pants, socks, 

shoes, waterproof gloves, protective eyewear and/or a dust/mist filtering respirator meeting National 

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

future products, the EPA may require additional/different PPE (e.g., chemical-resistant gloves), other 

than the standard described above, on a product-specific basis. 

Public comment

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

public to participate in major registration decisions before they occur. According to this new policy, the 

Agency intends to provide a public comment period prior to making a registration decision for, at

minimum, the following types of applications: new active ingredients; first food uses; first outdoor uses; 

first residential uses; and any registration decisions for which the Agency believes there may be 

substantial public interest.

Consistent with the new policy of making registration actions more transparent, Clonostachys rosea

strain 321U is subject to a 15-day comment period as a “new active ingredient” whose registration 

would result in a “first outdoor use.” While a final decision on registration is contingent upon review 

and consideration of public comments, the Agency presently believes that, based upon the preliminary 

risk assessment and information submitted in support of registration of Clonostachys rosea strain 321U, 

it is in the best interest of the public and the environment to issue the registration for Clonostachys rosea 

strain 321U. The basis for this preliminary decision can be found in the risk assessment for

Clonostachys rosea strain 321U, which is characterized throughout this BRAD and the associated 

reference documents written by Agency scientists. 

II. ACTIVE INGREDIENT OVERVIEW

Biological Name: Clonostachys rosea strain 321U previously known as Gliocladium 

roseum 321U.

Culture Deposit: The MPCA is referred to as Clonostachys rosea strain 321U. It is 

stored in the Forintek Culture Collection and at the Centrallbureau 

voor schimmelcultures (BS 226.48) and the American Type 

Culture Collection (ATCC 10406).
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OPP Chemical Code: 021000

Type of Pesticide: Microbial Pesticide – Fungicide

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

application rates, methods of application, formulation types, and 

target pests) regarding the proposed labeling for pesticide products 

containing this active ingredient.

III. REGULATORY BACKGROUND

A. Applications for Pesticide Product Registration

On March 25, 2012, William Stoneman, on behalf of AEF global, Inc., submitted applications for a 

manufacturing-use pesticide product, FPSolano® Technical 100% (EPA File Symbol: 89046-E), and an 

end-use pesticide product, FPSolano® (EPA File Symbol: 89046-R), to the U.S. Environmental 

Protection Agency (EPA) under section 3 of the Federal Insecticide, Fungicide, and Rodenticide Act 

(FIFRA). These pesticide products contain a new active ingredient, Clonostachys rosea strain 321U

previously known as Gliocladium roseum strain 321U. 

Clonostachys rosea strain 321U was the subject of a North American Free Trade Agreement joint 

registration review conducted by the Environmental Protection Agency Office of Pesticide Programs 

and the Health Canada Pest Management Regulatory Agency. The PMRA documents for registration of 

Clonostachys rosea strain 321U are under review. For further information, see http://www.hcsc.gc.ca/cps-spc/pest/part/consultations/index-eng.php.

PMRA was the leader for the health effects reviews and EPA for the ecological reviews. PMRA 

informed the applicant of deficiencies July 27, 2012 (letter from Marion Law to Andreja Preradov). The 

applicant responded to the deficiencies to both PMRA and EPA on October 11, 2012.

On August 14, 2012 (77 FR 48519, EPA-HQ-OPP-2012-0507), the EPA announced receipt of these 

applications to register pesticide products containing a new active ingredient and opened a 30-day public 

comment period pursuant to the provisions of FIFRA section 3(c)(4). No comments were received 

following this publication when the 30 day comment period ended. 

On March 5, 2013 PMRA informed the applicant of a second set of deficiencies arising out of the 

October 11, 2012 resubmission. The applicant responded on May 10, 2013. These data were subject to a 

second completness check and were found to be sufficient to proceed with full review.

The Pesticide Registration Improvement Renewal Act (PRIA II) date, originally scheduled for April 23, 

2013, was renegotiated to April 30, 2014. One study, submitted to PMRA, was not part of EPA’s data 

package. When it was submitted to EPA, the PRIA date was renegotiated to June 9, 2014 to allow time 

for its review and incorporation into the database for this MPCA.

B. Food Tolerance Exemption

No tolerance or exemption from the requirement of a tolerance is associated with or required for this 

regulatory action as only non-food uses are currently proposed for Clonostachys rosea strain 321U. 
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Accordingly, this action is outside the scope of the Food Quality Protection Act (FQPA). There are no 

CODEX maximum residue levels established for Clonostachys rosea strain 321U.

IV. RISK ASSESSMENT

In the Federal Registers of October 26, 2007 (72 FR 60988) and August 30, 2012 (77 FR 52610), the 

EPA issued Final Rules on the data requirements to support registration of microbial pesticides and 

updated the definition for microbial pesticides. The data and information evaluated for this BRAD were 

considered in light of these requirements.

The classifications that are found for each data submission are assigned by Agency 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 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 Code of Federal Regulations (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 the acute toxicity data requirements, Toxicity Categories are assigned based on the hazard(s) 

identified from studies and/or other information submitted to the EPA in support of a pesticide 

registration. The active ingredient or particular product is classified into Toxicity Category I, II, III, or 

IV, where Toxicity Category I indicates the highest toxicity and Toxicity Category IV indicates the 

lowest toxicity.

A. Product Analysis Assessment (40 C.F.R. § 158.2120)

 

The applicant fulfilled most of the product analysis data requirements for Clonostachys rosea strain 

321U except for the storage stability data requirement. Acceptable studies include product chemistry and 

composition, analysis of samples, and physical and chemical characteristics. Storage stability of 

FPSolano® (manufactured at the AEF Global Inc. site) has been evaluated over a period of three months 

after storage at temperatures of -20°C, 4°C and 20°C using the method in accordance with both PMRA’s 

and the EPA’s microbial pesticides data requirements. The applicant has indicated that additional 

storage stability testing will be conducted at 6, 9, 12 and 24 months. These data must be submitted 

within 1 year of registration based on the potential that testing has been ongoing while the application 

was being processed.

The label for FPSolano® proposes storage conditions of 4°C for up to four months. Based on the data 

submitted, FPSolano® continues to meet the guarantee after three months of storage at 4°C. However, 

in the absence of data, the default storage period for wettable powder formulations is 6 months at 4°C.

Refer to Tables 1 and 2 in Appendix A for a brief summary of the data requirements, including both 

generic and product-specific information. 
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B. Human Health Assessment (40 C.F.R. § 158.2140)

1. Toxicity

All toxicology data requirements for non-food use of Clonostachys rosea strain 321U have been 

satisfied.

Tier II and Tier III studies were not required for Clonostachys rosea strain 321U based on the lack of 

acute toxicity/pathogenicity in the Tier I studies.

Refer to Tables 3 (generic - for Technical) and 4 (product specific - for EP) in Appendix A for a brief 

summary of the studies and information reviewed for the acute toxicology data requirements. 

a. Endocrine Disruptors

As required by the Administrator under FFDCA section 408(p), the EPA has developed the Endocrine 

Disruptor Screening Program (EDSP) and has begun to implement the screening program that is to be 

used to test all pesticides in order to determine whether certain substances (including pesticide active 

and other ingredients) may have an effect in humans or wildlife similar to an effect produced by a 

“naturally occurring estrogen, or other such endocrine effects as the Administrator may designate.” 

FFDCA section 408(p)(4) authorizes the Administrator, by order, to exempt from the requirements of 

the Estrogenic Substances Screening Program a biologic substance or other substance if a determination 

is made that the substance is not anticipated to produce any effect in humans similar to an effect

produced by a naturally occurring estrogenic substance. 

Between October 2009 and February 2010, the EPA issued test orders/data call-ins for the first group of 

67 chemicals, which contains 58 pesticide active ingredients and 9 inert ingredients. Clonostachys rosea 

strain 321U is not among the group of 58 pesticide active ingredients on the initial list to be screened 

under the EDSP. 

The EPA believes that Clonostachys rosea strain 321U, the active ingredient described in this document, 

likely is a substance that would not produce any effect in humans similar to an effect produced by a 

naturally occurring estrogenic substance. As such, pursuant to Section 408(p)(4), the EPA will 

determine in the future whether it can exempt Clonostachys rosea strain 321U from the requirements of 

the Section 408(p) EDSP. In the event the EPA does determine to exempt this substance from the EDSP, 

an order will be issued. 

For further information on the status of the EDSP, the policies and procedures, the list of 67 chemicals, 

the test guidelines and the Tier 1 screening battery, please visit our website at http://www.epa.gov/endo/.

2. Federal Food, Drug, and Cosmetic Act (FFDCA) Considerations

No tolerance or exemption from the requirement of a tolerance is associated with or required for this 

regulatory action as only non-food uses are currently proposed for Clonostachys rosea strain 321U. 

Accordingly, this action is outside the scope of the Food Quality Protection Act (FQPA). There are no 

CODEX maximum residue levels established for Clonostachys rosea strain 321U.
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3. Occupational Exposure and Risk Characterization

Handler exposure to Clonostachys rosea strain 321U is not expected to pose any undue risk. Regardless,

the EPA is requiring appropriate personal protective equipment and precautionary statements on 

pesticide product labels to mitigate any potential risks to pesticide handlers due to prolonged and/or 

repeated exposures. Handlers working with Clonostachys rosea strain 321U for uses as labeled are 

directed to wear a long-sleeved shirt, long pants, socks, shoes, waterproof gloves, protective eyewear, 

and/or a dust/mist filtering respirator meeting NIOSH standards of at least N-95, R-95, or P-95. For 

future products, the EPA may require additional/different PPE (e.g., chemical-resistant gloves), other 

than the standard described above, on a product-specific basis. 

C. Environmental Assessment (40 C.F.R. § 158.2150)

1. Nontarget Organism Toxicology 

All nontarget toxicology data requirements for Clonostachys rosea strain 321U have been satisfied. 

Adequate Tier I nontarget organism data/information are available to support registration of

Clonostachys rosea strain 321U. The EPA found that guideline studies and scientific rationale 

submitted by the applicant are sufficient to fulfill the nontarget organism data requirements and for risk 

assessment purposes. Further testing of nontarget organisms at higher tier levels (i.e., Tiers II, III, and 

IV) is not required. The EPA performed an environmental risk assessment, based on information 

provided by the applicant, and determined that no effects are anticipated for nontarget species exposed 

to Clonostachys rosea strain 321U as a result of the proposed labeled applications.

Refer to Table 5 in Appendix A for a brief summary of the review of the data/information submitted for 

these data requirements. 

For a summary of the generic data requirements described in sections IV(C)(1), refer to Table 7 in 

Appendix A.

2. Ecological Exposure and Risk Characterization 

Clonostachys rosea is a cosmopolitan microorganism. It is purported to be very common in soil and on 

organic matter; however, since it has not been the subject of much field research, field collections of this 

fungus are limited. In Canada, isolates of this species that have been deposited in culture collections 

have been collected from British Columbia, Alberta, Ontario and Quebec. Moreover, Clonostachys 

rosea is common in a wide range of habitats in tropical, temperate, subarctic, and desert regions of the 

world. It was reported in cultivated, grassland, woodland, forest, heathland, freshwater, and coastal soils, 

particularly those of neutral to alkaline pH, and is not known to produce any adverse health or 

environmental effects (Pugh and Dickinson, 1965; Foley and Deacon, 1985; Sutton et al., 1997). 

3. Environmental Fate Data 

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

and for nontarget organism risk assessment for Clonostachys rosea strain 321U, further testing at higher 

tier levels (i.e., Tiers II, III, and IV) is not required.
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4. Threatened and Endangered Species Assessment

The EPA has determined that no effects are anticipated for nontarget species exposed to Clonostachys 

rosea strain 321U as a result of the proposed labeled applications. Therefore, a “No Effect” 

determination is made for direct and indirect effects to federally listed endangered and threatened 

(“listed”) species and for their designated critical habitat. 

V. ENVIRONMENTAL JUSTICE

The EPA seeks to achieve environmental justice—the fair treatment and meaningful involvement of all 

people regardless of race, color, national origin, or income—with respect to the development, 

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

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

disproportionate share of the negative environmental consequences resulting from industrial, municipal, 

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

and policies. Meaningful involvement means that (1) potentially affected community residents have an 

appropriate opportunity to participate in decisions about a proposed activity that will affect their 

environment and/or health; (2) the public’s contribution can influence the regulatory agency’s decision; 

(3) the concerns of all participants involved will be considered in the decision-making process; and (4) 

the decision-makers seek out and facilitate the involvement of those potentially affected. The EPA has 

this goal for all communities and persons across the United States. 

To help address potential environmental justice issues, during the 15-day public participation comment 

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

location, cultural practices, or other factors, may have atypical, unusually high exposure to Clonostachys 

rosea strain 321U, compared to the general population. The EPA received no public comments on this 

particular matter.

For additional information regarding environmental justice issues, please visit the EPA’s website at 

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

VI. RISK MANAGEMENT DECISION

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

determinations are made:

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

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

 requirements of FIFRA;

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

 the environment; AND

(4) When used in accordance with widespread and commonly recognized 

 practice, it will not generally cause unreasonable adverse effects on the 

 environment.

To satisfy criterion 1, the EPA understands that Clonostachys rosea strain 321U is well-known to have 
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fungicidal capabilities. The EPA has no knowledge that would contradict the claims made on the

FPSolano® biological fungicide labels, and the proposed pesticide products are not expected to cause 

unreasonable adverse effects on the environment when used according to their respective label 

instructions. Criterion 2 is satisfied by the product labels, as well as the data and information presented 

in this document. It is believed that the Clonostachys rosea strain 321U pesticide products will not cause 

any unreasonable adverse effects on the environment, and FPSolano® biological fungicide (end-use 

pesticide product), in particular, is likely to control certain fungi as claimed, satisfying criterion 3. 

Criterion 4 is satisfied in that the Clonostachys rosea strain 321U pesticide products are not expected to 

cause unreasonable adverse effects when used according to label instructions. Therefore, the EPA 

believes FPSolano® Technical and FPSolano® biological fungicide, containing the Clonostachys rosea

strain 321U as a new active ingredient, are eligible for registration under FIFRA section 3(c)(5) for the

labeled uses. 

VII. ACTIONS REQUIRED OF THE REGISTRANT

A. Final Printed Labeling

Before releasing pesticide products containing Clonostachys rosea strain 321U for shipment, the 

registrant is required to provide appropriate final printed labeling to the EPA.

B. Terms of Registration

The following items will be terms of the FPSolano® Technical (EPA File Symbol: 89046-E) 

registration:

1. Discussion of Formation of Unintentional Ingredients (OCSPP) Harmonized Guideline 885.1300:

a) Confirmatory analysis for production of toxic secondary metabolites using the appropriate 

controls and samples, and information on the stability of potential toxins in the environment and 

at temperatures typical for kiln-drying.

b) Microbial contaminant analysis and colony forming unit (CFU) count determination for a 

minimum of five batches of TGAI and EP produced at each of the alternative manufacturing and 

formulating sites

2. Storage Stability (OCSPP Harmonized Guideline 830.6317): Within one year of registration, AEF 

Global, Inc., must provide the EPA with the results of a one-year storage stability study on 

FPSolano® Technical

The following items will be terms of the FPSolano® (EPA File Symbol: 89046-R) registration: 

1. Storage Stability (Harmonized Guideline 830.6317): Within one year of registration, AEF Global, 

Inc., must provide the EPA with the results of a one-year storage stability study on FPSolano® if 

the applicant wishes to extend the length of storage time for FPSolano®. The label for FPSolano®

proposes storage conditions of 4°C for up to four months. The applicant has indicated that 

additional storage stability testing will be conducted at 6, 9, 12 and 24 months. Based on the 

assumption that storage stability studies have been ongoing while the application was being 

processed, these data must be submitted within one year of registration.
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C. Reporting of Adverse Effects and Hypersensitivity Incidents

Notwithstanding the information stated in the previous sections, it should be clearly understood that 

certain specific data are required to be reported to the EPA as a requirement for maintaining the federal 

registration for a pesticide product. A brief summary of these types of data are described below.

Reports of all incidents of adverse effects to the environment must be submitted to the EPA under the 

provisions stated in FIFRA section 6(a)(2). Additionally, all incidents of hypersensitivity (including both 

suspected and confirmed incidents) must be reported to the EPA under the provisions of 40 C.F.R. § 

158.2140(d).

VIII. GLOSSARY OF ACRONYMS AND ABBREVIATIONS

A/cm2acre per square centimeter

a.i. active ingredient

BRAD Biopesticides Registration Action Document

C.F.R. Code of Federal Regulations

cfu colony-forming units

cfu/kg colony-forming units per kilogram

cfu/mL colony-forming units per milliliter

cm centimeter

EC environmental concentration

EC50 half maximal effective concentration

EDSP Endocrine Disruptor Screening Program

EP end-use pesticide product

EPA United States Environmental Protection Agency 

FFDCA Federal Food, Drug, and Cosmetic Act

FIFRA Federal Insecticide, Fungicide, and Rodenticide Act

FR Federal Register

g/A grams per acre

g/lb grams per pound

g/m

2

grams per square meter

lb/A pounds per acre

lb/ft3pounds per cubic foot

LC50 median lethal concentration. A statistically derived concentration of a substance that can be 

expected to cause death in 50% of test animals. It is usually expressed as the weight of substance 

per weight or volume of water, air, or feed (e.g., mg/L, mg/kg, or ppm).

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

in 50% of the test animals when administered by the route indicated (oral, dermal, or

inhalation). It is expressed as a weight of substance per unit weight of animal (e.g., mg/kg).

μg/μL micrograms per microliter

μg/mL micrograms per milliliter

mg/cm2 milligrams per square centimeter

mg/kg milligrams per kilogram

mg/L milligrams per liter

mg/mL milligrams per milliliter

mL/L milliliters per liter

mm millimeter
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MP manufacturing-use pesticide product

MPCA microbial pest control agent

MRID No. Master Record Identification Number

ng nanogram

NIOSH National Institute for Occupational Safety and Health

NOEC no observed effect concentration

OPP Office of Pesticide Programs

PC Code Pesticide Chemical Code

PPE personal protective equipment

ppm parts per million

QC quality control

TGAI technical grade of the active ingredient

U.S. United States

USDA United States Department of Agriculture

USFWS United States Fish and Wildlife Service

IX. BIBLIOGRAPHY

A. Studies Submitted to Support the Clonostachys rosea strain 321U, previously known as 

Gliocladium roseum strain 321U, Pesticide Product Registrations

MRID Citation Reference

48782601 Preradov, A. (2012) FPSolanoTM Gliocladium roseum Strain 321U: Product 

Characterization and Analysis. Unpublished study prepared by AEF Global, Inc. 78p.

48782602 Preradov, A. (2012) FPSolanoTM Glicoladium roseum Strain 321U: Characterisation of 

the MPCA (A.I) Product Identity, Chemical Identity. Unpublished study prepared by AEF 

Global, Inc. 17p.

48782603 Preradov, A. (2012) FPSolanoTM Gliocladium roseum Strain 321U: Manufacturing 

Methods and Quality Assurance, Preservation and Maintenance of Production Strain. 

Unpublished study prepared by AEF Global, Inc. 23p.

48782604 Preradov, A. (2012) FPSolanoTM Gliocladium roseum Strain 321U: Storage Stability. 

Unpublished study prepared by AEF Global, Inc. 7p.

48782605 Preradov, A. (2012) FPSolanoTM Gliocladium roseum Strain 321U: Fulfillment of Data 

Requirement with Public Literature and/or Scientific Rationale: Acute Oral 

Toxicity/Pathogenicity. Unpublished study prepared by AEF Global, Inc. 9p.

48782606 Preradov, A. (2012) FPSolanoTM Gliocladium roseum Strain 321U: Fulfillment of Data 

Requirement with Public Literature and/or Scientific Rationale: Acute Pulmonary

Infectivity and Toxicity. Unpublished study prepared by AEF Global, Inc. 10p.
Clonostachys rosea strain 321U previously known as Gliocladium roseum strain 321U Page 14 of 29

Biopesticides Registration Action Document DRAFT

48782607 Monds, K. (2010) FPSolanoTM Gliocladium roseum Strain 321U: Intraperitoneal (IP) 

Injection Toxicity/Pathogenicity Study: Final Report. Project Number: 13910/10. 

Unpublished study prepared by Stillmeadow, Inc. 23p.

48782608 Preradov, A. (2012) FPSolanoTM Gliocladium roseum Strain 321U: Fulfillment of Data 

Requirement with Public Literature and/or Scientific Rationale Hypersensitivity Incidents. 

Unpublished study prepared by AEF Global, Inc. 5p.

48782609 Younger, C. (2010) FPSolanoTM Gliocladium roseum Strain 321U: Acute Oral Toxicity

Study in Bobwhite Quail: Final Report. Project Number: 13909/10. Unpublished study

prepared by Stillmeadow, Inc. 19p.

48782610 Preradov, A. (2012) FPSolanoTM Gliocladium roseum Strain 321U: Fulfillment of Data 

Requirement with Public Literature and/or Scientific Rationale Wild Mammal Testing, 

Tier I. Unpublished study prepared by AEF Global, Inc. 8p.

48782611 Brunner, S. (2010) FPSolanoTM Gliocladium roseum: Microbial Pest Control Agent 

(MPCA) Freshwater Fish Test Oncorhynchus mykiss (Rainbow Trout): Final Report. 

Project Number: 13946/10. Unpublished study prepared by Stillmeadow, Inc. 15p.

48782612 Brunner, S. (2010) FPSolanoTM Gliocladium roseum: Microbial Pest Control Agent 

(MPCA) Freshwater Aquatic Invertebrate Test with Daphnia magna: Final Report. 

Project Number: 13947/10. Unpublished study prepared by Stillmeadow, Inc. 18p.

48782613 Preradov, A. (2012) FPSolanoTM Gliocladium roseum Strain 321U: Fulfillment of Data 

Requirement with Public Literature and/or Scientific Rationale Nontarget Aquatic Plants. 

Unpublished study prepared by AEF Global, Inc. 7p.

48782614 Preradov, A. (2012) FPSolanoTM Gliocladium roseum Strain 321U: Fulfillment of Data 

Requirement with Public Literature and/or Scientific Rationale Terrestrial Plants. 

Unpublished study prepared by AEF Global, Inc. 7p.

48782615 Preradov, A. (2012) FPSolanoTM Gliocladium roseum Strain 321U: Fulfillment of Data 

Requirement with Public Literature and/or Scientific Rationale: Terrestrial Arthropods. 

Unpublished study prepared by AEF Global, Inc. 6p.

48782616 Preradov, A. (2012) FPSolanoTM Gliocladium roseum Strain 321U: Fulfillment of Data 

Requirement with Public Literature and/or Scientific Rationale: Acute Oral Toxicity - Rat, 

Acute Inhalation Toxicity - Rat. Unpublished study prepared by AEF Global, Inc. 10p.

48782617 Kuhn, J. (2010) FPSolanoTM Gliocladium roseum: Acute Dermal Toxicity in Rats: Final 

Report. Project Number: 13954/10. Unpublished study prepared by Stillmeadow, Inc. 

11p.
Clonostachys rosea strain 321U previously known as Gliocladium roseum strain 321U Page 15 of 29

Biopesticides Registration Action Document DRAFT

48782618 Preradov, A. (2012) FPSolanoTM Gliocladium roseum Strain 321U: Fulfillment of Data 

Requirement with Public Literature and/or Scientific Rationale: Primary Eye Irritation. 

Unpublished study prepared by AEF Global, Inc. 11p.

48782619 Kuhn, J. (2010) FPSolanoTM Gliocladium roseum: Acute Dermal Irritation Study in 

Rabbits: Final Report. Project Number: 13955/10. Unpublished study prepared by

Stillmeadow, Inc. 11p.

48782620 Stoneman, W. (2012) FPSolanoTM Gliocladium roseum Strain 321U: Reference Volume 

for Fulfillment of Data Requirement with Public Literature and/or Scientific Rationale. 

Project Number: 13955/10. Unpublished study prepared by AEF Global, Inc. 158p.

48978900 AEF Global Inc. (2012) Submission of Pesticide Use Data in Support of the Registration 

of FPSolanoTM. Transmittal of 1 Study.

48978901 Bourassa, A. (2012) FPSolanoTM: Gliocladium roseum Strain 321U: Response to 

Deficiency Letter Sent on July 27, 2012 by Mario Law, PMRA. Project Number: 

101/1094 2012/1160. Unpublished study prepared by AEF Global Inc. 255p.

49172001 Bourassa, A. (2013) FPSolano and FPSolano Technical: Gliocladium roseum Strain 

321U: Response to Deficiency Letter Sent on March 5, 2013 by Marion Law, PMRA. 

Unpublished study prepared by AEF Global, Inc. 1381 p.

49351601 Brummer, S. (2011) Gliocladium roseum. FINAL REPORT: Microbial Pest Control 

Agent (MCPA): Freshwater Aquatic Invertebrate Test with Daphnia magna. Project 

Number: 14415/10. Unpublished study prepared by Stillmeadow, Inc. 28p.

B. Environmental Protection Agency Risk Assessment Memoranda

U.S. EPA. 2014. Clonostachys rosea strain 321U. Memorandum from S. Borges, to S. Bacchus, dated 

May 13, 2014

C. Other References

USDA. 1994. Extension Service, National Agricultural Pesticide Impact Assessment Program.

U.S. EPA. 2012b. Chapter 7 of the Label Review Manual (Precautionary Statements). Available from 

http://www.epa.gov/oppfead1/labeling/lrm/chap-07.pdf.

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

Pest Control Agents (OPPTS 885.2000). Available from 

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

USFWS. 2013. Endangered Species. Available from http://www.fws.gov/endangered/.

World Health Organization. 2011. Guidelines for Drinking-water Quality (Fourth Edition). Available 
Clonostachys rosea strain 321U previously known as Gliocladium roseum strain 321U Page 16 of 29

Biopesticides Registration Action Document DRAFT

from http://whqlibdoc.who.int/publications/2011/9789241548151_eng.pdf.
Clonostachys rosea strain 321U previously known as Gliocladium roseum strain 321U Page 17 of 29

Biopesticides Registration Action Document DRAFT

APPENDIX A. MICROBIAL PESTICIDES DATA REQUIREMENTS 

(40 .F.R. PART 158 – SUBPART V)

A. Product Analysis

The applicant fulfilled most of the product analysis data requirements (except for the storage stability

data requirement) for Clonostachys rosea strain 321U, including product chemistry and composition, 

analysis of samples, and physical and chemical characteristics, by acceptable guideline studies (Tables 1 

and 2, Appendix A).

The original submission package referred to the MPCA as Gliocladium roseum strain 321U. 

Gliocladium roseum was reclassified as Clonostachys rosea, because G. roseum differed from the type 

species of Gliocladium, G. penicillioides, in morphology, ecology, teleomorph, and DNA sequence data. 

Therefore, the MPCA is to be referred to as Clonostachys rosea strain 321U.

Gliocladium catenulatum has also been transferred to the genus Clonostachys and is known as C. rosea 

f. catenulata. The relationship between Gliocladium and Clonostachys is therefore considered close. 

Furthermore, the Gliocladium genus is closely related to the Trichoderma genus. For example, G. 

virens has been reclassified as T. virens. As a result, information on various Gliocladium and 

Trichoderma species are considered in this review. The MPCA is permanently stored in liquid nitrogen 

in the Forintek Culture Collection and has also been deposited at the Centrallbureau voor 

schimmelcultures (BS 226.48) and the American Type Culture Collection (ATCC 10406).

The taxonomic designation of the microbial pest control agent (MPCA) is as follows (MRID 49172001):

Kingdom Fungi

Phylum Ascomycota

Class Sordariomycetes

Order Hypocreales

Family Bionectriaceae

Genera Clonostachys

Species rosea

Strain 321U

Product identity

Clonostachys rosea strain 321U can be distinguished from species belonging to the closely related 

Gliocladium genus by morphology. Sequencing of the internal transcribed spacer (ITS) can be used to 

distinguish the active ingredient from other Clonostachys species and other strains of C. rosea. 

Unintentional Ingredient analysis

Results of microbial contaminant testing for five batches of TGAI and five batches of EP (manufactured 

at the AEF Global Inc. site) were provided (MRID 49172001). Analyses of the total Mesophiles, total 

coliforms, fecal coliforms, fecal Enterococci and Staphylococci were acceptable (<10 cfu/mL for the 

Technical and <10 cfu/g for the EP). Although testing for Salmonella and Shigella species is not 

routinely conducted as part of the manufacturing process/quality assurance protocol, testing was carried 

out on five batches of EP using Health Canada protocols MFHPB-20 and MFLP-25 that include an 

enrichment step (MRID 49172001). Results provided for five batches of the TGAI and EP demonstrated 

that they were absent.
Clonostachys rosea strain 321U previously known as Gliocladium roseum strain 321U Page 18 of 29

Biopesticides Registration Action Document DRAFT

These microbial contaminant analysis data submitted for batches of FPSolano produced at the primary

manufacturing and formulating site were acceptable for the registration of the pesticides. Additional 

microbial contaminant analysis data on batches produced at the secondary manufacturing and 

formulating sites will be required.

B. Biological Properties of the MPCA(s):

1. Natural Occurrence of the Microorganism: 

Clonostachy rosea has a cosmopolitan distribution. It is common in a wide range of habitats in tropical, 

temperate, sub-arctic and desert regions of the world. It has been reported from cultivated grassland and 

woodland, forest, heathland, freshwater and coastal soils. This fungus has been frequently associated 

with cysts of Heterodera sp., Globodera sp. and other nematodes in soil and with sclerotia of Sclerotnia 

sclerotorium, Phymatotrichum omnivorum, Rhizoctonia solani, Botryis sp., Verticillium sp. and other 

soil fungi and plant materials (MRID 49172001). 

2. Target Organisms:

The MPCA is intended as an antisapstain product for control of blue stain and mould on freshly cut 

lumber and logs in hardwoods and softwoods. Target organisms on which the MPCA has been tested 

include Ophiostoma piceae 387E, Aureobasidium pullulans 132S, Ceratocystis adiposa 251E, 

Alternaria alternata 2G, Cladosporium sphaerospermum 806B, Hormonema dematioides 742D, 

Rhinocladiella atrovierens 135E, O. minus 864A, Phialophora botulispora 707C and O. piliferum 55B 

(MRID 48978901).

The end-use product, FPSolano, is mixed with an alkaline solution prior to the wood being treated. The 

wood staining fungi are susceptible to alkaline conditions while the MPCA is unaffected by high pH. 

The alkaline solution displaces wild moulds and staining fungi from taking up their natural ecological 

niches on sawn wood products and logs allowing the MPCA to become established before the pest 

species. The MPCA metabolises simple, non-structural nutrients present in woody plant materials, 

thereby minimizing nutrients for staining fungi (MRID 48978901).

The antagonistic behaviour of C. rosea/G. roseum towards other fungi has also been attributed to 

mycoparasitism, substrate competition, enzymatic activity (e.g., β1-3 glucanases,, chitinases) and 

induced resistance. Moreover, production of toxic secondary metabolites may also exert an antibiotic 

effect (MRID 49172001). 

3. Host Range:

The host range of C. rosea includes Acacia, Acer, Beta, Buxus, Cucumis, Daucus, Fragaria, 

Liquidambar, Lycopersicon, Metrosideros, Phaseolus, Phoenix, Pinus, Poncirus, Psilotum, Quercus, 

Rhizophora, Trifolium, citrus and glycine (MRID 48978901).
Clonostachys rosea strain 321U previously known as Gliocladium roseum strain 321U Page 19 of 29

Biopesticides Registration Action Document DRAFT

TABLE 1. Summary of Product Analysis Data Submitted for FPSolanoTM Technical [the Technical Grade of the Active 

Ingredient (TGAI)], Clonostachys rosea strain 321U and Its End-Use Pesticide Product (EP), FPSolano® (40 C.F.R. § 

158.2120)

Harmonized 

Guideline 

Number 

Data 

Requirement

Results MRID No.

or Other 

TGAI EP Reference

885.1100 Product 

Identity

Submitted data fulfill the 

requirement for product identity. 

Submitted data fulfill the requirement 

for product identity. 

(EP contains a minimum of 5 x 108

CFU/gram).

48782602

885.1200 Manufacturing 

Process Submitted data fulfill the requirement for manufacturing process. 48782603

885.1250

Deposition of 

a Sample in a 

Nationally 

Recognized 

Culture 

Collection

The MPCA is referred to as 

Clonostachys rosea strain 321U. 

It is stored in the Forintek Culture 

Collection and at the 

Centrallbureau voor 

schimmelcultures (BS 226.48) 

and the American Type Culture 

Collection (ATCC 10406).

N/A 48782602

885.1300

Discussion of 

Formation of 

Unintentional 

Ingredients

Submitted data fulfill the requirement for discussion of formation of 

unintentional ingredients. 48782603

885.1400 Analysis of 

Samples Submitted data fulfill the requirement for analysis of samples. 48782603

885.1500 Certification 

of Limits N/A

Limits listed on the confidential 

statement of formula are adequate. 48782603

TABLE 2. Physical and Chemical Characteristics for FPSolano® Technical [the Technical Grade of the Active Ingredient 

(TGAI)], Clonostachys rosea strain 321U and Its End-Use Pesticide Product (EP), FPSolano® (40 C.F.R. § 158.2120)

Harmonized 

Guideline 

Number

Data Requirement

Results MRID No.

or Other 

Reference TGAI EP

830.6302 Color Not reported Not reported 48978901

48782602 830.6303 Physical State Liquid Wettable powder

830.6304 Odor mild Not reported

830.6313

Stability to Normal and 

Elevated Temperatures, 

Metals, and Metal Ions

N/A N/A

830.6317 Storage Stability

Based on the data submitted, FPSolano® continues to meet the 

guarantee after three months of storage at 4°C. However, in the 

absence of data, the default storage period for wettable powder 

formulations is 6 months at 4°C. The label for FPSolano 

proposes storage conditions of 4°C for up to four months. The 

applicant has indicated that additional storage stability testing 

will be conducted at 6, 9, 12 and 24 months. Based on the 

assumption that storage stability studies have been ongoing 

while the application was being processed, these data must be 

submitted within one year of registration.

48782604

48978901

830.6319 Miscibility N/A
Clonostachys rosea strain 321U previously known as Gliocladium roseum strain 321U Page 20 of 29

Biopesticides Registration Action Document DRAFT

TABLE 2. Physical and Chemical Characteristics for FPSolano® Technical [the Technical Grade of the Active Ingredient 

(TGAI)], Clonostachys rosea strain 321U and Its End-Use Pesticide Product (EP), FPSolano® (40 C.F.R. § 158.2120)

Harmonized 

Guideline 

Number

Data Requirement

Results MRID No.

or Other 

Reference TGAI EP

N/A

N/A

830.6320 Corrosion Characteristics N/A N/A

830.7100 Viscosity N/A

Not required because EP is not a 

liquid form of microbial pesticide 

(refer to test note #5 of 40 C.F.R. 

§ 158.2120(d)). 

830.7300

Density/Relative 

Density/Bulk Density 

(Specific Gravity)

0.43

Soluble in water

48978901

C. Acute mammalian toxicology

Tier I mammalian toxicology data/information are sufficient for the registration of Clonostachys rosea 

strain 321U (see Tables 3 and 4 below). Three studies were conducted for the acute dermal, acute 

intraperitoneal injection and primary dermal irritation data requirements for the technical. The 

infectivity/pathogenicity endpoints were not addressed by the studies conducted for the acute dermal and 

acute injection guidelines. The test substances used in these studies were not representative of the 

technical or the end-use product (i.e., low level of active ingredient and no formulation ingredients 

added). Furthermore, the test substances used in these studies were liquid formulations whereas the enduse product is a wettable powder which may pose a greater potential for eye irritation.

In lieu of conducting acute oral and acute pulmonary infectivity/toxicity studies, the applicant has 

provided waiver rationales based on the results of the three above-mentioned studies, in addition to the 

mouse IP batch analysis studies, a literature search, the practices and design features employed in antisapstain facilities and the nature of the formulation ingredient. A similar rationale, the findings of a 

review paper and the use of PPE were found acceptable to waive testing for the primary eye irritation 

study. Appropriate PPE is required to mitigate potential skin sensitization that may be due to substances 

produced by all microorganisms that can elicit allergic reactions after repeated exposure to high 

concentrations. 

Requests to waive toxicology testing for the EP were accepted based on the foregoing studies and 

rationales submitted. No hypersensitivity incidents, including immediate-type or delayed-type reactions 

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

rosea strain 321U. Any future hypersensitivity incidents must be reported to the EPA (refer to test note 

#3 of 40 C.F.R. § 158.2140(d)).

A cell culture study is not required because Clonostachys rosea strain 321U is not a virus (refer to test 

note #4 of 40 C.F.R. § 158.2140(d)).
Clonostachys rosea strain 321U previously known as Gliocladium roseum strain 321U Page 21 of 29

Biopesticides Registration Action Document DRAFT

TABLE 3. Summary of Tier 1 Toxicology Data Submitted for FPSolano® Technical [the Technical Grade of the Active 

Ingredient (TGAI)], Clonostachys rosea strain 321U (40 C.F.R. § 158.2140)

Harmonized Guideline 

Number and Data 

Requirement

Summary of review 

MRID 

Number

885.3050

(waiver request)

Acute Oral 

Toxicity/Pathogenicity

(waiver request based on IP, 

dermal, mouse IP batch 

analysis and nature of 

formulation ingredient)

Waiver rationale based on the results of an acute IP injection, acute dermal toxicity, 

dermal irritation and mouse IP batch analysis studies, a literature search, the 

practices and design features employed in anti-sapstain facilities and the nature of 

the formulation ingredient. In lieu of acute oral pathogenicity testing, the registrant 

submitted a scientific rationale to waive testing. 

No signs of toxicity were noted in rats acutely exposed by IP injection to a 5.0×106

CFU/animal dose of C. rosea strain 321U. Infectivity was not assessed in this study 

although no signs of pathogenicity were observed. 

Results from a mouse IP batch analysis were also submitted in which no signs of 

adverse effects were observed amongst mice dosed with at least 106 CFU of C. rosea 

strain 321U via IP injection. 

Clonostachys rosea strain 321U was not toxic or irritating via the dermal route 

although the test material used was lower in toxicity than both the TGAI and the EP. 

As well, there were no incidents of pathogenicity/infectivity or other deleterious 

effects to mammalian species due to Clonostachys or Gliocladium reported in the 

public scientific literature. 

There were a number of cases of infections or adverse effects in 

immunocompromised individuals due to species of the related genus Trichoderma. 

Toxicological studies on toxins and enzymes associated with Trichoderma species 

used purified forms of these substances whose levels are unlikely to be encountered 

under the proposed manufacturing process of use scenario. In addition to the 

toxicology database, the registrant also cited the design and practices employed at 

anti-sapstain facilities that are designed to minimize exposure. The formulation 

ingredient does not pose a concern. The EP is not intended for use on food or feed 

crops.

WAIVER ACCEPTEDTOXICITY CATEGORY III - Classification: 

Acceptable

48782605

885.3100

(14-Day study)

Acute Dermal 

Toxicity/Pathogenicity

LD50 > 5050 mg/kg bw; test material 1x106cfu/ml is lower than technical 

(2x108cfu/ml) or EP (5x108cfu/g). 

Materials and methods:

Tested Sprague-Dawley Rat -5/sex, 24-hour dermal exposure, with C. rosea strain 

321U (1.0×106 CFU/mL) at dose of 5050 mg/kg bw (4.82 mL/kg bw) and animals 

observed for 14 days. Observations for mortality and clinical/behavioural signs of 

toxicity made three times on day of dosing and at least once daily thereafter. 

Individual body weights recorded just prior to dosing and on Days 7 and 14. 

Results:

There were no mortalities noted throughout the study period. All animals appeared 

normal and gained weight. There were no abnormalities upon necropsy.

LOW TOXICITY- viability of test substance is lower than that of TGAI or EP and 

there were no formulation ingredients in the test substance.

TOXICITY CATEGORY IV -Classification: Supplemental

48782617
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Biopesticides Registration Action Document DRAFT

TABLE 3. Summary of Tier 1 Toxicology Data Submitted for FPSolano® Technical [the Technical Grade of the Active 

Ingredient (TGAI)], Clonostachys rosea strain 321U (40 C.F.R. § 158.2140)

Harmonized Guideline 

Number and Data 

Requirement

Summary of review 

MRID 

Number

885.3150

(waiver request)

Acute Pulmonary 

Toxicity/Pathogenicity

Waiver Rationale based on mouse IP (48782607); 4 hr dermal irritation study in 

rabbits (48782619); acute dermal (48782617); public literature, nature of ingredients 

in formulation and low exposure potential due to the application method. In lieu of 

acute oral pathogenicity testing, the registrant submitted a scientific rationale to 

waive testing. 

No signs of toxicity were noted in rats acutely exposed by IP injection to a 5.0×106

CFU/animal dose of C. rosea strain 321U. Infectivity was not assessed in this study 

although no signs of pathogenicity were observed. 

Results from a mouse IP batch analysis were also submitted in which no signs of 

adverse effects were observed amongst mice dosed with at least 106 CFU of C. rosea 

strain 321U via IP injection. 

Clonostachys rosea strain 321U was not toxic or irritating via the dermal route 

although the test material used was lower in toxicity than both the TGAI and the EP. 

As well, there were no incidents of pathogenicity/infectivity or other deleterious 

effects to mammalian species due to Clonostachys or Gliocladium reported in the 

public scientific literature. 

There were a number of cases of infections or adverse effects in 

immunocompromised individuals due to species of the related genus Trichoderma. 

Toxicological studies on toxins and enzymes associated with Trichoderma species 

used purified forms of these substances whose levels are unlikely to be encountered 

under the proposed manufacturing process of use scenario. In addition to the 

toxicology database, the registrant also cited the design and practices employed at 

anti-sapstain facilities that are designed to minimize exposure. The formulation 

ingredient does not pose a concern. 

SUPPLEMENTAL as infectivity has not been assessed via any route of 

exposure and inhalation will be a primary source of exposure to occupational 

workers. Appropriate PPE is required to mitigate any potential for inhalation 

exposure. 

WAIVER ACCEPTED

TOXICITY CATEGORY III - Classification: Supplemental

48782606.

885.3200

(21-Day study)

Acute Injection 

Toxicity/Pathogenicity

At an IP dose of 5.0 × 106 CFU/animal, 1 month old Sprague Dawley rats were 

observed for 21 days. Infectivity was not assessed although no signs of 

pathogenicity were noted. Replacement study waived based an acute IP injection, 

acute dermal toxicity, dermal irritation and mouse IP batch analysis studies, a 

literature search, the practices and design features employed in anti-sapstain 

facilities and the nature of the formulation ingredient. 

Materials and methods:

1) C. rosea strain 321U (5×106 CFU/mL); (15/sex); IP injection; 1 mL/animal

(equivalent to 5.0×106 CFU/animal); 3 animals sacrificed 1 hour after dosing to very 

dose administered

2) Inactivated test substance; (5/sex); IP injection; 1 mL/animal (inactivated 

equivalent of 5×106 CFU/animal)

3) Untreated control (5/sex)

48782607

49172001
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Biopesticides Registration Action Document DRAFT

TABLE 3. Summary of Tier 1 Toxicology Data Submitted for FPSolano® Technical [the Technical Grade of the Active 

Ingredient (TGAI)], Clonostachys rosea strain 321U (40 C.F.R. § 158.2140)

Harmonized Guideline 

Number and Data 

Requirement

Summary of review 

MRID 

Number

Observations: Mortality and adverse effects observations were made three times on 

Day 0 and daily thereafter. Body weights were measured on Day 0 prior to dosing 

and on Days 7, 14 and 21. Necropsies were performed at the end of the study on 

animals in the treatment group. Organ weights were recorded for all animals.

Results: Test substance does not appear to be toxic. Infectivity was not assessed 

although no signs of pathogenicity were observed. 

There were no mortalities at any point of the study. All animals appeared normal 

throughout. No adverse effects were attributed to the test substance. No significant 

differences between groups in body weights, body weight gains or organ weights. 

No observable abnormalities noted at necropsy.

TOXICITY CATEGORY IV - Classification: Acceptable.

870.2500

Primary Dermal Irritation

A dermal irritation study was conducted for the technical. This study is required for 

the EP for labeling purposes to determine appropriate PPE to mitigate exposure risk 

to workers.

Material and methods:

2 ♂; 1 ♀ New Zealand white Rabbit treated with 0.5 mL/animal Clonostachys rosea 

strain 321U (1.0×106 CFU/mL) for 4-hour dermal exposure. Erythema and edema 

formation and any other dermal defects or irritation were scored at 1, 24, 28 and 72 

hours after unwrapping.

Results:

MIS=0/8; MAS=0/8.

This dose is less than the technical and EP is a wettable powder. Guideline requires 

testing with EP.

TOXICITY CATEGORY IV - Classification: Supplemental

48782619

Hypersensitivity incidents

The applicant reported that no hypersensitivity incidents, including immediate-type 

or delayed-type reactions of humans and domestic animals, occurred during 

research, development, or testing of the TGAI or MP. Any future hypersensitivity 

incidents must be reported to the EPA (refer to test note #3 of 40 C.F.R. § 

158.2140(d)). 

48782608

885.3500

Cell culture

Not required because Clonostachys rosea strain 321U is not a virus (refer to test note #4 

of 40 C.F.R. § 158.2140(d)).

N/A

Table 4 includes a summary of the reviews of the requests to waive testing for the Clonostachys rosea

strain 321U End-Use Pesticide Product (EP), FPSolano® (40 C.F.R. § 158.2140)

TABLE 4. Summary of Tier 1 Toxicology Data Submitted for Clonostachys rosea strain 321U End-Use Pesticide

Product (EP), FPSolano® (40 C.F.R. § 158.2140)

Data Requirement Results

MRID 

Number

870.1100

(waiver request)

Waiver rationale based on results of an acute IP injection, acute dermal toxicity, dermal 

irritation and mouse IP batch analysis studies, a literature search, the practices and design 

features employed in anti-sapstain facilities and the nature of the formulation ingredient. 
Clonostachys rosea strain 321U previously known as Gliocladium roseum strain 321U Page 24 of 29

Biopesticides Registration Action Document DRAFT

TABLE 4. Summary of Tier 1 Toxicology Data Submitted for Clonostachys rosea strain 321U End-Use Pesticide

Product (EP), FPSolano® (40 C.F.R. § 158.2140)

Data Requirement Results

MRID 

Number

Acute Oral Toxicity FPSolano is not intended for use on food or feed crops (refer to test note #5 of 40 C.F.R. 

§ 158.2140(d)).

WAIVER ACCEPTED

TOXICITY CATEGORY III - Classification: Acceptable

48782616

870.1200

(waiver request)

Acute Dermal 

Toxicity

Waiver rationale based on MRID 48782617; safety of ingredients in formulation; use of 

PPE and low exposure potential. LD50 > 5050 mg/kg bw; test material 1x106cfu/ml is 

lower than technical (2x108cfu/ml) or EP (5x108cfu/g); low toxicity; limited exposure 

via application method; appropriate PPE required, (refer to test note #5 of 40 C.F.R. § 

158.2140(d)).

WAIVER ACCEPTED

TOXICITY CATEGORY IV - Classification: Supplemental

48782617

870.1300

(waiver request)

Acute Inhalation 

Toxicity

Based on pulmonary study, low exposure due to method of application in dip tanks 

outdoors, appropriate mask with NIOSH prefix N-95.

WAIVER ACCEPTED

TOXICITY CATEGORY III - Classification: Supplemental 

48782616

870.2400

(waiver request)

Acute Eye Irritation

Waiver rationale based on results the results of IP injection, dermal irritation, dermal 

toxicity and IP batch analysis studies, the findings of a review paper and the use of PPE; 

the label must include use of PPE for eye protection based on dermal irritation and 

dermal toxicity studies. 

WAIVER ACCEPTED

TOXICITY CATEGORY III - Classification: Acceptable

48782618

Sensitization

A dermal sensitization study was not conducted. All microorganisms are recognized as 

being able to produce substance that can elicit allergic reactions after repeated exposure 

to high concentrations. Labelling must include appropriate PPE for potential sensitizing 

effects of microbial products must be included in the label.

Potential sensitizer - Classification: Acceptable

48782608

870.2500

(waiver request)

Primary Dermal 

Irritation

In lieu of a dermal irritation study on the end-use product, FPSolano, the applicant 

provided a waiver rationale based on the nature of the formulation ingredient, the 

absence of toxin production by the active ingredient and the requirement for personal 

protective equipment to minimize dermal exposure. A primary dermal irritation study 

was conducted for the technical at a dose less than the technical and EP is a wettable 

powder. Although the claim that the active ingredient does not produce toxins was 

determined to be inconclusive, the waiver request was considered to be acceptable based 

on the identity and general safety of the formulation ingredient and the PPE 

requirements. See 870.2500 in TGAI table 1; MRID 48782619). 

WAIVER ACCEPTED

TOXICITY CATEGORY IV - Classification: Acceptable

48782619

Hypersensitivity 

incidents

Published literature reported two incidents related to Gliocladium that were not very 

clearly linked directly to this pesticide active ingredient. The applicant reported that no 

hypersensitivity incidents, including immediate-type or delayed-type reactions of 

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

TGAI or MP. Any future hypersensitivity incidents must be reported to the EPA (refer to 

test note #3 of 40 C.F.R. § 158.2140(d)). 

49172001

Tiers II and III studies were not required for of Clonostachys rosea strain 321U based on the assessment 

of the submissions for the fungus in the Tier I studies.
Clonostachys rosea strain 321U previously known as Gliocladium roseum strain 321U Page 25 of 29

Biopesticides Registration Action Document DRAFT

D. Ecological Effects

Environmental Exposure

The proposed use pattern is expected to limit exposure to nontarget organisms. The a.i. will be confined 

to dip tanks used for treatment, and is not expected to result in exposure under labeled use. Birds, wild 

mammals, and terrestrial plants are not expected to be exposed to C. rosea strain 321U in dip tanks, and 

would be expected to have very limited exposure to the a.i. that is present on freshly treated wood stored 

at the treatment facility. Nontarget insects and honey bees may be incidentally exposed to freshly treated 

lumber, though this exposure is also expected to be very limited. Aquatic organisms may be exposed if 

the treatment solution is released into the environment, such as with splash during treatment or wash-off 

of the treatment during rain events. Language is present on the label to limit exposure in aquatic 

environments.

Summary of Nontarget Effects Data

Table 5 provides the status of the nontarget organism data requirements for the active ingredient, 

Clonostachys rosea strain 321U, required under 40 CFR§ 158.2150. AEF Global submitted guideline 

studies to address toxicity/pathogenicity in birds, freshwater fish, and freshwater invertebrates, and 

submitted scientific rationale to address toxicity/pathogenicity in wild mammals, nontarget insects and 

honey bees, and terrestrial and aquatic plants. Data Evaluation Records for these studies and rationale 

were completed through joint review with Canada’s Pesticide Regulatory Management Agency

(PMRA). EPA determined that the information provided is sufficient to satisfy the nontarget organism 

data requirements for this new a.i. and its proposed uses. Additional testing at higher tiers is not required 

for the proposed uses.

Table 5. Summary of Tier 1 Ecological Effects data submitted to comply with requirements published in 40 

CFR § 158.2150 to support the registration of products containing the a.i., Clonostachys rosea strain 321U. 

OCSPP 

Guideline 

Number

Data Requirement Results MRID No.

885.4050 Avian Oral Toxicity

The LD50 in Northern bobwhite (Colinus virginianus) was 

determined to be > 5mL/kg bw (of test material containing 

106cfu/mL). The study had several deficiencies, including 

viability of the test material, which was less than that of the 

technical product.

Classification: Supplemental

48782609

885.4150 Wild Mammal 

Toxicity/Pathogenicity

Acceptable scientific rationale indicates that adverse effects 

to mammals would not be expected. This rationale is 

sufficient for assessing risks to wild mammals. 

WAIVER ACCEPTED

Classification: Acceptable

48782610

885.4100 Avian Inhalation 

Toxicity/Pathogenicity

Not required; the nature of the microbial pesticide does not 

indicate potential pathogenicity to birds. N/A
Clonostachys rosea strain 321U previously known as Gliocladium roseum strain 321U Page 26 of 29

Biopesticides Registration Action Document DRAFT

Table 5. Summary of Tier 1 Ecological Effects data submitted to comply with requirements published in 40 

CFR § 158.2150 to support the registration of products containing the a.i., Clonostachys rosea strain 321U. 

OCSPP 

Guideline 

Number

Data Requirement Results MRID No.

885.4200 Freshwater Fish 

Toxicity/Pathogenicity

In a 30-day study, mortality was >50% in rainbow trout 

(Oncorhyncus mykiss) exposed to Clonostachys rosea strain 

321U at 104cfu/mL, and an LC50 could not be established. 

Use pattern and proposed label language is expected to 

minimize aquatic exposure.

WAIVER ACCEPTED

Classification: Supplemental

48782611

885.4240 Freshwater Invertebrate 

Toxicity/Pathogenicity

In a 7-day study, mortality was >50% in Daphnia magna

exposed to Clonostachys rosea strain 321U at nominal 

concentration of 108cfu/mL (likely actually <106), and an 

LC50 could not be established. Use pattern and proposed 

label language is expected to minimize aquatic exposure.

Classification: Supplemental

48782612

885.4240 Freshwater Invertebrate 

Toxicity/Pathogenicity

In a 21-day study, the EC50 for survival was determined to 

be 5.5 x 105cfu/mL, the IC50 and NOEC for reproduction 

were 5.8 x 105and 5.0 x 105cfu/mL, respectively, and the 

IC50 for growth was determined to be > 5.0 x 105cfu/mL. 

Identity, purity, viability, and stability of the test substance 

were not demonstrated.

Classification: Supplemental

49351601

885.4280 Estuarine/Marine Fish 

and Invertebrate Testing

Not required; significant exposure in marine or estuarine 

environments is not expected due to the use pattern. N/A

885.4340

885.4380

Nontarget Insect 

Testing

Honey Bee Testing

Scientific rationale was sufficient to show that exposure to 

nontarget insects and honey bees is expected to be minimal 

based on the intended use pattern and proposed label 

language.

WAIVER ACCEPTED

Classification: Acceptable

48782615

885.4300 Nontarget Plant Testing

Scientific rationale was sufficient to show that adverse 

effects to terrestrial and aquatic plants are not expected due 

to the proposed use pattern and cited data showing a lack of 

effects in some terrestrial plants.

WAIVER ACCEPTED

Classification: Acceptable

48782613

48782614

Tiers II, III, and IV

Not required for Clonostachys rosea based on the acceptability of the scientific rationale and studies provided for 

Tier I.

Ecological Effects Risk Characterization 

Birds and Mammals

Exposure to birds and mammals is expected to be minimal because the intended use pattern is limited to 
Clonostachys rosea strain 321U previously known as Gliocladium roseum strain 321U Page 27 of 29

Biopesticides Registration Action Document DRAFT

treatment of wood contained in dip tanks. The label also contains sufficient language to limit 

environmental release and exposure to birds and mammals. Studies that were submitted did not show 

toxicity or pathogenicity in these animals; however, the studies were determined to have limited use for 

risk assessment. Since exposure is expected to be minimal, adverse effects to birds and mammals are not 

expected as a result of the proposed use of C. rosea strain 321U, as labeled.

Nontarget Insects and Honeybees

Data in the public literature indicate that Clonostachys rosea is a potential entomopathogen (Toledo et 

al. 2006, Vega et al. 2008). Insects may be exposed if they contact treated wood; however, while most 

of this exposure is likely to be incidental, some insects may be attracted to freshly treated wood as a 

potential water source while it is still wet. Since freshly treated wood will be limited to these treatment 

facilities and not widespread in the environment, such exposure is expected to be minimal and to have 

little impact on nontarget insect and honey bee populations. Therefore, adverse effects to nontarget 

insects and honey bees are not expected. 

Rationale presented by AEF Global to address nontarget data requirements included a proposal to 

include a label statement requiring users to cover freshly treated wood. This statement was part of 

EPA’s and PMRA’s consideration in acceptance of this rationale. While exposure to nontarget insects 

and honey bees may be limited, requiring users to cover freshly treated wood would reduce exposure 

further and also any uncertainty about risk due to the potential for effects on insects. It is recommended 

to include this requirement on the label.

Freshwater Fish and Invertebrates

Studies submitted in support of this registration indicated adverse effects on survival in freshwater fish 

and survival, reproduction, and growth in freshwater invertebrates with exposure to C. rosea strain 

321U. Exposure in aquatic environments could occur through splashing during treatment, spills, 

washing of treatment equipment, or wash-off of treated materials during rain events. Such exposure is 

expected to be limited, since these events would only occur at the lumber yard where the wood is 

treated, and these sites are likely limited within any landscape. Therefore, adverse effects to freshwater 

fish and invertebrates are not expected due to minimal exposure. 

The label contains language that is expected to sufficiently limit exposure through these types of 

releases into the environment. Rationale submitted as a supplement to the freshwater fish and 

invertebrate studies provided information on clean-up of spills, including application of a 5% bleach 

solution for one hour to the spill location. While the label does acknowledge that spills and splash are 

the responsibility of the applicator, it is recommended that the label provide information to the user on 

proper clean-up of spills so that environmental exposure is minimized. Label language requiring freshly

treated lumber to be covered was also proposed in this rationale, and was considered in EPA’s and 

PMRA’s acceptance of this rationale. This language is also recommended, since this would prevent 

wash-off of the C. rosea strain 321U. 

Marine/Estuarine Fish and Invertebrates

Exposure in marine/estuarine environments is not expected to be significant. Use of the EP is limited to 

facilities that cut or chip wood, and the product is applied by use of a dip tank. Exposure in nearby

aquatic areas is expected to be limited, and any exposure in marine or estuarine sites would be further 
Clonostachys rosea strain 321U previously known as Gliocladium roseum strain 321U Page 28 of 29

Biopesticides Registration Action Document DRAFT

minimized by additional dilution within these larger water bodies. Therefore, adverse effects are not 

expected in marine/estuarine fish or invertebrates as a result of the use of C. rosea strain 321U, as 

proposed.

Terrestrial and Aquatic Plants

Exposure to terrestrial and aquatic plants is also expected to be limited as a result of the use pattern and 

limited exposure. Some data were also submitted showing no observed adverse effects in cabbage, 

lettuce, and tomato plants. Label statements that limit any potential run-off from treated lumber would 

further reduce exposure in aquatic environments. Based on the use pattern limiting exposure and 

expected lack of effects, risk to terrestrial and aquatic plants is not anticipated as a result of the proposed 

uses of C. rosea strain 321U, as labeled.

Threatened and Endangered Species Assessment 

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

rosea strain 321U as a result of the proposed labeled uses, effects to federally listed threatened and 

endangered species and their designated critical habitats are also not expected. Therefore, a “No Effect” 

determination is made for direct and indirect effects to federally listed threatened and endangered 

species and their designated critical habitats for the proposed use of Clonostachys rosea strain 321U. 

Conclusions

Adverse effects to nontarget organisms are not expected as a result of the proposed use of C. rosea strain 

321U. Rationale submitted to support the registrations of products containing this a.i. proposed label 

language requiring that users cover freshly treated wood to prevent exposure to nontarget organisms. 

This language is not on the latest proposed label for the EP, but is recommended for inclusion on the 

label reduce any potential exposure to nontarget aquatic organisms, insects, and honey bees and any 

uncertainty with these risk conclusions. The label should also include guidance on proper clean-up of C. 

rosea strain 321U that may spill or splash during treatment.

References

Toledo, A.V., Virla, E., Humber, R.A., Paradell, S.L., Lopez Lastra, C.C. 2006. Journal of Invertebrate 

Pathology 92: 7-10.

Vega, F.E., Posada, F., Aime, M.C., Pava-Ripoll, M., Infante, F., and Rehner, S.A. 2008. 

Entomopathogenic fungal endophytes. Biological Control 46: 72-82. 
Clonostachys rosea strain 321U previously known as Gliocladium roseum strain 321U Page 29 of 29

Biopesticides Registration Action Document DRAFT

APPENDIX B. PESTICIDE PRODUCTS

EPA Reg. 

Number

Registration 

Name

Percent 

Active 

Ingredient

Formulation 

Type

Use 

Site(s)

Method(s) 

of 

Application

Application Rate(s) Target Pest

89046-2

FPSolano®

Technical

100% (3.8 x 

109CFU/ 

gallon)

Technical

liquid 

formulation

N/A

89046-1

FPSolano®

(End-use 

Product)

20%

5 x 108 CFU/

gram 

End Use –

Wettable 

powder

Sawn 

wood 

products 

and logs

Apply by 

diptank 

application 

technology. 

Do not tank 

mix with any 

pesticides.

Mix 7 ounces of 

FPSolano® per 26 

US gallon of water 

containing 4% w/v 

sodium carbonate and 

1% w/v sodium 

bicarbonate e.g. per 

26 US gallon of 

water for example: 

add 7 ounces of 

FPSolano®, 8.8 lbs 

of sodium carbonate 

and 2.2 lbs of sodium 

bicarbonate. 

Complete with up to 

per 26 US gallons of 

water to make final 

suspension 

containing 3.8x109

cfu/quart of 

Clonostachys rosea

strain 321U.

Biological 

protectant 

for treatment 

of green 

lumber 

against 

staining 

fungi (e.g. 

blue stain 

fungi) and 

molds

1 Go to www.regulations.gov. Conduct a search for Docket ID Number EPA-HQ-OPP-2012-0507. 

2

See http://oaspub.epa.gov/apex/pesticides/f?p=PPLS:1.