### BIOPESTICIDES REGISTRATION ACTION DOCUMENT



## Tobacco mild green mosaic tobamovirus strain U2

Pesticide Chemical (PC) Code: 056705

U.S. Environmental Protection Agency Office of Pesticide Programs

Bio pesticides and Pollution Prevention Division

DRAFT

– November 14 , 2014Tobacco mild green mosaic tobamovirus strai n U2 (TMGMV U2) Page 2 of 31

Biopesticides Registration Action Document



### TABLE OF CONTENTS

I. EXECUTIVE SUMMARY

................................ ................................ ................................ ......................... 4

II. ACTIVE ING REDIENT OVERVIEW

................................ ................................ ................................ ..... 6

III. REGULATORY BACKGROUN D

................................ ................................ ................................ ............ 6

A. Application for Pesticide Product Registration

................................ ................................ ............. 6

B. Food Tolerance Exemption

................................ ................................ ................................ ............. 7

IV. RISK ASSESSMENT

................................ ................................ ................................ ................................ .. 7

A. Pr oduct Analysis A ssessment (40 CFR § 158.2120)

................................ ................................ ...... 8

B. Hu man Health Assessment (40 CFR § 158.2140)

................................ ................................ .......... 8

C. Environmental Assessment (40 CFR § 158.2150)

................................ ................................ ........ 12

V. ENVIRONMENTAL JUSTIC E

................................ ................................ ................................ ............... 16

VI. RISK MANAGEMENT DECI SION

................................ ................................ ................................ ........ 17

VII. ACTIONS REQUIRED OF THE REGISTRANT

................................ ................................ ................. 18

VIII. GLOSSARY OF ACRONYMS AND ABBREVIATIONS

................................ ................................ .... 19

IX. BIBLIOGRAPHY

................................ ................................ ................................ ................................ ...... 20

A. Studies Submitted to Support the Tobacco mild green mosaic tobamovirus strain U2

Pesticide Product Registration

................................ ................................ ................................ ...... 20

B. Environmental Protection Agency Risk Assessment Memoranda

................................ ............. 21

C. Other References

................................ ................................ ................................ ............................ 22

APPENDIX A. MICROBI AL PESTICIDES DATA R EQUIREMENTS

................................ ........................ 25

APPENDIX B. PESTICI DE PRODUCTS

................................ ................................ ................................ ........... 31Tobacco mild green mosaic tobamovirus strai n U2 (TMGMV U2) Page 3 of 31

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### BIOPESTICIDES REGISTRATION ACTION DOCUMENT TEAM

Office of Pesticide Programs (OPP) Biopesticides and Pollution Prevention Division

Microbial Pesticides Branch

Science Reviews

Shannon Borges Environmental Effects Joel V. Gagliardi , Ph.D. Product Analysis, Human Health

, and

Environmental Effects

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

Regulatory

ShaRon Carlisle Acting Associate Chief, Microbial Pesticides

Branch

Jeannine Kausch Regulatory Action Leade r Kimberly Nesci Chief, Microbial Pesticides BranchTobacco mild green mosaic tobamovirus strai n U2 (TMGMV U2) Page 4 of 31

Biopesticides Registration Action Document

I. EXECUTIVE SUMMARY

Background

In May 2013

, BioProdex , Inc. (BioProdex)

, submitted an application for an end

- use pesticide product (SolviNix ® LC, EPA File Symbol : 81179

- G ) to the U .S. Environmental Protection Agen cy (EPA) under section 3 of the Federal Insecticide, Fungicide, and Rodenticide Act (FIFRA)

. This pesticide product

contain s a new active ingredient: Tobacco mild green mosaic tobamovirus strain U2 (TMGMV U2)

.

TMGMV is a tobamovirus, a type of plant virus , and is known as a pathogen of tobaccos ( Nicotiana spp.), peppers ( Capsicum spp.), and about 20 other plant species ( Brunt et al ., 1996; Font et al ., 2009; Green and Kim, 1991; ICTVdB Management, 2006; Wetter, 1989). According to literature provided by

BioProdex

, TMGMV probably occurs in all tropical and subtropical regions in the world where tree tobacco ( Nicotiana glauca ), a natural host to this virus, occurs (Brunt et al ., 1996; Wetter, 1989). I n the U.S. specifically, this virus occurs in California (Bald and Goodchild, 1960; Bodaghi et al ., 2000 and 2004), Florida (Baker and Adkins, 2004; Baker and Zettler, 1988; Zettler and Nagel, 1983), Georgia (McKinney, 1952; Wetter, 1984), Illinois (Knigh t et al ., 1962; Miller and Thornberry, 1958), Ohio (Letschert et al ., 2002), South Carolina (McKinney, 1952; Wetter, 1984), and Wisconsin (Johnson, 1947; Siegel and Wildman, 1954 ; Weber, 1951 ).

BioProdex isolated TMGMV U2 in Florida from horticultur al materials and identified the virus by its host range and genomic sequence. Data demonstrate that TMGMV U2 infects and kills tropical soda apple ( Solanum viarum ). Tropical soda apple is a thorny plant native to South America (Argentina, Brazil, and Parag uay), is designated as a Federal Noxious Weed under the Plant Protection Act in the

U.S. , and crowds out native plant species and forage for livestock in the southeastern U.S. ( UF

– IFAS, 2008; USDA, 2014). In light of TMGMV U2’s herbicida l capabilities, B ioProdex proposed to register an end

- use pesticide product to control tropical soda apple in pastures and wooded areas that are fenced to contain grazing animals

.

The EPA ’s s cientists reviewed product analysis, toxicology, and nontarget organism data and information ( 40 CFR §§ 158.2120, 158.2140, and 158.2150, respectively) submitted to support the registration of the proposed TMGMV U2 pesticide product

. Overall, such data and information are adequate for risk assessment purposes , fulfill the current micro bial pesticide data requirements

, and
support registration of the proposed pesticide product under FIFRA section 3(c)(5)

.

Product Analysis

BioProdex fulfilled the product analysis data requirements for TMGMV U2 , including product chemistry and composition , analysis of samples, and physical and chemical characteristics, by submitting acceptable guideline studies.

Toxicology

BioProdex submitted a dequate mamm alian toxicology information to support the TMGMV U2 pesticide product

. Based on BioProdex ’s scientif ic ra tionale, the EPA waived acute toxicity/pathogenicity and cell culture testing for this active ingredient. The rationale included references to scientific literature and showed that plant viruses are ubiquitous in plants and food derived from plants

, a re not known or expected to cause toxicity, pathogenicity, or infectivity in mammals, and are unable to infect animal

cells. Moreover, BioProdex reported that no hypersensitivity incidents occurred during research, development, or testing of TMGMV U2

. In l ight of the acceptability of the toxicological scientificTobacco mild green mosaic tobamovirus strai n U2 (TMGMV U2) Page 5 of 31

Biopesticides Registration Action Document

rationale and literature provided by BioProdex and the absence of hypersensitivity incidents, the EPA is not requiring testing at higher tiers (i.e., Tiers II and III)

.

Tolerance Exemption

The EPA concludes that there is a reasonable certainty that no harm will result to the U.S. population, including infants and children, from aggregate exposure to residues of TMGMV U2. The EPA does not
expect dietary (food and drinking water) or other non

- occupati onal risks from use of TMGMV U2 as an active ingredient in the proposed pesticide product. V irus

- infected food plants have always been a part of the human diet and domestic animal food supply (Beemster and de Bokx, 1987; Dewan and Pearson, 1995; Jones et a l ., 1934; McKinney, 1929; Palukaitis, 1991; P rovvidenti and Gonsalves, 1984)

. T o date, there have been no reports of adverse human or animal health effects associated with consumption of plant viruses in food. Fur thermore, since applications of TMGMV U2 ar e limited to pastures and wooded areas that are fenced to contain grazing animals, increased exposures of the active ingredient to the general population through food and drinking water and in residential settings are not expected.

Should TMGMV U2 be prese nt in food or drinking water, supporting toxicological information indicate s that human exposure to t his plant virus would not result in adverse effects.

Occupational Exposure Despite the low toxicological profile of TMGMV U2

, the EPA is requiring baselin e protective clothing

for individuals who, due to their occupation, may be exposed to the active ingredient for prolonged periods and/ or multiple times

. Individuals working with TMGMV U2 to treat tropical soda apple are directed to wear a long

- sleeved shir t, long pants, socks, shoes, waterproof gloves, and 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 TMGMV U2 pesticide products, the EPA may requir e additional protective clothing (e.g., eyewear ) , other than the standard described above, on a product

- specific basis.

Nontarget Organisms The EPA found that scientific rationale and nontarget plant studies submitted by BioProdex are sufficient to fulf

ill 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 a n environmental risk assessment based on informati on provided by BioProdex. Provided that limited use sites and application methods, mitigating label language, and certain manufacturing process steps are in place, the EPA has determined that u nreasonable adverse effects to nontarget organisms are
not expe cted from the proposed pesticidal uses of TMGMV U2

. Moreover, the EPA made a “No Effect” determination for direct and indirect effects to federally listed

, threatened and endangered species and their designated critical habitats resulting from these same p roposed pesticidal uses.

Public Participation

On Octob er 1, 2009, the EPA announced a policy to provide a more meaningful opportunity for the public to participate in major registration decisions before they occur. According to this policy, the EPA provi des a public comment period prior to making a registration decision for the following types of applications: new active ingredients; first food uses; first outdoor uses; first residential uses; or any other registration actions for which the EPA believes t here may be significant public interest.

Consistent with the policy of making registration actions more transparent, the proposed pe sticide product containing TMGMV U2

, a new active ingredient, will be subject to a comment period. In

addition to contain ing a new active ingredient, the registration of th is pesticide product would result inTobacco mild green mosaic tobamovirus strai n U2 (TMGMV U2) Page 6 of 31

Biopesticides Registration Action Document

the first food and outdoor use s for TMGMV U2

. The docket identification number, which is associated

with th is registration action and can be accessed through http://www.regulations.gov

, is EPA

- HQ

- OPP

201 3

- 0 759 . The following documents are available for comment in EPA

- HQ

- OPP

- 201 3

- 0 759 : (1) draft Tobacco mild green mosaic tobamovirus strain U2 Biopesticides Regi stration Action Do cument (BRAD) and (2) draft label for the end

- use pesticide product SolviNix ® LC (EPA File Sym bol : 81179

- G ). While a final decision on registration is contingent upon review and consideration of public

comments, the EPA presently believes that, based upon the risk assessment and info rmation submitted in support of SolviNix ® LC

, it is appropriate to issue this registration . The basis for this preliminary

decision can be f

ound in the risk assessment for TMGMV U2 , which is characterized throughout this

BRAD.

II. ACTIVE INGREDIENT OVERVIEW

Biological Name: Tobacco mild green mosaic tobamovirus strain U2

Culture Deposit: American Type Culture Collection (ATCC) in Manassas, Virginia

under Accession Number PV

- 1200

OPP Chemical Code: 056705

Type of Pesticide: Microbial Pesticide

– Herbicide

See Appendix B for specific information (e.g., use sites, application rates, methods of application, formulation type , and target pest ) regarding the proposed p est icide product containing this active ingredient.

III. REGULATORY BACKGROUND

A. Application for Pesticide Product Registration

In May 2013

, BioProdex (address: 3131 NW 13 th Street #54A; Gainesville, Florida 32609

- 2183 )

submitted an application to reg ister a n end

- use pesticide product SolviNix ® LC (EPA File Symbol:

81179

- G ) under FIFRA section 3. On January 1 7, 201 4 ( 7 9 FR 3203 )

, the EPA announced receipt of this application to register a pesticide product containing a new active ingredient and opened a 3 0

- day public comment period pursuant to the provisions of FIFRA section 3(c)(4). In response to this Notice of

R eceipt, the EPA received one comment that expressed concern ov er the lack of information provide d in the Federal Register document and generally opposed both pesticide product applications (i.e., the

SolviNix ® LC applicatio n and an application for an antimicrobial pesticide product ) summarized in that

document . Speci fically, the anonymous commenter cited concern for “conventional production practices” and the introduction of “new chemical agents” during these practices.

With publication of the Notice of R eceipt, the EPA met the statutory requirement, as indicated in the previous paragraph , to inform the public of applications for pesticide products containing a new active ingredient, such as SolviNix ® LC , with basic information (e.g., name s and proposed uses of the activeTobacco mild green mosaic tobamovirus strai n U2 (TMGMV U2) Page 7 of 31

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ingredients)

. At that time, the EPA had not y et completed its review of the data and information submitted by BioProdex . Pursuant to its authority under FIFRA, the EPA has now conducted a comprehensive assessment of TMGMV U2 and concluded that this herbicide is not expected to cause any unreasonable adverse effects to human health or the environment. Further, the EPA emphasizes that

TMGMV U2 is not a conventional pesticide. Conventional pesticides generally consist of synthetic materials, may affect a broad spectrum of nontarget organisms, and may be inherently more toxic. Instead, t he E PA considers TMGMV U2, a naturally occurring plant virus, to be a biopesticide

. A s described in section I of this document, this biop esticide will be used to control tropical soda apple, a noxious weed that adversely affects native plant species and forage for livestock in the southeastern U.S.

B. Food Tolerance Exemption

Concurrent with BioProdex’s registration application and under Federal Food, Drug, and Cosmetic Act (FFDCA) section 408(d), Interregional Research Project Number 4 (IR

- 4) submitted a petition to establish an exemption from the requirement of a tolerance for TMGMV U2 ( PP 3E8181 ). In the Federal Register of Febru ary 21, 2014 ( 79 FR 9870 )

, the EPA announced that IR

- 4 proposed to establish an exemption from the requirement of a tolerance for residues of the herbicide TMGMV U2 in or on all

commodities of crop groups 17 (grass forage, fodder, and hay group) and 18 (nongrass animal feeds group) and opened a 30

- day comment period. The EPA received no comments on this publication.

IV. RISK ASSESSMENT

In the Federal Register s of October 26, 2007 ( 72 FR 60988 )

, and August 30, 2012 ( 77 FR 52610 )

, the

EPA issued Final Rule s on the data requi rements 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 the EPA ’s 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

as sessment. 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 : upgradeable ”). If a study is rated as “supplemental: upgradeable ,” the EPA 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 t he study is not required by 40

C FR P art 158. Both “ acceptable” and “supplemental ” studies may be used in the risk assessment process as appropriate. An “unacceptable ” rating indicates that new data must be submitted.

For the acu te toxicity data requirements, Toxicity C ategories 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 (see section II .B. of U.S. EPA ( 201 4 d ) )

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Biopesticides Registration Action Document

A. Product Analysis Assessment ( 40 CFR § 158.2120 )

BioProdex fulfilled a ll product analysis data requirements for TMGMV U2 . Refer to Tables 1 and 2 in Appendix A for a summary of the data requirements, including both generic and product

- specific

information.

B. Human Health Assessment ( 40 CFR § 158.2140 )

1. Toxicity

BioProdex fulfilled a ll toxicology data requirements for TMGMV U2

. Acceptable Tier I mammalian toxicology scientific rationale and literature support registration of the TMGMV U2 pesticide product

. The EPA is not requiring Tier II and Tier III studies for TMGMV U2 based on the acceptability of the information provided by BioProdex for Tier I

.

For a summary of the generic toxicology data requirements described in sections IV(B)(1)(a) and IV(B)(1)(b) of this document , as well as additional product

- specific data submitted to supp ort the individual registration , refer to Table 3 in Appendix A

.

a. Acute Toxicity/Pathogenicity

– Tier I

Acute Oral Toxicity/Pathogenicity ( Harmonized Guideline 885.3050; Master Record Identification

Number s (MRID No s .) 467416

- 02, 469187

- 01, 479232

- 01, and 480296

- 01 ) : To fulfill this data requirement, BioProdex submitted supporting scientific rationale and public literature tha t show plant viruses, including TMGMV U2, are found in food ingested daily by humans and animals and that no known adverse effects or deaths have occurred in any species as a result of such dietary exposures. The EPA believes that this information indicates that this virus would not be toxic, infective, or pathogenic through the oral route of exposur e and, as a result, has waived acute oral toxicity/pathogenicity testing

for TMGMV U2.

Acute Pulmonary Toxicity/Pathogenicity ( Harmonized Gui deline 885.3150; MRID No s

. 467416

- 02,

469187

- 01, 479232

- 01, and 480296

- 01 ) : To fulfill this data requirement, BioP rodex submitted supporting scientific rationale and public literature tha t show plant viruses, including TMGMV U2, are ubiquitous in plants and are not known or expected to cause acute pulmonary toxicity, pathogenicity, or infectivity in mammals. Upon cons ideration of all this information, the EPA has waived acute pulmonary toxicity/pathogenicity testing for TMGMV U2.

Acute Injection Toxicity/Pathogenicity (Harmonized Guideline 885.3200) : Not required because

TMGMV U2 is a virus (refer to test note #2 of 4 0 CFR § 158.2140(d) ).

Hypersensitivity Incidents ( Harmonized Guideline 885.3400 ; MRID Nos. 467416

- 02, 469187

- 01,

479232

- 01, and 480296

- 01 ) : BioProdex reported that n o hypersensitivity incidents, including

immediate

- type or delayed

- type reactions of hum ans and domestic animals

, occurred during research, development, or testing of TMGMV U2 . Any future hypersensitivity incidents must be reported to the EPA (refer to test note #3 of 40 CFR § 158.2140(d))

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Cell Culture ( Harmonized Guideline 885.3500 ; MRID No s

. 467416

- 02, 469187

- 01, 479232

- 01, and

480296

- 01 ) : To fulfill this data requirement, BioProdex submitted scientific rationale and supported it with public literature. Tobamoviruses are unable to infect animal cells since the cell surface plays an import ant role in infection of animal cells; during infection, animal viruses interact specifically with receptors on the animal cell surface. Plant viruses, on the other hand, lack recognition for these receptors and only enter plant cells through open wounds o r via cell

- to

- cell transfer through intercellular connections (Fraenkel

- Court et al ., 1988). There is one report in the literature of growing Tobacco

mosaic virus (TMV; a different tobamovirus from the one that is the subject of this action) in cultured,

i mmune

- suppressed, monkey kidney cell lines (Atherton, 1968); however, this was an artificial system and does not indicate that plant viruses can normally replicate in or infect animal cells. Upon consideration of all this information, the EPA has waived ce ll culture testing for TMGMV U2.

b. Acute Toxicology and Subchronic Toxicity/Pathogenicity

– Tier II; Reproductive Fertility Effects, Carcinogenicity, Immunotoxicity, and Infectivity/Pathogenicity Analysis

– Tier III

The EPA is not requiri ng Tier II and Tier III studies for TMGMV U2 based on the acceptability of the information provided by BioProdex for Tier I.

c. Endocrine Disruptors

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

Disru ptor 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 ingr edients) 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 ex empt 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. A second list of

chemicals identi fied for EDSP screening was published on June 14, 2013, 1 and includes some pesticides scheduled for registration revie w and chemicals found in water. TMGMV U2 is not among the group of pesticide active ingredients on the list s to be screened under the EDSP

.

The EPA believes that TMGMV U2 , the active ingredient described in this document, is a substance that

would not likely produce any effect in humans similar to an effect produced by a naturally occurring estrogenic substance. As such, pursuant to Sectio n 408(p)(4), the EPA will determine in t he future whether it can exempt TMGMV U2 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.

1 See http://www.regulations.gov/#!documentDetail;D=EPA

- HQ

- OPPT

- 2009

- 0477

- 0074 for the second list of chemicals.Tobacco mild green mosaic tobamovirus strai n U2 (TMGMV U2) Page 10 of 31

Biopesticides Registration Action Document

For further informati on on the status of the EDSP, the policies and procedures, the list of chemicals, future lists, the test guidelines and the Tier 1 screening battery, please visit our websit e at http://www.epa.gov/endo/

.

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

Section 408(c)(2)(A)(i) of FFDCA allows the EPA to establish an exemption from the requirement for a tolerance (the legal limit for a pesticide chemical residue in or on a food) only if the EPA determ ines that the exemption is “safe.” Section 408(c)(2)(A)(ii) of FFDCA defines “safe” to mean that “there is a reasonable certainty that no harm will result from aggregate exposure to the pesticide chemical residue, including all anticipated dietary exposure s and all other exposures for which there is reliable information.” This includes exposure through drinking water and in residential settings but does not include occupational exposure. Pursuant to FFDCA section 408(b)(2)(B), in establishing or maintaining in effect an exemption from the requirement of a tolerance, the EPA must take into account the factors set forth in FFDCA section 408(b)(2)(C), which require the EPA to give special consideration to exposure of infants and children to the pesticide chemic al residue in establishing a tolerance exemption, and to “ensure that there is a reasonable certainty that no harm will result to infants and children from aggregate exposure to th e pesticide chemical residue . . . . ” Additionally, FFDCA section 408(b)(2)( D) requires that the EPA consider “available information concerning the cumulative eff ects of [a particular pesticide’ s] . . . residues and other substances that have a common mechanism of toxicity.”

The EPA performs a number of analyses to determine the r isks from aggregate exposure to pesticide residues. First, the EPA determines the toxicity of the pesticide . Second, the EPA examines exposure to the pesticide through food, drinking water, and other exposures that occur as a result of pesticide use in

res idential settings.

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

relevant information on TMGMV U2 , and considered its validity, completeness, and reliability, as well as the relationship of this inform ation to human risk. The EPA also considered available information concerning the variability of the sensitivities of major identifiable subgroups of consumers, including

infants and children.

Based on the acute toxicity/pathogenicity information discussed previously and presented in Table 3 in Appendix A , t he data required for a FFDCA risk assessment for TMGMV U2 have been fulfilled.

a. Aggregate Exposure

In examining aggregate exposure, FFDCA section 408 directs the EPA to consider available information concerning exposures from the pesticide residue in food and all other non

- occupational exposures, including drinking water from ground water or surface water and exposure through pesticide use in gardens, lawns, or buildi ngs (residential and other indoor uses).

Food Exposure and Risk Characterization : Virus

- infected food plants have always been a part of the human diet and domestic animal food supply (Beemster and de Bokx, 1987; Dewan and Pearson, 1995;

Jones et al ., 1934 ; McKinney, 1929; Palukaitis, 1991; Provvidenti and Gonsalves, 1984). Most plants may be infected by at least one virus, and components of plant viruses are often found in the produce of crop plants. Even plants that show no disease symptoms are often foun d to be infected with virusesTobacco mild green mosaic tobamovirus strai n U2 (TMGMV U2) Page 11 of 31

Biopesticides Registration Action Document

( Fulton, 1986; Jones et al ., 1934 ). In addition, a common agricultural practice used since the 1920s for protection against viral disease involves intentionally inoculating healthy plants with a mild form of a

virus in order t o prevent infection by a more virulent form ( Fulton, 1986 ). A great deal of information supports the ubiquitous appearance of plant viruses in foods and, to date, there have been no reports of adverse human or animal health effects associated with consumpt ion of plant viruses in food. Furthermore, applications of TMGMV U2, which are limited to pastures and wooded areas that are fenced to contain grazing animals, are not expected to result in increased exposures of the active ingredient to the general popula tion. The only residues anticipated on food with this limited use site are on grass, grass hay, and nongrass animal feeds (e.g., alfalfa and clover). These residues on grass, grass hay, and nongrass animal feeds would be incidental to applications to the t arget plant (tropical soda apple), given that grass and the nongrass animal feeds are not hosts for TMGMV U2 and that TMGMV U2 cannot infect or replicate in grass or in the nongrass animal feeds. In the remote likelihood that

TMGMV U2 is present i n food, s upporting scientific literature indicates that no toxicity, pathogenicity, or infectivity is likely to occur with any food exposure resulting from the use of this microbial herbicide when applied in accordance with label directions and good agricultural pr actice s (see section IV(B)(1)(a)

of this document and Table 3 in Appendix A ).

Drinking Water Exposure and Risk Characterization : TMGMV U2 is not intended for use in drinking water. In the unlikely event that TMGMV U2 reaches water consumed by humans (e.g., through spray drift), for the reasons enumerated previously, the EPA concludes that no toxicity, pathogenicity, or infectivity is likely to occur with any drinking water exposure resulting from the use of this microbial

herb icide when applied in accordance with label directions and good agricultural practices (see section IV(B)(1)(a) of this document and Table 3 in Appendix A ).

Non

- occupational, Residential Risk Characterization : Dermal or inha lation exposure to the g eneral population due to use of TMGMV U2 is not likely to occur, based on the limited use site and application methods. Moreover, the general population, including infants and children, are exposed to plant viruses daily in food wit h no known adverse effects ever being reported. Therefore, the EPA concludes that,

should there be non

- occupational, non

- dietary exposures to TMGMV U2, such exposures would pose no risks to the general population, including infants and children ( see sectio n IV(B)(1)(a) of this document

and Table 3 in Appendix A ).

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

Section 408(b)(2)(D)(v) of FFDCA requires that, when considering whether to establish, modify, or revoke a tolerance, the EPA consider “available information concerning the cumulative effects of [a particular pesticide’s] . . . residues and other substances that have a common mechanism of toxicity.”

The EPA has not found TMGMV U2 to share a common mechanism of toxicity with any other substances, and TMGMV U2 does not appear to produce a toxic metabolite. For the purposes of the tolerance action, there fore, the EPA has assumed that TMGMV U2 does not have a common mechanism of toxicity with ot her substances. Following from this, therefore, the EPA concludes that there are no

cum ulative effects associated with TMGMV U2 that need to be considered. For information regarding
the EPA’s efforts to determine chemicals that have a common mechanism of t oxicity and to evaluate the cumulative effects of such chemicals, see the EPA’s website at http://www.epa.gov/pesticides/cumulative

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c. Determination of Safety for the U.S. Population, Infants an d Children

U.S. Population : For all of the reasons discussed previously, there is reasonable certainty that no harm will result to the U.S. population, including infants and children, from aggregate exposure to residues of

TMGMV U2. This includes all anti cipated dietary exposures and all other exposures for which there is

reliable information.

Infants and Children : FFDCA section 408(b)(2)(C) provides that the EPA shall apply an additional tenfold (10X) margin of safety for infants and children in the case of threshold effects to account for prenatal and postnatal toxicity and the completeness of the database on toxicity and exposure, unless the

EPA determines based on reliable data that a different margin of safety will be safe for infants and

children. Th is additional margin of safety is commonly referred to as the Food Quality Protection Act Safety Factor. In applying this provision, the EPA either retains the default value of 10X or uses a different additional safety factor when reliable data available t o the EPA support the choice of a different

factor. Humans, including infants and children, have been exposed to plant viruses through food, where they are commonly found, with no known or reported adverse effects. As discussed ab ove, the EPA has

concluded that TMGMV U2 is not toxic, pathogenic, or infective to mammals, including infants and children. Because there are no threshold levels of concern to infants, children, and adults when TMGMV U2 is used according to label directions and good agricultural pr actices, the EPA concludes that no

additional margin of safety is necessary to protect infants and children.

3 . Occupational Exposure and Risk Characterization

Worker exposure to TMGMV U2 is not e xpected to pose any undue risk. Regardless, the EPA is

re quiring appropriate protective clothing and precautionary statements on the pesticide product label to mitigate any potential risks to pesticide handlers and applicators due to prolonged and/ or multiple

exposures. Individuals working with TMGMV U2 to treat tropical soda apple are directed to wear a long

- sleeved shirt, long pants, socks, shoes, waterproof gloves, and a dust/mist filtering respirator meeting NIOSH standards of at least N

- 95, R

- 95, or P

- 95. For future TMGMV U2 pesticide products, the EPA may r equire additional protective clothing (e.g., eyewear), other than the standard described above, on a product

- specific basis.

4 . Human Health Risk Characterization

The EPA considered human exposure to TMGMV U2 in light of the standard for registration i n FIFRA

and the re levant safety factors in FFDCA. When the TMGMV U2 pesticide product is applied as a post

emergent herbicide to tropical soda apple in accordance with the EPA’s approved labeling, the EPA has

determined that no unreasonable adverse effects to human health are expected and that there is a reasonable certainty that no harm will result to the U.S. population, including i nf ants and children.

C. Environmental Assessment ( 40 CFR § 158.2150 )

The EPA found that scientific rationale and nontarget plant studies submitted by BioProdex are sufficient to fulfill the nontarget organism data requirements and for risk assessment purp oses. Further testing of nontarget organisms at higher tier levels (i.e., Tiers II, III, and IV) is not required. The EPA performed a n environmental risk assessment based on information provided by BioProdex . Provided that limited use sites and application methods, mitigating label language, and certain manufacturing processTobacco mild green mosaic tobamovirus strai n U2 (TMGMV U2) Page 13 of 31

Biopesticides Registration Action Document

steps are in place, the EPA has determined that u nreasonable adverse effects to nontarget organisms are not expected from the proposed pesticidal uses of TMGMV U2

. Moreover, the EPA mad e a “No Effect”

determination for direct and indirect effects to federally listed

, threatened and endangered species and their designated critical habitats resulting from these same proposed pesticidal uses.

For a summary of the generic data requiremen ts described in sections IV(C)(1) of this document , refer to

Table 4 in Appendix A

.

1. Ecological Exposure and Risk Characterization

The active ingredient in SolviNix ® LC is TMGMV U2, a n aturally occurring plant virus. T MGMV occurs widely in the U.S., primarily in Solanaceae (nightshade family) plants and especially in tree tobacco ( Nicotinia glauca ), some commercial tobacco ( Nicotinia tabacum ), and pepper ( Capsicum spp.)

varieties. This virus is mechanically transmitted and spreads by physical contact; it is not vector

transmitted or seedborne.

SolviNix ® LC is intended for use against a single target weed (tropical soda apple, Solanum viarum ) in

which TMGMV U2 elicits a lethal hypersensitive response that produces wiltin g and death of the entire tropical soda apple plant, including the roots. Use sites are restricted to pastures and wooded areas that

are fen ced to contain grazing animals, and applications are to be made only by backpack sprayer or a dedicated wiper system

. Use according to the proposed label is expected to minimize exposure and therefore risk to nontarget organisms.

a. Birds (Harmonized Gu idelines 885.4050 and 885.4100) (MRID Nos. 467416

- 02 and

478955

- 03 )

TMGMV U2 is a plant virus and is not known to i nfect or produce toxicity in birds. Due to the application methods and low use rates, direct avian ex posure is expected to be extremely limited.

SolviNix ® LC is to be applied prior to tropical soda apple flowering, which would prevent any exposure

via frui t. BioProdex ’s search of the public literature found no in formation on adverse effects of

TMGMV or any other tobamovirus on avian species. Furthermore, TMGMV was not found on any list

of avian pathogens or in a volume on avian viruses (Ritchie, 1995)

. Chic kens can be immunized with purified tobamoviruses for the production of antisera without developing infection or toxic reactions. The optimal temperature for replication of tobamoviruses in plants generally ranges from 24oC to 35oC and, in tests conducted b y BioProdex , typical hypersensitive death of tropical soda apple did not develop in plants maintained at ˃32oC (normal avian body temperature is approximately 40oC to 42oC).

b. Wild Mammals (Harmonized Guideline 885.4150) (MRID Nos. 467416

- 02 and

478955

- 03)

TMGMV U2 is a plant virus and is not known to infect or produce toxicity in mammals. Due to the

app lication methods and low use rates, direct exposure to wild mammals is expected to be extremely

limited. Mammals are not known to eat the tropical soda apple foliage, which is covered with sharp thorns, and the pre

- flowering application of SolviNix ® LC kil ls the plant p rior to formation of the fruit.

BioProdex ’s search of the public literature found no in formation on adverse effects of TMGMV or any other to bamovirus on mammalian species , and TMGMV was not found on any list of mammalian pathogens or in a sta ndard reference on plant viruses (Hull, 2002 ). Wild mammals, particularlyTobacco mild green mosaic tobamovirus strai n U2 (TMGMV U2) Page 14 of 31

Biopesticides Registration Action Document

herbivores, are naturally exposed to plant viruses such as TMGMV a nd other tobamovirus es without any adverse effects. Rabbits, mice, goats, and horses can be immunized with purified tobamoviruses for antiserum production without causing toxic reactions or infection . The optimal temperature for replication of tobamoviruses in plants generally ranges from 24oC to 35oC and, in tests conducted by

BioProdex , typical hypersensitive death of tropical soda apple did not develop in plants m aintained at ˃32oC (normal mammalian body temperature is approximately 37 oC).

c. Freshwater Fish (Harmonized Guideline 885.4200) (MRID Nos. 467416

- 02 and

478955

- 03)

TMGMV U2 is a plant virus and is not kn own to infect or produce toxicity in fish. SolviNix ® LC will not be applied in aquatic sites, since tropical soda apple does not grow or survive in wetlands. The low application rates and methods of application will limit the amount of pesticide product re aching nontarget sites, making direct exposure to freshwater fish extremely limited. No aquatic plant has been found to be susceptible to TMGMV

. BioProdex ’s search of the public literature found no in formation on

adverse effects of TMGMV or any other tobam ovirus on fish. Furthermore , TMGMV was not found on any list of fish pathogens, and no mention of toxicity to fish was found in either of two standard reference books (Wolf, 1988; Woo and Bruno, 1999). There are no reports of iso lation of TMGMV from

water from streams, ponds, or lakes. BioProdex cond ucted a study in which purified TMGMV U2 was diluted with sterile deionized water, tap water, or lake water stored at room temperature. Nicotiana sylvestris plants (a local lesion host to TMGMV U2) were inoculat ed with the stored solutions after 19 days and monthly thereafter for one year. Local lesions were counted 3 to 5 days after each inoculation. At the end of one year, the number of lesions/inoc ulation ranged from 0 to 38 for TMGMV U2 in tap water, 1 to 168 for TMGMV U2 in deionized water, and 0 to 2 for TMGMV U2 in lake water.

BioProdex concluded that TMGMV U2 is unlikely to persist and remain infective in natural waters where microbial activity and other factors contribute to its degradation.

d. Freshwa ter Invertebrates (Harmonized Guideline 885.4240) (MRID Nos. 467416

- 02 and

478955

- 03)

TMGMV U2 is a plant virus and is not known to infect or produce toxicity in freshwater aquatic

invertebrates . SolviNix ® LC will not be applied in aquatic sites, sinc e tropical soda apple does not grow or survive in wetlands. The low application rates and methods of application will limit the amount of pesticide product reaching nontarget sites, making direct exposure to fr eshwater aquatic invertebrates extremely limit ed. No aquatic plant has been found to be susceptible to TMGMV. BioProdex ’s search of the public literature found no information on adverse effects of TMGMV or any other tobamovirus on freshwater aquatic invertebrates . Furthermore, TMGMV w as not found on a ny list of invertebrate pathogens. There are no reports of isolation of TMGMV from water from streams, ponds, or lakes , and

BioProdex ’s testing (see freshwater fish, directly above) indicates that TMGMV U2 is unlikely to persist

and remain infective in nat ural waters

.

e. Estuarine/Marine Fish and Invertebrates (Harmonized Guideline 885.4280) (MRID No s. 467416

- 02 and 478955

- 03 )

TMGMV U2 is a plant virus and is not known to infect or produce toxicity in estuarine/marine fish and

invertebrates. Solvi Nix ® LC will not be applied in aquatic sites, since tropical soda apple does not grow or survive in wetlands. The low application rates and methods of application will limit the amount ofTobacco mild green mosaic tobamovirus strai n U2 (TMGMV U2) Page 15 of 31

Biopesticides Registration Action Document

pesticide product reaching nontarget sites, making direct exposure t o estuarine/marine fish and invertebrates extremely limited. No aquatic plant has been found to be susceptible to TMGMV.

BioProdex ’s search of the public literature found no information on adverse effects of TMGMV or any other t obamovirus on estuarine/mari ne fish and invertebrates. Furthermore, TMGMV was not found on any list of fish or invertebrate pathogens. There are no reports of isolation of TMGMV from water from
streams, po nds, or lakes, and BioProdex ’s testing (see freshwater fish, directly above) in dicates that

TMGMV U2 is unlikely to persist and remain infective in natural waters.

f. Nontarget Plants (Harmonized Guideline 885.4300) (MRID No s

. 467416

- 04, 478955

- 04,

491377

- 01, and 492315

- 01 )

In a nontarget plant study, over 400 plant spe cies and varieties representing 61 families were tested to

de termine their susceptibility to TMGMV U2. Seedlings w ere hand

- inoculated with either TMGMV U2 or a sodium phosphate buffer control and observed over time for hypersensitive lesions on the inocula ted leaves and for systemic symptoms on new growth. Additionally, new growth was tested for TMGMV U2 using enzyme

- linked immunosorbent assay (ELISA). Of the plants tested, 57.70% were immune (no symptoms or infection), 11.03% were resistant (hypersensitive local lesions), 8.74% were asymptomatic but positive in ELISA, 14.94% were susceptible (mild, systemic chlorosis), and 7.59% were highly susceptible (severe damage or death). All the highly susceptible plants were members of the Solanaceae family. Only 2% of plants (seven species representing five unrelated families) outside the Solanaceae family were susceptible to TMGMV U2.

Since TMGMV U2 and other plant viruses can infect certain nontarget plants, BioProdex has agreed to

limit SolviNix ® LC’s use sites to pastures and wooded areas that are fe nced to contain grazing animals

and the methods of application to backpack sprayers and dedicated wiper systems to reduce exposure. Further, the SolviNix ® LC label bears (and must continue to bear) the following mit igating language :

(1) “ The virus in SolviNix ® LC has no natural means of spread other than by physical and abrasive contact between infected and healthy plants. A remote possibility is through workers (via contaminated clothes, shoes, hands) and contaminated application equipment. To prevent unwanted virus spread, avoid contact with skin and clothing. Wash skin with soap and water and clothes with laundry detergent. See below instructions for washing application equipment. Thorough washing will prevent spread. ”

(2) “Stop any pruning or mechanical activities in treated areas, such as mowing or tillage, 48 hours before and after application of SolviNix ® LC.”

(3) “May infect some non

- woody flowering plants, including Solanaceous crop plants (eggplant, pepper, tobacco, a nd tomatillo).”

(4) “Apply only when tropical soda apple plants are actively growing.”

(5) “Do not apply within 300 feet of greenhouse and grafting operations.”

(6) “Spray directly at the target plant

– Avoid any crops and nontarget plants.”

(7) “Must not re

- use the s pray equipment for other uses without thorough cleaning as specified.”Tobacco mild green mosaic tobamovirus strai n U2 (TMGMV U2) Page 16 of 31

Biopesticides Registration Action Document

(8) “Do not apply this product through any type of irrigation system.”

(9) “For any surviving treated tropical soda apple ( Solanum viarum ) plants, re

- apply according to label

instructions. If there are still surviving plants, consider other mechanical or chemical means of control. Consult local extension recommendations and contact the registrant. The registrant will notify EPA under FIFRA 6(a)(2) for assessment of pesticide resistance in trop ical soda apple. ”

Fin ally, BioProdex is implementing particular manufacturing process steps to ensure tha t no other plant

viruses are introduced into SolviNix ® LC.

g

. Nontarget Insects (Harmonized Guideline 885.4340) and Honey Bees (Harmonized Guideline 885.4380) (MRID No s. 467416

- 02 and 478955

- 03 )

TMGMV U2 is a plant virus and is not known to infect or produce toxicity in insects. Nontarget insects, including honey bees, are naturally exposed to plant viruses, includ ing tobamoviruses. BioPro dex ’s search of the public literature found no information on adverse effects of TMGMV or any other tobamovirus on insects or honey bees. TMGMV was not found in any list of insect or honey bee pathogens or in a standard reference on plant virology (Hull, 2 002). A study cited by BioProdex report ed

that bumble bees transmitted TMV from tomato to tomato under greenhouse conditions, but there was no mention of toxicity to the insect. Due to the application methods and low us e rates, direct exposure of

TMGMV U2 to nontarget insects and honey bees i s expected to be extremely limited.

h

. Conclusions for Nontarget Organisms

Based on the expected lack of exposure and data demonstrating no effects, as discussed above, unreasonable adverse effects to nontarget organisms are not anticipated as a result of the proposed use patterns of the SolviNix ® LC pesticide product , containing TMGMV U2 , as labeled.

2. Environmental Fate Data

As t he rationale , literature, and data provided are sufficient to fulfill the Tie r I nontarget organism data requirements and allow for nontarget organism risk asse ssment for TMGMV U2 , the EPA is not requiring testing at higher tier levels (i.e., Tiers II, III, and IV)

.

3

. Threatened and Endangered Species Assessment

Since the EPA has determined that no effects are anticipated to nontarget species from TMGMV U2 in

SolviNix ® LC as a result of the proposed labeled use s, effects to federally listed threatened and endangered species and their designated critical habitats are also not e xpected. 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 pesticidal use s of TMGMV U2 , as labeled.

V. ENVIRONMEN TAL JUSTICE

The EPA seeks to achieve environmental justice

— the fair treatment and meaningful involvement of allTobacco mild green mosaic tobamovirus strai n U2 (TMGMV U2) Page 17 of 31

Biopesticides Registration Action Document

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 comm ercial 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 d ecision

- 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, the EPA seeks 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 TMGMV U2 , compared to the general population. Please comment if you are aware of any subpopulations t hat may have atypical, unusually high exposure compared to the general population.

For additional information regarding environmental justice issues, please visit the EPA’s web site at http://www.epa.gov/compliance/environmentaljustice/index.html

.

V I

. 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 s uch 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 environm ent; 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 TMGMV U2 is well known to ha ve herbicidal capabili ties

. The EPA has no knowledge that would contradict the claims made on the SolviNix ® LC label, and this pesticide product is not expected to cause unreasonable adverse effects on the

environment when used according to its label instr uctions. Criterion 2 is satisfied by the product label, as well as the data and information presented in this document. It is believed that the TMGMV U2 pesticide product will not cause any unreasonable adverse effects on the environment, and SolviNix ® LC

is likel y to control tropical soda apple, satisfying criterion 3. Criterion 4 is satisfied in that the TMGMV

U2 pesticide product is not expected to cause unreasonable adverse effects when used according to label instructions. Therefore, the EPA believes S olviNix ® LC

, containing TMGMV U2 as a new active ingredient, is eligible for registration under FIFRA section 3(c)(5) for the labeled uses.Tobacco mild green mosaic tobamovirus strai n U2 (TMGMV U2) Page 18 of 31

Biopesticides Registration Action Document

V I I

. ACTIONS REQUIRED OF THE REGISTRANT

A. Final Printed Labeling

Bef ore releasing the pesticide product cont aining TMGMV U2 for shipment, BioProdex is required to provide appropriate final printed labeling to the EPA

.

B. Terms of Registration

No additional data are being required as terms of the SolviNix ® LC registration.

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 CFR § 158.2140(d).Tobacco mild green mosaic tobamovirus strai n U2 (TMGMV U2) Page 19 of 31

Biopesticides Registration Action Document

VIII. GLOSSARY OF ACRONYMS AND ABBREVIATIONS

ATCC American Type Culture Collection

BR AD Biopesticides Registration Action Document

CFR Code of Federal Regulations

EDSP Endocrine Disruptor Screening Program

ELISA enzyme

- linked immunosorbent assay

EP end

- use pesticide product

EPA United States Envi ronmental Protection Agency

FFDCA Federal F ood, Drug, and Cosmetic Act

FIFRA Federal Insecticide, Fungicide, and Rodenticide Act

FR Federal Register

MOE margin of exposure

MRID No. Master Record Identification Number

NIOSH National Institute for Occupational Safety and Health

OPP Office of Pestici de Programs

PC Code Pesticide Chemical Code

PP Pesticide Petition

TGAI technical grade of the active ingredient

TMGMV Tobacco mild green mosaic tobamovirus

TMGMV U2 Tobacco mild green mosaic tobamovirus strain U2

TMV Tobacco mosaic virus

UF

– IFAS Univer sity of Florida

– Institute of Food and Agricultural Sciences

U.S. United States

USDA United States Department of AgricultureTobacco mild green mosaic tobamovirus strai n U2 (TMGMV U2) Page 20 of 31

Biopesticides Registration Action Document

IX. BIBLIOGRAPHY

A. Studies Submitted to Support the Tobacco mild green mosaic tobamovirus strain U 2 Pesticide

Product Registration

MRID No. Study Information

467416

- 02 Braverman M. 2005. Group C

– Toxicology Data Waiver Request s: Solvinix. Project

Number: 036 4B. Unpublished study prepared by Interregional Research Project No. 4, 18

pages.

467416

- 03 Braverman M. 2005. Group D

– Nontarget Data Waiver Request: Solvinix. Project

Number: 03754B. Unpublished study prepared by Interregional Research Project No. 4, 69

pages.

467416

- 04 Braverman M. 2005. Group D

– Nontarget Plant Studies: Solvinix. Proj ect Number:

0374B. Unpublished study prepared by Interregional Research Project No. 4, 29 pages.

467588

- 01 Braverman M. 2005

. Group A

– Product Analysis Test Guidelines: Solvinix ( Amended

Report ) . Unpublished study pr epared by Interregional Research Proj ect No. 4, 24 pages.

469187

- 01 Braverman M. 2006. Group C

– Toxicology Data Waiver Requests: Solvinix. Project

Number: 0364B. Unpublished study prepared by Interregional Research Project No. 4, 22

pages.

478955

- 01 Braverman M . 20 09

. Solvinix: Group A

– Product Analysis Test Guidelines

(Amendment)

. Project Number: IR/4/PR/NO/0364B. Unpublished study prepared by Interregional Research Project No. 4, 33 pages.

478955

- 03 Braverman M. 2009. Solvinix: Group D

– Nontarget Data Waiver Request (Amendment).

Pro ject Number: IR/4/PR/NO/0364B. Unpublished study prepared by Interregional Research Project No. 4, 102 pages.

478955

- 04 Braverman M. 2009. Solvinix: Group D

– Nontarget Plant Studies (Amendment). Project

Number: IR/4/PR/NO/0364B. Unpublished study prepar ed by Interregional Research Project No. 4, 27 pages.

479232

- 01 Braverman M. 2009. Group C

– Toxicology Data Waiver Requests: Solvinix. Project

Number: 0364B. Unpublished study prepared by Interregional Research Project No. 4, 18

pages.

480296

- 01 Brave rman M. 2010. Group C

– Toxicology Data Waiver Requests: Solvinix

(Amendment). Project Number: 0364B. Unpublished study prepared by Interregional Research Project No. 4, 18 pages.Tobacco mild green mosaic tobamovirus strai n U2 (TMGMV U2) Page 21 of 31

Biopesticides Registration Action Document

MRID No. Study Information

491377

- 01 Charudattan R, Braverman M. 2013. Solvinix: Tobacco Mild Green M osaic Virus. Project

Number: 0364B. Unpublished study prepared by BioProdex, Inc., and Interregional Research Project No. 4, 451 pages.

492315

- 01 Braverman M. 2013, Supplement Information. Amendment #1 to MRID No. 491377

- 01:

Tobacco Mild Green Mosaic Vir us (Administrative Volume). Project Number: IR/4/PR/NO/0364B. Unpublished study prepared by Interregional Research Project No. 4, 110 pages.

494002

- 01 Charudattan R, Braverman M. 2014

. Solvinix, Tobacco Mild Green Mosaic Tobamovirus:

TMGMV U2. Project Nu mber: 0364B. Unpublished study prepared by BioProdex , Inc., and Interregional Research Project No. 4, 90 pages.

494291

- 01 Charudattan R, Braverman M. 2014

. Solvinix, Tobacco Mild Green Mosaic Tobamovirus:

TMGMV U2

– Supplement to MRID Nos. 491377

- 01, 491 377

- 02, 492315

- 01, and

494002

- 01. Project Number: 0364B . Unpublished study prepared by BioProdex, Inc., and Interregional Research Project No. 4, 7 pages.

B. Environmental Protection Agency Risk Assessment Memoranda

U.S. EPA. 201 4 a. SolviNix TM LC Contai ning Tobacco Mild Green Mosaic Virus, ATCC PV

- 1200. Memorandum from J.V. Gagliardi, Ph.D. through J.L. Kough, Ph.D. to J. Kausch, dated March 13,

2014.

U.S. EPA. 2014b. SolviNix TM LC Containing Tobacco Mild Green Mosaic Virus, ATCC PV

- 1200.

Memorandu m from J.V. Gagliardi, Ph.D. through S. Borges to J. Kausch, dated July 2,

2014.

U.S. EPA. 2014c. SolviNix TM LC Containing Tobacco Mild Green Mosaic Virus, ATCC PV

- 1200. Memorandum from J.V. Gagliardi, Ph.D. through J.L. Kough, Ph .D. to J. Kausch, date d July 3,

2014.

U.S. EPA. 2010

. SolviNix TM LC Containing Tobacco Mild Green Mosaic Virus, ATCC PV

- 1200

. Memorandum from J.V. Gagliardi, Ph.D. through J.L. Kough, Ph.D. to J. Kausch

, dated N ovember 9, 2010

.

U.S. EPA. 2006 a. Solvinix Containing TM GMV (Tobacco Mild Green Mosaic Tobamovirus)

. Memorandum from J

. V. Gagliardi, Ph.D . through J.L. Kough, Ph.D. to R

. Edelstein

, Ph.D., dated May 23, 2006

.

U.S. EPA. 2006b. Review of Environmental Effects Data and Justifications for Data Waivers Submitte d in Support of BioProdex, Inc.’s Solvinex [sic] Bioherbicide, Which Contains the Active Ingredient TMGMV. Memorandum from T. Milofsky through Z. Vaituzis, Ph.D. to R. Edelstein, Ph.D., dated June 21, 2006.Tobacco mild green mosaic tobamovirus strai n U2 (TMGMV U2) Page 22 of 31

Biopesticides Registration Action Document

U.S. EPA. 20 06c. Replacement Data Waive r Reviews for Solvinix Containing TMGMV (Tobacco M ild Green Mosaic Tobamovirus) . Memorandum from J.V. Gagliardi , Ph.D. throug h J.L. Kough, Ph.D. to R. Edelstein, Ph.D.

, dated September 7, 2006

.

C. Other References

Atherton JG. 1968. Formation of to bacco mosaic virus in an animal cell culture. Archiv fur die gesamte Virusforschung 24:406

– 418.

Baker C, Adkins S. 2004. Peppers, Tomatoes, and Tobamoviruses. Plant Pathology Circular No. 400, May/June 2000, Division of Plant Industry, Florida Departme nt of Agriculture & Consumer Services, Gainesville, Florida 32608.

Baker CA, Zettler FW. 1988. Viruses infecting wild and cultivated species of the Commelinaceae. Plant

Disease 72:513

– 518.

Bald JG, Goodchild DJ. 1960. Tobacco mosaic virus in Nicotiana glauca

. Phytopathology 50:497

– 499.

Beemster ABR, de Bokx JA. 1987. Survey of properties and symptoms. In: de Bokx JA, van der Want JPH. Viruses of Potatoes and Seed Potato Production. Wageningen: Pudoc, pgs. 84

– 93.

Bodaghi S, Mathews DM, Dodds JA. 2 004. Natural incidence of mixed infections and experimental cross protection between two genotypes of Tobacco mild green mosaic virus. Phytopathology

94:1337

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Bodaghi S, Yassi MNA, Dodds JA. 2000. Heterogeneity in the 3’

- terminal untranslated reg ion of tobacco mild green mosaic tobamoviruses from Nicotiana glauca resulting in variants with three or six pseudoknots. Journal of General Virology 81:577

– 586.

Braverman M. 2010. Administrative Materials for Solvinix from M. Braverman of Interregion al Research Project No. 4 (Volume 1 of 2) (Dated March 10, 2010).

Brunt AA, Crabtree K, Dallwitz MJ, Gibbs AJ, Watson L, Zurcher EJ (eds.). 1996. Plant Viruses Online: Descriptions and Lists from the VIDE Database (Tobacco mild green mosaic tobamovirus ).

Version: 20 th August 1996. Available from http://pvo.bio

- mirror.cn/descr801.htm

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Dewan C, Pearson MN. 1995. Natural field infection of garlic by garlic yellow streak virus in the

Pukekohe area of New Zealand and associated problems with the introduction of new garlic

cultivars. New Zealand Journal of Crop and Horticultural Science 23:97

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Font MI, Córdoba

- Sellés MC, Cebrián MC, Herrera

- Vásquez JA, Alfaro

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annuum in Tunisia. Plant Disease 93:761.

Fraenkel

- Court H, Kimball PC, Levy JA. 1988. Virology, 2 nd edition. Prentice Hall, Englewood Cliffs, NJ (Virus cellular receptor s and cell membrane changes, pgs. 299

– 300).Tobacco mild green mosaic tobamovirus strai n U2 (TMGMV U2) Page 23 of 31

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Fulton R. 1986. Practices and precautions in the use of cross protection for plant virus disease control. Annual Review of Phytopathology 24:67

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Green SK, Kim JS. 1991. Characteristics and control of virus es infecting peppers: A literature review. Tech. Bull. No. 18, Asian Vegetable Research and Development Center, P.O. Box 205, Taipei.

Hull R. 2002. Matthews ’ Plant Virology, 4 th Edition. Academic Press, San Diego, California.

ICTVdB Management. 2006

. 00.071.0.01.011. Tobacco mild green mosaic virus. In: ICTVdB

– The

Universal Virus Dat abase, version 4. Büchen

- Osmond C. (Ed .), Columbia University , New York.

Available from http://ictv db.bio

- mirror.cn/ICTVdB/00.071.0.01.011.htm

.

Johnson J. 1947. Virus attenuation and the separation of strains by specific hosts. Phytopathology

37:822

– 837.

Jones L, Anderson E, Burnett G. 1934. The latent virus of potatoes. Journal of Phytopathology 7 :93

– 115.

Knight CA, Silva DM, Dahl D, Tsugita A. 1962. Two distinctive strains of tobacco mosaic virus. Virology 16:236

– 243.

Letschert B, Günter A, Lesemann D

- E, Willingmann P, Heinze C. 2002. Detection and differentiation of serologically cross

- re acting tobamoviruses of economical importance by RTPCR and RT

- PCR

RFLP. Journal of Virological Methods 106:1

– 10.

McKinney HH. 1952. Two strains of tobacco mosaic virus, one of which is seed

- borne in an etch

immune pungent pepper. Plant Disease Reporte r 36:184

– 187.

McKinney HH. 1929. Mosaic diseases in the Canary Islands, West Africa, and Gibraltar. Journal of Agricultural Research 39:557

– 578.

Miller PM, Thornberry HH. 1958. A new viral disease of tomato and pepper. Phytopathology 48:665

– 669.

Pal ukaitis P. 1991. Virus

- mediated genetic transfer in plants. In: Levin M, Strauss H. Risk Assessment in Genetic Engineering. New York: McGraw

- Hill, pgs. 140

– 162.

Provvidenti R, Gonsalves D. 1984. Occurrence of zucchini yellow mosaic virus in cucurbits fr om Connecticut, New York, Florida, and California. Plant Disease 68:443

– 446.

Ritchie BW. 1995. Avian Viruses

– Function and Control. Wingers Publishing, Inc., Lake Worth, Florida, pgs. 2, 468, 469, and 484

– 515.

Siegel A, Wildman SG. 1954. Some natura l relationships between strains of tobacco mosaic virus. Phytopathology 44:277

– 282.Tobacco mild green mosaic tobamovirus strai n U2 (TMGMV U2) Page 24 of 31

Biopesticides Registration Action Document

UF

– IFAS. 2008. Solanum viarum Dunal. In: Identification and Biology of Nonnative Plants in Florida’s Natural Areas (2 nd Edition, Pub. SP 257). Available from http://plants.ifas.ufl.edu/misc/pdfs/SP257/Solanum_viarum(SP257

- 125).pdf

.

USDA. 2014. Plants Species Profiles

– Tropical Soda Apple (Last Modified July 1, 2014). Availa ble

from http://www.invasivespeciesinfo.gov/plants/tropicalsoda.shtml

.

U.S. EPA. 2014d . Chapter 7 of the Label Review Manual (Precautionary Statements) (Revised July 2014) . Av ailable from http://www2.epa.gov/sites/production/files/2014

- 07/documents/chapter7_revised_final_0714.pdf

.

Weber PVV. 1951. Inheritance of a necro tic

- lesion reaction to a mild strain of tobacco mosaic virus. Phytopathology 41:593

– 609.

Wetter C. 1989. Tobacco mild green mosaic virus. In: AAB Description of Plant Viruses No. 351.

Available from http://www.dpvweb.net/dpv/showdpv.php?dpvno=351

.

Wetter C. 1984. Serological identification of four tobamoviruses infecti ng pepper. Plant Disease

68:597

– 599.

Wolf K. 1988. Fish Viruses and Fish Viral Diseases. Comstock Publishing Association , Cornell University Press, Ithaca, New York.

Woo PTK, Bruno DW. 1999. Fish Diseases and Disorders, Volume 3, Viral, Bacterial and Fungal Infections. CABI Publishing, Oxon, United Kingdom.

Zettler FW, Nagel J. 1983. Infection of cultivated gesneriads by two strains of tobacco mosaic virus.

Plant Disease 67:1123

– 1125.Tobacco mild green mosaic tobamovirus strai n U2 (TMGMV U2) Page 25 of 31

Biopesticides Registration Action Document

A PPENDIX A. MICROBIAL PESTICIDES DATA REQUIREMENTS

(40 CFR PART 158

– SUBPART V )

TABLE 1. Product Analysis Data Requirements for the Technical Grade of the Active Ingredient (TGAI)
, TMGMV U2
, and Its End

use Pesticide Product (E P)
, SolviNix ® LC (40 CFR § 158.2 120 )

Harmonized

Guideline

Number Data Requirement Results MRID No. TGAI E P

885.1100 Product Identity N/A Submitted data fulfill the requirement for pr oduct identity. SolviNix ® LC

contains 2.9 % by weight TMGMV U 2 (contains a minimum of 1.25 x 10 1 6

virions per fluid ounce ). 467588

- 01

478955

- 01

885.1200 Manufacturing Process Submitted data fulfill the requirement for manufacturing process. 467588

- 01

47 8955

- 01

491377

- 01

494002

- 01

494291

- 01

885.1250 Deposition of a Sample in

a Nationally Recognized

Culture Collection TMGMV U2 is on deposit with the American Type Culture Collection in Manassas, Virginia

under Accession Number PV

1200. N/A 467588

- 01

478955

- 01

885.1300 Discussion of Formation

of Unintentional Ingredients Submitted data fulfill the requirement for discussion of formation of unintentional ingredients. 467588

- 01

478955

- 01

885.1400 Analysis of Samples Submitted data fulfill the requirement for analysis of samples. 467588

- 01

478955

- 01

491377

- 01

492315

- 01

885.1500 Certification of Limits N/A Limits listed on the confidential

statement of formula are adequate/acceptable. 478955

- 01

491377

- 01Tobacco mild green mosaic tobamovirus strai n U2 (TMGMV U2) Page 26 of 31

Biopesticides Registration Action Document

TABLE 2 . Physical and Chemical Charac teristics for the Technical Grade of the Active Ingredient (TGAI), TMGMV U2 , and Its End

u se Pesticide Product ( E P), SolviNix ® LC (40 C F R § 158.2120)

Harmonized

Guideline Number Data Requirement Results MRID No. or Other Reference TGAI E P

830.6302 Co lor

See EP column. 1 Light green (birefringent) Braverman (2010) 830.6303 Physical State Liquid

830.6304 Odor No odor

830.6313 Stability to Normal and Elevated

Temperatures, Metals, and Metal

Ions Stable for greater than three

years , but the SolviNi x ® LC

label will bear a date mandating that the product be sold or used one year from the day of the herbicide’s assay and batch

certification. Braverman (2010)

478955

- 01

491377

- 01

492315

- 01

494002

- 01 830.6317 Storage Stability

830.6319 Miscibility N/A Water soluble

Braverman (2010) 830.6320 Corrosion Characteristics Not corrosive

830.7000 pH See EP column. 1 5.0

830.7100 Viscosity N/A 1 poise

830.7300 Density/Relative Density/Bulk

Density (Specific Gravity) See EP column. 1 8.35 pounds/gallon

1 According to 40 CFR § 158.2120, these data are required for t he TGAI

. BioProdex provided data on the EP instead of the TGAI. The EPA

considers these EP data to be acceptable because no separate TGAI is produced in this case and because the TGAI is equivalent to the EP . See 40 CFR § 158.30 and 40 CFR § 158.120(b)(5)(iii)

.Tobacco mild green mosaic tobamovirus strai n U2 (TMGMV U2) Page 27 of 31

Biopesticides Registration Action Document

T ABLE 3

. Toxicology Data Requirements for the Technical Grade of the Active Ingredient (TGAI), TMGMV U2 , and Its End

- u se Pesticide Product ( E P), SolviNix ® LC (4 0 CFR § 158.2140)

Harmonized

Guideline

N umber Data Requirement Results MRID N o. TGAI E P

Tier I

885.3050 Acute Oral

Toxicity/Pathogenicity Testing w aived based on BioProdex ’s scientific rationale, which included

references to scientific literature an d showed that plant viruses, including

TMGMV U2, are found in food ingested daily by humans and animals

with no known adverse effects or

deaths as a result of such dietary

exposures

. Classification: Acceptable

N/A 467416

- 02

469187

- 01

479232

- 01

480296

- 01

885.3150 Acute Pulmonary

Toxicity/Pathogenicity Testing w aived based on BioProdex ’s scientific rationale, which included

references to scientific literature and showed that plant viruses, including

TMGMV U2, are ubiquitous in plants

and are not known or e xpected to cause acute pulmonary toxicity, pathogenicity, or infectivity in

mammals. Classification: Acceptable

885.3200 Acute Injection

Toxicity/Pathogenicity Not required because TMGMV U2 is a virus (refer to test note #2 of 40 CFR § 158.2140(d)). N/A

885.3400 Hypersensitivity

Incidents BioProdex 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/E P. Any future hypersensitivity incidents must be reported to the EPA (refer to test note #3 of 40 CFR § 158.2140(d)).

467416

- 02

469187

- 01

479232

- 01

480296

- 01 885.3500 Cell Culture Testing w aived based on BioProdex ’s scientific rationale, which included

references to s cientific literature and showed that plant viruses, including

TMGMV U2, enter plant cells only through open wounds or by cell

- to

- cell

transfer and are unable to infect animal cells because they lack the cell surface binding site recognition common to

anima l viruses. Classification: Acceptable N/ATobacco mild green mosaic tobamovirus strai n U2 (TMGMV U2) Page 28 of 31

Biopesticides Registration Action Document

T ABLE 3

. Toxicology Data Requirements for the Technical Grade of the Active Ingredient (TGAI), TMGMV U2 , and Its End

- u se Pesticide Product ( E P), SolviNix ® LC (4 0 CFR § 158.2140)

Harmonized

Guideline

N umber Data Requirement Results MRID N o. TGAI E P

870.1100 Acute Oral Toxicity

N/A Testing waived based on BioProdex ’s scientific rationale, which included

references to scientific literature and showed that plant viruses, including

TMGMV U2, are found in food ingested daily by humans and animals with no

known adverse effects or deaths as a result of such dietary exposures , and

because this formulation contains an inert ingredient that is not expected to be of toxicological concern (refer to test note

#5 of 40 CFR § 158.2140(d)). Classification: Acceptable

TOXICITY CATEGORY III

480296

- 01 870.1200 Acute Dermal

Toxicity Testing waived based on BioProdex ’s scientific rationale, which included

references to scientific literature and showed that plant viruses, incl uding

TMGMV U2, are ubiquitous in plants and are not known or expected to cause acute dermal toxicity in mammals, and

because this formulation contains an inert ingredient that is not expected to be of toxicological concern (refer to test note

#5 of 40 CFR § 158.2140(d))

. Classification: Acceptable

TOXICITY CATEGORY III

870.1300 Acute Inhalation

Toxicity Testing waived based on BioProdex ’s scientific rationale, which included

references to scientific literature and showed that plant viruses, including

TM GMV U 2, are ubiquitous in plants and are not known or expected to cause acute inhalation toxicity in mammals, and

because this formulation contains an inert ingredient that is not expected to be of toxicological concern (refer to test note #5 of 40 CFR § 1 58.2140(d))

. Classification: Acceptable

TOXICITY CATEGORY IIITobacco mild green mosaic tobamovirus strai n U2 (TMGMV U2) Page 29 of 31

Biopesticides Registration Action Document

T ABLE 3

. Toxicology Data Requirements for the Technical Grade of the Active Ingredient (TGAI), TMGMV U2 , and Its End

- u se Pesticide Product ( E P), SolviNix ® LC (4 0 CFR § 158.2140)

Harmonized

Guideline

N umber Data Requirement Results MRID N o. TGAI E P

870.2400 Acute Eye Irritation

N/A Testing waived based on BioProdex ’s scientific rationale, which included

references to scientific literature and showed that plant viruses, including

TMGMV U2 , are ubiquitous in plants and are not known or expected to cause acute eye irritation in mammals, and

because this formulation contains an inert ingredient that is not expected to be of toxicological concern (refer to test note #5 of 40 CFR § 158.2140(d))

. Classification: Acceptable

TOXICITY CATEGORY III 469187

- 01

479232

- 01

480296

- 01

870.2500 Primary Dermal

Irritation Testing waived based on BioProdex ’s scientific rationale, which included

references to scientific literature and showed that plant viruses , including

TMGMV U2, are ubiquitous in plants and are not known or expected to cause primary dermal irritation in mammals,

and because this formulation contains an inert ingredient that is not expected to be of toxicological concern (refer to test note

#5 of 40 CFR § 158.2140(d))

. Classification: Acceptable

TOXICITY CATEGORY III

Tiers II and III Not required for TMGMV U2 based on the acceptability of the sc ientific rationale and lite rature provided by BioProdex for Tier I

.Tobacco mild green mosaic tobamovirus strai n U2 (TMGMV U2) Page 30 of 31

Biopesticides Registration Action Document

TABLE 4 . No ntarget Organism Toxicity and Environmental Fate Data Requirements for the Technical Grade of the Active Ingredient (TGAI), TMGMV U2 (40 CFR § 158.2150)

Harmonized Guideline

Number Data Requirement Results MRID No.

Tier I

885.4050 Avian Oral Toxicity

Te sting waived based on BioProdex ’s scientific rationale, which included references to scientific literature and showed that plant viruses, including

TMGMV U2, are not known or expected to cause toxicity or pathology to any organism other than plants and pos sibly insect vectors. Classification: Acceptable 467416

- 02

478955

- 03 885.4100 Avian Inhalation

Toxicity/Pathogenicity

885.4150 Wild Mammal

Toxicity/Pathogenicity

885.4200 Freshwater Fish

Toxicity/Pathogenicity

885.4240 Freshwater Invertebrate

Tox icity/Pathogenicity

885.4280 Estuarine/Marine Fish and Invertebrate Testing

885.4300 Nontarget Plant Testing Some plants are susceptible to TMGMV U2

. Limited use sites and application methods, mitigating label language, and certain manufacturing proc ess steps will minimize risk to nontarget plants. Classification: Acceptable 467416

- 04

478955

- 04

491377

- 01

492315

- 01

885.4340 Nontarget Insect Testing Testi ng waived based on BioProdex ’s scientific rationale, which included references to scientific

lite rature and show ed that plant viruses, including

TMGMV U2, are not known or expected to cause toxicity or pathology to any organism other than plants and possibly insect vectors. Classification: Acceptable 467416

- 02

478955

- 03 885.4380 Honey Bee Testing

Tiers II, III, and IV Not required for TMGMV U2 based on the acceptability of the scientific rationale

, literature , and data provided by BioProdex for Tier I

.Tobacco mild green mosaic tobamovirus strai n U2 (TMGMV U2) Page 31 of 31

Biopesticides Registration Action Document

APPENDIX B. PESTICIDE PRODUCTS

EPA File Symbol Registration

Name Percent age

Active Ingredient Formulation

Type Use Site(s) Method(s) of Application Application Rate (s) Target Pest

81179

- G SolviNix ® LC 2.9% End Use

– Liquid Pastures and

wooded areas that

are fenced to contain grazing

animals High

- pressure

backpack sprayer and

hand

- held

herbicide applicators 1.5 milliliters of

SolviNix ® LC per 1 gallon of water

Tropical soda apple ( Solanum viarum ) 10 mL of SolviNix ® LC per 6.5 gallons of

water

35 milliliters of

SolviNix ® LC per 23 gallons of water