BIOPESTICIDES REGISTRATION ACTION DOCUMENT



## Pseudomonas fluorescens strain CL145A

Pesticide Chemical (PC) Code: 006533

U.S. Environmental Protection Agency Office of Pesticide Programs Biopesticid es and Pollution Prevention Division

Version # 2

J u ly 2 9 , 2011Pseudomonas fluorescens s train CL145A Page 2 of 35 Biopesticides Registration Action Document

TABLE OF CONTENTS

I. OVERVIEW

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

II. ACTIVE INGREDIENT OV ER VIEW

................................ ................................ ............ 5

III. REGULATORY BACKGROUN D

................................ ................................ ................... 5

A. Pesticide Emergency Exemption s

................................ ................................ ................................ ........... 5

B. Applications for Pesticide Registration

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

C. Food Tolerance Exemption

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

IV. RISK ASSESSMENT

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

A. Product Analysis Assessment (40 CFR § 158.2120)

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

B. Human Health Assessment (40 CFR § 158.2140)

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

C. Environmental Assessment (40 CFR § 158.2150)

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

V. ENVIRONMENTAL JUSTIC E

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

VI. RISK MANAGEMENT DECISION

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

VII. ACTIONS REQUIRED BY THE REGISTRANT

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

VIII. GLOSSARY OF ACRONYMS AND ABBREVIATIONS

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

IX. BIBLIOGRAPHY

................................ ................................ ................................ ............ 23

A. Studies Submitted to Support the Pseudomonas fluorescens strain CL145A

................................ ... 23

B. Environmental Protection Agency Risk Assessment Memoranda

................................ ..................... 24

C. Other References

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

APPENDIX A . MICROBIAL PESTICI DES DATA REQUIREMENT S

............................ 26

APPENDIX B. PESTICI DE PRODUCTS

................................ ................................ ............... 35Pseudomonas fluorescens s train CL145A Page 3 of 35 Biopesticides Registration Action Document

BIOPESTICIDES REGISTRATION ACTION DOCUMENT TEAM

Office of Pesticide Pro grams (OPP) Biopesticides and Pollution Prevention Division

Microbial Pesticides Branch

Science Reviews

Joel V. Gagliardi , Ph.D. Product Analysis, Human Health

John L. Kough, Ph.D. Product Analysis, Human Health Gail Tomimatsu, Ph.D. E nvironmental Effects

Mike Mendelsohn Environmental Effects

Regulations

Sheryl K. Reilly, Ph.D. Chief, Microbial Pesticides Branch Alan Reynolds Team Leader, Microbial Pesticides Branch

Ann Sibold Regulatory Action Leader

Jeannine Kausch Regulatory Action LeaderPseudomonas fluorescens s train CL145A Page 4 of 35 Biopesticides Registration Action Document

I. OVERV IEW

The active ingredient, Pseudomonas fluorescens strain CL145A, was isolated from river mud in the northeastern United States and found to have molluscicidal properties. In particular, when

live or killed Pseudomonas fluorescens strain CL145A is applied to water structures infested with quagga mussels ( Dreissena bugensis ) and zebra mussels ( Dreissena polymorpha ), these mussels ingest and filter the substance, which then degrades the epithelial lining of digestive tubules in their digestive glands and kil ls them

. Given this bacterium’s molluscicidal capabilities, Marrone Bio Innovations proposed to register both a man ufacturing

- use pesticide product, MOI

401 TGAI (EPA Reg. No. 84059

- 4) , and an end

- use pestic ide product, MOI

- 401 EP (EPA Reg.

No. 84059

- 5 ), c ontaining Pseudomonas fluorescens strain CL145A. MOI

- 401 EP will be applied to water in enclosed and other confined static or flowing water infrastructures (e.g., water storage chambers and tanks, pipes, general plumbing and equipment, and other water conv eyance structures associated with civil infrastructure) to control the aforementioned invasive mussels

.

Environmental Protection Agency (EPA) scientists have reviewed product analysis, toxicology, and nontarget organism data and information (40 Code of F ederal Regulations (CFR) §§ 158.2120, 158.2140, and 158.2150, respectively) submitted to support the registration of the two Pseudomonas fluorescens strain CL145A pesticide products. Overall, such data and information

allow for unconditional, time

- limited registration under Section 3(c)(5) of the Federal Insecticide, Fungicide, and Rodenticide Act (FIFRA)

.

On October 1, 2009, EPA a nnounced a policy to provide a more meaningful opportunity for the public to participate in major registration decisions before they occur. According to this policy, EPA intends to provide a public comment period prior to making a registration decision for, at minimum, the following types of applications: new active ingredients; first food uses; first outdoor uses; first residenti al uses; or any other registration actions for which EPA believes there may be significant public interest.

Notwithstanding that the current Pseudomonas fluorescens strain CL145A actions involve a new active ingredient, first outdoor use, and first food u se under the policy described above, EPA believes that it is in the best interests of the public and the environment to issue the registrations

for the Pseudomonas fluorescens strain CL145A pesticide products without delay. Acceptable acute toxicity data f or Pseudomonas fluorescens strain CL145A demonstrated that it is either Toxicity Category III or Toxicity Category IV ( the lowest toxicity c lassifications), while an acceptable acute pulmonary toxicity/pathogenicity study indicated that Pseudomonas fluores cens strain CL145A is not toxic, infective, and/or pathogenic. EPA believes it is unlikely that Pseudomonas fluorescens strain CL145A will cause s ignificant adverse effects to nontarget organisms

, when use d in enclosed and other confined static or flowing water infrastructures

, provided all of the following occur: (1) discharge is prohibited in the vicinity of federally listed endangered or threatened species, (2) the calculated concentration of MOI

- 401 EP in receiving

waters is limited to concentrations at or below 100 parts per million (ppm), and (3) treatment monitoring ensur es that the measured concentration is at or below 1 ppm at a pre

- determined location 200 meters from discharge into flowing or static water. Thus, given that Pseudomonas fluorescens s train CL145A has low toxicity and presents little risk to nontarget organisms, when

used and monitored as indicated above, EPA concludes that it is in the best interests of the public and the environment to issue the registrations for the Pseudomonas fluor escens strain CL145A pesticide products without delay.Pseudomonas fluorescens s train CL145A Page 5 of 35 Biopesticides Registration Action Document

Consistent with the policy of making registration actions more transparent, however, EPA is issuing these registrations with an initial period of one

- year and, concurrent with their issuance,

p roviding a 30

- day comment public comment period on these time

- limited registrations. EPA is registering these products as time

- limited registrations, with the understanding that public comments could bring to light new information or concerns that could in form EPA’ s initial

decision. Any subsequent action taken by EPA will be made in the context of information received during the public comment period.

II. ACTIVE INGREDIENT OVERVIEW

Biological Name: Pseudomonas fluorescens strain CL145A

Culture Depos it: American Type Culture Collection in Manassas, Virginia

under Accession Number 55799.

OPP Chemical Code: 0 06533

Type of Pesticide: Microbial Pesticide

– Molluscicide

See Appendix B for specific informati on (e.g., use sites, application rate s , methods of application, formulation type s , and target pests) regarding the registered pesticide products containing this active ingredient.

III. REGULATORY BACKGROUND

A. Pesticide Emergency Exemption s

On Nove mber 12, 2009, EPA announced receipt of a quarantine exemption request from the United States Department of Interior’s Bureau of Reclamation to use Pseudomonas fluorescens strain CL145A to treat dams, water distribution (e.g., canals, pipes and plumbing), water treatment, water pumping facilities, irrigation and power generation facilities infested with invasive quagga and zebra mussels and associated reservoirs, water

- holding marinas and watercraft, recreational facilities (e.g., beaches and boat launches) , fish hatcheries and fish protection facilities (e.g., fish ladders and screens) ( 74 FR 58287 ). On August 26, 201 0 , EPA granted a quarantine exemption , pursuant to section 18 of the Federal Insecticide, Fungicide and

Rodenticide Act

, to the United States Department of Interior’s Bureau of Reclamation for use of the unregistered active ingredient

, Pseudomonas fluorescens strain CL145A, in three dams (Hoover, Davis and Parker) on th e lower Colorado River to control invasive zebra and quagga mussels. The exemption expires on August 26, 2013.Pseudomonas fluorescens s train CL145A Page 6 of 35 Biopesticides Registration Action Document

B . Applications for Pesticide Registration

On December 3, 2008, Marrone Organic Innovations (now Marrone Bio Innovations) (address:

2121 S econd Street, Suite B

- 107, Davis, CA 95618) submitted applications for registration of the technical product, MOI 401 TGAI (EPA File Symbol 84059

- U), and the end

- use product, MOI 401 EP (EPA File Symbol 84059

- L), containing the microbial active ingredient, Pseudomonas fluorescens strain CL145A, for use as a molluscicide applied to water to control zebra and quagga mussels. Notice of Receipt was published in the Federal Register on March 16, 2009 ( 74

FR 11098 ). No comments were received. For more information, see the following docket at www.regulations.gov : EPA

- HQ

- OPP

- 2009

- 0102.

C . Food Tolerance Exemption

Concurrent with its registration application s and under Federal Food, Drug, and Cosmetic Act (FFDCA) section 408(d), Marrone Organic Innovations (now Marrone Bio Innovations) submitted a petition to establish an exemption from the requirement of a tolerance for Pseudomonas fluorescens str ain CL145A

. In the Federal Register of March 16, 2009 ( 74 FR

11100 ), EPA announced that Marrone Organic Innovations proposed to establish an exemption from the requirement of a tol erance for residues of the microbial molluscicide

, Pseudomonas fluorescens strain CL145A

, in or on all food commodities and opened a 30

- day comment period. EPA received one comment in response to this publication and a response is provided in the preamble of the Pseudomonas fluorescens strain CL145A Final Rule.

IV. RISK ASSESSMENT

In the Federal Register of October 26, 2007, EPA issued a Final Rule on the data requirements to support registration of microbial pesticides and updated the definition for microbial pesticides ( 72 FR 61002 )

. The rule became effective on December 26, 2007. The data and information evaluated for this BRAD were considered in light of these requirement s.

The classifications that are found for each data submission are assigned by EPA science

reviewers and are an indication of the usefulness of the information contained in the documents for risk assessment. A rating of “ acceptable ” indicates the study is scientifically sound and is

useful for risk assessment. A “ supplemental ” rating indicates the data provide some information that can be useful for risk assessment. The studies may have certain aspects determined not to be scientifically acceptable (“ suppl emental : upgradeable ”). If a study is rated as “supplemental: upgradeable ,” 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

oft en 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 need to be submitted.

For the acu te toxici ty data requirements, Toxicity C ategories are assigned based on the hazard(s) identified from studies and/or other infor mation submitted to 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.Pseudomonas fluorescens s train CL145A Page 7 of 35 Biopesticides Registration Action Document

A. Product Analysis Assessment ( 40 CFR § 158.2120 )

Most of the product analysis data requirements for Pseudomonas fluorescens strain CL145A

have been fulfilled . Only a storage stability study is outstandin g , and this information must be submitted to EPA by January 29, 2012. Refer to Tables 1, 2, 3, and 4 in Appendix A for a brief summary of the data requirements, including both generic and product

- specific information.

B. H uman Health Assessment ( 40 CFR § 158.2140 )

1. Toxicity

All toxicology data requirements for P seudomonas fluorescens strain CL145A have been

fulfilled

. Acceptable Tier I mammalian toxicology data and information support registration of

the Pseudomonas fluorescens strain CL145A pesticide products

. Furthermore, Tier II and Tier III

studies were not r equired for Pseudomonas fluorescens strain CL145A based on the lack of acute toxicity/pathogenicity in the Tier I studies.

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

- specific data submitted to support the individual registrations, refer to Tables 5 and 6 in Appendix A

.

a. Acute Toxicity/Pathogenicity

– Tier I

Acute Oral Toxicity/Pathogenicity (O ffice of Chemical Sa fety and Pollution Prevention (O CSPP ) Guideline 885.3050; Master Record Identification Number ( MRID No. ) 47 7494

- 03 ) : Rationale for an acute oral toxicity/pathogenicity waiver request stated that Pseudomonas fluorescens is considered a ubiquitous inhabitant of soil and water and is found on the surface and roots of a variety of plant types, including those of food plants consumed raw. Pseudomonads and, in particular, Pseudomonas fluorescens are considered a benign part of the regular human diet commonly occu rring on the surface of leafy green vegetables and other food stuffs . Additionally, an acute oral toxicity study conducted on rats (MRID No. 476402

- 02) found the no observed adverse effect level ( NOAEL ) and median lethal dose ( LD 50 ) were greater than 5,00 0 milligrams per kilogram ( mg/kg ) , corresponding to greater than 2.35 x 10 10 colony

- forming units per kilogram ( CFU/kg ) (Toxicity Category IV). Sufficient information was supplied and deemed acceptable. Thus, the requirement for acute oral toxicity/pathoge nicity testing was waived.

Acute Pulmonary Toxicity/Pathogenicity (OCSPP Gui deline 885.3150; MRID No. 482767

- 02 ) : No animals died during this study

, and there were no abno rmalities seen during necropsy.

Adverse effects (emaciation and reduced activity) we re noted in three females dosed with live

test substance only on day 7 with recovery after day 14. Significantly lower organ weights were noted in treated males (mesenteric lymph nodes and kidneys) and females (spleens) when comparing inactivated test subs tance treated to control groups. No signs of toxicity were noted in

ina ctivated test substance treated or untreated rats. Weight losses by day 7 were seen in three

females dosed with live test substance, notably in two also showing emaciation on day 7. Tes t substance clearance was established by day 21 from all tissues

, and no signs of infectivity or pathogenicity were noticed. Pseudomonas fluorescens strain CL145A does not appear to be toxic, infective, and/or pathogenic to rats when dosed intratracheally at 3.4 x 10 8 colony

- formingPseudomonas fluorescens s train CL145A Page 8 of 35 Biopesticides Registration Action Document

units (CFU)/rat. This study was rated acceptable.

Acute Injection Toxicity/Pathogenicity (Intravenous)

– Rat (OCSPP Guideline 885.3200; MRID

No. 476402

- 07 ) : An acute injection toxicity and pathogenicity (intravenous) study show ed high mortality in the test animals and it was unclear whether the Pseudomonas or a toxin was responsible for the mortality seen. A pattern of clearance for Pseudomonas fluorescens strain CL145A was demonstrated in the study. Given this, and lower mortal ity with freshly prepared test substance, lack of toxicity with autoclaved culture, and results from other toxicological tests, we conclude that there is adequate information to perform a risk assessment on Pseudomonas fluorescens strain CL145A. Moreover, this exposure pattern is not relevant to the registered uses. This study was rated supplemental.

Hypersensitivity Incidents (OCSPP Guideline 885.3400; MRID No. Not Applicable ) : No hypersensitivity incidents, including immediate

- type or delayed

- type reacti ons of humans and

domestic animals that occurred during research, development, or testing of Pseudomonas fluorescens strain CL145A

, were reported by the applicant . Any future hypersensitivity incidents must be reported per OCSPP Guideline 885.3400.

Cell C ulture (OCSPP Guideline 885.3500) : This study is not required because Pseudomonas fluorescens strain CL145A is not a virus (refer to test note #4 of 40 CFR § 158.2140(d)).

b. Acute Toxicology and Subchronic Toxicity/Pathogenicity

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

– Tier III

Tier II and Tier III studies were not required for Pseudomonas fluorescens strain CL145A based on the lack of acute toxicity/pathogenicity in the Tier I studies.

c. Endocrine Disruptors

As required under FFDCA section 408(p), EPA has developed the Endocrine Disruptor Screening Program (EDSP) to determine whether certain substances (including pesti cide 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.” The EDSP employs a two

- tiered approach to mak ing the statutorily required determinations. Tier 1 consists of a battery of 11 screening assays to identify the potential of a chemical substance to interact with the estrogen, androgen, or thyroid (E, A, or T) hormonal systems. Chemicals that go through Tier 1 screening and are found to have the potential to interact with E, A, or T hormonal systems will proceed to the next stage of the EDSP wh ere EPA will determine which, if any, of the Tier 2 tests are necessary based on the available data. Tier 2

testi ng is designed to identify any adverse endocrine

- related effects caused by the substance, and establish a quantitative relationship between the dose and the E, A, or T effect.

Between October 2009 a nd February 2010, EPA issued test orders/data call

- ins fo r the first group of 67 chemicals, which contains 58 pesticide active ingredients and 9 inert ingredients. This list of chemicals was selected based on the potential for human exposure through pathways such as food and water, residential activity, and cert ain post

- application agricultural scenarios. This list should not be construed as a list of known or likely endocrine disruptors.Pseudomonas fluorescens s train CL145A Page 9 of 35 Biopesticides Registration Action Document

Pseudomonas fluorescens strain CL145A is not among the group of 58 pesticide active ingredients on the initial list to be scr eened under the EDSP. Under FFDCA section 408(p), EPA must screen all pesti cide chemicals. Accordingly, EPA anticipates issuing future EDSP

orders/data call

- ins for all pesticide active ingredients.

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: http://www.epa.gov/endo/

.

2

. F ederal Food, Drug, and C osmetic Act (FFDCA) Considerations

S ection 408(c)(2)(A)(i) of FFDCA allows EPA to establish an exemption from the requirement for a tolerance (the legal limit for a pesticide chemical resi due in or on a food) only if EPA determines that the exemption is “safe.” Section 408(c)(2)(A)(ii) of FF DCA defines “safe” to mean that “there is a reasonable certainty that no harm will result from aggregate exposure to the pesticide chemical residue, including all anticipated dietary exposures and all other exposures for

which there is reliable information .” This includes exposure through drinking water and in residential settings, but does not include occupational exposure. Pursuant to section 408(c)(2)(B) of FFDCA, in establishing or maintaining in effect an exemption from the requirement of a tolerance, EPA must take into account the factors set forth in section 408(b)(2 )(C) of FFDCA, which require EPA to give special consideration to exposure of infants and children to the pesticide chemical residue in establishing a tolerance and to “ensure that there i s a reasonable certainty that no harm will result to infants and children from aggregate exposure to the pesticide chemical residue....” Additionally, section 408(b)(2)(D) of FFDCA requires that EPA consider “available information concerning the cumulative effects of [a particular pesticide's]

. . . residues and other substances that have a common mechanism of toxicity.”

EPA performs a number of analyses to determine the risks from aggregate exposure t o pesticide residues. First, EPA determines the tox icity of pe sticides. Second, EPA examines exposure to the pesticide through food, drinking water, and through other exposures that occur as a result of pesticide use in residential settings.

Consistent with s ection 408(b)(2)(D) of FFDCA, EPA has reviewed the availabl e scientific data and other relevant information, and considered its validity, completeness, and reliability and the relationship of thi s information to human risk. EPA has also considered available information concerning the variability of the sensitiviti es of major identifiable subgroups of consumers, including infants and children.

Based on the acute toxicity/pathogenicity data and information discussed previously and presented in Table 5 in Appendix A , t he data required fo r a FQPA risk assessment for Pseudomonas fluorescens strain CL145A have been fulfilled

.

a. Aggregate Exposure

In examining aggregate exposure, section 408 of FFDCA directs EPA to consider available information concerning exposures from the pesticide r esidue in food and all other nonoccupational exposures, including drinking water from ground water or surface water andPseudomonas fluorescens s train CL145A Page 10 of 35 Biopesticides Registration Action Document

exposure through pesticide use in gardens, lawns, or buildings (residential and other indoor uses).

Dietary Exposure and Risk Character ization : Exposure to this microbial active ingredient through food is expected to be minimal. Pseudomonas fluorescens strain CL145A end

- use products are n ot labeled for direct application to food crops

. Rather, Pseudomonas fluorescens

strain CL145A will be applied to water in enclosed and other confined static or flowing water

infrastructures to control zebra and quagga mussels

. The treatment areas are limi ted to completely enclosed pipe or water conveyance syste ms or concrete chambers with defined inlet s or outlet s

. Nevertheless

, water drawn downstream from points of application (e.g., pump stations and irrigation systems) and used to irrigate or wash food crops may contain Pseudomonas fluores cens strain CL145A

. C oncentrations of Pseudomonas fluorescens strain CL145A will be diluted as water flows past points of application and thus will rapidly decrease

. A lso, natural degradation (e.g., environmental factors such as ultraviolet light, nutrient depletion and bacterial grazing/predation by protist s and others ) and manmade filtering operations w ould greatly lower the overall level of the pesticide after application ( U.S. EPA 1996, 2004 ). Furthermore, Pseudomonas fluorescens is considered ubiquito us in soil and water and is commonly associated with plants , including food plants consumed raw ; thus, this microorganism is already part of the

normal human diet ( Garrity et al . 2005; Murray et al . 2003; OECD 1997 ). Exposure to Pseudomonas fluorescens strain CL145A through food, whi ch has come into contact with treated irrigation or wash waters, is not expected to exceed background levels of similar Pseudomonads already present in the human diet ( Garrity et al . 2005; Murray et al . 2003; OECD 1997 ).

N one theless, i n the unlikely event that this microbial pesticide is present on food, the acute oral toxicity and pathogenicity data/ information demonstrate d that no toxicity, infectivity and/or pathogenicity is likely to occur w ith any exposure level of Pseudomonas fluorescens strain

CL145A

.

Drinking Water Risk Characterization : Much like food exposure, d rinking water exposure is expected to be negligible for similar reasons: (1) c oncentrations of Pseudomonas fluorescens

strain CL145A will be diluted as water flows past points of application

, ( 2) natural degradation (e.g., environmental factors such as ultraviolet light, nutrient depletion and bacterial grazing/predation by protists and others ) of the microbial active ingredient w ill occur, and ( 3)

flocculation and filtering at water treatment plants w ill further inactiv ate and decrease levels of Pseudomonas fluorescens strain CL145A ( U.S. EPA 1996, 2004 ). Additionally, Pseudomonas fluorescens is al ready present naturally in soil, in water

, and on plants , thereby making it a part of

the normal human diet ( Garrity et al . 2005; Murray et al . 2003; OECD 1997 ). Exposure to Pseudomonas fluorescens strain CL145A through drinking water is not expected to exceed background level s of similar Pseudomonads already present in the human diet ( Garrity et al

. 2005; Murray et al . 2003; OECD 1997 ). N one theless, i n the unlikely event that this microbial pesticide is present on food, the acute oral toxicity and pathogenicity data/ information

demonstrate d that no toxicity, infectivity and/or pathogenicity is likely to occur w ith any exposure level of Pseudomonas fluorescens strain CL145A

.

Nonoccupational, Residential Risk Characterization : Dermal and inhalation non

- occupational
exposure to Pseudomonas fluorescens strain CL145A is expected to be minimal to non

- existent. Pseudomonas fluorescens strain CL145A end

- use products are labeled for application to use

sites

— enclosed and other confined static or flowing water infrastructures infested with zebra and/or quagga mussels

— that are not considered residential areas.Pseudomonas fluorescens s train CL145A Page 11 of 35 Biopesticides Registration Action Document

Although d ermal exposure to Pseudomonas fluorescens strain CL145A may occur when water from a treated dam or industrial facility is discharged to surface water and is subsequently used by a community water system in a residential area

, s uch exposure is expected to be minimal due

to dilution , natural degradation, and filtering at water treatment plants ( U.S. EPA 1996, 2004 ). Moreover, acute dermal toxicity and primary dermal irritation tests d emonstrated that Pseudomonas fluorescens strain CL145A is not toxic and is practically non

- irritating via the dermal route of exposure (see Table 5 of Appendix A )

.

Inhalation exposure to Pseudomonas fluorescens strain CL145A is not anticipated with the

labeled (i.e., water

- based) molluscicide use

. If inhalation exposure to Pseudomonas fluorescens

strain CL145A were to occur in g ardens, lawns, or buildings (i.e., residential areas)

, such exposure would not exceed EPA ’ s level of concern given th e acute inhalation toxicity and acute pulmonary toxicity/pathogenicity tests that demonstrated Pseudomonas fluorescens strain

CL145A ’ s lack of toxicity, pathogenicity and/or infectivity (see Table 5 of 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 exemption, EPA consider “available information concerning the cumulative effects of [a particular pesticide’s] residues and other substances that have a common mechanism of toxicity.”

EPA has not found Pseudomonas fluoresce ns strain CL145A to share a common mechanism of toxicity with any other substances. Pseudomonas fluorescens strain CL145A affects gut function in the target molluscs and does not produce similar toxic response in the other species tested. For the purposes of this tolerance action, therefore, EPA has assumed that Pseudomonas fluorescens strain CL145A does not have a common mechanism of toxicity with other substances. Following from this, therefore, EPA concludes that there are no cumulative effects associate d with Pseudomonas fluorescens strain CL145A that need to be considered. For information regarding EPA’s efforts to determine which chemicals have a common mechanism of toxicity and to evaluate the cumulative effects of such chemicals, see EPA’s website at http://www.epa.gov/pesticides/cumulative

.

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

FFDCA section 408(b)(2)(C) provides that EPA shall assess the available infor mation about consumption patterns among infants and children, special susceptibility of infants and children to pesticide chemical residues, and the cumulative effects on infants and children of the residues

and other substances with a common mechanism of toxicity. In addition, FFDCA section 408(b)(2)(C) provides that 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 th e database on toxicity and exposure unless EPA determines that a different margin of safety will be safe for infants and children. This additional margin of safety is commonly referred to as the Food Quality Protection Act Safety Factor. In applying this

p rovision, EPA either retains the default value of 10X or uses a different additional safety factor when reliable data available to EPA support the choice of a different factor.Pseudomonas fluorescens s train CL145A Page 12 of 35 Biopesticides Registration Action Document

Based on the acute toxicity and pathogenicity data/information discussed above , 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 the residues of Pseudomonas fluorescens strain CL145A. This includes all anticipated dietary exposures and all other exposures for which there is reliable information. EPA has arrived at this conclusion because, considered collective ly, the data (e.g., lack of toxicity noted for oral, dermal, and inhalation routes of exposure) available on Pseudomonas fluorescens strain CL145A do not demonstrate toxic, pathoge nic, and/or infective potential to sensitive populations from exposure to this microbial pest control agent. Thus, there are no threshold effects of concern and, as a result, an additional margin of safety is not

necessary.

3. Occupational Exposure and R isk Characterization

The acute pulmonary toxicity/pathogenicity showed clearance by day 21 and no signs of toxicity, pathogenicity and/or infectivity. Due to high mortality in the intravenous toxicity/pathogenicity study, respiratory protection is necessa ry when handling and applying undiluted Pseudomonas fluorescens strain CL145A to sites listed on the label, including confined water structures in

dams and industrial sites . Exposures to dilute end

- use product would be low hazard

, and this

assessment is su pported by the available no observed adverse effect level of greater than 0.225 milligrams per liter (mg/L) and 1.1 x 10 6 CFU/L noted in the pulmonary toxicity study. Exposure to discharges of treated water from the treatment site is not expected to be si gnificant.

4. Human Health Risk Characterization

EPA considered human exposure to Pseudomonas fluorescens strain CL145A in light of the

standard for registration in FIFRA and the relevant safety factors in FFDCA. A determination has

been made that no unr easonab le adverse effects to the U.S. population in general, and to infants and children in particu lar, will result when Pseudomonas fluorescens strain CL145A pesticide products are used in accordance with EPA

- approved labeling.

C. Environmental Assessme nt ( 40 CFR § 158.2150 )

1. Ecological Effects Hazards Assessment

Non t arget Organism Effects

T able 7 (see Appendix A ) shows that P seudomonas fluorescens strain CL145A had no effect at 100 ppm on fish, aquatic invertebrates, fresh and salt water mussels, and mallard ducks.

E xposures , however, were acute and the level of irradiation used on the t est material in studies

with aquatic invertebrates and mallard ducks was significantly greater than that used on the

manufactured MOI

- 401 EP. Non

- irradiated cells caused up to 89% mortality in treated fish. The

Pseudomonas fluorescens strain CL145A used in the MOI

- 401 EP product is irradiated to significantly reduce the number of viable Pseudomonas fluorescens strain CL145A cells. In

submitted non target studies, this irradiation was observed as having the effect of eliminating observed toxicity while maintaining efficac y against targeted zebra mussels.Pseudomonas fluorescens s train CL145A Page 13 of 35 Biopesticides Registration Action Document

2. Exposure Assessment

– MOI

- 401 EP Use, EEC Calculations, and Pre

- treatment Site

Notification Information

a. Use Sites

According to the draft label dated July 26, 2011

, MOI

- 401 E P lists the following use sites:

Treatment sites for MOI

- 401 EP: Treatments of MOI

- 401 EP are limited to enclosed and other confined static or flowing water infrastructures infested wit h zebra and/or quagga mussels. Treatment area must be a completely enclosed pipe or water conv eyance system or concrete
chamber w ith a defined inlet or outlet. Application of MOI

- 401 EP assumes the full flow and volume of water through the treated pipe or water conveyance system or concrete chamber with a defined

inlet or outlet. Enclosed and other confined static or flowing water infrastructures include water storage chambers and tanks, pipes, general plumbing and equipment, and other water conveyance structures associated with civil infrastructure such as the cooling water systems and contained wa ter storage chambers of power plants, pump stations, irrigation systems, industrial and manufacturing facilities (e.g. au tomobile and steel), and dams. Treatment of open water, such as infested ponds, lakes, reservoirs, rivers and streams, or other unconfi ned aquatic systems, with MOI

- 401 EP is prohibited.

b. EEC Calculations, Mitigation Factors, and Reports

The pre

- release calculated concentration of MOI

- 401 EP active ingredient in receiving waters must be at or below 100 ppm (Refer to SOP #MBI 20.16.0, July 26, 2011) [or when the PDM model is used, the concentration of concern of 100 ppm will not be exceeded on any calendar day]. Treatment monitoring must ensure that the measured concentration of active ingredient will be at or below 1

ppm at a pre

- dete rmined location 200 meters from discharge into flowing or static water (Refer to

SOP # MBI

- G

- 20.15.1 , July 26, 2011 and SOP # MBI

- G

- 20.17.1 , July 26, 2011).

Marrone Bio Innovations will provide the following pre

- treatment notification information to

EPA 30 days prior to treatment. If calculations show that expected environmental concentrations will exceed 100 ppm or that the facility may be within 5 miles of the critical habitat of federal endangered or threatened species, Marrone Bio Innovations will consu lt EPA for further guidance and obtain EPA concurrence on application to the site prior to treatment.

Type of facility

Facility a ddress and contact person details

Endangered and Threatened Species Site Review

– Marrone Bio Innovations will review the Endangered Species List (http://www.fws.gov/endangered/) for any endangered or threatened terrestrial or aquatic organisms and their critical habitats, if any, and submit the results of the review. The dates of the search must also be included, since the

da tabase is periodically updated. In addition, the notification will confirm that critical habitats of endangered or threatened species are outside of a 5

- mile radius of the treatment site

.

Sampling locations for application monitoring

Water intake loca tion (and water body)Pseudomonas fluorescens s train CL145A Page 14 of 35 Biopesticides Registration Action Document

Application Information

--

Treated system flow rate (e.g. gallon/day, m 3 /hr) or volume (L)

Application concentration (mg a.i./L)

Application time (min)

Calculated total kg/site/day

Water body and location of discharge

River flow rate from nearest USGS gauging station. In locations with no USGS gauging station, flow rate will be attained from an independent and scient ifically validated source (e.g., state and federal agencies and local non

profits such as Streamkeepers)

.

Calculations and calculated expected environmental discharge concentration per treatment application rate, treated water flow rate, and receiving water body dilution information

Model assumptions and model summary

Model input data

Model output da ta to include a minimum of the expected environmental concentration within 200 yards of the discharge zone

PDM print out when the PDM model is used

Treatment monitoring

, via measurements described in SOP s # MBI

- G

- 20.15. 1 , July 26, 2011

, and

# MBI

- G

- 20.17. 1 , July 26, 2011 , are required to ensure that the EEC is measured at or below 1 ppm

at a pre

- determined location 200 meters from discharge into flowing or static water

. In treatment monitoring, if the downstream turbidity concentration exceeds expected envi ronme ntal concentration

of 1 mg a.i./ L based on correlation between turbidity and dry cell weight per SOP # MBI

- G

- 20.15.1

, Marrone Bio Innovations or the applicator will take corrective action. Marrone Bio Innovations or the applicator will p erform addition al downstream turbidity monitoring after every thirty minutes of any corrective action. If measurements continue to show an exceed a nce of the expected

environmental concentration of 1 mg a.i./L after 1.5 hours, M arrone Bio Innovations or the applicator wil l turn off treatment metering pump. Any exceed a nce of the expected environmental

concentration wil l be reported by Marrone Bio Innovations to the U.S. EPA Microbial Pesticides

Branch Chief with in 24 hours by calling (703) 308

- 8269 and by e mail addressed to

r eilly. s heryl@epa.gov

.

On a quarterly basis, M arrone Bio Innovations will submit a report to EPA that includes the recorded

measurement data

. The report will be organized by facility and present a data summary per facility.

The di scharge monitoring data will pre sent the turbidity and Pseudomonas f luorescens concentrations during and after tre atment. The turbidity correlation and application monitoring will present data demonstrating the in

- facility treatment concentration.Pseudomonas fluorescens s train CL145A Page 15 of 35 Biopesticides Registration Action Document

Per SOP # MBI

- G

- 20.16.1, Sit e Selection Criteria and Calculation of Expected Environmental Concentration for MOI

- 401 EP , pre

- release calculations will be conducted as follows:

Calculated total kg/site/day released  B = (X)(O)(T) (4 kg/gal)(l 0 - 6 mg/L)

B = R elease of material via bl owdown water to surface water (kg/site/day)

X = Application concentration of material in water (mg a.i. /L) T = Application time (minutes)

O = Treated syste m flow rate (e.g. gallon/day, m 3 /hr) or volume (L)

Below are the three conditions under which data w ill be presented in the pre

- treatment notification

documentation:

1) For facilities with NPDES permits discharging to flowing waters that are included in the EFAST V 2.0 Probabilistic Dilution Model (PDM), M arrone Bio Innovations will run the PDM model and p rovide a printout of the PDM site

- specific results page. PDM is a screening

- level exposure assessment tool developed by EPA to model chemical releases from point sources to flowing surface waters. For this analysis, the PDM component within EPA's Exposure and Fate Assessment Screening Tool Version 2.0 (E

- FAST2) will

be used. 1

PDM uses detailed U.S. Geological Survey (USGS) stream flow data and facility

s pecific data from National Pollutant Discharge Elimination System (NPDES) permits to model chemical re leases from actual facilities. For a modeling period of a given number of days, PDM calculates probability distribution of the chemical

concentration in the receiving stream, and then estimates the number of days during

which the in

- stream chemical conc entration is expected to exceed a concentration of concern (COC). PDM counts a day as having an exceedance of a COC if the COC is exceeded for any part of a 24

- hour day. As a screening

- level model, PDM outputs do not include the duration, location, or a erial extent of exceedances.

Inputs to PDM include facility NPDES number, pre

- treatment release (i.e., loading rate), post

- treatment release, number of release days, and COCs. Input values and assumptions used for this analysis are discussed below.

1 The E

- FAST2 model is available from EPA at http://www.epa.gov/opptintr/exposure/pubs/efastdl.htm and

documentation is available at http://www.epa.gov/opptint r/exposure/pubs/efast2man.pdf

.Pseudomonas fluorescens s train CL145A Page 16 of 35 Biopesticides Registration Action Document

2) For facilities discharg ing to flowing waters that curre ntly do not hold an NPDES permit or for facilities in which the discharge water body does not have a USGS gauging station and therefore is not included in the EFAST V 2.0 PDM, M arrone Bio Innovatio ns will conduct and document mass balance calculations to determine the expected

environmental concentration as follows:

C
c F
c + C R F R = C T F T

C
c = Applied concentration of product in cooling water. The maximum label amount is

0. 026 oz. a.i./gallon (200 mg a.i

. / L).

F
c = Treated volumetric flow r ate (Liters/hour, gallons/minute ).

C R = Concentra tion of MOI

- 401 EP in the river prior to point discharge. This value should be zero as there should not be produ ct already existing in th e wat er.

F R = River volumetric flow rate prior to point discharge from cooling water systems (Liters/hour, gallons/minute).

C T = Concentration after dilution with river water. This should be less than 0.00013

oz. a.i. /gallon (1 mg a.i. /L) per MOI

- 401 EP label requirements

.

F T = Total river volumetric flow rate and volume of discharge water.

These calculations and assumptions will be presented in the pre

- treatment notification

documentation. Documented information will include the following : water body and location of discharge, river flow rate from nearest USGS gauging station. In locations with no USGS gauging station, flow rate will be attained from an independent and scientifically validated source (e.g.

, state and federal agenc ies and local non

- profits such as Streamkeepers ). Documented information will also include calculations and calculated expected environmental discharge concentration per treatment application rate, treated water flow rate, and receiving water body dilution information

3) For facilities discharging to still waters (harbors, lakes, forebays), M arrone Bio

Innovations will request diffusion and dilution models developed specifically for the facility and that have been certified to be representative of actual disc harge dilution.

M arrone Bio Innovations will use the models that facilities currently discharging under NPDES permits have previously developed and validated to determine the expected environmental concentration within 200 y ar ds of the discharge point.

3

. Background Product Analysis

a. P. fluo rescens strain CL145A and MOI

- 401 EP

The active ingredient was identified as Pseudomonas fluorescens , a member of biovar I, and named strain CL145A. This isolate originated from a river mud sample in the northea stern United States. Subsequent testing showed degradation of the epithelial lining of digestive tubules in the digestive gland, specifically in quagga and zebra mussels, upon filtering live or killedPseudomonas fluorescens s train CL145A Page 17 of 35 Biopesticides Registration Action Document

P seudomonas fluorescens strain CL145A

, while other epit helial cell s in the mussel digestive gland and t issues elsewhere in the mussels appear unaffected. The registrant points out that the tubule epithelium is the “primary location for nut rient uptake within the mussel.” There is no

evidence of infection from ingestion of Pseudomonas fluorescens strain CL145A though a hemolysin may play a role in this tissue specific activity. The Pseudomonas fluorescens strain

CL145A used in the MOI

- 401 EP product is irradiated to significantly reduce the number of

viable P seu domonas fluorescens strain CL145A cells. In submitted non target studies, this irradiation has the effect of eliminating observed toxicity while maintaining efficacy against target mussels.

b. Pseudomonas fluorescens

Pseudomonas fluorescens is consider ed ubiquitous in soil and water and is found commonly associated with plants in high numbers, including on food plants consumed raw. The Manual of Clinical Microbiology, 8 th edition, states “ Pseudomonas spp. have a worldwide distribution with a predilectio n for moist environments. They are found in water and soil and on plants, including fruits and vegetables.”

Several peer

- reviewed scientific literature reports concerning the ecological and biological profile of Pseudomonas fluorescens were provided in the OECD Consensus Document on Information Used in the Assessment of Environmental Applications involving Pseudomonas

. 2

General Considerations

“

Pathogenicity to animals P. fluorescens can infect a wide range of animals including horses (Sarasola et al., 1992), chickens (Lin et al., 1993), marine turtles (Glazebrook and Campbell, 1990), and many fish and invertebrate species. However, because it is unable to grow at elevated temperatures like that of the human body (Palleroni, 1992a), it is unlikely to be more than a rare opportunistic pathogen for warm

- blooded animals.

P. fluorescens is considered a secondary invader of damaged fish tissues, but may also be a primary pathogen (Roberts and Horne, 1978; Stoskopf, 1993). The species causes bacterial tail rot and can affect freshwater and saltwater fishes throughout the world (Stoskopf, 1993). P. fluorescens releases extracellular proteases upon invasion of the fish host (Li and Flemming, 1968), and morbidity can be quite high. Outbreaks of bacterial tail

rot have been reported in goldfish, silver carp, bighead, tench, grass carp, black carp, golden shiner, rainbow trout, European eels, paradise fish, and other labyrinth fishes. Egusa (1992) reviewed Pseudomonas diseases in fish and indicated that, in the Unite d States, the disease termed Pseudomonas septicemia, due to infection by bacteria related

to P. fluorescens (AFS

- FHS, 1975), is seen in comparatively large numbers in warm

- water fish.

Adverse effects associated with P. fluorescens in fish species often ap pear to be linked to stress from transportation or cultivation of fish. For example, P. fluorescens has been associated with disease in the cultivation of rainbow trout, Oncorhynchus mykiss (Barros et al., 1986), Atlantic salmon, Salmo salar (Carson and

Sc hmidtke, 1993), chinook salmon, Oncorhynchus tshawytscha (Newbound et al., 1993), sea bream, Evynnis japonica (Kusuda et al., 1974), bighead carp, Aristichthys nobilis , and silver carp, Hypophthalmichthys molitrix (Petrinec et al., 1985), catfish and carp (Gatti and Nigelli, 1984), tench (Ahne et al., 1982), and tilapia species (Okaeme, 1989; Miyashita, 1984; Miyazaki et al., 1984).

Barker et al. (1991) found that exposure of high numbers of P. fluorescens to egg surfaces of rainbow trout ( Oncorhynchus myk iss ) during the initial stages of incubation poses a threat to egg survival. Conversely, P. fluorescens was not pathogenic when injected into brown trout (Smith and Davey, 1993) or silver

2 http://www.rebeca

- net.de/downloads/OECD%20Consensus%20document%20pseudomonas..pdfPseudomonas fluorescens s train CL145A Page 18 of 35 Biopesticides Registration Action Document

mullet fish ( Mugil curema ) (Alvarez and Conroy, 1987). P. fluorescen s has also been implicated in pathogenicity to

some invertebrates. James and Lighthart (1992) determined an LC50 for the 1st, 2nd, 3rd and 4th instar larvae of a coleopteran insect ( Hippodamia convergens ) (4.8 x 109, 2.8 x 1010, 3.9 x 109, and 3.2 x1011 CF U/ml, respectively)

and concluded that P. fluorescens is a weak bacterial pathogen. P. fluorescens has also been reported to be associated with pathogenicity in the mosquitoes Culex quinquefasciatus

, Anopheles stephensi , and Aedes aegypti (Murty et al., 19 94) and in the field slug Deroeras reticulatum (Wilson et al., 1994). However, Genthner et al. (1993) studied the effects of P. fluorescens on eastern oysters ( Crassostrea virginica ) and found no signs of infectivity or pathogenicity.

Pathogenicity to pla nts P. fluorescens is generally considered to be a saprophyte rather than a plant pathogen (Bradbury, 1986), although Ormrod and Jarvis (1994) considered it to be an opportunistic pathogen causing soft rot in plants. P. fluorescens biovar 2 ( P. marginalis ) , however, is actively pectinolytic, causing soft rot of various plants, and is considered a plant pathogen (Tsuchiya et al., 1980; Hildebrand, 1989; Membre and Burlot, 1994; Brock et al., 1994). Bradbury (1986) recognised three pathovars in P. marginalis which cause soft rot in a wide range of vegetables and other plants.

A number of studies have reported adverse effects associated with P. fluorescens and plants (Gaudet et al., 1980; Anson, 1982; Hwang et al., 1989; Richardson, 1993; Ozaktan and Bora, 199 4). Tranel et al. (1993) found that P. fluorescens strain D7 inhibited root growth of downy brome ( Bromus tectorum ) by production of a phytotoxin. Sellwood et al. (1981) confirmed pathogenicity experimentally for an atypical P. fluorescens biotype

I on chi cory plants and suggested that the group P. fluorescens does not solely comprise saprophytes. However, other studies have found no adverse effects on plants from inoculations with P. fluorescens (Arsenijevic, 1986; Arsenijevic and Balaz, 1986; Surico and S cala, 1992).

At present, the epidemiology of pathogenic strains of P. fluorescens is not well understood (Hildebrand, 1994). Liao (1991) has cautioned that, at present, it is not known for sure that P. fluorescens strains are non

- pathogens and do not

caus e deleterious effects on plants. Liao indicated that the pel gene encoding production of pectate lysase (an enzyme which contributes ability to cause soft rot in plants) is well conserved in fluorescent pseudomonads, and may exist and remain repressed in c ertain strains or species which exhibit non

- pectolytic phenotypes under laboratory conditions. Liao further indicated that saprophytic strains of P. fluorescens can be induced to become pathogenic and raised a concern about the safety of using the species as a biocontrol agent. “

4. Environmental Assessment Conclusions

P seudomonas fluorescens strain CL145A had no effect at 100 ppm on fish, aquatic invertebrates,

fresh and salt water musse ls, and mallard ducks. E xposures , however, were acute and the level of

irradiation used on the test material in studies on aquatic invertebrates and mallard ducks was significantly greater than that used for the manufactured MOI

- 401 EP. Non

- irradiated cells of

Pseudomonas fluorescens strain CL145A caused up to 89% mortality in treated fish. The

Pseudomonas fluo rescens strain CL145A used in the MOI

- 401 EP product is irradiated to significantly reduce the number of viable Pseudomonas fluorescens strain CL145A cells. In

submitted non target organism studies, this irradiation was observed as having the effect of

elim inating observed toxicity in nontarget organisms while maintaining efficacy against target ed

zebra mussels ( Dreissena polymorpha ).

It is unlikely that MOI

- 401 EP will cause substantial adverse effects to non target organisms during a one

- year limited regi stration for use in completely enclosed pipe or water conveyance
system s or concrete chamber s with a defined inlet or outlet in enclosed and other confined static or flowing water infrastructures provided (1) discharge is prohibited in the vicinity of fede rally

listed endangered or threatened species (verified via http://www.fws.gov/endangered/ and other

sources ), (2) the calculated concentration of MOI

- 401 EP in receiving waters is limited to

concentrations at o r below 100 ppm (Refer to SOP #MBI 20.16. 1 , July 26, 2011) and (3)Pseudomonas fluorescens s train CL145A Page 19 of 35 Biopesticides Registration Action Document

treatment monitoring ensures that the measured concentration is at or below 1 ppm at a pre

determined location 200 meters from discharge into flowing or static water (Refer to SOP

# MBI

- G

- 2 0.15.1 , July 26, 2011 and SOP # MBI

- G

- 20.17.1 , July 26, 2011).

In order to support an assessment for longer term use, Marrone Bio Innovations should consult

with EPA and address the irradiation and test substance equivalency issue, ensure maximum

hazard dose testing, and provide test substance enumeration within the following toxicity or toxicity/pathogenicity studies:

885.4050

- Avian Oral

885.4200

- Freshwater Fish Testing

885.4240

- Freshwater Aquatic Invertebrate Testing

885.4280

- Estuarine and Mar ine Animal Testing

885.4340

- Nontarget Insect Testing

885.4650

- Aquatic Invertebrate Range Testing

885.4700

- Fish Life Cycle Studies

850.1055

– Bivalve Acute Toxicity Test (Embryo

- Larval)

850.1025

– Oyster Acute Toxicity Test (Shell Deposition)

V. EN VIRONMENTAL JUSTICE

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 environmen tal 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 abou t 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) t he decision

- makers seek out and facilitate the involvement of those potentially affected. EPA has this goal for all communities and persons

across the United States.

To help address potential environmental justice issues, EPA seeks information on any gro ups or segments of the population who, as a result their location, cultural practices, or other factors, may have atypical, unusually high exposure to Chromobacterium subtsugae strain PRAA4

- 1 T

, compared to the general population. Please comment if you are aware of any subpopulations that may have atypical, unusually high exposure compared to the general population.

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

.Pseudomonas fluorescens s train CL145A Page 20 of 35 Biopesticides Registration Action Document

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 ma de:

(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 Pseudomonas fluores cens strain CL145A pesticide products have well

known properties. EPA has no knowledge that would contradict the claims made on the MBI

- 401

TGAI and MBI

- 401 EP label s, and such products are not expected to cause unreasonable adverse

effects on the environm ent when used according to the label instructions. Criterion 2 is satisfied by the current product label s, as well as the data and information presented in this document. It is

believed that the Pseudomonas fluorescens strain CL145A pesticide products will not cause any
unreasonable adverse effects on the environment, and MBI

- 401 EP (end

- use product) , in particular, is likely to control zebra and quagga mussels as claimed, satisfying criterion 3.

Criterion 4 is satisfied in that the Pseudomonas fluorescens strain CL145A pesticide products are not expected to cause unreasonable adverse effects when used according to label instructions. Therefore, MBI

- 401 TGAI and MBI

- 401 EP, containing Pseudomonas fluorescens strain

CL145A as a new active ingredient, are elig ible for registration under FIFRA section 3(c)(5) for

the labeled uses.

V I I

. ACTIONS REQUIRED BY THE REGISTRANT

A. Final Printed Labeling

Before releasing pesticide products containing Pseudomonas fluorescens strain CL145A for shipment, the registra nt is required to provide appropriate final printed labeling to EPA

.

B. Terms of Registration

As a term of the MBI

- 401 TGAI registration, the registrant must submit a storage stability study (OCSPP 830.6317) within 6 months of registration (by January 29 , 2012). For the MBI

- 401 EP registration, the registrant must submit storage stability (OCSPP 830.6317) and corrosion

characteristics (OCSPP 830.6320) data within 6 months of registration (by January 29, 2012).

C. Reporting of Adverse Effects and Hypers ensitivity Incidents

Notwithstanding the information stated in the previous sections, it should be clearly understoodPseudomonas fluorescens s train CL145A Page 21 of 35 Biopesticides Registration Action Document

that certain, specific data are required to be reported to 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 EPA under the provisions stated in FIFRA section 6(a)(2). Additionally, all incidents of hypersensitiv ity (including both suspected and confirmed incidents) must be reported to EPA under the provisions of 40 CFR § 158.2140(d).Pseudomonas fluorescens s train CL145A Page 22 of 35 Biopesticides Registration Action Document

VIII. GLOSSARY OF ACRONYMS AND ABBREVIATIONS

BPPD Biopesticides and Pollution Prevention Division

BRAD Biopesticides Regi stration Action Document

CFR Code of Federal Regulations

c fu colony

- forming unit (s) cfu/kg colony

- forming units per kilogram

cfu/mL colony

- forming unit s per milliliter

cP centipoise

EDSP Endocrine Disruptor Screening Program

EP end

- use product

EPA Environm ental Protection Agency (the “Agency”)

FFDCA Federal Food, Drug, and Cosmetic Act

FIFRA Federal Insecticide, Fungicide, and Rodenticide Act FQPA Food Quality Protection Act

FR Federal Register g/ mL gram per milliliter

LC 50 median lethal concentration. A s tatistically 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).

LD 50 median leth al 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). mg/kg milligram s per kilogram mg/L milligram s per liter

MP manufacturing

- use product

MRID No. Master Record Identification Number

NIOSH National Institute for Occupational Safety and Health

OCSPP Office of Chemical Safety and Pollution Prevent ion

OPP Office of Pesticide Programs

PC Code Pesticide Chemical Code

ppm parts per million

TGAI technical grade of the active ingredientPseudomonas fluorescens s train CL145A Page 23 of 35 Biopesticides Registration Action Document

I X. BIBLIOGRAPHY

A. Studies Submitted to Support the Pseudomonas fluorescens strain CL145A Pesticide

Product Registrations

476402

- 02 Kuhn J. 2008. Acute Oral Toxicity Study (UDP) in Rats : MOI 401 Pseudomonas fluorescens

– Final Report. Project Number: 11593/08. Unpublished study prepared by Stillmeadow, Inc., 10 pages.

476402

- 03 Kuhn J. 2008. Acu te Dermal Toxicity Study in Rats: MOI 401 Pseudomonas fluorescens

– Final Report. Project Number: 11594/08. Unpublished study prepared by Stillmeadow, Inc., 12 pages.

476402

- 04 Carter L. 2008. Acute Inhalation Toxicity Study in Rats: MOI 401 Pseudomonas f

luorescens

– Final Report. Project Number: 11595/08. Unpublished study prepared by Stillmeadow, Inc.

, 17 p ages

.

476402

- 05 Kuhn J. 2008. Acute Eye Irritation Study in Rabbits: MOI 401 Pseudomonas fluorescens

– Final Report. Project Number: 11596/08. Unpubl ished study prepared by Stillmeadow, Inc.

, 17 p ages

.

476402

- 06 Kuhn J. 2008. Acute Dermal Irritation Study in Rabbits: MOI 401 Pseudomonas fluorescens

– Final Report. Project Number: 11597/08. Unpublished study prepared by Stillmeadow, Inc.

, 10 p ages

.

4 76402

- 07 Monds K. 2008. Acute Intravenous Toxicity/Pathogenicity Study in Rats with a Microbial Pest Control Agent (MPCA): MOI 401 Pseudomonas fluorescens

– Final Report. Project Number: 12048/08. Unpublished study prepared by Stillmeadow, Inc., 27 pages.

476402

- 08 Fletcher C. 2008. Avian Acute Oral Toxicity Test with Killed Cells of Pseudomonas fluorescens (Pf

- CL145A) in Mallard Ducks ( Anas platyrhynchos ). Project Number: 212/003/04. Unpublished study prepared by Genesis Midwest Laboratories, 40 pages.

476402

- 09 Molloy D, Mayer D. 2008. Fish ( Salmo trutta ) Acute Toxicity Test Summary,

Freshwater: MOI 401 Pseudomonas fluorescens , Unpublished study prepared by Marrone Organic Innovations, Inc., 6 pages.

476402

- 10 Molloy D. 2008. Aquatic Invertebrate Acut e Toxicity Test, Freshwater Daphnids:

MOI 401 Pseudomonas fluorescens . Unpublished study prepared by Marrone Organic Innovations, Inc., 8 pages.

476402

- 16 Pitts K. 2008. Waiver Request for Microbial Pesticide Data Requirements: Pseudomonas fluorescens CL1 45A. Unpublished study prepared by Marrone Organic Innovations, Inc., 7 pages.Pseudomonas fluorescens s train CL145A Page 24 of 35 Biopesticides Registration Action Document

477494

- 03 Pitts K. 2009. Amended Waiver Request for Microbial Pesticide Data Requirements: MOI 401 TGAI. Unpublished study prepared by Marrone Organic Innovations, Inc., 8 pag es.

477494

- 05 Molloy D, Mayer D, Pitts K. 2009. Addendum to Ecological Toxicity Studies Regarding Live Pf Cells Contained in Each Treatment Dose: MOI 401 Pseudomonas fluorescens

– Final Report. Unpublished study prepared by Marrone Organic Innovations, In c., 6 pages.

479177

- 03 Mayer D. 2009. MOI 401 Pseudomonas fluorescens CL145A: Blue Mussel ( Mytilus edulis ) Acute Toxicity Tests. Project Number: MOI/401/TGAI. Unpublished study prepared by New York State Museum, 7 pages.

479177

- 05 Mayer D. 2009. MOI 401 Pseudomonas fluorescens CL145A: Daphnia magna Acute Toxicity Tests. Project Number: MOI/401/TGAI. Unpublished study prepared by New York State Museum, 15 pages.

479177

- 06 Mayer D. 2009. MOI 401 Pseudomonas fluorescens CL145A : Sunfish ( Lepomis

macrochirus ) Acute Toxicity Tests. Project Number: MOI/401/TGAI. Unpublished study prepared by New York State Museum, 9 pages.

479177

- 07 Mayer D. 2009. MOI 401 Pseudomonas fluorescens CL145A: Brown Trout ( Salmo trutta )

– Acute Toxicity Tests. Project Number: MOI/401/ TGAI. Unpublished study prepared by New York State Museum, 9 pages.

479 267

- 01 Koivunen M. 2009

. Product Chemistry for MOI

- 401 TGAI . Unpublished study

pr epared by Marrone Bio Innovations

, Inc., 187 p ages

.

479267

- 02 Koivunen M. 2009. Product Chemistry f or MOI

- 401 EP. Unpublished study prepared by Marrone Bio Innovations, Inc., 123 pages.

481619

- 01 Koivunen M. 20 10

. Updated Product Chemistry for MOI

- 401 TGAI

. Project

Number: MBI/M19/2009/V2. Unpublished study prepared by Marrone Bio

Innovations

, Inc., 4 4 pages.

482767

- 02 Monds K. 2010. Acute Pulmonary Toxicity/Pathogenicity Study in Rats: MOI

- 40 1

– Final Report. Project Number: 14246/10. Unpublished study prepared by Stillmeadow, Inc., 34 pages.

485133

- 01 Pitts K. 2011. Response to Tier I Micro bial Pesticide Data Requirements to MOI

401 EP. Unpublished study prepared by Marrone Bio Innovations, Inc., 11 pages.

B. Environmental Protection Agency Risk Assessment Memoranda

U.S. EPA. 20 1 0a

. Pseudomonas fluorescens strain CL145A / MOI

- 401

. M emorandum from J.V. Gagliardi , Ph.D. and J. L. Kough, Ph.D. to A

. Sibold dated April 27, 2010

.Pseudomonas fluorescens s train CL145A Page 25 of 35 Biopesticides Registration Action Document

U.S. EPA. 2010b. Pseudomonas fluorescens strain CL145A / MOI

- 401. Memorandum from J.V. Gagliardi , Ph.D. and J.L. Kough, Ph.D. to A. Sibold dated December 2 1, 2010.

U.S. EPA. 2011a. MOI

- 401 EP containing Pseudomonas fluorescens strain CL145A / MOI

- 401. Memorandum from J.V. Gagliardi, Ph.D. and J.L. Kough, Ph.D. to A. Sibold dated June 28, 2011.

U.S. EPA. 2011b. MOI

- 401 EP containing Pseudomonas fluorescens strain CL145A / MOI

- 401; Environmental Assessment for Limited Release from Confined Water Operations. Memorandum from M. Mendelsohn and G.S. Tomimatsu, Ph.D. to A. Sibold dated July 28, 2011.

C. Other References

Garrity GM, Bell JA, Lilburn T, editors. 2005. “Pseudomonadales” in Systematic Bacteriology. 2nd ed. New York (NY): Springer. Bergey’s Manual of

Murray PR, Baron E, Jorgensen JH, Pfaller MA, Yolken RH, editors. 2003. Manual of Clinical Biology. 8th ed. Washington (DC): ASM Press.

Organ isation for Economic Co

- operation and Development. 1997. Consensus Document on Information Used in the Assessment of Environmental Applications Involving

Pseudomonas . Available from http://www.rebeca

net.de/downloads/OECD%20Consensus%20document%20pseudomonas..pdf

.

U.S. EPA. 2004. Primer for Municipal Wastewater Treatment Systems. EPA 832

- R

- 04

- 001.

U.S. EPA. 1996. Microbial Pesticide Test Guidelines

– Bac kground for Residue Analysis of Microbial Pest Control Agents (OPPTS 885.2000). Available from http://www.epa.gov/ocspp/pubs/frs/publications/Test_Guidelines/seri es885.htm

.Pseudomonas fluorescens s train CL145A Page 26 of 35 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) /Manufacturing

- Use Product ( MP )
, MOI

- 401 T GAI (40 CFR § 158.2 120 )

OCSPP

G uideline

N umber Data Requirement Results MRID No. TGAI MP

885.1100 Product Identity Not applicable Submitt ed data fulfill the requirement for product identity. MOI

- 40 1 TGAI

contains 100

. 00 % by weight

Pseudomonas fluor escens strain CL145A cells and spent fermentation media (contains no less than 1.0 x

10 14 CFU/mL of Pseudomonas fluorescens strain CL145A cells)

. 479267

- 01

885.1200 Manufacturing Process Submitted data fulfill the requirement for manufacturing

process

. 4 79267

- 01

481619

- 01

Not a pplicable Deposition of a Sample in a Nationally Recognized Culture

Collection Pseudomonas fluorescens

strain CL145A is on deposit

with the American Type

Culture Collection in Manassas, Virginia under

Accession Number 55799

. Not ap plicable 479267

- 01

885.1300 Discussion of Formation

of Unintentional Ingredients Submitted data fulfill the requiremen t for discussion of formation of unintentional ingredients

. 479267

- 01

885.1400 Analysis of S amples Submitted data fulfill the requiremen t for analysis of samples. 479267

- 01

8 85 .1 50 0 Certification of L imits Not applicable Limits listed o n the

confidential statement of

formula are adequate/a cceptable. 479267

- 01

481619

- 01Pseudomonas fluorescens s train CL145A Page 27 of 35 Biopesticides Registration Action Document

TABLE 2. Product Analysis Data Requirements for the End

- Use Product ( E P), M OI

- 401 EP (40 CFR § 158.2120)

OCSPP Guideline Number Data Requirement Results MRID No.

885.1100 Product Identity Submitt ed data fulfill the requirement for product identity. M OI

- 401 EP contains

14

. 6 0 0 % by weight Pseudomonas fluore scens strain CL145A cells ( contains

no more than 1.0 x 10 5 CFU/mL of Pseudomonas fluorescens strain CL145A

cells ). 479267

- 02

885.1200 Manufacturing Process Submitted data fulfill the requirement for manufacturing process. 479267

- 02

Not applicable Deposi tion of a Sample in a Nationally Recognized Culture

Collection Not applicable Not applicable

885.1300 Discussion of Formation of Unintentional Ingredients Submitted data fulfill the requirement for

discussion of formation of unintentional ingredients. 479 267

- 02

885.1400 Analysis of Samples Submitted data fulfill the requirement for analysis of samples. 479267

- 02

885.1500 Certification of Limits Limits listed on the confidential statement of formula are adequate/acceptable. 479267

- 02

481619

- 01

TABLE 3

. Physical and Chemical Characteristics for the Technical Grade of the Active Ingredient (TGAI)/Manufacturing

- Use Product (MP), M OI

- 401 TGAI (40 CFR § 158.2120)

OCSPP

G uideline

N umber Data Requirement Results MRID No. TGAI MP

830.6302 Color Brown Not applicable 479267

- 01

830.6303 Physical State Liquid Not applicable 479267

- 01

830.6304 Odor Sweet, earthy Not applicable 479267

- 01

830.6313 Stability to Normal and

E levated Temperatures, Metals,

and Metal I ons Observations at 4°C and 25°C suggest th e TGAI is stable. Not applicable 479267

- 01

830.6317 Storage Stability Observations at 4°C and 25°C suggest the TGAI/MP is stable.

As a term of the M OI

- 401 TGAI registration, EPA will require the results of a one
- year storage stability study be

submitted w ithin six months (by January 29, 2012)

. 479267

- 01

481619

- 01

830.6319 Miscibility Not applicable Not required because MOI

- 401

TGAI is not an e mulsifiable form of microbial pesticide (refer to test note #2 of 40 CFR § 158.2120(d)). Not applicable

830.6320 Corrosion

Characteristics Not applicable Not corrosive to commercial packaging. 479267

- 01

830.7000 pH 7.0 Not applicable 479267

- 01

830.7100 Viscosity Not applicable 3.8 cP 479267

- 01

830.7300 Density/Relative Density/Bulk Density

(Specific Gravity) 1.0 1 g/mL Not applicable 479267

- 01Pseudomonas fluorescens s train CL145A Page 28 of 35 Biopesticides Registration Action Document

TABLE 4. Physical and Chemical Characteristics for the End

- Use Product (EP), M OI

- 401 EP (40 CFR § 158.2120)

OCSPP Guideline Number Data Requirement Results MRID No.

830.6302 * Color Brown 479267

- 02

830.6303* Physica l State Liquid 479267

- 02

830.6304* Odor Sweet, earthy 479267

- 02

830.6313 Stability to Normal and Elevated Temperatures, Metals, and Metal Ions Not applicable Not applicable

830.6317 Storage Stability No data available

. As a term of the

M OI

- 401 EP regist ration, EPA will require the results of a one
- year storage stability and corrosion characteristics study (ongoi ng) be

submitted within six months (by January 29, 2012)

. 479267

- 02

481619

- 01

830.6319 Miscibility Miscible in water 479267

- 02

830.6320 Corrosi on Characteristics Not expected to be corrosive to packaging

materials. As a term of the M OI

- 401 EP registration, EPA will require the

results of a one
- year storage stability and corrosion characteristics study (ongoing) be submitted within six

months (by January 29, 2012)

. 479267

- 02

481619

- 01

830.7000* pH 7.1 479267

- 02

830.7100 Viscosity 63 cP 479267

- 02

830.7300* Density/Relative Density/Bulk Density (Specific Gravity) 1.0 5 g/mL 479267

- 02

* According to 40 CFR § 158.2120, these data are only required f or the technical grade of the active ingredient. Since Marrone Bio Innovations included this information with its application for M OI

- 401 EP, it is summarized appropriately in this table

.Pseudomonas fluorescens s train CL145A Page 29 of 35 Biopesticides Registration Action Document

T ABLE 5

. Toxicology Data Requirements for the Technical Gra de of the Active Ingredient (TGAI) /Manufacturing

- Use Product (MP)
, M O I

- 401 TGAI (40 CFR § 158.2140)

OCSPP Guideline

N umber Data Requirement Results MRID N o.

TGAI M P

Tier I

885.3050 Acute Oral Toxicity/Pathogenicity Waived based o n the following rati onale: (1) Pseudomonas fluorescens strain

CL145A ’s natural occurrence in soil, in water, and on the surface and roots of a variety of plant types (including those of raw foods); (2) pseudomonads being considered a benign part of the regular human diet; and (3) the results of MRID

No

. 476402

- 02

. Classification: Acceptable Not applicable 477494

- 03

885.3150 Acute Pulmonary Toxicity/Pathogenicity An acute pulmonary toxicity and pathogenicity study demonstrated that Pseudomonas fluorescens strain CL145A

was not toxic, infective, and/or pathogenic to rats when dosed intratracheally at 3.4 x

10 8 CFU/rat. T his study partially replaces

MRID No. 477494

- 03 and upgrade s MRID

No. 476402

- 04 to Toxicity Category III. Classification: Acceptable Not applicable 482767

- 02

885.3200 Acute Injection Toxicity/Pathogenicity (Intravenous) An acute injection toxicity and pathogenicity (intravenous) study showed high mortality in the test animals and it was

unclear whether the Pseudomonas or a toxin was responsible for the mortali ty seen. A pattern of clearance for Pseudomonas fluorescens strain CL145A was demonstrated in the study. Given this, and lower mortality with freshly prepared test substance, lack of toxicity with autoclaved culture, and results from other toxicological

te sts, we conclude that there is adequate information to perform a risk assessment on Pseudomonas fluorescens strain CL145A.

Classification: Supplemental Not applicable 476402

- 07

885.3400 Hypersensitivity

Incidents No hypersensitivity incidents, includin g immediate

- type or delayed

- type reactions of humans and domestic animals that

occurred durin g research, development, or testing of the

TGAI /MP , were reported by the applicant. Any future hypersensitivity incidents must be reported per OCSPP Guideline

885. 3400. Not applicable

885.3500 Cell Culture Not required because Pseudomonas fluorescens

strain CL145A is not a virus (refer to test note #4 of 40 CFR § 158.2140(d)). Not applicable Not applicable

870.1100 Acute Oral Toxicity Not applicable NOAEL and LD 5 0 > 5,000 mg/kg (or > 2.35 x 10 10
CFU/kg) Classification: Acceptable

TOXICITY CATEGORY I V 476402

- 02Pseudomonas fluorescens s train CL145A Page 30 of 35 Biopesticides Registration Action Document

T ABLE 5

. Toxicology Data Requirements for the Technical Gra de of the Active Ingredient (TGAI) /Manufacturing

- Use Product (MP)
, M O I

- 401 TGAI (40 CFR § 158.2140)

OCSPP Guideline

N umber Data Requirement Results MRID N o.

TGAI M P

870.1200 Acute Dermal Toxicity Not applicable LD 50 > 5,050 mg/kg (or > 2.38

x 10 10 CFU/kg) Classification: Acceptable

TOXICITY CATEGORY I V 476402

- 03

870.1 300 Acute Inhalation Toxicity Not applicable NOAEL and LC 50 > 0.225 mg/L (or > 1.1 x 10 6 CFU/L) Classification: Acceptable

TOXICITY CATEGORY I I

(upgraded to TOXICITY

CATEGORY III with results

from MRID No. 482767

- 02) 476402

- 04

870.2400 Acute Eye Irritati on Not applicable MOI

- 401 TGAI was practically

non

- irritating to the eyes of

rabbits. Irritation symptoms cleared by 48 hours. Classification: Acceptable

TOXICITY CATEGORY I V 476402

- 05

870.2500 Primary Dermal

Irritation Not applicable MBI

- 203 TGAI was pra ctically

non

- irritating to the skin of

rabbits. Irritation symptoms cleared by 24 hours. Classification: Acceptable

TOXICITY CATEGORY I V 476402

- 06

Tiers II and III Not required for Pseudomonas fluorescens strain CL145A based on the lack of acute toxicit y/pathogenicity in the Tier I

studies.Pseudomonas fluorescens s train CL145A Page 31 of 35 Biopesticides Registration Action Document

TABLE 6 . Toxicology Data Requirements for the End

- Use Product (EP), M OI

- 401 EP (40 CFR § 158.2140)

OCSPP Guideline Number Data Requirement Results MRID No.

885.3050 Acute Oral Toxicity/Pathoge nicity Not applicable Not applicable

885.3150 Acute Pulmonary Toxicity/Pathogenicity Not applicable Not applicable

885.3200 Acute Injection Toxicity/Pathogenicity Not applicable Not applicable

885.3400 Hypersensitivity Incidents No hypersensitivity inci dents, including

immediate

- type or delayed

- type
reactions of humans and domestic

animals that occurred during research, development, or testing of the EP, were reported by the applicant. Any future hypersensitivity incidents must be reported per OCSPP Guid eline

885.3400. Not applicable

885.3500 Cell Culture Not applicable Not applicable

870.1100 Acute Oral Toxicity Waived based on the following rationale: (1) Pseudomonas fluorescens strain CL145A’s natural occurrence in soil, in water, and on the surface and roots of a variety of plant types (including those of raw foods); (2) pseudomonads being considered a benign part of the regular human diet; (3) the inert ingredients being present in diluted form and not expected to be of toxicological concern; and (4 ) the results of MRID No. 476402

- 02. Classification: Acceptable

TOXICITY CATEGORY I V 476402

- 16

870.1200 Acute Dermal Toxicity Waived based on the following rationale: (1) Pseudomonas fluorescens strain CL145A’s natural occurrence in soil, in water, and on the surface and roots of a variety of plant types (including those of raw foods); (2) potential for low dermal exposure; (3) the inert ingredients being present at low concentrations and not

considered to be dermal toxins or irritants; and (4) the resu lts of MRID No.

476402

- 03. Classification: Acceptable

TOXICITY CATEGORY IV 485133

- 01

870.1300 Acute Inhalation Toxicity Waived based on the following rationale: (1) Pseudomonas fluorescens strain CL145A’s natural occurrence in soil, in water, and on th e surface and roots of a variety of plant types (including those of raw foods); (2) the inert ingredients being present in diluted form and not expected to be of toxicological concern; and (3) the results of MRID No. 476402

- 04. Classification: Acceptabl e TOXICITY CATEGORY III 476402

- 16Pseudomonas fluorescens s train CL145A Page 32 of 35 Biopesticides Registration Action Document

TABLE 6 . Toxicology Data Requirements for the End

- Use Product (EP), M OI

- 401 EP (40 CFR § 158.2140)

OCSPP Guideline Number Data Requirement Results MRID No.

870.2400 Acute Eye Irritation Waived based on the following rationale: (1) Pseudomonas fluorescens strain CL145A’s natural occurrence in soil, in water, and on the surface and roots of a variety of plant types (including those of raw foods); (2) potential for low eye exposure; (3) the inert ingredients being present at low concentrations and not considered to be eye irritants at such concentrations; and (4) the results of

MRID No. 476402

- 05. Classification: Acceptable

T OXICITY CATEGORY IV 485133

- 01

870.2500 Primary Dermal Irritation Waived based on the following rationale: (1) Pseudomonas fluorescens strain CL145A’s natural occurrence in soil, in water, and on the surface and roots of a variety of plant types (including those of raw foods); (2) potential for low dermal exposure; (3) the inert ingredients being present at low concentrations and not

considered to be dermal irritants at such concentrations; and (4) the results of

MRID No. 476402

- 0 6

. Classification: Accep table

TOXICITY CATEGORY IV 485133

- 01Pseudomonas fluorescens s train CL145A Page 33 of 35 Biopesticides Registration Action Document

TABLE 7. Summary of Non

- Target Organism Studies for Irradiated and Live Cells of Pseudomonas fluorescens strain CL145A

MRID Number(s) Nontarget Species Tested Study Rating Treatments and Level Treatment

Time % Mortality Reported on Tested Nontarget Species % Mortality Reported on

Zebra Mussels

47917707

47640209

47749405 a Brown Trout Supplemental/unacceptable since

not tested at a maximum hazard dose and/or no biological endpoints were observed in the tested sp ecies at treatment levels of irradiated cells. However, the

study is scientifically sound. 100 ppm live Pf

- CL 145

A cells

100 ppm irradiated Pf

- CL

145 A cells via gamma

irradiator NYS

- DOH 0.84 kGy 2.3 x 10 2 cfu/ml 72 hour 39 days post treatment

@ 6° C

89 % w/ live cells

3% w/ irradiated cells

1% w/ untreated

control 11 days post treatment

@ 20°C

95% w/ live cells

97% w/ irradiated cells

4% w/ untreated control

68 days post treatment

@ 5°C

73% w/ live cells

71% w/ irradiated cells

0% w/ untreated control

47917706 Sunfish Supplemental/unacceptable since

not tested at a maximum hazard dose and/or no biological endpoints were observed in the tested species at treatment levels of irradiated cells. However, the

study is scientifically sound. 100 ppm live Pf

- C L 145 A cells and 100 ppm

irradiated Pf

- CL 145 A

cells.

Level of irradiation not reported. 72 hour 11 days post treatment

@ 20°C

47% w/ live cells

7% w/ irradiated cells

4% w/ untreated

control 11 days post treatment

@ 20°C

95% w/ live cells

96% w/ irr adiated cells

0% w/ untreated control

47640210

47917705

47749405 a Daphnia

magna Unacceptable since test material not defined in the study report and

not tested at a maximum hazard dose and/or no biological endpoints were observed in the tested species a t treatment levels. However, the study is scientifically sound. 200 ppm irradiated Pf

- CL

145 A cells via electron beam ionizing radiation

at 9.5 kGy 1.7 x 10 - 2
cfu/ml

Significantly Greater

Irradiation than MOI

Product 48 hour Daphnia

24 hour

Zebra

Mussel 8 days post treatment

@ ~ 23°C

5% w/ irradiated cells

6% w/ untreated

control 9 days post treatment

@ ~23°C

93% w/ irradiated cells

2% w/ untreated controlPseudomonas fluorescens s train CL145A Page 34 of 35 Biopesticides Registration Action Document

TABLE 7. Summary of Non

- Target Organism Studies for Irradiated and Live Cells of Pseudomonas fluorescens strain CL145A

MRID Number(s) Nontarget Species Tested Study Rating Treatments and Level Treatment

Time % Mortality Reported on Tested Nontarget Species % Mortality Reported on

Zebra Mussels

47917703 Blue Mussel (marine) Supplemental/unacceptable since

not tested at a maximum hazard

d ose and no biological endpoints

were observed in the tested species at treatment levels.

However, the study is scientifically sound. 100 ppm live Pf

- CL 145 A

cells 120 hours (5 days) 14 days post treatment

@ 20°C

1% w/ live cells

4% w/ untreated

control 14 days post treatment

@ 20°C

100% w/ live cells

0% w/ untreated control

47917704 Unionid

Mussel (fresh water) The study is classified as Supplemental because it is scientifically sound. However, it is Unacceptable for regulatory risk assessment purpose s, because no biological endpoints or adverse

effects were determined. Live CL 145 A cells at 100 or 200 ppm. 0%

- In four laboratory

tests and in a small mesocosm study using six species of

freshwater native

- NY unionid mussels, there

was no evidence of

adverse effects or mortality following

24

- , 48

- , or 72

- hr exposure periods to

live cells of Ps. fluorescens Strain

CL145A at 100 or 200

ppm. 98

- 100%

- Contrastingly, there were high mortalities of

zebra mussels ( Dreissena polymorpha ) at the same dosages f or 24

- or 48

- hr exposures to the bacteria.

47640208

47749405 a Mallard Ducks Supplemental. Upgradeable to acceptable upon submission of acceptable information re: method of inactivation and cell viability. Irradiated via electron beam ionizing radiation

at 9.5 kGy. 0.83 cfu/mg in a cell paste.

Significantly Greater

Irradiation than MOI

Product A 14 day limit dose by oral gavage of “killed”

cells of

Pf

- CL145A

demonstrated an acute

oral LD 50 and NOEL > 2,000 mg/kg.

a Italicized MRID No. 477494

- 05 cites it alicized text in the column labeled “Treatments and Level.” The summary style format in this document was determined Unaccept able because some of the reported data results conflicted with individual study reports.Pseudomonas fluorescens s train CL145A Page 35 of 35 Biopesticides Registration Action Document

APPENDIX B. PESTICIDE PRODUCTS

EPA Regi stration

Number Registration

Name Percentage

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

84059

- 4 M OI

- 4 0 1

TGAI 100.00% Technical N/A N/A N/A N/A

84059

- 5 MOI

- 401 EP 14.600% End Use

– Liquid Encl osed and

other confined static or flowing

water
infrastructures Injection using

chemical metering pumps Up to 0.026 ounce/gallon active ingredient for up to 24 hours

total per month Zebra and quagga mussels