BIOPESTICIDES REGISTRATION ACTION DOCUMENT

Pseudomonas fluorescens strain CL145A

Pesticide Chemical (PC) Code: 006533

U.S. Environmental Protection Agency

Office of Pesticide Programs

Biopesticides and Pollution Prevention Division

Version #2

July 29, 2011
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TABLE OF CONTENTS

I. OVERVIEW ....................................................................................................................... 4

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

III. REGULATORY BACKGROUND ................................................................................... 5

A. Pesticide Emergency Exemptions...........................................................................................................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 JUSTICE...................................................................................... 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 PESTICIDES DATA REQUIREMENTS............................ 26

APPENDIX B. PESTICIDE PRODUCTS............................................................................... 35
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BIOPESTICIDES REGISTRATION ACTION DOCUMENT TEAM

Office of Pesticide Programs (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. Environmental 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 Leader
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I. OVERVIEW

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 kills them. Given this bacterium’s molluscicidal capabilities, 

Marrone Bio Innovations proposed to register both a manufacturing-use pesticide product, MOI401 TGAI (EPA Reg. No. 84059-4), and an end-use pesticide product, MOI-401 EP (EPA Reg. 

No. 84059-5), containing 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 conveyance 

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 Federal 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 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, 

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 residential 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 use 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 for Pseudomonas fluorescens strain CL145A demonstrated that it is either 

Toxicity Category III or Toxicity Category IV (the lowest toxicity classifications), while an 

acceptable acute pulmonary toxicity/pathogenicity study indicated that Pseudomonas fluorescens

strain CL145A is not toxic, infective, and/or pathogenic. EPA believes it is unlikely that 

Pseudomonas fluorescens strain CL145A will cause significant adverse effects to nontarget 

organisms, when used 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 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. Thus, given that Pseudomonas 

fluorescens strain 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 fluorescens strain CL145A 

pesticide products without delay.
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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, 

providing 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 inform 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 Deposit: American Type Culture Collection in Manassas, Virginia 

under Accession Number 55799. 

OPP Chemical Code: 006533

Type of Pesticide: Microbial Pesticide – Molluscicide

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

use sites, application rates, methods of application, 

formulation types, and target pests) regarding

the registered pesticide products containing this 

active ingredient.

III. REGULATORY BACKGROUND

A. Pesticide Emergency Exemptions

On November 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, 2010, 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 the lower Colorado River to control invasive zebra and quagga 

mussels. The exemption expires on August 26, 2013. 
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B. Applications for Pesticide Registration

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

2121 Second 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 applications 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 strain 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 tolerance 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 requirements.

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

reviewers and are an indication of the usefulness of the information contained in the documents 

for risk assessment. A rating of “acceptable” indicates the study is scientifically sound and is 

useful for risk assessment. A “supplemental” rating indicates the data provide some information 

that can be useful for risk assessment. The studies may have certain aspects determined not to be 

scientifically acceptable (“supplemental: 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 

often state that the study is not required by 40 CFR Part 158. Both “acceptable” and 

“supplemental” studies may be used in the risk assessment process as appropriate. An 

“unacceptable” rating indicates that new data need to be submitted.

For the acute toxicity data requirements, Toxicity Categories are assigned based on the 

hazard(s) identified from studies and/or other information 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. 
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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 outstanding, 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. Human Health Assessment (40 CFR § 158.2140)

1. Toxicity

All toxicology data requirements for Pseudomonas 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 required for Pseudomonas fluorescens strain CL145A based on the lack of acute 

toxicity/pathogenicity in the Tier I studies.

For a comprehensive summary of the generic toxicology data requirements described in sections 

IV(B)(1)(a) and IV(B)(1)(b), as well as 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 (Office of Chemical Safety and Pollution Prevention (OCSPP) 

Guideline 885.3050; Master Record Identification Number (MRID No.) 477494-03): 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 occurring 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 (LD50) were greater than 5,000 milligrams 

per kilogram (mg/kg), corresponding to greater than 2.35 x 1010 colony-forming units per 

kilogram (CFU/kg) (Toxicity Category IV). Sufficient information was supplied and deemed 

acceptable. Thus, the requirement for acute oral toxicity/pathogenicity testing was waived.

Acute Pulmonary Toxicity/Pathogenicity (OCSPP Guideline 885.3150; MRID No. 482767-02): 

No animals died during this study, and there were no abnormalities seen during necropsy. 

Adverse effects (emaciation and reduced activity) were 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 substance treated to control groups. No signs of toxicity were noted in 

inactivated 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. Test 

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 108 colony-forming 
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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 showed 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 mortality 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 reactions 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 Culture (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;

 Reproductive 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 pesticide active 

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

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

designate.” The EDSP employs a two-tiered approach to making 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 where EPA 

will determine which, if any, of the Tier 2 tests are necessary based on the available data. Tier 2 

testing 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 and February 2010, EPA issued test orders/data call-ins for 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 certain post-application agricultural scenarios. This list 

should not be construed as a list of known or likely endocrine disruptors.
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Pseudomonas fluorescens strain CL145A is not among the group of 58 pesticide active 

ingredients on the initial list to be screened under the EDSP. Under FFDCA section 408(p), EPA 

must screen all pesticide 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. Federal Food, Drug, and Cosmetic Act (FFDCA) Considerations

Section 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 residue in or on a food) only if EPA 

determines 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 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 is 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 to pesticide 

residues. First, EPA determines the toxicity of pesticides. 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 section 408(b)(2)(D) of FFDCA, EPA has reviewed the available scientific data 

and other relevant information, and considered its validity, completeness, and reliability and the 

relationship of this information to human risk. EPA has 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 data and information discussed previously and 

presented in Table 5 in Appendix A, the data required for 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 residue in food and all other 

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

Dietary Exposure and Risk Characterization: Exposure to this microbial active ingredient 

through food is expected to be minimal. Pseudomonas fluorescens strain CL145A end-use 

products are not 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 limited to 

completely enclosed pipe or water conveyance systems or concrete chambers with defined inlets

or outlets. 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 

fluorescens strain CL145A. Concentrations of Pseudomonas fluorescens strain CL145A will be 

diluted as water flows past points of application and thus will rapidly decrease. Also, natural 

degradation (e.g., environmental factors such as ultraviolet light, nutrient depletion and bacterial 

grazing/predation by protists and others) and manmade filtering operations would greatly lower

the overall level of the pesticide after application (U.S. EPA 1996, 2004). Furthermore, 

Pseudomonas fluorescens is considered ubiquitous 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, which 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). 

Nonetheless, in the unlikely event that this microbial pesticide is present on food, the acute oral 

toxicity and pathogenicity data/information demonstrated that no toxicity, infectivity and/or 

pathogenicity is likely to occur with any exposure level of Pseudomonas fluorescens strain 

CL145A.

Drinking Water Risk Characterization: Much like food exposure, drinking water exposure is

expected to be negligible for similar reasons: (1) concentrations 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 will occur, and (3) 

flocculation and filtering at water treatment plants will further inactivate and decrease levels of 

Pseudomonas fluorescens strain CL145A (U.S. EPA 1996, 2004). Additionally, Pseudomonas 

fluorescens is already 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 levels of similar Pseudomonads already present in the human diet (Garrity et al. 

2005; Murray et al. 2003; OECD 1997). Nonetheless, in the unlikely event that this microbial 

pesticide is present on food, the acute oral toxicity and pathogenicity data/information 

demonstrated that no toxicity, infectivity and/or pathogenicity is likely to occur with 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.
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Although dermal 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, such 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 demonstrated 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 gardens, lawns, or buildings (i.e., residential areas), such

exposure would not exceed EPA’s level of concern given the 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 fluorescens 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 associated 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 information 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 the 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 

provision, 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.
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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 collectively, 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, pathogenic, 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 Risk 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 necessary 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 supported by the available no observed adverse effect level of greater than 0.225 

milligrams per liter (mg/L) and 1.1 x 106 CFU/L noted in the pulmonary toxicity study. 

Exposure to discharges of treated water from the treatment site is not expected to be significant.

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 unreasonable adverse effects to the U.S. population in general, and to infants 

and children in particular, will result when Pseudomonas fluorescens strain CL145A pesticide 

products are used in accordance with EPA-approved labeling.

C. Environmental Assessment (40 CFR § 158.2150)

1. Ecological Effects Hazards Assessment

Nontarget Organism Effects

Table 7 (see Appendix A) shows that Pseudomonas fluorescens strain CL145A had no effect at 

100 ppm on fish, aquatic invertebrates, fresh and salt water mussels, and mallard ducks. 

Exposures, however, were acute and the level of irradiation used on the test 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 nontarget studies, this irradiation was observed as having the effect of eliminating 

observed toxicity while maintaining efficacy against targeted zebra mussels. 
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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 EP 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 with zebra and/or quagga mussels. 

Treatment area must be a completely enclosed pipe or water conveyance system or concrete 

chamber with 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 water 

storage chambers of power plants, pump stations, irrigation systems, industrial and manufacturing 

facilities (e.g. automobile and steel), and dams. Treatment of open water, such as infested ponds, 

lakes, reservoirs, rivers and streams, or other unconfined 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-determined 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 consult EPA for further 

guidance and obtain EPA concurrence on application to the site prior to treatment. 

Type of facility 

Facility address 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 

database 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 location (and water body) 
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Application Information --

Treated system flow rate (e.g. gallon/day, m3

/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 scientifically validated source (e.g., state and federal agencies and local nonprofits 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 data 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 SOPs #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 environmental 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 perform additional downstream turbidity monitoring after every thirty minutes of 

any corrective action. If measurements continue to show an exceedance of the expected 

environmental concentration of 1 mg a.i./L after 1.5 hours, Marrone Bio Innovations or the 

applicator will turn off treatment metering pump. Any exceedance of the expected environmental 

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

Branch Chief within 24 hours by calling (703) 308-8269 and by email addressed to

reilly.sheryl@epa.gov.

On a quarterly basis, Marrone 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 discharge monitoring data will present the turbidity and Pseudomonas fluorescens 

concentrations during and after treatment. The turbidity correlation and application monitoring will

present data demonstrating the in-facility treatment concentration.
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Per SOP #MBI-G-20.16.1, Site 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)(l0-6 mg/L)

B = Release of material via blowdown water to surface water (kg/site/day)

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

T = Application time (minutes)

O = Treated system flow rate (e.g. gallon/day, m3

/hr) or volume (L)

Below are the three conditions under which data will 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), Marrone Bio Innovations will run the 

PDM model and provide 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

specific data from National Pollutant Discharge Elimination System (NPDES) 

permits to model chemical releases 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 concentration 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 aerial 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/opptintr/exposure/pubs/efast2man.pdf.
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2) For facilities discharging to flowing waters that currently 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, Marrone Bio Innovations will 

conduct and document mass balance calculations to determine the expected 

environmental concentration as follows:

CcFc + CRFR = CTFT

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

 0.026 oz. a.i./gallon (200 mg a.i./L).

Fc = Treated volumetric flow rate (Liters/hour, gallons/minute).

CR = Concentration of MOI-401 EP in the river prior to point discharge. This value

 should be zero as there should not be product already existing in the water.

FR = River volumetric flow rate prior to point discharge from cooling water systems

 (Liters/hour, gallons/minute).

CT = 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.

FT = 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 agencies 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), Marrone Bio 

Innovations will request diffusion and dilution models developed specifically for the 

facility and that have been certified to be representative of actual discharge dilution. 

Marrone 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 yards of the discharge point. 

3. Background Product Analysis 

a. P. fluorescens 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 northeastern 

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 killed 
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Pseudomonas fluorescens strain CL145A, while other epithelial cells in the mussel digestive 

gland and tissues elsewhere in the mussels appear unaffected. The registrant points out that the 

tubule epithelium is the “primary location for nutrient 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 Pseudomonas fluorescens strain CL145A cells. In submitted nontarget studies, this 

irradiation has the effect of eliminating observed toxicity while maintaining efficacy against 

target mussels. 

b. Pseudomonas fluorescens

Pseudomonas fluorescens is considered 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, 8th edition, states “Pseudomonas spp. have a worldwide distribution with 

a predilection 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 United 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 appear 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 

Schmidtke, 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 mykiss) 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..pdf
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mullet fish (Mugil curema) (Alvarez and Conroy, 1987). P. fluorescens 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 CFU/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., 1994) 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 plants

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, 1994). 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 chicory 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 Scala, 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 

cause 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 certain 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

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

fresh and salt water mussels, and mallard ducks. Exposures, 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 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 nontarget organism studies, this irradiation was observed as having the effect of 

eliminating observed toxicity in nontarget organisms while maintaining efficacy against targeted 

zebra mussels (Dreissena polymorpha).

It is unlikely that MOI-401 EP will cause substantial adverse effects to nontarget organisms 

during a one-year limited registration for use in completely enclosed pipe or water conveyance 

systems or concrete chambers 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 federally 

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 or below 100 ppm (Refer to SOP #MBI 20.16.1, July 26, 2011) and (3) 
Pseudomonas fluorescens strain CL145A Page 19 of 35

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treatment monitoring ensures that the measured concentration is at or below 1 ppm at a predetermined 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).

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 Marine 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. ENVIRONMENTAL JUSTICE

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

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

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

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

disproportionate share of the negative environmental consequences resulting from industrial, 

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

environmental programs and policies. Meaningful involvement means that (1) potentially 

affected community residents have an appropriate opportunity to participate in decisions about a 

proposed activity that will affect their environment and/or health; (2) the public’s contribution 

can influence the regulatory agency’s decision; (3) the concerns of all participants involved will 

be considered in the decision-making process; and (4) the decision-makers seek out and facilitate 

the involvement of those potentially affected. 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 groups 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 PRAA4-1T

, 

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 strain CL145A Page 20 of 35

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VI. RISK MANAGEMENT DECISION

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

following determinations are made:

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

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

 requirements of FIFRA;

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

 the environment; AND

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

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

 environment.

To satisfy criterion 1, the Pseudomonas fluorescens strain CL145A pesticide products have wellknown properties. EPA has no knowledge that would contradict the claims made on the MBI-401

TGAI and MBI-401 EP labels, and such products are not expected to cause unreasonable adverse 

effects on the environment when used according to the label instructions. Criterion 2 is satisfied 

by the current product labels, 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 eligible for registration under FIFRA section 3(c)(5) for 

the labeled uses. 

VII. ACTIONS REQUIRED BY THE REGISTRANT

A. Final Printed Labeling

Before releasing pesticide products containing Pseudomonas fluorescens strain CL145A for 

shipment, the registrant 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 Hypersensitivity Incidents

Notwithstanding the information stated in the previous sections, it should be clearly understood 
Pseudomonas fluorescens strain CL145A Page 21 of 35

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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 hypersensitivity 

(including both suspected and confirmed incidents) must be reported to EPA under the 

provisions of 40 CFR § 158.2140(d).
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VIII. GLOSSARY OF ACRONYMS AND ABBREVIATIONS

BPPD Biopesticides and Pollution Prevention Division

BRAD Biopesticides Registration Action Document

CFR Code of Federal Regulations

cfu colony-forming unit(s)

cfu/kg colony-forming units per kilogram

cfu/mL colony-forming units per milliliter

cP centipoise

EDSP Endocrine Disruptor Screening Program

EP end-use product

EPA Environmental Protection Agency (the “Agency”)

FFDCA Federal Food, Drug, and Cosmetic Act

FIFRA Federal Insecticide, Fungicide, and Rodenticide Act

FQPA Food Quality Protection Act

FR Federal Register

g/mL gram per milliliter

LC50 median lethal concentration. A statistically derived concentration of a 

substance that can be expected to cause death in 50% of test animals. It is 

usually expressed as the weight of substance per weight or volume of water, 

air, or feed (e.g., mg/L, mg/kg, or ppm).

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

to cause death in 50% of the test animals when administered by the route 

indicated (oral, dermal, or inhalation). It is expressed as a weight of 

substance per unit weight of animal (e.g., mg/kg).

mg/kg milligrams per kilogram

mg/L milligrams 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 Prevention

OPP Office of Pesticide Programs

PC Code Pesticide Chemical Code

ppm parts per million

TGAI technical grade of the active ingredient
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IX. 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. Acute 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 

fluorescens – Final Report. Project Number: 11595/08. Unpublished study 

prepared by Stillmeadow, Inc., 17 pages.

476402-05 Kuhn J. 2008. Acute Eye Irritation Study in Rabbits: MOI 401 Pseudomonas 

fluorescens – Final Report. Project Number: 11596/08. Unpublished study 

prepared by Stillmeadow, Inc., 17 pages.

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 pages.

476402-07 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 Acute 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 CL145A. Unpublished study prepared by Marrone 

Organic Innovations, Inc., 7 pages. 
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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 pages.

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, Inc., 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.

479267-01 Koivunen M. 2009. Product Chemistry for MOI-401 TGAI. Unpublished study 

prepared by Marrone Bio Innovations, Inc., 187 pages.

479267-02 Koivunen M. 2009. Product Chemistry for MOI-401 EP. Unpublished study 

prepared by Marrone Bio Innovations, Inc., 123 pages.

481619-01 Koivunen M. 2010. Updated Product Chemistry for MOI-401 TGAI. Project 

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

Innovations, Inc., 44 pages.

482767-02 Monds K. 2010. Acute Pulmonary Toxicity/Pathogenicity Study in Rats: MOI-401 

– Final Report. Project Number: 14246/10. Unpublished study prepared by 

Stillmeadow, Inc., 34 pages. 

485133-01 Pitts K. 2011. Response to Tier I Microbial Pesticide Data Requirements to MOI401 EP. Unpublished study prepared by Marrone Bio Innovations, Inc., 11 pages.

B. Environmental Protection Agency Risk Assessment Memoranda

U.S. EPA. 2010a. Pseudomonas fluorescens strain CL145A / MOI-401. Memorandum from 

J.V. Gagliardi, Ph.D. and J.L. Kough, Ph.D. to A. Sibold dated April 27, 2010.
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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 21, 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. 

Organisation for Economic Co-operation and Development. 1997. Consensus Document on 

Information Used in the Assessment of Environmental Applications Involving 

Pseudomonas. Available from http://www.rebecanet.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 – Background for Residue Analysis of 

Microbial Pest Control Agents (OPPTS 885.2000). Available from 

http://www.epa.gov/ocspp/pubs/frs/publications/Test_Guidelines/series885.htm.
Pseudomonas fluorescens strain CL145A Page 26 of 35

Biopesticides Registration Action Document

APPENDIX 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 TGAI (40 CFR § 158.2120)

OCSPP 

Guideline 

Number 

Data Requirement

Results

MRID No.

TGAI MP

885.1100 Product Identity Not applicable Submitted data fulfill the 

requirement for product 

identity. MOI-401 TGAI 

contains 100.00% by weight 

Pseudomonas fluorescens

strain CL145A cells and 

spent fermentation media

(contains no less than 1.0 x 

1014 CFU/mL of 

Pseudomonas fluorescens

strain CL145A cells).

479267-01

885.1200 Manufacturing Process Submitted data fulfill the requirement for manufacturing 

process.

479267-01

481619-01

Not applicable 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 applicable 479267-01

885.1300 Discussion of Formation 

of Unintentional 

Ingredients

Submitted data fulfill the requirement for discussion of 

formation of unintentional ingredients.

479267-01

885.1400 Analysis of Samples Submitted data fulfill the requirement for analysis of samples. 479267-01

885.1500 Certification of Limits Not applicable Limits listed on the 

confidential statement of 

formula are 

adequate/acceptable.

479267-01

481619-01
Pseudomonas fluorescens strain CL145A Page 27 of 35

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TABLE 2. Product Analysis Data Requirements for the End-Use Product (EP), MOI-401 EP (40 CFR § 158.2120)

OCSPP Guideline Number Data Requirement Results MRID No.

885.1100 Product Identity Submitted data fulfill the requirement for 

product identity. MOI-401 EP contains 

14.600% by weight Pseudomonas 

fluorescens strain CL145A cells (contains 

no more than 1.0 x 105 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 Deposition 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.

479267-02

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), MOI-401 TGAI (40 CFR § 158.2120)

OCSPP 

Guideline 

Number 

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 

Elevated

Temperatures, Metals,

and Metal Ions

Observations at 4°C 

and 25°C suggest the 

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 MOI-401 TGAI registration, EPA will 

require the results of a one-year storage stability study be 

submitted within six months (by January 29, 2012).

479267-01

481619-01

830.6319 Miscibility Not applicable Not required because MOI-401 

TGAI is not an emulsifiable 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)

1.01 g/mL Not applicable 479267-01
Pseudomonas fluorescens strain CL145A Page 28 of 35

Biopesticides Registration Action Document

TABLE 4. Physical and Chemical Characteristics for the End-Use Product (EP), MOI-401 EP 

(40 CFR § 158.2120)

OCSPP Guideline Number Data Requirement Results MRID No.

830.6302* Color Brown 479267-02

830.6303* Physical 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 

MOI-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

830.6319 Miscibility Miscible in water 479267-02

830.6320 Corrosion Characteristics Not expected to be corrosive to packaging 

materials. As a term of the MOI-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)

1.05 g/mL 479267-02

* According to 40 CFR § 158.2120, these data are only required for the technical grade of the active 

 ingredient. Since Marrone Bio Innovations included this information with its application for MOI-401 EP, it is

 summarized appropriately in this table.
Pseudomonas fluorescens strain CL145A Page 29 of 35

Biopesticides Registration Action Document

TABLE 5. Toxicology Data Requirements for the Technical Grade of the Active Ingredient (TGAI)/Manufacturing-Use 

Product (MP), MOI-401 TGAI (40 CFR § 158.2140)

OCSPP Guideline 

Number

Data Requirement Results MRID No.

TGAI MP

Tier I

885.3050 Acute Oral 

Toxicity/Pathogenicity

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; 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 

108 CFU/rat. This study partially replaces 

MRID No. 477494-03 and upgrades 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 mortality 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 

tests, 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, including immediate-type or 

delayed-type reactions of humans and domestic animals that 

occurred during 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 LD50 > 5,000 

mg/kg (or > 2.35 x 1010

CFU/kg)

Classification: Acceptable

TOXICITY CATEGORY IV

476402-02
Pseudomonas fluorescens strain CL145A Page 30 of 35

Biopesticides Registration Action Document

TABLE 5. Toxicology Data Requirements for the Technical Grade of the Active Ingredient (TGAI)/Manufacturing-Use 

Product (MP), MOI-401 TGAI (40 CFR § 158.2140)

OCSPP Guideline 

Number

Data Requirement Results MRID No.

TGAI MP

870.1200 Acute Dermal Toxicity Not applicable LD50 > 5,050 mg/kg (or > 2.38 

x 1010 CFU/kg)

Classification: Acceptable

TOXICITY CATEGORY IV

476402-03

870.1300 Acute Inhalation 

Toxicity

Not applicable NOAEL and LC50 > 0.225 

mg/L (or > 1.1 x 106 CFU/L) 

Classification: Acceptable

TOXICITY CATEGORY II 

(upgraded to TOXICITY 

CATEGORY III with results 

from MRID No. 482767-02)

476402-04

870.2400 Acute Eye Irritation Not applicable MOI-401 TGAI was practically 

non-irritating to the eyes of 

rabbits. Irritation symptoms 

cleared by 48 hours.

Classification: Acceptable

TOXICITY CATEGORY IV

476402-05

870.2500 Primary Dermal 

Irritation

Not applicable MBI-203 TGAI was practically 

non-irritating to the skin of 

rabbits. Irritation symptoms 

cleared by 24 hours.

Classification: Acceptable

TOXICITY CATEGORY IV

476402-06

Tiers II and III

Not required for Pseudomonas fluorescens strain CL145A based on the lack of acute toxicity/pathogenicity in the Tier I 

studies.
Pseudomonas fluorescens strain CL145A Page 31 of 35

Biopesticides Registration Action Document

TABLE 6. Toxicology Data Requirements for the End-Use Product (EP), MOI-401 EP (40 CFR § 158.2140)

OCSPP Guideline Number Data Requirement Results MRID No.

885.3050 Acute Oral 

Toxicity/Pathogenicity

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 incidents, 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 Guideline 

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 IV

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 results 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 the 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: Acceptable

TOXICITY CATEGORY III

476402-16
Pseudomonas fluorescens strain CL145A Page 32 of 35

Biopesticides Registration Action Document

TABLE 6. Toxicology Data Requirements for the End-Use Product (EP), MOI-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

TOXICITY 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. 476402-06. 

Classification: Acceptable

TOXICITY CATEGORY IV

485133-01
Pseudomonas fluorescens strain 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

47749405a Brown Trout

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-CL 145 

A cells

100 ppm irradiated Pf-CL 

145 A cells via gamma 

irradiator NYS-DOH 0.84 

kGy 2.3 x 102 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-CL 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/ irradiated cells

0% w/ untreated control

47640210

47917705

47749405a

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 at 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 control
Pseudomonas fluorescens strain 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 

dose and no biological endpoints 

were observed in the tested 

species at treatment levels. 

However, the study is 

scientifically sound.

100 ppm livePf-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 purposes, 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 for 24- or 48-hr 

exposures to the bacteria.

47640208

47749405a 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 LD50 and NOEL > 

2,000 mg/kg.

a Italicized MRID No. 477494-05 cites italicized text in the column labeled “Treatments and Level.” The summary style format in this document was determined Unacceptable 

because some of the reported data results conflicted with individual study reports.
Pseudomonas fluorescens strain CL145A Page 35 of 35

Biopesticides Registration Action Document

APPENDIX B. PESTICIDE PRODUCTS

EPA 

Registration 

Number

Registration 

Name

Percentage 

Active 

Ingredient

Formulation 

Type

Use Site(s) Method(s) of 

Application

Application Rate Target Pest(s)

84059-4 MOI-401

TGAI

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

84059-5 MOI-401 EP 14.600% End Use –

Liquid

Enclosed 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