Document ID: EPA-HQ-OPP-2009-0509-0007
Agency: epa
Document Type: Supporting & Related Material
Title: 
Posted Date: 2011-04-13T04:00Z

Docket Number EPA-HQ-OPP-2009-0509
                                       
                           Streptomyces Strain K61 
                      Registration Review Final Decision
                                   Case 6066
                                       
                                   PC 129069

                                  March 2011

          Approved by: ________________________                      
                                       
                          Keith A. Matthews, Director
                Biopesticides and Pollution Prevention Division
                                       
                Date:    ______________________               
                                       
                                       
                                       
                                       
                                       
                                       
                                       
                                       
                               Table of Contents
                      Streptomyces Strain K61 (Case 6066)
                                       
I.  INTRODUCTION	2
A.  Streptomyces Strain K61 General Background	2
B.  Streptomyces Strain K61 Registered Pesticide Products	2
II.  SCIENTIFIC ASSESSMENT	2
A.  Product Analysis (40 CFR § 158.2120)	2
B.  Human Health	2
1.	Acute Toxicity  -  Tier I (40 CFR § 158.2140)	2
2.	Subchronic Toxicity, Developmental Toxicity, Mutagenicity, and Immunotoxicity (Tier II); Chronic Testing (Tier III)	2
3.	FQPA Assessment:  Dietary Exposure and Risk Characterization	2
4.  Occupational Exposure and Risk Characterization	2
5.  Endocrine Disruptors	2
6.  Human Health Risk Characterization	2
C.  Environmental Assessment	2
1.  Effects on Nontarget Organisms  -  Tier I (40 CFR § 158.2060)	2
2.  Environmental Fate, Ecological Exposure, and Environmental Expression  -  Tiers II and III (40 CFR § 158.2060)	2
3.  Endangered Species Assessment	2
4.  Environmental Risk Characterization	2
D.  Product Performance (Efficacy) (40 CFR § 158.2070)	2
E.  Incidents	2
F.  Public Comments	2
G.  Water Quality	2
H.  Trade Irritants	2
I.  Environmental Justice	2
III.  FINAL REGISTRATION REVIEW DECISION	2
IV.  BIBLIOGRAPHY	2
V.  GLOSSARY OF ACRONYMS AND ABBREVIATIONS	2
VI.  APPENDIX	2
 

Streptomyces Strain K61  (Case 6066) Registration Review Team

Human Health and Environmental Effects 
Joel Gagliardi (BPPD)
Zigfridas Vaituzis (BPPD)

Risk Management
Anna Gross (BPPD)

Office of General Counsel
Philip Ross, Esq. 

I.  INTRODUCTION

 	This document is the Environmental Protection Agency's (EPA or the Agency) Final Registration Review Decision for Streptomyces Strain K61 and is being issued in accordance with 40 CFR §§ 155.57 and 155.58.  A registration review decision is the Agency's determination whether a pesticide meets, or does not meet, the standard for registration in the Federal Insecticide, Fungicide, and Rodenticide Act (FIFRA).  With regard to registered pesticides containing Streptomyces Strain K61, EPA determined that the FIFRA standard of registration has been met.  For further information on Streptomyces Strain K61, additional documents can be found in EPA's public docket (EPA-HQ-OPP-2009-0509) at www.regulations.gov.  

	FIFRA, as amended by the Food Quality Protection Act (FQPA) of 1996, mandated the continuous review of existing pesticides.  All pesticides distributed or sold in the United States must generally be registered by EPA, based on scientific data showing that they will not cause unreasonable risks to human health or the environment when used as directed on product labeling.  The registration review program is intended to make sure that, as the ability to assess risk evolves and as policies and practices change, all registered pesticides continue to meet the statutory standard of no unreasonable adverse effects to human health or the environment.  Changes in science, public policy, and pesticide use practices will occur over time.  Through the registration review program, the Agency periodically reevaluates pesticides to make sure that as change occurs, products in the marketplace can be used safely.  Information on this program is provided at: http://www.epa.gov/oppsrrd1/registration_review/. 

	In 2006, the Agency implemented the Registration Review program pursuant to FIFRA section 3(g).  EPA will review each registered pesticide every 15 years to determine whether it continues to meet the FIFRA standard for registration.   

	In accordance with 40 CFR § 155.50, the Agency formally initiated registration review for Streptomyces Strain K61 (Case 6066).  The following timeline highlights significant events that have occurred during the registration review of Streptomyces Strain K61: 

   * September 2009  -  Publication of the Streptomyces griseoviridis Strain K61 Summary Document in the docket (EPA-HQ-OPP-2009-0509) for a 60-day public comment period.  The Summary Document also included the preliminary work plan (PWP).  No comments were received.
   * October 2009  -  A request by the registrant, Verdera Oy, to change the name of the a.i. to Streptomyces K61 was approved as part of a product formulation amendment.  This change was made due to an inability to positively identify this species as Streptomyces griseoviridis.  Both the label and Confidential Statement of Formula (CSF) were updated with the revised a.i. name.  In order to harmonize both product registrations containing this a.i., Verdera Oy will also be required, as part of the registration process, to update the label and CSF for Mycostop(R) G Biofungicide (EPA Reg. No. 64137-9).  This change affects only the nomenclature of the a.i.  It does not have an impact on any of the previously completed risk assessments, which are still valid because all of the submitted studies were conducted using the same test material that is now registered as Streptomyces Strain K61. 
   * January 2010  -  Publication of the Streptomyces Strain K61 Final Work Plan, which stated that the most recent exposure and risk assessments still supported the registration of the currently registered pesticide products containing Streptomyces Strain K61 and that these products appeared to have met the requirement of registration review under 40 CFR § 155.50.  The Final Work Plan also documented the modification of the active ingredient name designation from Streptomyces griseoviridis Strain K61 to Streptomyces Strain K61.
   * July 2010  -  Publication of the Streptomyces Strain K61 Proposed Decision for a 60-day public comment period.  No comments were received.  The document proposed a final registration review decision affirming that the data on file continue to adequately support the registered pesticides containing Streptomyces Strain K61.
   * March 2011 - Publication of the Streptomyces Strain K61 Registration Review Final Decision, which formally determined that registered pesticides containing Streptomyces Strain K61 meet the current standards of registration under FIFRA.
   
	The information evaluated to support Streptomyces Strain K61, as published in the Summary Document, continues to support the active ingredient as summarized herein.  The status of this and other registration review cases is available at: http://www.epa.gov/oppsrrd1/registration_review/reg_review_status.htm.  Additional information on Streptomyces Strain K61 is available on the Biopesticides and Pollution Prevention Division's website in the form of a succinct fact sheet (http://www.epa.gov/oppbppd1/biopesticides/ingredients/factsheets/factsheet_129069.htm).  
			
  	On October 26, 2007, the Agency issued a Final Rule in the Federal Register (FR) on the data requirements to support registration of biochemical and microbial pesticides and updated definitions for both biochemical and microbial pesticides (72 FR 61002).  This rule became effective on December 26, 2007.  The data and information summarized below were considered in light of these requirements.  
	A.  Streptomyces Strain K61 General Background
	Streptomyces Strain K61 is a naturally occurring soil bacterium initially isolated from peat in Finland.  It is believed to act against disease-causing fungi in at least two ways:  by colonizing plant roots to deprive disease organisms of space and nourishment; and by producing antifungal compounds.  It is closely related to the bacterial species that produces the antibiotic streptomycin.  Streptomyces Strain K61 forms light colored colonies consisting of brown substrate mycelium, beige aerial mycelium, and long rectiflexible smooth spore chains.  As a microbial active ingredient, products containing Streptomyces Strain K61 are registered for control of seed, root and stem rot, and to prevent wilt of ornamentals, vegetables and tree and forest seedlings caused by Fusarium, Alternaria, and Phomopsis.  Streptomyces Strain K61 also suppresses root rots of Pythium, Phytophthora and Rhizoctonia in greenhouse plants and is used as a seed treatment for seed or soil borne damping off and early root rot.  Streptomyces Strain K61 is approved for use on all raw agricultural commodities and can be applied to seeds, soil, roots and transplants, or as a dip or spray.

      Based on data submitted to the Agency, Streptomyces Strain K61 is not expected to infect humans or to cause harmful effects if ingested.  EPA determined, however, that users may experience mild eye and skin irritation as well as adverse effects on lungs.  Current product labels specify required personal protective equipment (PPE).  Adverse human health or ecological risks are not expected as a result of exposure to this active ingredient when used as directed on product labels.  Moreover, the assessment conducted in the course of this Registration Review and included in the docket indicates that all data requirements were adequately addressed in the course of Streptomyces Strain K61 registration.  
      B.  Streptomyces Strain K61 Registered Pesticide Products
	There are currently two Streptomyces Strain K61 end use products (EPs) registered with EPA, both of which are listed in the table below.  There are no registered manufacturing use products (MUPs) for Streptomyces Strain K61.

                                 EPA Reg. No.
                               Registration Name
                                 Company Name
                                Current Status
% Streptomyces (a.i.)
                                    64137-5
                           Mycostop[(R)] Biofungicide
                                  Verdera Oy
                        Conditionally Registered (1993)
                                      35%
                                    64137-9
                          Mycostop[(R)] G Biofungicide
                                  Verdera Oy
                               Registered (2002)
                                      4%

II.  SCIENTIFIC ASSESSMENT

	A.  Product Analysis (40 CFR § 158.2120) 
      
      The data and information submitted to support applications for registration for the active ingredient Streptomyces Strain K61 were reassessed and found sufficient to fulfill current product chemistry data requirements (40 CFR § 158.2120) for all the use patterns described above.  Adequate product analysis data for Streptomyces Strain K61 were found in the submissions and reviews associated with the original registration of Mycostop[(R)] (EPA Reg. No. 64137-5) in 1993 and in the reassessment documents associated with this registration review.  A list of supporting product chemistry studies (including MRIDs) can be found within the `Studies Supporting Registration Review of Streptomyces Strain K61' in the bibliography.  Mycostop[(R)] G (EPA Reg. No. 64137-9) was determined to be substantially similar to Mycostop[(R)].  The manufacturer submitted new product chemistry data to support the product's registration but relied on previously submitted Mycostop[(R)] data to support all other data requirements.  A summary of the the specific physical and chemical characteristics of Streptomyces Strain K61 is found in Table 1 of the Appendix. 
      
	B.  Human Health

      1. Acute Toxicity  -  Tier I (40 CFR § 158.2140) 

      The acute toxicity data submitted with the original registration of Mycostop(R) (EPA Reg. No. 64137-5), as well as the justification to bridge to that data in support of the Mycostop(R) G (EPA Reg. No. 64137-9) registration, were reassessed and found sufficient to address current acute toxicity data requirements (40 CFR § 158.2140).  Acute toxicity results are discussed below.  A summary of the Streptomyces Strain K61 acute toxicity profile, categorized by guideline, is available in Table 2 of the Appendix.  A complete list of the acute toxicity studies can be found in the Bibliography.

   *    Acute Oral Toxicity/Pathogenicity (Office of Chemical Safety and Pollution Prevention (OCSPP) Guideline 885.3050)  -    Rats (21 males, 21 females):  MRID 418211-12.  The data show no clinically significant signs in rats.  Streptomyces Strain K61 was not detected in kidney, brain, liver, lungs, spleen, mesenteric lymph nodes, or in blood or urine samples and cleared from the feces and cecum following day two.  Necropsy studies showed no significant signs of abnormalities.  There were significant reductions in the weights of dosed males compared to controls at all study times and in the dosed females on day 8 and day 22.  In the absence of pyrogenic effects or any sign of infectivity, however, the test organism was not toxic, infective or pathogenic to rats.    

   *    Acute Injection Toxicity/Pathogenicity (OSCPP Guideline 885.3200)  -  Mice (38 males, 39 females):  MRID 418211-15.  An acute intraperitoneal injection of 5000 mg of the test article/Kg killed all males and females within 1 day of treatment.  1000 mg/Kg and 500 mg/Kg doses also caused some deaths within 2 days of dosing.  This indicates a toxic response to the test article rather than an infectious process, which was confirmed by the mortality caused by the heat-killed test article.  The toxin appears to be heat stable.  Adverse clinical signs returned to normal by day 5 in surviving mice except for one female with a distended abdomen and two with enlarged spleens.  The spleens showed no sign of the test article.  The LD50 for intraperitoneal injection was estimated to be 1306 mg/Kg for males, 870 mg/Kg for females and 1042 mg/Kg for both sexes combined.  

   *    Acute Pulmonary Toxicity/Pathogenicity (OCSPP Guideline 885.3150)  -  Rats (23 male, 23 female):  MRID 418211-14.  Generally, instillation of bacterial spores into rat lungs will result in a small number of deaths (which was seen in this study).  Also, large numbers of spores are usually recovered from the lungs shortly after dosing  -  with successively lower numbers of spores seen at later time intervals and concurrent increases in the spores isolated from the caecal contents  -  as a result of clearance from the lungs via the trachea and subsequent ingestion.  The present study was not typical in this regard.  The authors reported that microscopic examination of the dosing preparation showed densely packed inter-meshing mycelia in aggregations of 100 mm to 200 mm, whereas spores normally have dimensions of 1 mm x 2 mm.  They concluded that the size of the mycelial masses would not penetrate into the peripheral lungs and would be rapidly cleared from the airways by the pulmonary mucociliary escalator system.  Evidence regarding the infectivity/pathogenicity of the test organism is, therefore, inconclusive since only small numbers of viable organisms appeared to have reached the lower lungs.  Since there was no proliferation of the organism by the end of the study and because of the large particle size of the preparation, however, the authors concluded that Streptomyces Strain K61 would not present an infectious hazard under field conditions.  

   *    Acute Pulmonary Toxicity/Pathogenicity (OCSPP Guideline 885.3150)  -  Rats (200 male, 200 female):  MRIDs 424415-01, 424415-02.  Streptomyces Strain K61 caused death in 54% of treated male and 48% of treated female rats with 45% (male) and 40% (female) mortality occurring within the first 2 days after intratracheal instillation.  This high death rate may be due partially to the large size of the test organism; Streptomyces Strain K61 caused sufficient mortality to provoke concern about pulmonary exposure to significant amounts of the bacterium.  Dosage was 0.1 mL of a 3.46 x 10[8] CFU/mL suspension.  

   *    Acute Eye Irritation (OCSPP Guideline 870.2400)  -  Rabbits (6 males):  MRID 418211-16.  Small numbers of spores of Streptomyces Strain K61 were found in the treated eyes of all 6 rabbits at 24 and 72 hours post-treatment, but none thereafter.  Instillation of Streptomyces Strain K61 (>10[7] viable spores per eye) elicited transient, mild conjunctival irritation.  

   *    Acute Dermal Toxicity/Pathogenicity (OCSPP Guideline 885.3100)  -  Rabbits (5 males. 5 females):  MRIDs 418211-13, 422980-08.  Streptomyces Strain K61 was not toxic to rabbits when a single 2 g/Kg dose was administered dermally.  Rabbit body weights were not affected by an exposure of 2 g/Kg of Streptomyces Strain K61.  

   *    Primary Dermal Irritation (OCSPP Guideline 870.2500)  -  Guinea Pigs (20 females): MRID 418211-17.  Moderate reactivity (55% of responders) of guinea pig skin to Mycostop[(R)] (EPA Reg. No. 64137-5) was recorded on day 2 following challenge of sensitized animals.  While no determination was made of the amount of viable Streptomyces Strain K61 spores used in any of the animal exposures and the dose formulations records are inconsistent, the basic conclusion that the end use product is a skin sensitizer is still valid.  Mycostop[(R)] is a moderate skin sensitizer.  Dosage was 0.1 mL of a 3% w/v dilution of Mycostop[(R)] in distilled water. 
  
      2. Subchronic Toxicity, Developmental Toxicity, Mutagenicity, and Immunotoxicity (Tier II); Chronic Testing (Tier III) 

      Tier II and Tier III studies were not required for Streptomyces Strain K61 based on the lack of acute toxicity observed in the Tier I studies for the EPs.

      3. FQPA Assessment:  Dietary Exposure and Risk Characterization
         	
	Section 408(c)(2)(A)(i) of the Federal Food, Drug, and Cosmetic Act (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 it 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 the Agency 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.
      On the basis of acute toxicity data discussed above and summarized by guideline in Table 2 in the Appendix, the data requirements required for FQPA risk assessments for Streptomyces Strain K61 have been satisfied.
      
      a.  Aggregate Exposures
      
      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 non-occupational exposures, including drinking water from ground water or surface water and exposure through pesticide use in gardens, lawns, or buildings (residential and other indoor uses).

      i. Dietary Exposure and Risk Characterization
      
      Dietary exposure to Streptomyces Strain K61 is expected to be minimal.  As the mode of action of Streptomyces Strain K61 is through root colonization, the majority of applications are to seeds and soil.  Certain foliar applications are permitted for the purposes of suppressing Botrytis infection and promoting growth; however, direct applications to crops are highly diluted and residues are not expected to persist.  Regardless, any risk associated with dietary exposure is expected to be insignificant.  Streptomyces Strain K61 is a naturally occurring microbe found in soils throughout the world, and there are no known reports of any deleterious effects associated with its consumption.  Additionally, the acute toxicity data on file with the Agency confirm its lack of acute toxicity and support an exemption from the requirement of a tolerance (40 CFR § 180.1120).  

	ii. Drinking Water Risk Characterization

      Exposure to Streptomyces Strain K61 via drinking water when the pesticide is used is not likely to be greater than current/existing exposures.  Although Streptomyces Strain K61 is found naturally, it does not thrive in aquatic environments. There are no aquatic use sites for the pesticide, so exposure in drinking water is not expected. In addition, there is no evidence of adverse effects from oral, dermal, or inhalation exposure to this microbial agent (refer to the acute toxicity summaries above and in Table 2 of the Appendix).  Data on file with the Agency confirm the lack of oral toxicity/pathogenicity of Streptomyces Strain K61 and continue to support its exemption from the requirement of a tolerance.
      iii. Non-Occupational, Residential Risk Characterization 

      There are no residential uses for Streptomyces Strain K61.  Non-occupational exposures are not expected; in the event of accidental exposure, no non-occupational risks are anticipated.  The acute toxicity data on file with the Agency show a lack of acute toxicity for all routes of exposure.  
       
	b.  Cumulative Effects

      Section 408(b)(2)(D)(v) of the FFDCA requires the Agency to consider the cumulative effects of exposure to Streptomyces Strain K61 and to other substances that have a common mechanism of toxicity.  These considerations include the possible cumulative effects of such residues on infants and children.  No mechanism of toxicity in mammals has been identified for Streptomyces Strain K61; therefore, no cumulative effect with other related compounds is anticipated.  Because the data available demonstrate a low toxicity potential for Streptomyces Strain K61, the likelihood of adverse dietary effects is expected to be minimal.   
      
	c.  Determination of Safety for 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) requires EPA to apply an additional ten-fold 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, unless EPA determines that a different margin of safety will protect infants and children.  Margins of exposure (safety), which are often referred to as uncertainty factors, are incorporated into EPA's risk assessment either directly, through the use of a margin of exposure analysis, or by using uncertainty (safety) factors in calculating a dose level that poses no appreciable risk.

	Based on the available acute toxicity data for Streptomyces Strain K61, along with the expected exposure profile, EPA believes that there is reasonable certainty that no harm will result from aggregate exposure to the United States population, including infants and children, to the residues of Streptomyces Strain K61.  This includes all anticipated dietary exposures and all other exposures for which there is reliable information.  The Agency has arrived at this conclusion because the submitted data available on Streptomyces Strain K61 do not demonstrate significant toxic effects on mammals.  Thus, there are no threshold effects of concern and, as a result, the provision requiring an additional margin of safety does not apply.  Further, the considerations of consumption patterns, special susceptibility, and cumulative effects do not apply to pesticides without a demonstrated significant adverse effect.
  
      d.  Food Tolerance Exemptions

	On April 21, 1993, a Notice of Filing was published in the Federal Register for Streptomyces sp. strain K61: exemption from the requirement of a tolerance [50 FR 21403].  Under 40 CFR 180.1120, the biological pesticide Streptomyces sp. strain K61 is exempted from the requirement of a tolerance in or on all raw agricultural commodities when used as a fungicide for the treatment of seeds, cuttings, transplants, and plants of agricultural crops in accordance with good agricultural practices.  
			
	4.  Occupational Exposure and Risk Characterization

        Due to the results of the acute pulmonary toxicity study, EPA is concerned about repeated exposure of workers, farm personnel or other agriculturists to Streptomyces Strain K61.  Though the primary applications of Streptomyces Strain K61 are either soil-incorporated or seed treatments, foliar applications are permitted, which increases the probability of repeated respiratory exposure.  Mycostop[(R)] was also found to be a moderate skin sensitizer.  To mitigate the effects of occupational exposure to this a.i., standard personal protective equipment (PPE) is required by the labels for both Mycostop[(R)] and Mycostop[(R)] G, including long-sleeved shirt and long pants, waterproof gloves, and shoes plus socks.  In addition, a dust/mist respirator or a NIOSH-approved respirator is required by the labels along with a four hour restricted-entry interval (REI) for both products. 
      
      5.  Endocrine Disruptors
	As required by FIFRA and FFDCA, EPA reviews numerous studies to assess potential adverse outcomes from exposure to chemicals.  Collectively, these studies include acute, subchronic and chronic toxicity, including assessments of carcinogenicity, neurotoxicity, developmental, reproductive, and general or systemic toxicity. These studies include endpoints which may be susceptible to endocrine influence, including effects on endocrine target organ histopathology, organ weights, estrus cyclicity, sexual maturation, fertility, pregnancy rates, reproductive loss, and sex ratios in offspring.  For ecological hazard assessments, EPA evaluates acute tests and chronic studies that assess growth, developmental and reproductive effects in different taxonomic groups.  As part of its registration decision, EPA reviewed these data and selected the most sensitive endpoints for relevant risk assessment scenarios from the existing hazard database.  However, as required by FFDCA section 408(p), Streptomyces Strain K61 is subject to the endocrine screening part of the Endocrine Disruptor Screening Program (EDSP). 
      EPA has developed the 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 dose-response relationship between the dose and the E, A, or T effect.
      Under FFDCA section 408(p), the Agency must screen all pesticide chemicals.  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.  Streptomyces Strain K61 is not among the group of 58 pesticide active ingredients on the initial list to be screened under the EDSP.  Accordingly, EPA will issue future EDSP orders/data call-ins, requiring the submission of EDSP screening assays for Streptomyces Strain K61.  For further information on the status of the EDSP, the policies and procedures, the list of 67 chemicals, future lists, the test guidelines and the Tier 1 screening battery, please visit our website:  http://www.epa.gov/endo/.
		6.  Human Health Risk Characterization 

		The human hazard and exposure assessments for Streptomyces Strain K61 indicate that the risks to human health are negligible to non-existent when products containing Streptomyces Strain K61 are used in accordance with their respective labels.  These assessments are considered complete and current, and they satisfy the standards of registration review.  All microbial pesticide toxicology data requirements applicable to human health effects determinations for Streptomyces Strain K61 were considered and are fulfilled. 

      C.  Environmental Assessment

      1.  Effects on Nontarget Organisms  -  Tier I (40 CFR § 158.2060) 

      Based on data and information submitted to support applications for registration, as well as the associated assessments evaluating those data, all microbial pesticide nontarget organism and environmental fate data requirements for the Streptomyces Strain K61 were considered and are fulfilled.  Ecological data submitted for the registration of Mycostop(R) and cited for the registration of Mycostop(R) G are discussed below.  (Additional testing on nontarget insects, marine/estarine animals, and nontarget plants was waived during registration because the conditions under which such tests are required were not met.)  A summary of the Streptomyces Strain K61 ecological effects is available in Table 3 of the Appendix.  A complete list of the nontarget organism and environmental fate studies can be found in the bibliography.
      
   *    Avian Oral Toxicity/Pathogenicity (OCSPP Guideline 885.4050)  -  Mallard (50 birds in 10 treatment groups of 5 each): MRID 418211-20.  No mortalities occurred in any of the control groups (negative and sterile filtrate) or among the treated birds (2500 mg/Kg or approximately 2.45 x 10[9] cfu/Kg per day for five days).  All birds were normal in appearance and behavior throughout the test period.  There were no apparent effects on body weight or feed consumption between the control and the treated groups.  All birds were euthanized using carbon dioxide at the termination of the study and then subjected to gross necropsy.  The results were not found to be remarkable except for two male birds in the control groups which had single plaques in their air sacs.  In the treatment group, one male bird was small, with reduced muscle mass and feather loss on the rump.  Two males and one female were found to have plaques on their air sacs, and a second female had plaques on and in the lungs and a somewhat pale spleen.  The study is scientifically sound and demonstrated an LD50 > 2,500 mg/Kg.  This indicates that Streptomyces Strain K61 is not toxic or pathogenic to waterfowl.  The study fulfils EPA Guideline requirements for an avian oral pathogenicity/toxicity test.  

   *    Avian Oral Toxicity/Pathogenicity (OCSPP Guideline 885.4050)  -  Northern Bobwhite (50 birds in 10 treatment groups of 5 each): MRID 418211-19.  No mortalities occurred in either of the control groups (negative or attenuated) and 2 occurred among the Streptomyces treated birds (2500 mg/Kg or approximately 2.45 x 10[9] cfu/Kg per day for five days).  Due to the nature of lesions observed in the necropsy, neither death was considered to be treatment-related.  All other birds were normal in appearance and behavior throughout the test period, except for one bird that exhibited limping, lethargy and a ruffled appearance.  These conditions were not thought to be treatment-related.  There were no apparent effects on body weight or feed consumption between the control and the treated groups.  All birds were euthanized using carbon dioxide at the termination of the study and then subjected to gross necropsy.  The results were not found to be remarkable.  The study was scientifically sound and demonstrated an LD50 > 2,500 mg/Kg.  This indicates that Streptomyces Strain K61 is not toxic or pathogenic to upland game birds.  The study fulfilled EPA Guideline requirements for an avian oral pathogenicity/toxicity test.  

   *    Freshwater Fish Toxicity/Pathogenicity (OCSPP Guideline 885.4200)  -  Rainbow trout (80 fish): MRID 418211-21.  Five-gallon glass jars holding 15 liters of solution were used as the exposure chambers.  Groups of 10 fish were exposed to a control (water) and five test concentrations, plus a killed-spore concentration equivalent to the high test level.  One hundred percent mortality occurred in the killed spore group (4.0 x 10[4] CFU/mL) at day 16, with 70% and 90% mortality in the 2.0 x 10[4] CFU/mL group at day 25 and 26 respectively.  Due to sporadic mortality, an LC50 could not be calculated but was estimated to be greater than 40.8 mg/L (4.0 x 10[8] CFU/mL) at 7, 14, and 21 days and greater than 10.2 mg/L at 30 days.  A NOAEC was determined to be 5.1 x 10[3] CFU/mL and there was no observed infectivity after 30 days.  The possibility of toxicity was indicated by the killed spore and the 2.0 x 10[4] test concentration results, but a lack of toxicity at the 4.0 x 10[4] test concentration made this interpretation questionable. (Problems with nonspecific toxicity when testing fungal preparations have been observed in the past and have been related to the autoclaving process used to kill the fungal spores for the killed spore control.)  All trout were normal in appearance and behavior throughout the test period.  The dissolved oxygen ranged from 8.0 to 10.2 mg/L in the renewed solutions and 4.8 to 10.6 mg/L in the expired solutions, pH ranged from 7.6 to 8.4 in the renewed solutions and 7.1 to 8.3 in the in the expired solutions, and the temperature ranged from 11 to 13°C in both solutions.  This study showed sporadic fish mortality and failed to demonstrate a definitive LC50 value due to the absence of a dose-response trend with the observed mortality.  The study results were not adequate for making a risk assessment.  

   *    Freshwater Fish Toxicity/Pathogenicity (OCSPP Guideline 885.4200)  -  Rainbow trout (70 fish): MRID 430903-01.  The 30-day static renewal toxicity/pathogenicity study of Mycostop[(R)] (EPA Reg. No. 64137-5) to rainbow trout (Oncorhynchus mykiss) was conducted by Kemira Agro Oy.  Single replicates of 10 fish each were exposed to Streptomyces Strain K61 concentrations of 0.25 x 10[5], 0.50 x 10[5], 1.0 x 10[5], 2.0 x 10[5], and 4.0 x 10[5] CFU/mL in dilution water, plus a dilution water control and a killed-spore treatment of 4.0 x 10[5] CFU/mL.  The dietary dose in the food offering was 10% of the water exposure concentration.  Water chemistry measurements of dissolved oxygen, pH, and temperature indicated that overall, adequate water quality was maintained throughout the term of the experiment.  Gentle aeration was supplied to all test chambers to maintain the dissolved oxygen levels.  The day 1, day 2, day 3, and day 4 LC50 values were 2.8 x 10[5], 1.6 x 10[5], 1.2 x 10[5], and 0.8 x 10[5] CFU/mL, respectively.  The concentration-response line slopes for days 1-4 were 15, 7.7, 7.7, and 7.7, respectively.  The slope of the day 5 through day 30 concentration-response line was 15.  The 30-day NOAEC was 0.50 x 10[5] CFU/mL based on the lack of mortality, statistically significant growth effects, and sublethal/abnormal effects at this concentration.  All surviving fish ate normally and showed an increase in length and weight on day 30 as compared to the representative group on day 0.  No lesions or tissue discolorations were noted in any surviving fish at the end of the study.  

   *    Freshwater Aquatic Invertebrate Toxicity/Pathogenicity (OCSPP Guideline 885.4240)  -  Daphnia (100 Daphnia magna in 5 treatment groups): MRID 418211-22.  A 21-day static renewal study was conducted to determine the toxicity and/or pathogenicity of Streptomyces Strain K61 spores to Daphnia magna.  A control group was tested against four treatment groups, including a maximum nominal dose exposure of 1.0 x 10[6] CFU/mL and low and median doses of 10[4] and 10[5] CFU/mL, plus a dose of 1.0 x 10[6] CFU/mL of heat-killed spores.  No significant differences (P>0.05) were found between the control and the exposure levels for reproduction and time to first brood.  All of the offspring produced during the study appeared normal.  Mean survival was 100% for control and low level concentration, 75% for the middle concentration, and 0% for the high concentration and killed spore treatment.  Mortality from killed spore treatments has been reported from previous studies and is thought to be caused by creating a toxic environment through the autoclaving process, which could have contributed to the effects observed in this test.  A day 21 EC50 was calculated to be approximately 190 mg/L (1.9 x 10[5] CFU/mL).  Daphnid growth was found to be significantly affected at the middle concentration (1 x 10[5] CFU/mL).  The NOAEC was found to be 1.0 x 10[4] CFU/mL, the LOEC was 1.0 x 10[5] CFU/mL, and the MATC was found to be 3.2 x 10[4] CFU/mL.  The mean temperatures of the test solutions were 20.6 + 0.5ºC for the old and 20.5 + 0.5ºC for the new solutions.  The pH values for the renewed solutions ranged from 7.8 to 8.5 (mean = 8.3 + 0.2) and for the expired solutions ranged from 4.8 to 8.8 (mean = 8.2 + 0.9).  Dissolved oxygen concentrations in the renewed solutions ranged from 8.0 to 8.5 mg/L (94% to 100% saturation) at 21ºC (mean = 8.3 + 0.2 mg/L) and in the expired solutions ranged from 0.7 to 9.5 mg/L (8% to 112% saturation) at 21ºC (mean = 7.1 + 2.4 mg/L).  The study is scientifically sound and demonstrated an EC50 = 190 ppm (1.9 x 10[5] CFU/mL).  This indicates that Streptomyces Strain K61 is not toxic or pathogenic to Daphnia magna.  The study fulfils EPA Guideline requirements for an acute toxicity test for an aquatic invertebrate.

   *    Toxicity/pathogenicity to Honey Bees (OCSPP Guideline 885.4380)  -  (25 bee pupae per treatment): MRID 418211-23.  Three treatment levels, representing 240, 760 and 2400 ppm at a specific activity of 9.8 x 10[8] cfu/g of diet, were tested along with an attenuated control and a negative control.  Two replicates were tested at each concentration with 25 bees per replicate.  Mortalities occurred in both of the control groups (negative and attenuated) and in all 3 of the treatment groups.  The mortalities in the negative and attenuated control groups were 30% and 62%, respectively, while those in the 240, 760, and 2400 ppm concentrations averaged 20%, 26% and 28%, respectively.  The pattern of mortality was not found to be dose responsive and did not appear to be treatment-related.  The LC50 was determined to be greater than 2400 ppm and the NOAEC was 2400 ppm, which was the highest concentration tested.  Due to nonspecific mortality in the controls, however, the study is considered supplemental.  The results were reliable enough to indicate that MYCOSTOP[(R)] is not toxic or pathogenic to the honey bee.  This study also satisfies the data requirements for nontarget insect testing (OCSPP Guideline 885.4340).   
      2.  Environmental Fate, Ecological Exposure, and Environmental Expression  -  Tiers II and III (40 CFR § 158.2060)  

	Environmental fate data requirements (i.e., Tiers II, III, and IV) were not triggered for Streptomyces Strain K61 because results of the nontarget toxicity/pathogenicity Tier I studies did not demonstrate hazard to nontarget organisms.
      
      3.  Endangered Species Assessment 

      Through a re-evaluation of the ecological effects data and EPA reviews of Streptomyces Strain K61, the Agency has determined that adverse effects will not occur to nontarget organisms as a result of the labeled applications of Streptomyces Strain K61.  In addition, Streptomyces Strain K61 is a naturally occurring soil bacterium that suppresses the growth of plant pathogenic fungi.  Reduction or loss of these fungi will not affect habitat or other resources required by threatened and endangered species.  

      Since EPA has determined that no effects are anticipated for any nontarget species exposed to Streptomyces Strain K61 as a result of the labeled applications, effects to federally-listed threatened and endangered species and their designated critical habitats are also not expected. As a result of these analyses, the Agency has determined that registered uses of Streptomyces Strain K61 will have "no effect" on endangered or threatened terrestrial or aquatic species as listed by the U.S. Fish and Wildlife Service (USFWS) and the National Marine Fisheries Services (NMFS) and will not destroy or adversely modify designated critical habitat.  The `Endangered Species Assessment for Streptomyces Strain K61' can be found in the Registration Review docket EPA-HQ-OPP-2009-0509.

      4.  Environmental Risk Characterization

		Based on the reviews of the aforementioned nontarget organism and environmental fate information, and the endangered species assessment, the Agency has determined that unreasonable adverse effects to nontarget organisms or the environment are not likely to result when products containing Streptomyces Strain K61 are used in accordance with their respective labels.  These assessments are considered complete and current, and they satisfy the standards of registration review. 

      D.  Product Performance (Efficacy) (40 CFR § 158.2070)

	Product performance data must be developed for all microbial pesticides to ensure efficacy when used in accordance with labeling directions and commonly accepted pest control practices.  The Agency waived all requirements to submit such efficacy data unless the pesticide product bears a claim to control public health pests, although the Agency reserves the right to require, on a case-by-case basis, submission of efficacy data for any pesticide product registered or proposed for registration.  The Streptomyces Strain K61 pesticide products are fungicides intended for agricultural use and do not have labeled uses for public health pests.  The Agency has not required submission of efficacy data to support any of the claims on the current labels.  

 	E.  Incidents

	Two instances of hypersensitivity were reported by Verdera Oy prior to EPA registration.  The first incident was reported in 1986, the details of which are included in MRID No. 418211-18.  One part-time summer trainee at the Espoo Research Centre in Finland had symptoms of a mild alveolar reaction to Mycostop(R) (EPA Reg. No. 64137-5).  The alveolar reaction could have been due to exposure to biological dust from Mycostop[(R)], though there were indications that the incident might have been avoided by careful use of the safety equipment that was recommended and provided.  In 1991, a research associate in a lab in Fort Collins, CO, experienced dizziness, faintness, difficulty breathing and dilated pupils after treating seed with Mycostop[(R)].  The individual had a history of allergies and asthma and was not wearing the required face mask when exposed.  The symptoms disappeared in approximately one hour, and no medical attention was sought.  These incidents do not have any bearing on the hazard or risk assessed pertaining to the use of registered Streptomyces Strain K61 pesticide products in accordance with their product labels.  Both individuals recovered completely, and no other employees working with Mycostop(R) have reported any problems.  Label language on Streptomyces Strain K61 products is appropriate and sufficient to protect users/handlers of these products.  

The Office of Pesticide Programs' Incident Data System (IDS) indicates no other reports of adverse human, domestic animal, or environmental incidents for products containing Streptomyces Strain K61. In addition, no incidents were reported in response to the Agency request for such information in the Streptomyces griseoviridis Preliminary Work Plan and Summary Document. 
      
      F.  Public Comments

      In accordance with 40 CFR § 155.50, the Agency formally initiated registration review for Streptomyces Strain K61 in September 2009, with the opening of a docket (Docket Number EPA-HQ-OPP-2009-0509) and the issuance of a Summary Document for a 60-day public comment period.  The Agency received no comments in response to the Streptomyces griseoviridis Summary Document and the initial opening of the docket.  A Federal Register (FR) Notice, published on July 28, 2010, announced the placement of the Streptomyces Strain K61 Proposed Decision document into the docket, and initiated a 60-day public comment period to allow for consideration of any new information in advance of a Final Decision.  EPA did not receive any comments during the 60-day public comment period for the Streptomyces Strain K61 registration review proposed decision. 

	G.  Water Quality

	Streptomyces Strain K61 has not been identified as a cause of impairment for any water bodies listed as impaired under Section 303(d) of the Clean Water Act, based on information provided at: http://iaspub.epa.gov/tmdl_waters10/attains_nation_cy.cause_detail_303d?p_cause_group_id=885.  In addition, no Total Maximum Daily Loads (TMDLs) have been developed for Streptomyces Strain K61, based on information provided at:http://iaspub.epa.gov/tmdl_waters10/attains_nation.tmdl_pollutant_detail?p_pollutant_group_id=885&p_pollutant_group_name=PESTICIDES.  More information on impaired water bodies and TMDLs can be found at http://www.epa.gov/owow/tmdl/.  The Agency solicited comments when the Summary Document was published in September 2009; however, no comments, data, or information regarding the existence of any water quality issues associated with Streptomyces Strain K61 were received. 

      H.  Trade Irritants

	Trade irritants are not expected for Streptomyces Strain K61.  Through the registration review process, the Agency is soliciting information on trade irritants and, to the extent feasible, will take steps toward facilitating irritant resolution.  Growers and other stakeholders are asked to comment on any trade irritant issues resulting from lack of Maximum Residue Levels (MRLs) or disparities between U.S. tolerances or exemptions from tolerance and MRLs or exemptions from MRLs in key export markets, providing as much specificity as possible regarding the nature of the concern.  Trade irritants are not expected for Streptomyces Strain K61because there are no U.S. tolerances for Streptomyces Strain K61, nor are there Codex MRL's.  The Agency did not receive any comments regarding the existence of any trade irritant issues associated with Streptomyces Strain K61 following issuance of the Summary Document.  
 
  	I.  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 -- in the development, implementation, and enforcement of environmental laws, regulations, and policies.  To help address potential environmental justice issues, the Agency sought information, as explained in the Summary Document, on groups or segments of the population who, as a result of their location, cultural practices, or other factors, may have atypical, unusually high exposure to registered pesticide products containing Streptomyces Strain K61 compared to the general population.  The Agency did not receive any comments.  At this time, EPA does not believe that use of the registered pesticide products containing Streptomyces Strain K61 will cause harm to or a disproportionate impact on at-risk communities.  

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

III.  FINAL REGISTRATION REVIEW DECISION

	The Agency has determined that no additional data are required at this time to support current registrations containing Streptomyces Strain K61.  The Agency has considered Streptomyces Strain K61 in light of the standard for registration and safety factors in FIFRA and FFDCA, as amended by FQPA.  EPA has found that there are not likely to be any unreasonable adverse effects to the U.S. population in general, and to infants and children in particular, or to nontarget organisms or the environment from the use of registered pesticide products containing Streptomyces Strain K61 when currently required label instructions are followed.  In addition, EPA has made a "No Effect" determination for endangered and threatened species, and their designated critical habitat, for Streptomyces Strain K61.  The Agency has found that it is not necessary to conduct a new risk assessment for this case and is, therefore, issuing a final decision in accordance with 40 CFR Section 155.53(c)(2) and 40 CFR Section 155.58.    

	In accordance with 40 CFR §§ 155.57 and 155.58, the Agency's final registration review decision is that the standards for Registration Review have been met and that the registrations of the two products containing Streptomyces Strain K61 be maintained.  

IV.  BIBLIOGRAPHY

      A. Studies Supporting Registration Review of Streptomyces Strain K61
41821100
Kemira Oy (1991) Submission of Chemistry and Toxicity Data in Support of Mycostop New Application Registration, and Petition for an Exemption from the Requirement of Tolerance. Transmittal of 23 studies.
41821101
Pulkkanen, H. (1990) Product Identity and Disclosure of Ingredients: Mycostop. Unpublished study prepared by Kemira Oy. 10 p.
41821102
Bjorkqvist, B.; Leppanen, L. (1989) Determination of the Active Ingredient of Streptomyces griseoviridis in Cucumber and Cabbage. Unpublished study prepared by Keira Oy. 10 p.
41821103
Bjorkqvist, B.; Leppanen, L.; Lahdenpera, M. (1990) Determination of Heptaenic Antibiotic Residues in Cucumber Fruit and Dill after Mycostop Treatment. Unpublished study prepared by Kemira Oy. 12 p.
41821104
Raatikainen, N. (1990) Determination of the Antifungal Antibiotic Produced by Mycostop in the Peat. Unpublished study prepared by University of Kuopio. 16 p.
41821105
Tuomisto, J. (1989) Summary Research Report. Unpublished study prepared by University of Kuopio. 8 p.
41821106
Pulkkanen, H. (1990) Manufacturing Process: Mycostop. Unpublished study prepared by Kemira Oy. 8 p.
41821107
Pulkkanen, H. (1990) Unintentional Ingredient Discussion: Mycostop. Unpublished study prepared by Kemira Oy. 7 p.
41821108
Pulkkanen, H. (1991) Mycostop (Streptomyces Griseoviridis): Determination of Viability. Unpublished study prepared by Kemira Oy. 6 p.
41821109
Pulkkanen, H. (1990) Certification of Limits: Mycostop. Unpub- lished study prepared by Kemira Oy. 7 p.
41821110
Snelson, A. (1990) Determination of Color, Physical State, Odor, Density, and pH of Mycostop: Lab Project Number: CO8074: 2. Unpublished study prepared by IIT Research Institute. 19 p.
41821111
Pulkkanen, H. (1991) Mycostop (Streptomyces Griseoviridis): Determination of Storage Stability. Unpublished study prepared by Kemira Oy. 6 p.
41821112
Hossack, D.; Baker, M.; Denton, S. (1989) Acute Oral Toxicity and Infectivity/Pathogenicity to Rats of Streptomyces griseoviridis (Mycostop): Lab Project Number: 89953D/RKY 114/0/AC. Unpub- lished study prepared by Huntingdon Research Centre Ltd. 28 p.
41821113
Liggett, M. (1989) Acute Dermal Toxicity to Rabbits of Streptomyces griseoviridis, (Mycostop): Lab Project Number: 89892D/RKY 114/1/ AC. Unpublished study prepared by Huntingdon Research Centre Ltd. 12 p.
41821114
Hossack, D.; Baker, M.; Denton, S. (1989) Acute Pulmonary Toxicity and Infectivity/Pathogenicity to Rats of Streptomyces Griseo- viridis (Mycostop): Lab Project Number: 89954D/RKY 114/0/AC. Unpublished study prepared by Huntingdon Research Centre Ltd. 31 p.
41821115
Sherwood, R. (1990) Acute Intraperitoneal Toxicity Limit Testing of Streptomyces griseoviridis, a Microbial Pesticide: Lab Project Number: LO8279. Unpublished study prepared by IIT Research Institute. 26 p.
41821116
Liggett, M.; Hossack, D.; Baker, M. (1989) Irritant Effects on the Rabbit Eye of Streptomyces griseoviridis (Mycostop): Lab Project Number: 891076D/RKY 114/4/SE. Unpublished study prepared by Huntingdon Research Centre Ltd. 15 p.
41821117
Lackenby, F. (1991) Skin Sensitization Study in the Guinea Pig (Magnusson-Kligman Maximization Test): Lab Project Number: 5383- 18R. Unpublished study prepared by Hazleton UK. 22 p.
41821118
Pulkkanen, H. (1990) Hypersensitivity Incident Reporting Mycostop. Unpublished study prepared by Kemira Oy. 4 p.
41821119
Beavers, J.; Smith, G. (1990) Mycostop (Streptomyces Griseoviridis) An Avian Oral Pathogenicity and Toxicity Study in the Bobwhite: Lab Project Number: 293-102. Unpublished study prepared by Wildlife International Ltd. 20 p.
41821120
Beavers, J.; Smith, G. (1990) Mycostop (Streptomyces Griseoviridis) An Avian Oral Pathogenicity and Toxicity Study in the Mallard: Lab Project Number: 293-103. Unpublished study prepared by Wildlife International Ltd. 20 p.
41821121
England, D. (1990) A 30-Day Static Renewal Toxicity and Pathogeni- city Evaluation on the Effects of Streptomyces griseoviridis to Rainbow Trout (Oncorhynchus mykiss): Lab Project Number: 38678. Unpublished study prepared by Analytical Bio-Chemistry Laboratories, Inc. 26 p.
41821122
England, D. (1990) A 21-Day Static Renewal Toxicity and Pathogenicity Evaluation on the Effects of Streptomyces griseovirivis to Daphnia magna: Lab Project Number: 38679. Unpublished study prepared by Analytical Bio-Chemistry Laboratories, Inc. 23 p.
41821123
Hoxter, K.; Smith, G. (1990) A Dietary Pathogenicity and Toxicity Study with the Honey Bee: Lab Project Number: 293-101. Unpublished study prepared by Wildlife International Ltd. 14 p.
42298000
E.R. Butts Intl., Inc. (1992) Submission of product chemistry and toxicity data to support the registration of the microbiocide Mycostop. Transmittal of 8 studies.
42298001
Mohammadi, O. (1992) Taxonomic Identification of Mycostop Biofungicide. Unpublished study prepared by Kemira Oy. 13 p.
42298002
American Cell Type Culture Collection (1983) Culture Identification (Streptomyces cultures): Lab Project Number: 564. Unpublished study. 11 p.
42298003
Franke, J. (1990) Streptomyces Characterization: Lab Project Number: SC 2344. Unpublished study prepared by American Type Cell Culture Collection. 13 p.
42298004
Kroppenstedt, R. (1992) Identification of Your Isolates Strain SS6 (90-551), SS61 (90-552), SS12 (91-469), and SS31 (91-470). Unpublished study prepared by Deustche Sammlung von Mikroorganismen und Zellkuluten GmbH. 61 p.
42298005
Mohammadi, O. (1992) Antibiotic Sensitivity of Streptomyces griseoviridis Strain SS61. Unpublished study prepared by Kemira Oy. 6 p.
42298006
Pulkannen, H. (1992) Manufacturing Process--Mycostop: Supplement to MRID 41821106. Unpublished study prepared by Kemira Oy. 10 p.
42298007
Pulkannen, H. (1992) Mycostop (Streptomyces griseoviridis) Determination of Storage Stability: Supplement to MRID 41821111. Unpublished study prepared by Kemira Oy. 7 p.
42298008
Liggert, M. (1989) Acute Dermal Toxicity to Rabbits of Streptomyces griseoviridis, Mycostop: Supplement to MRID 41821113. Unpublished study prepared by Huntingdon Research Centre Ltd. 5 p.
42441500
Kemira Oy. (1992) Submission of toxicity data to support the registration of MYCOSTOP biofungicide. Transmittal of 2 studies.
42441501
Mega, W. (1992) Sensitivity of Detection of Streptomyces griseoviridis for Toxicity/Pathogenicity Testing of Rats: Final Report: Lab Project Number: L08363. Unpublished study prepared by IIT Research Institute. 21 p.
42441502
Mega, W. (1992) Pulmonary Toxicity/Pathogenicity Testing of Streptomyces griseoviridis following Acute Intrachael Challenge in Rats: Final Report: Lab Project Number: L08363. Unpublished study prepared by IIT Research Institute. 146 p.
43090300
Kemira Agro Oy (1994) Submittal of Pathogenicity Data in Support of Registration of Mycostop Biofungicide. Transmittal of 1 study.
43090301
Bowman, J.; Sword, M. (1994) A 30-Day Static Renewal Toxicity and Pathogenicity Evaluation on the Effects of Mycostop (Streptomyces griscoviridis) to Rainbow Trout (Oncorhynchus mykiss): Final Report: Lab Project Number: 41072. Unpublished study prepared by Analytical Bio-Chemistry Laboratories, Inc. 287 p.
45511400
Kemira Agro OY (2001) Submission of Product Chemistry Data in Support of the Application for Registration of Mycostop G Biofungicide. Transmittal of 2 Studies.
45511401
Seiskari, P. (2001) Product Chemistry Data of Mycostop G Biofungicide: Lab Project Number: PCD MYCOSTOP G. Unpublished study prepared by Kemira Agro Oy. 27 p.
45511402
Klausner, K. (2001) Physical and Chemical Characteristics Data of Streptomyces K61 KF Formulation: Lab Project Number: 00-6000-G1. Unpublished study prepared by Toxikon Corp. 21 p.
45623100
Kemira Agro Oy (2002) Submission of Product Chemistry Data in Support of the Application for Registration of Mycostop G Biofungicide. Transmittal of 1 Study.
45623101
Seiskari, P. (2002) Product Chemistry Data (Addendum) of Mycostop G Biofungicide: Lab Project Number: PCD MYCOSTOP G. Unpublished study prepared by Kemira Agro Oy. 13 p.
47576800
Verdera Oy (2008) Submission of Product Chemistry Data in Support of the Amended Registration of Mycostop Biofungicide. Transmittal of 1 Study.
47576801
Seiskari, P. (2008) Updated Product Chemistry Data of Mycostop Biofungicide. Project Number: REPORT/162/2. Unpublished study prepared by Verdera Oy. 17 p.
47678000
Verdera Oy (2009) Submission of Product Chemistry Data in Support of the Amended Registration of Mycostop Biofungicide. Transmittal of 1 Study.
47678001
Korteniemi, M. (2009) Supplement to Updated Product Chemistry Data of Mycostop Biofungicide: Streptomyces griseoviridis Strain K61 @ 35%. Project Number: 162/2. Unpublished study prepared by Verdera Oy. 45 p.
47771900
Verdera Oy (2009) Submission of Product Chemistry Data in Support of the Amended Registration of Mycostop Biofungicide. Transmittal of 1 Study.
47771901
Seiskari, P. (2009) Second Supplement to Updated Product Chemistry Data of Mycostop Biofungicide. Project Number: 160/3. Unpublished study prepared by Verdera Oy. 14 p.

      B. Environmental Protection Agency Risk Assessment Memoranda
      
     Barsoum, I. 2002. Product Chemistry review of Mycostop G Biofungicide.  U.S. EPA, Washington, D.C. Memorandum dated April 1, 2002.
     
     Barsoum, I. 2002. Review of follow-up data submitted by Verdera Oy for product Mycostop G. U.S. EPA, Washington, D.C. Memorandum dated October 10, 2002.
     
     Barsoum, I. 2008. Review of an updated product chemistry study of the product Mycostop Biofungicide. U.S. EPA, Washington, D.C. Memorandum dated December 16, 2008.
     
     Barsoum, I. 2009. Review of supplemental data submitted by the registrant in response to a deficiency letter sent by the Agency regarding the registration of the product Mycostop Biofungicide.  U.S. EPA, Washington, D.C. Memorandum dated March 31, 2009.
     
     Schaffer, C. 1991.  SACB Review of Data Submitted by Kemira Oy to Support the Registration of Mycostop.  U.S. EPA, Washington, D.C. Memorandum dated September 28, 1991.
     
     Schaffer, C. 1992.  SACB Review of Data Submitted by Kemira Oy to Support the Registration of Mycostop.  U.S. EPA, Washington, D.C. Memorandum dated August 12, 1992.
     
     Schaffer, C. 1992. SAB Review of Acute Pulmonary Toxicity/Pathogenicity Data Resubmitted by Kemira Oy to Support the Registration of Mycostop. U.S. EPA, Washington, D.C. Memorandum dated September 17, 1992.
     
     Schaffer, C. 1992. SAB Review of Data Submitted by Kemira Oy to Support the Exemption from Tolerance for Mycostop. U.S. EPA, Washington, D.C. Memorandum dated November 3, 1992.
     
     Schaffer, C. 1992. SAB comment on the bridging of data from the resubmitted Acute Pulmonary Toxicity/Pathogenicity Data to the Acute Oral Toxicity/Pathogenicity study by Kemira Oy to Support the Registration of Mycostop. U.S. EPA, Washington, D.C. Memorandum dated November 10, 1992.
     
     Schaffer, C. 1993. SAB Review of Additional Information Regarding the Taxonomic Classification of Mycostop, Submitted for Registration by Kemira Oy.  U.S. EPA, Washington, D.C. Memorandum dated May 26, 1993.
     
     Schneider, W. 1990.  OPP Scientific Assessment of the January 12, 1990, Kemira Oy Notification of a Small Scale Field Test of a Non-indigenous Strain of Streptomyces griseoviridis.  U.S. EPA, Washington, D.C. Memorandum dated April 4, 1990.
     
     Sjoblod, R. 1988. Mammalian toxicology data requirements for registration of the microbial pesticide Mycostop. U.S. EPA, Washington, D.C. Memorandum dated December 16, 1988.
     
     Sjoblod, R. 1993. Mycostop Biofungicide:  Company Response to Pulmonary Exposure/Toxicity Issues. U.S. EPA, Washington, D.C. Memorandum dated January 15, 1993.
     
     Vaituzis, Z. 1991. Kemira Oy Notification of a Small Scale Field Test of a Non-indigenous Strain of Streptomyces griseoviridis.  U.S. EPA, Washington, D.C. Memorandum dated May 31, 1991.
     
     Wozniak, C. 2002. Review of the Mammalian Toxicity, Pathogenicity, Hypersensitivity and Irritation Potential of Streptomyces griseoviridis relative to establishment of an appropriate restricted entry interval. U.S. EPA, Washington, D.C. Memorandum dated July 18, 2002.

V.  GLOSSARY OF ACRONYMS AND ABBREVIATIONS 

APHIS	Animal and Plant Health Inspection Service
BPPD	Biopesticides and Pollution Prevention Division
BRADs	Biopesticides Registration Action Documents
°C 	degrees Celsius
CFR	Code of Federal Regulations
cfu 	colony-forming unit
cm	centimeter
EC50	half maximal effective concentration
EDSP	Endocrine Disruptor Screening Program
EEC	estimated environmental concentration.  The estimated pesticide concentration in an 
	environment, such as a terrestrial ecosystem.
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 	grams
g/kg	grams per kilogram
kg	kilograms
L 	liter
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, inhalation).  It is expressed as a weight of substance per unit weight of 
	animal (e.g., mg/kg).   
LOC	level of concern
LOEC	lowest observable effect concentration
mg	milligrams
mg/kg	milligrams per kilogram
mg/L	milligrams per liter
mL	milliliter
mL/kg	milliliters per kilogram
MRID Nos.	Master Record Identification Numbers
NOAEC	no observable adverse effect concentration
NOEC	no observable effect concentration
OECD	Organization for Economic Cooperation and Development
OPP	Office of Pesticide Programs
OCSPP 	Office of Chemical Safety and Pollution Prevention
PC Code	Pesticide Chemical Code
ppm	parts per million 
PWP 	preliminary work plan
SAR	systemic acquired resistance
TGAI 	technical grade of the active ingredient
TMDLs	Total Maximum Daily Loads
TSA	trypticase soy agar
USDA	United States Department of Agriculture

VI.  APPENDIX

TABLE 1. Product Analysis Data Requirements for Streptomyces Strain K61 (40 CFR § 158.2120)

                               Data Requirement
                               (OCSPP Guideline)
                                    Results

                                 MRID Numbers
                       Product Chemistry and Composition
                               Product Identity
                                  (885.1100)
Submitted data do not completely satisfy the requirement of product identity for Streptomyces Strain K61 (Mycostop[(R)])
                                   418211-01
                                   418211-02
                                   418211-03
                                   418211-04
                                   418211-05
                               Product Identity
                                  (885.1100)
Submitted data completely satisfy the requirement of product identity for Streptomyces Strain K61 (Mycostop[(R)] and Mycostop(R) G).
                                 Mycostop[(R)]:
                                   422980-01
                                   422980-02
                                   422980-03
                                   422980-04
                                   422980-05
                                Mycostop[(R)]G:
                                  455114-01 
                             Manufacturing Process
                                  (885.1200)
Submitted data do not completely satisfy the requirement of manufacturing process for Streptomyces Strain K61 (Mycostop[(R)])
                                   418211-06

                             Manufacturing Process
                                  (885.1200)
Submitted data completely satisfy the requirement of manufacturing process for Streptomyces Strain K61 (Mycostop[(R)] and Mycostop(R) G).
                                 Mycostop[(R)]
                                   422980-06
                                Mycostop[(R)]G:
                                   455114-01
     Deposition of a Sample in a Nationally Recognized Culture Collection
                               (Not applicable)
A deposit of Streptomyces Strain K61 has been made in both the American Type Culture Collection and at the German Collection of Microorganisms and Cell Cultures.
                                   422980-02
                                   422980-04
             Discussion of Formation of Unintentional Ingredients
                                  (885.1300)
Submitted data completely satisfy the requirement of discussion of formation of unintentional ingredients for Streptomyces Strain K61 (Mycostop[(R)] and Mycostop(R) G).

                                 Mycostop[(R)]
                                   418211-07
                                Mycostop[(R)]G:
                                   455114-01
                         Analysis and Certified Limits
                              Analysis of Samples
                                  (885.1400)
Submitted data completely satisfy the requirement of analysis of samples for Streptomyces Strain K61 (Mycostop[(R)] and Mycostop(R) G).
                                 Mycostop[(R)]
                                   418211-08
                                   475768-01
                                   476780-01
                                   477719-01
                                Mycostop[(R)]G:
                                   455114-01
                           Certification of Limits 
                                  (885.1500)
Submitted data completely satisfy the requirement of certification of limits for Streptomyces Strain K61 (Mycostop[(R)] and Mycostop(R) G).
                                 Mycostop[(R)]
                                   418211-00
                                   475768-01
                                Mycostop[(R)]G:
                                   455114-01
                     Physical and Chemical Characteristics
                                     Color
                                  (830.6302)
Mycostop[(R)]:  Hue: 2.5 Y, Value: 9, Chroma: 2
Mycostop(R) G:  Hue: 2.5 yr, Value: 8.5, Chroma: 4
                                   418211-10
                                   455114-02
                                Physical State
                                  (830.6303)
Mycostop[(R)]:  Brown-tan color, fine powdery solid.  Not free-flowing. 
Mycostop(R) G:  Free-flowing powder.
                                   418211-10
                                   455114-02
                                     Odor
                                  (830.6304)
Mycostop[(R)]:  Sweaty, fishy, putrid, bitter
Mycostop(R) G:  Smells of mold
                                   418211-10
                                   455114-02
     Stability to Normal and Elevated Temperatures, Metals, and Metal Ions
                                  (830.6313)
The Agency has not required that these data be submitted for Streptomyces Strain K61. 
                                Not applicable
                               Storage Stability
                                  (830.6317)
Mycostop[(R)]:  below +8°C, viability over 10[8] CFU/g for 6 months

Mycostop(R) G: +4°C, viable > 12 months; +28°C, viable for 6 months
                                   418211-11
                                   422980-07
                                   455114-02
                                  Miscibility
                                  (830.6319)
                                      N/A
                                Not applicable
                           Corrosion Characteristics
                                  (830.6320)
                                      N/A
                                Not applicable
                                      pH
                                  (830.7000)
Mycostop[(R)] (1%):  5.63
Mycostop(R) G (1%):  7.68
                                   418211-10
                                   455114-02
                                   Viscosity
                                  (830.7100)
                                      N/A
                                Not applicable
           Density/Relative Density/Bulk Density (Specific Gravity)
                                  (830.7300)
Mycostop[(R)]:  28.68 lb/ft[3]
Mycostop(R) G:  7.68 lb/ft[3]
                                   418211-10
                                   455114-02

TABLE 2. Toxicology Data Requirements for Streptomyces Strain K61 (40 CFR § 158.2140)

                               Data Requirement
                               (OCSPP Guideline)
                                    Results

                                  MRID Number
                                    Tier I
                       Acute Oral Toxicity/Pathogenicity
                                  (885.3050)
The data show no clinically significant signs in rats.  Streptomyces Strain K61 was not detected in kidney, brain, liver, lungs, spleen, mesenteric lymph nodes, or in blood or urine samples, and cleared from the feces and cecum following day two.  Necropsy studies showed no significant signs of abnormalities.  There were significant reductions in the weights of dosed males compared to controls at all study times and in the dosed females on day 8 and day 22.  In the absence of pyrogenic effects or any sign of infectivity, however, the test organism was not toxic, infective or pathogenic to rats.  
Classification: Acceptable
                                   418211-12
                    Acute Pulmonary Toxicity/Pathogenicity
                                  (885.3150)
Generally, instillation of bacterial spores into rat lungs will result in a small number of deaths (which was seen in this study).  Also, large numbers of spores are usually recovered from the lungs shortly after dosing  -  with successively lower numbers of spores seen at later time intervals and concurrent increases in the spores isolated from the caecal contents  -  as a result of clearance from the lungs via the trachea and subsequent ingestion.  The present study was not typical in this regard.  The authors reported that microscopic examination of the dosing preparation showed densely packed inter-meshing mycelia in aggregations of 100mm to 200 mm, whereas spores normally have dimensions of 1mm x 2 mm.  They concluded that the size of the mycelial masses would not penetrate into the peripheral lungs and would be rapidly cleared from the airways by the pulmonary mucociliary escalator system.  Evidence regarding the infectivity/pathogenicity of the test organism is, therefore, inconclusive, since only small numbers of viable organisms appeared to have reached the lower lungs.  Since there was no proliferation of the organism by the end of the study and because of the large particle size of the preparation, however, the authors concluded that Streptomyces Strain K61 would not present an infectious hazard under field conditions.  Classification: Uncceptable
                                   418211-14
                                   418211-15
                    Acute Pulmonary Toxicity/Pathogenicity
                                  (885.3150)
Streptomyces Strain K61 caused death in 54% of treated male and 48% of treated female rats with 45% (male) and 40% (female) mortality occurring within the first 2 days after intratracheal instillation.  This high death rate may partially be due to the large size of the test organism; Streptomyces Strain K61 caused sufficient mortality to provoke concern about pulmonary exposure to significant amounts of the bacterium.  Dosage was 0.1 mL of a 3.46 x 10[8] CFU/mL suspension.  Classification: Acceptable
                                   424415-01
                                   424415-02
                    Acute Injection Toxicity/Pathogenicity
                                  (885.3200)
An acute intraperitoneal injection of 5000 mg of the test article/Kg killed all males and females within 1 day of treatment.  1000 mg/Kg and 500 mg/Kg doses also caused some deaths within 2 days of dosing.  This indicates a toxic response to the test article rather than an infectious process, which was confirmed by the mortality caused by the heat-killed test article.  The toxin appears to be heat stable.  Adverse clinical signs returned to normal by day 5 in surviving mice except for one female with a distended abdomen and two with enlarged spleens.  The spleens showed no sign of the test article.  The LD50 for intraperitoneal injection was estimated to be 1306 mg/Kg for males, 870 mg/Kg for females and 1042 mg/Kg for both sexes combined. 
Classification: Acceptable
                                   418211-15
                          Hypersensitivity Incidents 
                                  (885.3400)
Two instances of hypersensitivity were reported by Verdera Oy prior to EPA registration.  The first incident was reported in 1986, the details of which are included in MRID No. 418211-18.  One part-time summer trainee at the Espoo Research Centre in Finland had symptoms of a mild alveolar reaction to Mycostop(R) (EPA Reg. No. 64137-5).  The alveolar reaction could have been due to exposure to biological dust from Mycostop[(R)], though there were indications that the incident might have been avoided by careful use of the safety equipment that was recommended and provided.  In 1991, a research associate in a lab in Fort Collins, CO experienced dizziness, faintness, difficulty breathing and dilated pupils after treating seed with Mycostop[(R)].  The individual had a history of allergies and asthma and was not wearing the required face mask when exposed.  The symptoms disappeared in approximately one hour and no medical attention was sought.  These incidents do not have any bearing on the hazard or risk assessed pertaining to the use of registered Streptomyces Strain K61 pesticide products in accordance with their product labels.  Both individuals recovered completely and no other employees working with Mycostop(R) have reported any problems.
                                   418211-18
                                 Cell Culture
                                  (885.3500)
Not required because Streptomyces Strain K61 is not a virus (refer to test note #4 of 40 CFR § 158.2140(d)).
                                Not applicable
                              Acute Oral Toxicity
                                  (870.1100)
                  Not applicable to the TGAI test substance.
                                Not applicable
                             Acute Dermal Toxicity
                                  (870.1200)
                  Not applicable to the TGAI test substance.
                                Not applicable 
                           Acute Inhalation Toxicity
                                  (870.1300)
                  Not applicable to the TGAI test substance.
                                Not applicable
                             Acute Eye Irritation
                                  (870.2400)
                  Not applicable to the TGAI test substance.
                                Not applicable
                           Primary Dermal Irritation
                                  (870.2500)
                  Not applicable to the TGAI test substance.
                                Not applicable
                                       
                               Tiers II and III
Not required for Streptomyces Strain K61 based on the lack of acute toxicity/pathogenicity in the Tier I studies.
                              Additional Studies
                           Acute Dermal Toxicity[*]
                                  (870.1200)
Streptomyces Strain K61 was not toxic to rabbits when a single 2 g/Kg dose was administered dermally.  Rabbit body weights were not affected by an exposure of 2 g/Kg of Streptomyces Strain K61.  
Classification: Acceptable
                                   418211-13
                                   422980-08
                            Acute Eye Irritation[*]
                                  (870.2400)
Small numbers of spores of Streptomyces Strain K61 were found in the treated eyes of all 6 rabbits at 24 and 72 hours post-treatment, but none thereafter.  Instillation of Streptomyces Strain K61 (>10[7] viable spores per eye) elicited transient, mild conjunctival irritation.  
Classification: Acceptable  -  Toxicity Category IV. 
                                   418211-16
                         Primary Dermal Irritation[*]
                                  (870.2500)
Moderate reactivity (55% of responders) of guinea pig skin to Mycostop[(R)] (EPA Reg. No. 64137-5) was recorded on day two following challenge of sensitized animals.  While no determination was made of the amount of viable Streptomyces Strain K61 spores used in any of the animal exposures and the dose formulations records are inconsistent, the basic conclusion that the end use product is a skin sensitizer is still valid.  Mycostop[(R)] is a moderate skin sensitizer.  Dosage was 0.1 mL of a 3% w/v dilution of Mycostop[(R)] in distilled water.  
Classification: Acceptable
                                   418211-17
* Not currently in the data requirements table in 40 CFR § 158.2140 for the TGAI test substance.

TABLE 3. Nontarget Organism and Environmental Fate Data Requirements for Streptomyces Strain K61 (40 CFR § 158.2150)

                               Data Requirement
                               (OCSPP Guideline)
                                    Results

                                  MRID Number
                                    Tier I
                              Avian Oral Toxicity
                                  (885.4050)
The avian oral LD50 was greater than 2,500 mg/kg body weight, based on results from tests on both mallards and northern bobwhites.  
Classification: Acceptable
                                   418211-20
                                   418211-19
                    Avian Inhalation Toxicity/Pathogenicity
                                  (885.4100)
Not required, as the nature of Streptomyces Strain K61 does not indicate potential pathogenicity to birds or relatedness to any known bird pathogens (refer to test note #3 of 40 CFR § 158.2150(e)).

                                Not applicable
                      Wild Mammal Toxicity/Pathogenicity
                                  (885.4150)
Not required for Streptomyces Strain K61 because the results of the tests required by 40 CFR § 158.2140 are adequate and appropriate for assessment of hazards to wild mammals (refer to test note #4 of 40 CFR § 158. 2150(e)).
                                Not applicable
                    Freshwater Fish Toxicity/Pathogenicity
                                  (885.4200)
This study showed sporadic fish mortality and failed to demonstrate a definitive LC50 value due to the absence of a dose-response trend with the observed mortality. The study results were not adequate for making a risk assessment. 
Classification: Unacceptable
                                   418211-21
                    Freshwater Fish Toxicity/Pathogenicity
                                  (885.4200)
The 30-day static renewal toxicity/pathogenicity study of Streptomyces Strain K61 to rainbow trout (Oncorhynchus mykiss) was conducted by Kemira Agro Oy.  Single replicates of 10 fish each were exposed to Streptomyces Strain K61 concentrations of 0.25x10[5], 0.50x10[5], 1.0x10[5], 2.0x10[5], and 4.0x10[5] CFU/mL in dilution water plus a dilution water control and a killed-spore treatment of 4.0x10[5] CFU/mL.  The dietary dose in the food offering was 10% of the water exposure concentration. The 30-day NOAEC was 0.50x10[5] CFU/mL based on the lack of mortality, statistically significant growth effects, and sublethal/abnormal effects at this concentration.  All surviving fish ate normally and showed an increase in length and weight on day 30 as compared to the representative group on day 0.  No lesions or tissue discolorations were noted in any surviving fish at the end of the study.
Classification: Acceptable 
                                   430903-01
                Freshwater Invertebrate Toxicity/Pathogenicity
                                  (885.4240)
A 21-day static renewal study was conducted to determine the toxicity and/or pathogenicity of Streptomyces Strain K61 spores to Daphnia magna.  A control group was tested against four treatment groups, including a maximum nominal dose exposure of 1.0 x 10[6] CFU/mL, low and median doses of 10[4] and 10[5] CFU/mL, plus a dose of 1.0 x 10[6] CFU/mL of heat-killed spores.  No significant differences (P>0.05) were found between the control and the exposure levels for reproduction and time to first brood.  All of the offspring produced during the study appeared normal.  Mean survival was 100% for control and low level concentration, 75% for the middle concentration, and 0% for the high concentration and killed spore treatment. Mortality from killed spore treatments has been reported from previous studies, and is thought to be caused by creating a toxic environment through the autoclaving process.  A day 21 EC50 was calculated to be approximately 190 mg/L (1.9 x 10[5] CFU/mL).  Daphnid growth was found to be significantly affected at the middle concentration (1 x 10[5] CFU/mL).  The NOAEC was found to be 1.0 x 10[4] CFU/mL, the LOAEC was 1.0 x 10[5] CFU/mL and the MATC was found to be 3.2 x 10[4] CFU/mL. This indicates that Streptomyces Strain K61 is practically non-toxic to Daphnia magna.  The study fulfils EPA Guideline requirements for an acute toxicity test for an aquatic invertebrate.
Classification: Acceptable
                                   418211-22
                            Nontarget Plant Testing
                                  (885.4300)
Not required for Streptomyces Strain K61 because the microbial pest control agent is not taxonomically related to known plant pathogens (refer to test note #7 of 40 CFR § 158. 2150(e)).
                                Not applicable
                               Honey Bee Testing
                                  (885.4380)
                                       
                           Nontarget Insect Testing
                                  (885.4340)
Three treatment levels, representing 240, 760 and 2400 ppm at a specific activity of 9.8 x 10[8] cfu/g of diet, were tested along with an attenuated control and a negative control.  Two replicates were tested at each concentration with 25 bees per replicate.  Mortalities occurred in both of the control groups (negative and attenuated) and in all 3 of the treatment groups.  The mortalities in the negative and attenuated control groups were 30% and 62%, respectively, while those in the 240, 760, and 2400 ppm concentrations averaged 20%, 26% and 28%, respectively.  The pattern of mortality was not found to be dose responsive and did not appear to be treatment related.  The LC50 was determined to be greater than 2400 ppm and the NOAEC was 2400 ppm, which was the highest concentration tested.  Due to nonspecific mortality in the controls, however, the study is considered supplemental.  The results were reliable enough to indicate that MYCOSTOP[(R)] is not toxic or pathogenic to the honey bee.  This study also satisfies the data requirements for nontarget insect testing (OCSPP Guideline 885.4340). 
Classification: Supplemental
                                   418211-23
                                       
                             Tiers II, III, and IV
Not required for Streptomyces Strain K61 based on the results of the nontarget toxicity/pathogenicity Tier I studies, which did not demonstrate adverse hazards.