UNITED STATES ENVIRONMEN TAL PROTECTION AGENCY WASHINGTON, D.C. 20460

OFFICE OF CHEMICAL SAFETY

AND POLLUTION PREVENTION

November 2, 2011

MEMORANDUM

SUBJECT: Environmental risk assessment for the FIFRA Section 3 registration of

Phoma macrostoma strain 94-44B (PC Code 080303; EPA Reg. Nos. 239- ETNE, 239-ETNG, 239-ENTU) for control of broadleaf weeds in turf and

ornamentals

FROM: Shannon Borges, Biologist / signed /

Microbial Pesticides Branch Biopesticides and Pollu tion Prevention Division, 7511P

TO: Kathleen Martin, Regulat ory Action Leader

Microbial Pesticides Branch Biopesticides and Pollu tion Prevention Division, 7511P

PEER REVIEW: Gail Tomimatsu, Ph.D., Plant Pathologist / signed /

Microbial Pesticides Branch Biopesticides and Polluti on Prevention Division, 7511P

The Scotts Company, LLC (hereafter “Scotts”), has submitted an application to register the microbial pesticide control agent (MPCA), Phoma macrostoma strain 94-44B, for control of preand post-emergent weeds in ornamentals and turf (including seed st arting, field nursery, landscape, and container uses), and on impervious surfaces, such as patios, driveways, and

sidewalks. Phoma macrostoma strain 94-44B is the active ingr edient in the manufacturing use product, Phoma Tech (EPA Reg. No. 239-ENTE) , and the end-use products, Phoma H (EPA Reg. No. 239-ENTG) and Phoma P (EPA Reg. N o. 239-ENTU). Each of these products is composed of the same materials, and contains 92% Phoma macrostoma strain 94-44B at a

minimum of 10 3 CFU/g. Phoma H is to be marketed to homeowners and other nonprofessional applicators, while Phoma P is to be marketed to professional us ers. This memorandum contains

BPPD’s environmental risk assessmen t for the proposed registration of Phoma macrostoma

strain 94-44B.

A. Introduction

Phoma macrostoma is a coelomycetous fungus that is comm only isolated from woody plants. It is also found incidentally on other plants (B oerema and Dorenbosch 1970, Boerema et al. 2004) and in soil and leaf litter (B engtsson et al. 2005, Newsham 2003). It has been isolated from2 plants (mainly trees) in severa l states within the U.S. (Fa rr and Rossman 2011), and is found worldwide (Aveskamp et al. 2008). It has not been reported as a widely prevalent phytopathogen in the contiguous U.S. (UGA 2011). Several species in the Phoma genus are considered to be relevant phytopathogens (Aveskamp et al. 2008). While Phoma macrostoma is generally regarded as a weak pa thogen or wound parasite (Caesar, et al., 2011, De Gruyter et al. 2002, Boerema et al. 2004, see also Chauha n and Pandey 1991, Shattock and Askew 1980, Williamson and Hargreaves 1979), there are several reports of Phoma macrostoma causing primary disease in plants (e.g., Caesar, et al., 2011, Baird 1993, Jamaluddin 1977, Kubota and Abiko 2002, Sidhu and Singh 1979). Many species of Phoma exist as saprobes in the absence of suitable host plants, but will switch to a pat hogenic existence when a suitable host plant is encountered (Aveskamp et al. 2008). A host list for Phoma macrostoma is available in Boerema and Dorenbosch (1970) and Farr and Rossman (2011), and the applicant has also provided a list of susceptible plants for P. macrostoma strain 94-44B, which is provided in Section C.1.

Phoma macrostoma strain 94-44B was initially isolated from diseased Canada thistle ( Cirsium

arvense L.), and has been developed as a microbial pesticide for broadleaf weeds in turf and ornamentals. Effects on plants include chlorotic leaf spots, necrosis, and severe bleaching of emerging leaves. These effects can be lethal, al though treated plants recove r in some cases. The

mode of herbicidal action is c onsidered bimodal; fungal mycelia infect plant root systems and produce phytotoxic macrocidins, which cause fo liar chlorosis and bleaching (Bailey, et al., 2010).

B. Summary of Nontarget Effects and Environmental Fate Studies

Table 1 provides the status of the data re quirements as published in 40 CFR § 158.2150 for the nontarget organism testing conducted on the active ingredient (TGAI), Phoma macrostoma strain 94-44B. Studies were submitted to satisfy data requirements for avian oral toxicity/pathogenicity, freshwate r fish and invertebrate toxici ty/pathogenicity testing, nontarget insect testing, honey bee testing, a nd nontarget plant testing (terrestri al and aquatic). Scientific rationale was submitted to satisfy the data require ment for wild mammal toxicity/pathogenicity testing. Evaluation of the studies and scientific rationale submitted to support the registration of

Phoma macrostoma strain 94-44B was completed by joint re view with the Health Canada Pest Management Regulatory Agency. Re levant information from the open peer-reviewed literature,

submitted studies and scientific rationale are included in environmental risk assessment. Data Evaluation Records (DERs) of th e scientific rationale and st udies are attached to this

memorandum.

The scientific evidence from peer-reviewed lite rature and the test data and information from submitted studies conducted by the applicant ar e sufficient to satisfy all Tier I nontarget organism data requirements and for nontarget or ganism risk assessment for the active ingredient (see Table 1). Additional data and informati on from published studies (e.g., Bailey et al. 2010, Bailey et al. 2011a, 2011b, Zhou et al. 2004) describe d the infection and colonization processes in plants and the environmental fate of Phoma macrostoma strain 94-44B. This information was determined sufficient to satisfy Tier II testing for the proposed uses, and are detailed below in

Section C.3 The Agency also sought respective opinions from the USDA-APHIS-PPQ, and a plant pathologist at the Cornell University concerning the regulated status of Phoma macrostoma as a plant pathogen, and its ecological relevance as a plant pathogen, relativ e to other pathogenic species of Phoma.

Table 1 . Summary of data submitted to comply with nontarget organism data requirements published in 40 CFR § 158.2150 for s upport of the registration of Phoma macrostoma strain 94-

44B. Data Requirement OCSPP (OPPTS)

Guideline No. Results Summary and Classification MRID No.

Avian oral toxicity/pathogenicity 885.4050 The 30-day oral LD 50 was > 2.22 × 10 5 CFU/kg bw/day.

Classification: Supplemental; adequate for

EPA risk assessment 1 479150-18

Avian inhalation toxicity/pathogenicity 885.4100 Not required; Phoma macrostoma is not known or shown to be an avian pathogen N/A

Wild mammal toxicity/pathogenicity 885.4150 Rationale states that P. macrostoma is ubiquitous and P. macrostoma 94-44B has been shown to decline in population after application. An acute oral toxicity study with laboratory rats estimated an LD 50 of > 10 8

CFU/animal, no adverse effects observed.

Classification: Acceptable rationale 479150-26

Freshwater fish toxicity/pathogenicity 885.4200 The 32-day LC 50 was >2.9 × 10 3 CFU/mL; no adverse effects were observed during the study.

Classification: Acceptable 479150-19

Freshwater

invertebrate toxicity/pathogenicity 885.4240 The 21-day EC 50 was > 3.78 x 10 2 CFU/mL. The NOAEC based on growth and reproduction was 2.71 × 10 3 CFU/mL.

Classification: Acceptable 479150-20

Estuarine/marine fish

and invertebrate testing 885.4280 Not required – significant concentrations are not expected to reach marine/estuarine environments as a result of the proposed uses. N/A

Nontarget insect testing 885.4340 Green lacewing : The 15-day LC 50 was > 1 ×

10 6 CFU/mL diet Lady bird beetle : The 12-day dietary LC 50 > 1 × 10 6 CFU/mL diet. Parasitic hymenoptera : The 16-day dietary

LC 50 > 1 × 10 6 CFU/mL.

Classification: All studies are acceptable 479150-23

479150-24

479150-22

Honey bee testing 885.4380 The 5-day LC 50 was >1 × 10 6 CFU/mL diet. The study was not long enough to assess pathogenicity.

Classification: Supplemental; adequate for

risk assessment 479150-254 Data Requirement OCSPP (OPPTS)

Guideline No. Results Summary and Classification MRID No.

Nontarget plant testing 885.4300 Aquatic ( Lemna gibba ) : The 7-day EC 50 = 1.51 × 10 4 CFU/mL; the NOAEC = 4.4 × 10 3

CFU/mL.

Terrestrial : A collection of studies showed

that P. macrostoma strain 94-44B is toxic/pathogenic to several species of terrestrial broadleaf plants.

Classification: Both studies are acceptable 479150-21

483184-01

1 Viability of the test substance was not confirmed, which is necessary for an “Acceptable” rating from PMRA. The study is acceptable for risk assessment.

C. Ecological Exposure and Environmental Risk Characterization

1. Terrestrial Animals and Plants

The EPs are to be applied pre- or post-emergence in spot or broadcast treatments to control various broadleaf weed species to ornamentals, turf, and impervious surfaces such as driveways and patios. Ornamental uses incl ude applications to flowers, rose s, ground covers, shrubs, trees, and ornamental grasses, as well as container grown plants, field grown nurseries, and landscapes, and the labels provide lists of sp ecific tolerant and susceptible or namental plants and grasses. Specific use sites for turf (e.g., sod farms, golf co urses) are not provided, and should be clarified on the label. Spot treatments are appl ied at a maximum rate of 2 grams/0.35 ft 2 (8-inch diameter circle), and broadcast app lications are made at a ma ximum rate of 8 lbs/1000ft 2 . The labeled viability of the Phoma macrostoma strain 94-44B for the EPs is 10 3 CFU/g product, therefore spot applications result in approximately 5.7 x 10 3 CFU/ft 2 and broadcast applications result in approximately 3.6 x 10 3 CFU/ft 2 at application. The applic ation methodology and equipment

must be clarified on the label.

Environmental Fate and Persistence

The applicant submitted studies from peer-reviewed journals as rationale to waive higher tier testing on nontarget plants. These studies pr ovided data and information regarding the

environmental fate of Phoma macrostoma strain 94-44B and macrocidins and its bimodal pesticidal activity as a wound pathogen and macroc idin release into col onized plant tissues or surrounding soil.

Zhou et al. (2004) reported the fate of several strains of Phoma macrostoma used as herbicides in the terrestrial environment, includi ng strain 94-44B. In this study, Phoma macrostoma (a different strain than the one propos ed for registration) was used to treat small field plots, and it was rarely detected in adjacent untreated areas 30 cm beyond the boundaries of the treated plots. The authors also noted the tendency of the test ma terial to drift, which was a likely reason it was found outside the plots. In soil, Phoma macrostoma strain 94-44B PCR dete ctions declined over time, but the number of viable propagules al so started to declin e. These trends of Phoma

macrostoma -PCR detections continued the following y ear, and susceptible species planted the following year did not show effects e xpected as a result of infection by Phoma macrostoma5 strain 94-44B. The authors noted that P

. macrostoma is considered as a poor competitor, and the ability for the fungus to grow is restricted in soils with highe r microbial activity.

Bailey et al. (2011b) show that m acrocidins are partially soluble in water, and may run off from areas of application under some conditions (e .g., with saturated soils). This work was inconclusive with respect to the ability of Phoma macrostoma to mobilize with runoff, but the

authors deduced from the available data that the fungus was less likely to be as mobile in soil as

the macrocidins. Phoma macrostoma also is not expected to move very far via natural dispersal once it is applied, since Phoma macrostoma is primarily dispersed by ra in splash (Bailey et al. 2010).

Both Zhou et al. (2004) and Baile y et al. (2011b) showed that Phoma macrostoma strain 94-44B colonizes both susceptible and re sistant plants. However, the in fection process and subsequent colonization of roots from susceptible plants appear s to be different from roots of resistant plants (Bailey, et al., 2011b). Phytotoxic macroc idins are produced only by mycelia of Phoma

macrostoma and not conidia (Graupner et al., 2006), although exce ss quantities may be exuded to the surrounding growth medium (Graupner et al., 2003). Granular formulations of P.

macrostoma strain 94-44B encourage root infecti on through root hairs and uptake of phloemmobile compounds by the plant roots after mycelia reach the rhizosphere s of treated plants (Bailey, et al., 2010). Fungal mycelia infect the inner tissues of the root core by breaking down internal cellular structures of susceptible pl ants (e.g., dandelion) and simultaneously producing additional phytotoxins, further dama ging the vascular system of sus ceptible plants. In contrast, resistant plants (e.g., barley) show restricted fungal grow th within the root and are asymptomatic of photobleaching, further supporting the combined e ffects of at least two modes of herbicidal activity (Bailey, 2010). Photobleachi ng has only been reported for susceptible plants where the mycelia were observed to colonize and disrupt the central vascul ar core of root tissues of susceptible plants. Therefore, it is the comb ined role of infection and the presence of macrocidins that have direct effects on the plant.

Colonized plants that survive may serve as reservoirs of Phoma macrostoma strain 94-44B that

would be available to cause future effects to su sceptible nontarget plants where conditions allow. However, these conditions would need to be main tained to promote persistence, since only an asexual life stage has been found for Phoma macrostoma (Bailey et al. 2010). Survival structures such as sclerotia have not been reported for Phoma macrostoma . Zhou et al. (2004) showed that after one year, the fungus could only be detected in 12% of non-susceptib le plant root samples from treated areas (in contrast to 5% of c ontrols). However, chlorosis (presumably from macrocidin) was seen in sunflowers planted into a treated site one year after treatment, although there may be other biological or environmental f actors that caused the chlorosis in sunflowers. (Bergstrom, 2011).

Environmental Fate Conclusions: The environmental fate data indicate that Phoma macrostoma strain 94-44B is unlikely to persis t at high levels for long periods of time or move great distances when applied to turf and ornamentals. The re viewers make the following conclusions with respect to environmental fate of Phoma macrostoma strain 94-44B:6 • The proposed labels include optional language for some uses that encourage users to apply the products on a calm day. Calm conditio ns (e.g., wind speed < 5 mph) should be required on the label for all applications. If these label instructions are followed, a relatively small amount of Phoma macrostoma strain 94-44B is expected to drift, and only to within the margins around the treated area.

• Bailey et al. (2011b) show that macrocidins may be mobilized by runoff when soils are saturated; therefore, some movement into ad jacent areas could occur via this process. Label language should be included that requir es the user to immediat ely water in the EP at application, but not to the level of soil saturation. If th is language is included, runoff into adjacent areas as a result of applications to turf and ornamentals is expected to be

minimized. Care should also be taken to not apply these products when soils are saturated (Bailey, et al., 2010).

• Significant runoff is expected with applicati ons to impervious surfaces; therefore, the reviewers recommend that these applications should be removed from the proposed

labels.

• Persistence is expected within treated areas with repeated applications. Studies show a declining trend after f our months and survival of suscep tible plants after one year has varied. Therefore, Phoma macrostoma strain 94-44B may persis t within treated areas.

Birds and Mammals

The primary route of exposure of birds and mammals to Phoma macrostoma strain 94-44B is likely through direct consumption of the EP. The avian LD 50 was determined > 2.22 × 10 5 CFU/kg bw/day for Northern bobwhite ( Colinus virginianus ), and no adverse effects were observed at this maximum hazard dose after 5 days of exposure at this dose and an additional 25 days of observation (total dose 1.1 x 10 6 CFU/kg bw). An acceptable LD 50 for laboratory rats

was determined to be > 1 x 10 8 CFU/animal, and no adverse eff ects were observed during the 21 day study (MRID 47915008). Based on these results, ad verse effects to birds and wild mammals as a result of the proposed applicatio ns of the EPs are not expected.

Nontarget Insects and Honey Bees

Some species of nontarget in sects could be exposed to Phoma macrostoma strain 94-44B through consumption of granules, consumption of treated plant tissues, or through cuticular

contact in treated soils. Since Phoma macrostoma is not reported as an insect pathogen,

cuticular contact with viable co nidia or mycelia of the fungal pr oduct is not expected to be a realistic route through which Phoma macrostoma strain 94-44B could inf ect an insect or other terrestrial arthropod.

Three species of beneficial nont arget insects were exposed to Phoma macrostoma strain 94-44B mycelia through the diet. These species were green lacewing ( Chrysoperla rufilabris ), lady bird beetle ( Hippodamia convergens ), and a parasitic wasp ( Pediobius foveolatus ). The LC 50 was determined to be > 1x 10 6 CFU/mL diet for each species, and significant adverse effects were not observed. Based on this testing, a dverse effects to nontar get insects are not e xpected to result from dietary exposure to Phoma macrostoma strain 94-44B.7 The submitted honey bee study showed that Phoma macrostoma strain 94-44B was not toxic to honey bees at a dose of 1 x 10 6 CFU/mL diet over five days. The study was determined to be supplemental because it was of short duration to allow for determination of pathogenicity. However, since exposure is expected to be low for bees, and because no reports have been found

in which Phoma macrostoma was identified as a bee or other in sect pathogen, adverse effects in bees are not expected to occur as a resu lt of the proposed labeled applications.

Nontarget Terrestrial Plants

Phoma macrostoma is considered to be a weak or wound pathogen, existing primarily as a saprobe. Summaries of efficacy stud ies with terrestrial species of plants were submitted to satisfy the nontarget plant data requirements (MRI D 48318401). These studies showed that many broadleaf plants were susceptible to Phoma macrostoma strain 94-44B, and susceptibility varied among species. Grass and related monocotyledonous species were not shown to be susceptible, but other types of monocot species were not te sted. More recent information from the open

literature noted that P. macrostoma strain 94-44B was pathogenic to many dicotyledonous plant species, but nonpathogenic to the sp ecies of monocots that were tested (Bailey et al., 2011a). Specifically, of the 75 plant species in 22 plant families studied, seven plant families were not adversely affected by P. macrostoma 94-44B (Salicaceae, Pinaceae, Iridaceae, Curcurbitaceae, Solanaceae (bell pepper, tomato), Linaceae, Graminae), whereas Phoma macrostoma 94-44B caused chlorosis and mortality in species from 16 plant families (Convolvulaceae, Lythraceae, Polygonaceae, Rubiaceae, Euphorbiaceae, Caryop hyllaceae, Brassicace ae, Amaranthaceae, Labiatae, Lamiaceae, Leguminosae/Fabaceae, Rosaceae, Plantaginaceae, Asteraceae, Balsminaceae, Solanaceae (petunia)). Most applications were pre-emergent, though postemergent applications were made in two of the st udies, and both lethal and sublethal effects were observed with both types of application. Thes e effects primarily in cluded chlorosis and mortality; some studies also measured biomass and emergence. Several studies examined the effects of multiple inoculum levels. Mortality and other effects were generally dose dependent in susceptible species, and sublethal effects (mainl y chlorosis) were observe d in some studies at the lowest rate tested (stated to be as low as 1/8x EP application rate). The study authors noted that chlorosis did not necessarily predict mortality.

The weight of evidence to date suggests that the micr obial active ingredient is a weak pathogen, affecting susceptible species of several plant families primaril y in the dicots (MRID 48318401; Bailey, et al., 2011a). It is also cons idered to be a poor competitor (Zhou, et al ., 2004) and soils with enhanced microbial activity wi ll likely restrict the ability of the biocontr ol fungus to grow.

Phoma macrostoma does not appear to be related to the soybean red blotch pathogen ( Phoma glycinicola ) or other more potent crop pathogens such as P. lingam (pathogenic to species in the Brassicaceae) (implied in Zhou et al., 2004; opi nion provided by Bergstrom, 2011). Therefore, the reviewers make the following conc lusions regarding nontarget plants:

• Adverse effects may occur to susceptible nont arget plants in areas treated with the mycoherbicide. This is of particular c oncern for rotational dico tyledonous plants that may be planted into a previously treated area (Bergstrom 2011).

• Effects to nontarget plants beyond the treate d sites are expected to be limited to immediately adjacent areas for applications to turf and ornamentals. Widespread losses of8 plants, such as might be seen with a br oad-spectrum chemical herbicide, are not expected; however, species of significant economic or ecological importance could be affected if they are susceptible. If the produc t is applied according to the label language recommended above, the species affected in th ese areas is expected to be very limited.

• Applications to impervious surfaces may result in significant effects to nontarget plants if

runoff occurs.

2. Aquatic Animals and Plants

Exposure in the Aquatic Environment

Since it is assumed the EP can drift or run off, it is possible Phoma macrostoma strain 94-44B or macrocidins can move to aquatic areas. The spot application for turf and ornamentals (2 g/8-in

circle or 5.7 x 10 3 cfu/ft 2 ) would produce the maximum possible concentration in water if applied continuously over a body of water. So, if 5.7 x 10 3 cfu/ft 2 were applied to the EPA Standard Wetland (a 1-ha body of water that is 15 cm de ep), this would produc e a concentration of Phoma

macrostoma strain 94-44B of 0.41 cfu/mL. This is below reported concentrations of Phoma species or total fungal counts containing Phoma species reported in some surface waters (e.g., Abdel Hameed et al. 2008, Pereira et al. 2009). Drif t and runoff would occur at a fraction of this maximum possible amount; ther efore, concentrations of Phoma macrostoma strain 94-44B and its macrocidins present at application are not lik ely to reach freshwater or marine/estuarine habitats in significant concentrations.

Freshwater fish

A 32-day study with rainbow trout ( Oncorhynchus mykiss ) showed that the LC 50 is > 2.9 x 10 3

CFU/mL (measured) for freshwater fish. The te st subjects in this study were exposed through

both the test water and the diet for the duration of the study, a nd no lethal or overt sublethal effects were observed. The exposure amount wa s well above the maximum hazard dose, and it

was determined that Phoma macrostoma strain 94-44B is not toxic or pathogenic to freshwater

fish. Since the test material contained a low amount of macrocidins, it is no t necessarily a test of the potential hazard of these metabolites to fish ; however, the study is acceptable for use in regulatory risk assessment. Based on the expe cted low exposure and the results of this study,

adverse effects to freshwater fi sh are not expected to result fr om the proposed applications of

Phoma macrostoma strain 94-44B.

Freshwater invertebrates

A 21- day study with Daphnia magna was submitted, which provided an estimated EC 50 of

>3.78 x 10 2 CFU/mL (measured). The NOAEC for re production and growth (length and dry weight) was 2.71 × 10 3 CFU/mL and the LOAEC was 4.22 × 10 3 CFU/mL. A dose-response relationship was not evident base d on mortality, reproduction, or growth. Effects were observed in the attenuated control group, such that mortality was 100% by the end of the test. The study authors noted that the test solutions at high con centrations, including the at tenuated control, were cloudy and that some of the test material settled on the bottom of the test vessels. This was also observed in the study with rainbow trout desc ribed above, and is not uncommon in aquatic9 testing with fungal MPCAs. It is likely that the effects observe d in this study are due to the turbidity resulting from high concentrations in the test water and not the te st material. However,

the test material contained a low amount of macrocidins, so the t oxicity of these metabolites was not necessarily tested. The measured values tested are not quite above the maximum hazard concentration suggested by OCSPP (OPPTS) guide lines for aquatic te sting with MPCAs; however, the EC 50 value is 920X the maximum possible concentration in water and the NOAEC is 1030X this concentration. Therefore, because of low expected exposure and the lack of toxicity/pathogenicity of the fungus, adverse effects to freshw ater invertebrates are not expected to result from the proposed labeled applications of Phoma macrostoma strain 94-44B.

Freshwater aquatic plants

A study with Lemna gibba showed that the 7-day EC 50 = 1.51 × 10 4 CFU/mL (measured) for

Phoma macrostoma strain 94-44B. The NOAEC was 4.4 × 10 3 CFU/mL and the LOAEC was

9.5 × 10 3 CFU/mL (measured). The test substance cau sed some necrotic fr onds and a significant

inhibition in frond number and biom ass at the higher tested concentrations as compared to that

seen in the lower test co ncentrations. Therefore, Phoma macrostoma strain 94-44B is likely to cause effects to aquatic plants if it is deposited in water in significant amounts. The measured

concentrations tested in the Lemna study, however, are above the maximum hazard dose of 413 CFU/mL, and are well above the maximum e nvironmental concentration of 0.413 CFU/mL. Therefore, since the concentration resulting from offsite movement is not expected to be significant, adverse effects to aqua tic plants as a result of the proposed labeled applications of

Phoma macrostoma strain 94-44B are not expected.

Marine/estuarine animals and plants

Phoma macrostoma strain 94-44B is not expected to re ach marine/estuarine environments in significant amounts. Therefore, ad verse effects to plants and anim als in this environment are not expected.

3. Open Literature Information for Nont arget Organisms Other Than Terrestrial

Plants

Literature searches were performed to determine if other effects have been reported that were not contained within the information provided by the registrant. A search was performed with the

Environmental Information Search service avai lable to the Agency. This search tool simultaneously queries the Agricola; Biosis Pr eviews; CAB Abstracts; Energy, Science, and Technology, General Science Abstracts, and Nati onal Technical Information Service databases within the years of 1926 – present. A search was performed with the term ‘ Phoma macrostoma ’ was performed and returned 67 records. No addi tional records of effects to nontarget organisms,

other than the effects to plants already reported were found. The search term ‘ Phoma ’ was also paired individually with the terms, ‘bird,’ ‘ma mmal,’ insect,’ ‘arthropod, ’ ‘arachnid,’ ‘aquatic,’ ‘bivalve,’ ‘invertebrate,’ ‘fish,’ ‘reptile,’ ‘amphibian,’ and ‘cru stacean,’ and no additional reports of toxicity or pathogenicity related to Phoma macrostoma were found for these taxa. Additionally, ‘macrocidin’ was searched within these databases, and no additional information was found as to its potential toxi city on nontarget organisms.10 4. Data Uncertainties

The test material for most of the nontarget studies (except for birds) contai ned very low levels of

macrocidins. BPPD concluded that risk of a dverse effects is low to nontarget insects and honeybees for the proposed applications of Phoma macrostoma strain 94-44B, but that the potential toxicity of macrocidin s present in the EPs is not known and presents some uncertainty in the risk assessment. An additional study with honeybee or other nonta rget insect species would clarify the potential hazard of macrocidins to these nontarg et species; however, additional data are not required at this time.

The environmental persistence of viable Phoma macrostoma is anticipated to be of relatively low risk in most turf environments as provided on the proposed labels. The level and relative persistence of macrocidins in the environment following applica tion(s) is also uncertain. Therefore, it is recommended that the labels be revised and clarified with respect to appropriate text to mitigate drift and runoff to adjacent lands that have plants of agricultural or conservation significance. Specific turf use sites should be clar ified on the label.

D. Threatened and Endangered Species Assessment

Direct Effects

Adverse effects on terrestrial vert ebrates, nontarget ins ects, and freshwater and marine/estuarine animals and plants are not expected to occur due to exposure resulting from all proposed labeled applications of Phoma macrostoma strain 94-44B. Therefore, a “No Effect” determination is made for direct effects to federally listed threat ened and endangered (liste d) species within these taxa as a result of all proposed applications as labeled.

While the proposed applications are not expected to result in widespread losses of nonlisted nontarget terrestrial plants, the potential for subl ethal or lethal effect s to individual listed terrestrial plants cannot be pr ecluded. The potential occurrence of listed plants on the treated areas may be unlikely, but it is unknown. Drift or runoff of the EP, especial ly with applications to impervious surfaces, may also deposit Phoma macrostoma strain 94-44B in areas containing listed plants. Even though exposure may not result in mortality, it can cause sublethal necrosis and chlorosis that may contribute to reduced survival and reproduction of a listed plant. Therefore, the proposed applications of Phoma macrostoma strain 94-44B have the potential to directly affect individual listed terrestrial plants. Further analys es are needed to determine the extent of effects and species affected.

Indirect Effects and Effect s to Critical Habitat

Effects to terrestrial vertebrate s, nontarget insects, and freshwat er and marine/estuarine animals and plants are not expected to occur as a result of proposed applications of Phoma macrostoma strain 94-44B. Therefore, indir ect effects to listed species de pendent on these taxa and related

effects to their Critical Habitat are not expected and a “No Eff ect” determination is made for

indirect and habitat effects to li sted species dependent on these taxa for all proposed applications.11 The proposed applications of Phoma macrostoma strain 94-44B are not expected to result in widespread effects to nonlisted te rrestrial plants, so indirect eff ects to listed generalist species dependent on plants are not e xpected, and EPA makes a “No Eff ect” determination for indirect and habitat effects to all listed species that do not have an obligat e relationship with any particular species of plant. Si gnificant effects may occur in certa in susceptible plant species, so applications of Phoma macrostoma strain 94-44B could affect a nonlisted (or listed) plant upon which a listed species depends exclusively for f ood or habitat. Theref ore, applications of Phoma

macrostoma strain 94-44B have the potential to cause in direct effects to listed species and also Critical Habitat for listed species with obligate relationships to susceptible plants. Further analyses will be necessary to determine th e extent of effects and species affected.

II. Environmental Assessment Conclusions

BPPD has performed an environmental risk assessment for Phoma macrostoma strain 94-44B based on the data provided by th e applicant and studies from the open literature, and has made the following conclusions:

1. Exposure of nontarget organisms to Phoma macrostoma strain 94-44B and/or macrocidins may occur in the terrestrial environment as a resu lt of the proposed applic ations. For turf and ornamental applications, exposure is expect ed to be limited to the treated site and immediately adjacent area if additional languag e is included on the label (see below). Longterm use in treated areas may lead to e ffects on susceptible rotational crop species. Significant offsite movement and nontarget expo sures are expected with applications to impervious surfaces (e.g., patios, driveways, sidewalks), and the reviewers recommend that these uses be removed from the proposed label.

2. Significant exposure of nontarget organisms to Phoma macrostoma strain 94-44B and/or macrocidins is not expected in aquatic environments.

3. Adverse effects to bird s, wild mammals, nontarget insects, honey bees, freshwater fish and invertebrates, marine/estuarine fish and invertebrates, and aq uatic plants are not expected.

4. Some species of nontarget plants may be a dversely affected wher e significant exposure

occurs.

• Nontarget plant effects may be significant fo r susceptible species in treated areas,

where its herbicidal effect is intende d, which may be a concern for susceptible rotational dicoty ledonous plants

• Effects to nontarget plants beyond the treate d sites are expected to be limited to immediately adjacent areas for applications to turf and ornamentals, and effects will be limited to more highly susceptible speci es. Additional label language is suggested below. If these are included in the label, effects on nontarget plan ts are expected to be very limited.

• Applications to impervious surfaces may result in significant effects to nontarget plants if runoff occurs.12 5. The following conclusions are made for listed species

• A “No Effect” determination is made for

o Direct effects to listed birds, wild ma mmals, insects, honey bees, freshwater fish and invertebrates, marine/estuarine fish and invertebrates, and aquatic plants.

o Indirect effects and effects on designat ed Critical Habitat for listed species dependent on birds, wild mammals, in sects, honey bees, freshwater fish and invertebrates, marine/estuarine fish a nd invertebrates, and aquatic plants.

o Indirect effects and effects on designated Cr itical Habitat for all listed species that do not have an obligate relationship with any particular species of plant.

• The proposed applications of Phoma macrostoma strain 94-44B have the potential to directly affect individual liste d terrestrial plants. The propos ed applications also have the potential to cause indirect effects to listed species and also Critical Habitat for listed species with obligate relationships to susceptible plants. Further analyses will be necessary to determine the extent of effects and species affected.

6. The following changes to the proposed label shou ld be made to reduce terrestrial nontarget

exposure:

• The EPs must be applied on calm days (e.g., wi nd speed is < 5 mph) to minimize drift

• The EPs must be immediately watered in with all applications to turf and ornamentals

• Applications to impervious surfaces should be removed from the label.

7. The Agency may require additional testing if us es are expanded, if formulations change for the end-use products, or if inci dents of adverse effects to nontarget organisms occur during commercial use of these products.

References Cited

Abdel Hameed, A.A., S. El Hawarry, and M.M. Kamel. 2008. Prevalence and distribution of

airborne and waterborne fungi and actinomycetes in the Nile river. Aerobiologia, 24: 231-240.

Aveskamp, M.M., J. De Gruyter , and P.W. Crous. 2008. Biology and recent developments in the systematic of Phoma , a complex genus of major quaran tine significance. Fungal Diversity,

31: 1-18.

Bailey, K.L., W.M. Pitt, J. Derby, S. Walter, W. Taylor, and S. Falk. 2010. Efficacy of Phoma

macrostoma , a bioherbicide, for control of dandelion ( Taraxacum officinale ) following simulated

rainfall conditions. Am. J. Plant Sci. Biotech., 4 (Special Issue 2): 35-42.

Bailey, K.L., W.M. Pitt, F. Leggett, C. Sh eedy, and J. Derby. 2011a. The effects of Phoma

macrostoma on nontarget plant and target weed speci es. Biol. Control, on-line publication available June 14, 2011.

Bailey, K.L., W.M. Pitt, F. Leggett, C. Sheedy, and J. Derby. 2011b. Determining the infection process of Phoma macrostoma that leads to bioherbicidal activ ity on broadleaved weeds. Biol. Control, 2011, doi: 10.1016/j.biocontrol.2011.06.019.13 Baird, R.E. 1993. First report of Phoma macrostoma causing leaf spot on fevertree. Plant Disease, 77: 1168.

Bengtsson, M., H. Green, N. Leroul, H.L. Pederse n, and J. Hockenhull. 2006. Effect of autumn application of urea on saprotrophic fungi in off-season leaf litter of sour cherry and evaluation of fungal isolates to reduce primary inoculum of Blumeriella jaapii . J. Plant Disease and Protection,

113: 107-112.

Bergstrom, G.C.2011. Email re: “Advice request fo r Phoma macrostoma”, sent 05/26/2011 from Gary Carlton Bergstrom ( gcb3@cornell.edu ) to J. Gagliardi ( gagliardi.joel@epa.gov )

Boerema, G.H., J. de Gruyter, M.E. Noordeloos, and M.E.C. Hamers. 2004. Phoma identification manual: differentiation of specifi c and infra-specific taxa in culture. CABI Publishing, Cambridge, MA. 470 pp.

Boerema, G.H., and M.M.J. Dorenbosch. 1970. On Phoma macrostomum Mont., a ubiquitous species on woody plants. Persoonia, 6: 49-58.

Caesar, A., R.T. Lartey, T.Caesar-TonThat, and A. Harris. 2011. First Report of a root and crown

disease of the invasive weed Lepidium draba caused by Phoma macrostoma . Plant Disease

DOI:10.1094/PDIS-02-11-0107. Po sted on line 09/07/2011.

Chauhan, S., and B.M. Pandey. 1991. Cercospora populina and Phoma macrostroma causing foliage diseases of Populus deltoides . Indian Phytopath. 44: 549-551.

De Gruyter, J., G.H. Boerema, and J.A. Van Der Aa. 2002. Contributions toward a monograph

of Phoma (Coelomycetes) VI – 2 Section Phyllostictoides: outline of its taxa. Persoonia, 18: 1-

53.

Farr, D.F., and A.Y. Rossman. 2011. Fungal Data bases, Systematic Mycology and Microbiology Laboratory, ARS, USDA. Retrieved N ovember 1, 2011, from /fungaldatabases/

Graupner, P.R., B.C. Gerwick, T.L. Siddall, A.W. Carr, E. Clancy, J.R. Gilbert, K.L. Bailey, and J.A. Derby. 2006. Chlorosis inducing phytotoxi c metabolites: new herbicides from Phoma

macrostoma

. In : Rimando, A.M., S.O. Duke (Eds). Natural Products for Pest Management. American Chemical Society, Washington, DC, pp.37-47.

Graupner, P.R., A. Carr, E. Clancy, J. Gilbert, K.L. Bailey, J-A. Derby, and B. C. Gerwick. 2003. The macrocidins: novel cyclic tetramic acids with herbicidal activity produced by Phoma

macrostoma . J. Nat. Prod., 66: 1558-1561.

Jamaluddin. 1977. A new leaf spot of Agathis caused by Phoma . Proc. Nat. Acad. Sci. India,

47: 71-72.

Kubota, M., and K. Abiko. 2002. Black rot of artichoke leav es caused by two Phoma species in Japan. J. Gen. Plant Pathol., 68: 208-211.14 Newsham, K.K. 2003. Response of saprot rophic fungal communities to declining SO 2 pollution

in the natural environment. Pedobiologia, 47: 77-84.

Pereira, V.J., M.C. Basilio, D. Fernandes, M. Domingues, J.M. Paiva, M.J. Benoliel, M.T. Crespo, and M.V. San Romao. 2009. Occurrence of filamentous fungi and yeasts in three different drinking water sources . Water Research, 43: 3813-3819.

Shattock, R.C., and S.J. Askew. 1980. Stem lesions, canker and dieback of Skimmia japonica

. Ann. Bot., 45: 535-540.

Sidhu J.S., and G. Singh. 1979. Leaf spot disease on Zizyphus mauritiana caused by Phoma

macrostoma in India. Plant Dis ease Reporter, 63: 678-681.

University of Georgia (UGA), Center for Inva sive Species and Ecosystem Health. 2011. Widely prevalent fungi of the United States. http://www.prev alentfungi.org/

Williamson, B., and A.J. Hargreaves. 1979. Fungi on red raspberry from lesions associated with feeding wounds of cane midge ( Resseliella theobaldi ). Ann. Appl. Biol., 91: 303-307.

Zhou, L., K.L. Bailey, and J. Derby. 2004. Plant colonization and enviro nmental fate of the biocontrol fungus Phoma macrostoma . Biol. Control, 30:634-644.