### UNITED STATES ENVIRONMENTAL PROTECTION AGENCY WASHINGTON, D.C. 20460



### MAR 0 4 2013 OFFICE OF CHEMICAL SAFETY AND POLLUTION PREVENTION



### MEMORANDUM SUBJECT:

FROM:

TO: Environmental Risk Assessment of Pasteuria spp. (Hoplolaimus galeatus nematode)- Ph 3 (PC 0 1 6457) for a Sec t ion 3 Registration of the Techni ca l Produ ct (EPA File Sym b ol 85004-RN ) and En d Use Product (EPA F ile Symbo l 85004

-RR) for Contro l ofthe Lance N ematode in Turf; Deci sion# 463287, DP Barcode# 404660 , Submission# 914496 (for technical); Decision # 463290, D P Barcode# 404664, Submission# 914500 (for end

-use); MRIDs 4 8787614 and 48787510

Jeannine Kausch , Regulatory Action L eader Microbial Pesticides Branch (7511P) B i opesticides and Pollution Prevention Divi sion


### I. Environmental Risk Assessment A. Introduction Macin t osh and Associates, Inc. has submitted on behalf of Pasteuria Bioscience, Inc. an application for a Section 3 registration of a technical product (Pasteuria spp. (Hoplolaimus galeatus nematode)

- Ph3) and end-use product (NewPro) containing a new active ingredient Pasteuria spp . (Hoplolaimus galeatus nematode)- Ph3. The end use product (EP) also contains Pasteuria usage- Bll, which is an active ingredient that was regi stered in 2009. The EP is proposaed for application as a liquid to turf to control both lance (Hoplolaimus galeatus) and sting (Belonolaimus longicaudatus) nematodes. Wi th the except ion ofPasteuria ramosa, Pasteuria spec ies are recognized as obligate parasites ofp lant-p arasitic nematodes (Chen and Dickson, 1998, Noel et al. 2005). Host reco rds show that Pasteuria spp. are found in nematodes in at least 80 countries on five continents, as well as on islands in the Atlantic , Pacific, and Indian Oceans (Chen and Dickson 1 998 , Cianci o et al. 1994 , Sturhan 1 988). Pasteuria spp. (Hoplolaimus galea/us nematode)- Ph3 exerts a p esticidal effect 

### on the lance nematode through paras i tism that ultimately results in 

### death.Because of its proposed u se patterns, expos ure of nontarget organi sms is possible with the proposed application of Pasteuria spp. (Hoplolaimus galeatus nematode)

- Ph3. Pasteuria Bioscience, I nc . has s ubmitted scientific rationale , c iting information from published literature, to satisfy data requirements for nontarget organism testing with the active ingredient (T GAI). T his memorandum contains BPPD 's ecological risk assessment for the prop o sed registrations of the technical product and EP containing Pasteuria spp. (Hoplolaimus galeatus nematode)- Ph3 as an active ingredient. B. Summary of N ontarget Effects Data and Evaluation of Scientific Rationale Ta ble 1 pro vi des the status of the data requirements fo r the TGAI. The information provided is suffi cient to satisfy th e Tier I nontarget organism data requirements for the technical and the EP
. Further testing of nontarget organisms at higher tier levels is not required. Evaluation of the scientific rationale submitted in support of the proposed registrations is contained in the attached Data Evalua tion Record

. Table 1. Summary of data submitted to comp ly with nontarget organism data requirements publi sh ed in 40 CFR 

### § 1 58.2150.
Data Requirement OPPTS Results Summary and Classification MRIDNo.• G uideline N o. • Avian ora l toxicity 885.4050 The rati onale provides su fficient information 48787614 • Freshwater fish 88 5

.4 2 00 to determ ine that toxicity / pathoge ni c ity t o tox ici ty/ pathogenicity these no ntarge t species is not expected. •Fre shw ater 885.424 0 C lass ification : Acceptable invertebrate toxic ity/pathogenic i ty • Nontar get insect 885.4340 testing • Ho n ey bee testing 885.4380 Avian inhalation 885.4100 Data are not required

. Nature of microbial N/A toxicity/pathogen ic ity pesticide does not indicate potentia l pathogenicity to birds or relatedness to any known b ird pathogens. Wild mammal 885.4150 Tests r e quired by 40 CFR § 158.2140 are N/A toxic i ty/ pathoge ni c i ty adequate and appropriate for assessment of hazards to wild mammals. Studies submitted with labo ratory rats indicate no adv e rse effects due to o ral exposure. Classification: Acceptable for wild mammal risk assessment

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## I Data Requirement OPPTS Results Summary and Classification MRIDNo.• Guideline No. Estuarine/marine fish 885.4280 Data not required. Pasteuria spp. N/A and inverteb rate (Hoplolaimus galeatus nematode)- Ph3 testing will not be applied directly to water and is not expected to enter marine /estuarine environments in amounts that would result in significant concentration s. Pasteuria spp. (Hoplolaimus galeatus nematode)- Ph3 is an obligate parasite of plant-parasit ic nemat odes inhabiting terr estrial soil. Nontarget plant testing 885.4300 Data not required. Pasteuria spp. N/A (Hoplolaimus galeatus nematode)

- Ph3 is not related to known plant pathogens , and adverse effects to plants are not expected. •copies of references Cited m the waiver rationales are contamed m MR1Ds 47426714 and 48787601. Additional rationale was sub mitted for the EP in MRID 48787510. Data were not specifically required on the EP, and the rationale was determined to be supplemental. C. Ecological Exposure and Risk Characterization 1. Terrestrial Animals and Plants The proposed EP is for use on turf, and is applied by directed spray to soil (pre-plant) or turf. Applications are to be followed with a sufficient amount of water to move the Pasteuria spp. (Hoplolaimus galeatus nematode)- Ph3 spores into the root zone. While this type of application reduce s the potential for exposure to many nontarget species , exposure to nontarget species consuming , contacting, or living in the soil will not necessarily be eliminated. Scientific rationale provides sufficient information to conclude that adverse effects are not expected in birds , non-target insects, and honey bees as a result of exposure to Pasteuria spp. (Hoplolaimus galeatus nematode)- Ph3 (MRID 48787614). Pasteuria spp. are widespread in their distribution throughout the world (Ciancio et al. 1994 , Ciancio et al. 1998 , Giblin-Davis et al. 1990 ; Sayre and Starr 1985 , Sturhan 1988 ). They are known only as parasites ofplantparasitic nematodes, with the exception ofP. ramosa that is known only to parasitize Daphnia spp . (Atibalentja et al 2000, Chen and Dickson 1998, Sayre et al. 1991 ). Therefore, Pasteuria spp. (Hoplolaimus galeatus nematode)- Ph3 is not expected to be toxic or pathogenic to birds , nontarget insects or honey bees if they are exposed as a result of the proposed labeled applications. An acute oral toxicity/pathogenicity study with laboratory rats (MRID 48787612) is available to evaluate the potential effects ofPasteuria spp. (Hoplolaimus galeatus nematode)

- Ph3 on wild mammals

. In the acute oral toxicity/pathogenicity study, laboratory rats were dosed with I mL oftest material (3 x 10 8 spores/mL) and were observed for 21 days. No signs of toxic or pathogenic effects were found as a result ofthe exposure. This study was determined to besupplemental because clearance and infectivity were not observed, since Pasteuria spp. cannot be cultured outside oftheir hosts. There is no reason that the test animals in these studies would not be adequate models with which to determine potential effects to wild mammals, so adverse effects to wild mammals are not expected as a result ofthe proposed labeled applications of Pasteuria spp. (Hoplolaimus galeatus nematode)- Ph3. Non-target plant testing is not required because Pasteuria species are not related to any known plant pathogen. Adverse effects on plants are not expected to result from proposed labeled applications of Pasteuria spp. (Hoplolaimus galeatus nematode)- Ph3

. Additionally, there are no reports oftoxicity or pathogenicity ofPasteuria spp. (Hoplolaimus galeatus nematode)

- Ph3 to nontarget terrestrial animals or plants. To support their data waiver rationale, a search was performed by the applicant within the Agricola and PubMed databases , along with several biological journals (i.e., Journal of Bacteriology, Applied and Environmental Microbiology, FEMS Microbiology Ecology, Soil Biology and Biochemistry) for the period of 1 980-2012. The search was conducted using "Pasteuria" as the search word, and no reports of acute toxicity d ata on birds , freshwater fish , freshwater invertebrates , nontarget insects, and honey bees. BPPD also conducted a search within the Thomson Reuters Web of Knowled ge Database , which simultaneously searches the**W~b** of Science, Current Contents Connect, Medline , and Journal Citation Reports databases. The search was performed with the term "Pasteu ria" coupled with each ofthe search terms of"bird," " mammal ," "invertebrate," "art hropod," "insect," " honey bee," "plant pathogen," and "phytopathogen" for all ofthe years available. N o

_

r eports of adverse effects ofPasteuria species in nontarget organisms were found in any ofthese searches. Any reports of effects to nontarget organisms related to Pasteuria species that are found in the future may trigger the need for testing; however , given the specificity of these bacteria for their hosts as is currently understood, such reports are not expected. Based on the above rationale and available data, adverse effects are not expected to occur to terrestrial animals or plants as a result of proposed labeled applications of Pasteuria spp. (Hoplolaimus ga/eatus nematode)- Ph3. Additionally, adverse effects to terrestrial nontarget organisms were not ident ified for Pasteuria usgae- B11, 1 and because of the specificity of Pasteuria spp. for their hosts, no interaction s between these bacteria are expected. Therefore, additional risk is not anticipated for terrestrial nontarget species resulting from proposed uses of the E P containing both Pasteuria spp. (Hoplolaimus galeatus nematode)

- Ph3 and Pasteuria usgae - Bll. 2. Aquatic Animals and Plants Applications of the EP are intended primarily to put Pasteuria spp. (Hoplolaimus galeatus nematode)

- Ph3 in the root zone of the treated plants, so runoff into aquatic areas is not likely to be significant. Pasteuria spp. are naturally found in soi l , so some natural presence ofPasteuria spp. (Hoplolaimus galeatus nemato de)- Ph 3 in surface waters is expected. T o date, no information is available on the size of naturally occurring populations ofPas t euria spp.

1 Memorandum from S

. Borges through Z
. V ituzis to J. Kausch, Subject: Environmental risk assessment for Pasteuria usgae .. .for control of sting nematodes in turf and strawberries, dated Apri18 , 2009. 

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# .. 

# 'Scientific rationale was submitted to fulfill data requirements for and support effects conclusions for freshwater aquatic organisms (MRI D 487876 14). The rationale provides suffic ient information to conclude that adverse effects are not expected in freshwater fish or freshwater invertebrates as a result of exposure to Pas teuria spp. (Hoplolaimus galeatus nematode)- Ph3. The rationale was similar to that presented for terrestrial organisms in that Pas teuria spp. are found in soils worldwide. Pasteuria ramosa is a closely

-related species that parasitizes Daphnia magna and other Daphnia spp. (Ebert et al. 1996). However, it is widely understood that all other known Pasteuria spp. are obligate parasites of soi l-dwelling nem atodes (Atibalentja et al. 2000, Chen and Dickson 1998 , Sayre et al. 1991 ). There are no available reports in the literature documenting pathogenicity or toxicity to freshwater fish or invertebrates as a result of exposure to Pasteuria spp. (Hoplolaimus galeatus nematode)

- Ph3

. A search similar to the one described above was performed for aquatic animals and plants. In addition to the search terms described above, the terms "fish , " "aquatic," "freshwater, " "estuarine, " and "marine" were also searched. No reports of adverse effects of Pasteuria spp. in aquatic nontarget organisms were found in any of these searches. As noted above , reports of adverse effects that may be found in the future may tri gger the need for testing ; however , none are anticipated due to the specificity ofPasteuria spp. for their nematode hosts. Based on the rationale and other information subm i tted, adverse effects to freshwater and marine/estuarine fish, invertebrates, and plants are not expected as a result of exposure to proposed labeled applications ofPasteuria spp. (Hoplolaimus galeatus nematode)

- Ph3. As noted above , due to the specificity ofPaste uria spp. for their hosts, no interactions between these bacteria are expected. Therefore , additional risk to aquatic nontarget species is not anticipated for the proposed uses ofthe EP containing both Pasteuria spp. (Hoplolaimus galeatus nematode)

- Ph3 and Pasteuria usgae - B 11. D. Threatened and Endangered Species Assessment Since EPA has determined that no effects are anticipated for any nontarget species exposed to Pasteuria spp. (Hoplolaimus galeatus nematode)- Ph3 as a result ofthe proposed labeled applications , effects to federally-listed threatened and endangered species and their designated critical habitats are also not expected. Therefore , a "No Effect" determination is made for direct and indirect effects to listed species and their designated critical habitats resulting from the proposed uses ofPasteuria spp. (Hoplolaimu s galeatus nematode)- Ph3 , as labeled. II. References Atibalentja , N., B. P. Jakstys, and G. R. Noel. 2004. Life cycle , ultras tructure , and host specificity ofthe North American isolate of Pasteuria that parasitizes the Soybean cyst nematode, Heterodera g l y cines. J. N ematol. 36: 171

- 180. Chen, Z. X., and D. W. Dickson

. 1998. Review ofPasteuria penetrans: biology, ecology, and biological control potential. J. Nematol. 30: 313

- 340.Ciancio , A., R. Bonsignore, N. Coclas, and F. Lamberti. 1994. Ho st re co rd s and spore morphometries of Pasteuria penetrans group parasites of nematodes. 

### J
. Invert. Pathol. 63: 260

- 267

. C iancio , A. , V. Farf an , E.C. Torres , and G. Grasso. 1998. Observations on a Pasteuria isolate parasitic o n Hoplolaimus galeatus in Peru. J. Nematol. 30:206-21 0

. Ebert, D., P. Rainey, T
. M. Embley , and D. Scholz. 1 996. Development, lif e cycle, ultrastructure, and phylogenetic position ofPasteuria ram osa M et chniko ff 1888: redi scovery of an obligate endoparasite ofDaphnia magna Straus. Philosophical Transactions of the Roy al Society of London series B. 351:1689-1701. Noe l, R.N., Atibalentja, N., and L. L. Domi er. 2005. Emended description ofPasteuria nishizawae. Int
. J. Systematics and Evolut. Mic robiol. 55: 1681-1685. Sayre, 

### R. M
. and M
. P. Starr. 1985. Pasteuriapenetrans (ex Thorn e, 1940) nom: rev. , comb. n , sp . m. , a mycelial and endospore-forming bacterium parasitic in plant-parasitic nematodes. Pro ceedings of the Helrninothological Society of Washington 52: 149-165. Sayre , R. M. , W. P
. Wergin, J
. M. Schmidt, and M.P. Starr. 199 1. Pasteuria nishizawae sp. nov. , a mycelial and endospore-forming bacterium parasitic on cyst nematodes of genera Heterodera and Globode ra. Res. Microbiol. 14 : 55 1-5 64

. Stur h an, D. 1988. New host and geographical records of nematode- parasitic bacteria of the Pasteuria penetrans group. Nematol ogica 3 4 : 350

- 356.