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)- Ph3 (PC 016457) for a Section 3 Registration of the Technical 

Product (EPA File Symbol 85004-RN) and End Use Product (EPA File 

Symbol 85004-RR) for Control of the Lance Nematode in Turf; Decision# 

463287, DP Barcode# 404660, Submission# 914496 (for technical); Decision 

# 463290, DP Barcode# 404664, Submission# 914500 (for end-use); MRIDs 

48787614 and 48787510 

Jeannine Kausch, Regulatory Action Leader 

Microbial Pesticides Branch (7511P) 

Biopesticides and Pollution Prevention Division 

I. Environmental Risk Assessment 

A. Introduction 

Macintosh 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 registered in 2009. The EP is 

proposaed for application as a liquid to turf to control both lance (Hoplolaimus galeatus) and 

sting (Belonolaimus longicaudatus) nematodes. 

With the exception of Pasteuria ramosa, Pasteuria species are recognized as obligate parasites 

of plant-parasitic nematodes (Chen and Dickson, 1998, Noel et al. 2005). Host records 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 1998, Ciancio et al. 1994, 

Sturhan 1988). Pasteuria spp. (Hoplolaimus galea/us nematode)- Ph3 exerts a pesticidal effect 

on the lance nematode through parasitism that ultimately results in death. 
Because of its proposed use patterns, exposure of nontarget organisms is possible with the 

proposed application of Pasteuria spp. (Hoplolaimus galeatus nematode) - Ph3. Pasteuria 

Bioscience, Inc. has submitted scientific rationale, citing information from published literature, 

to satisfy data requirements for nontarget organism testing with the active ingredient (TGAI). 

This memorandum contains BPPD's ecological risk assessment for the proposed registrations of 

the technical product and EP containing Pasteuria spp. (Hoplolaimus galeatus nematode)- Ph3 

as an active ingredient. 

B. Summary of Nontarget Effects Data and Evaluation of Scientific Rationale 

Table 1 provides the status of the data requirements for the TGAI. The information provided is 

sufficient to satisfy the 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 Evaluation Record. 

Table 1. Summary of data submitted to comply with nontarget organism data requirements 

published in 40 CFR § 158.2150. 

Data Requirement OPPTS Results Summary and Classification MRIDNo.• 

Guideline No. 

• Avian oral toxicity 885.4050 The rationale provides sufficient information 48787614 

• Freshwater fish 885.4200 to determine that toxicity/pathogenicity to 

toxicity/pathogenicity these nontarget species is not expected. 

•Freshwater 885.4240 Classification: Acceptable 

invertebrate 

toxicity/pathogenicity 

• Nontarget insect 885.4340 

testing 

• Honey bee testing 885.4380 

Avian inhalation 885.4100 Data are not required. Nature of microbial N/A 

toxicity/pathogenicity pesticide does not indicate potential 

pathogenicity to birds or relatedness to any 

known bird pathogens. 

Wild mammal 885.4150 Tests required by 40 CFR § 158.2140 are N/A 

toxicity/pathogenicity adequate and appropriate for assessment of 

hazards to wild mammals. Studies submitted 

with laboratory rats indicate no adverse 

effects due to oral exposure. 

Classification: Acceptable for wild 

mammal risk assessment 
.. 

~ 

I 

Data Requirement OPPTS Results Summary and Classification MRIDNo.• 

Guideline No. 

Estuarine/marine fish 885.4280 Data not required. Pasteuria spp. N/A 

and invertebrate (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 concentrations. Pasteuria spp. 

(Hoplolaimus galeatus nematode)- Ph3 

is an obligate parasite of plant-parasitic 

nematodes inhabiting terrestrial 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 submitted 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 

reduces 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 of plantparasitic nematodes, with the exception of P. 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 of Pasteuria 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 108 spores/mL) and were observed for 21 days. No signs of toxic or 

pathogenic effects were found as a result of the exposure. This study was determined to be 
supplemental because clearance and infectivity were not observed, since Pasteuria spp. cannot 

be cultured outside of their 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 of the 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 of toxicity or pathogenicity of Pasteuria 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 

1980-2012. The search was conducted using "Pasteuria" as the search word, and no reports of 

acute toxicity data on birds, freshwater fish, freshwater invertebrates, nontarget insects, and 

honey bees. BPPD also conducted a search within the Thomson Reuters Web of Knowledge 

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 

"Pasteuria" coupled with each ofthe search terms of"bird," "mammal," "invertebrate," 

"arthropod," "insect," "honey bee," "plant pathogen," and "phytopathogen" for all of the years 

available. No_reports of adverse effects of Pasteuria species in nontarget organisms were found 

in any of these 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 identified for Pasteuria usgae- B11,1 and because of the specificity of 

Pasteuria spp. for their hosts, no interactions between these bacteria are expected. Therefore, 

additional risk is not anticipated for terrestrial nontarget species resulting from proposed uses of 

the EP 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 soil, so some natural presence of Pasteuria 

spp. (Hoplolaimus galeatus nematode)- Ph3 in surface waters is expected. To date, no 

information is available on the size of naturally occurring populations of Pasteuria spp. 

1 Memorandum from S. Borges through Z. Vituzis to J. Kausch, Subject: Environmental risk assessment for 

Pasteuria usgae .. . for control of sting nematodes in turf and strawberries, dated Apri18, 2009. 

+ .. 

' 
Scientific rationale was submitted to fulfill data requirements for and support effects conclusions 

for freshwater aquatic organisms (MRID 48787614). The rationale provides sufficient 

information to conclude that adverse effects are not expected in freshwater fish or freshwater 

invertebrates as a result of exposure to Pasteuria spp. (Hoplolaimus galeatus nematode)- Ph3. 

The rationale was similar to that presented for terrestrial organisms in that Pasteuria 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 soil-dwelling nematodes (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 trigger the need for testing; 

however, none are anticipated due to the specificity of Pasteuria spp. for their nematode hosts. 

Based on the rationale and other information submitted, adverse effects to freshwater and 

marine/estuarine fish, invertebrates, and plants are not expected as a result of exposure to 

proposed labeled applications of Pasteuria spp. (Hoplolaimus galeatus nematode) - Ph3. As 

noted above, due to the specificity of Pasteuria 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 of the 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 of the 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 of Pasteuria spp. (Hoplolaimus galeatus nematode)- Ph3, as labeled. 

II. References 

Atibalentja, N., B. P. Jakstys, and G. R. Noel. 2004. Life cycle, ultrastructure, and host 

specificity of the North American isolate of Pasteuria that parasitizes the Soybean cyst 

nematode, Heterodera glycines. J. Nematol. 36: 171-180. 

Chen, Z. X., and D. W. Dickson. 1998. Review of Pasteuria penetrans: biology, ecology, and 

biological control potential. J. Nematol. 30: 313-340. 
Ciancio, A., R. Bonsignore, N. Coclas, and F. Lamberti. 1994. Host records and spore 

morphometries of Pasteuria penetrans group parasites of nematodes. J. Invert. Pathol. 63: 

260-267. 

Ciancio, A., V. Farfan, E.C. Torres, and G. Grasso. 1998. Observations on a Pasteuria isolate parasitic 

on Hoplolaimus galeatus in Peru. J. Nematol. 30:206-210. 

Ebert, D., P. Rainey, T. M. Embley, and D. Scholz. 1996. Development, life cycle, 

ultrastructure, and phylogenetic position of Pasteuria ramosa Metchnikoff 1888: rediscovery 

of an obligate endoparasite of Daphnia magna Straus. Philosophical Transactions of the 

Royal Society of London series B. 351:1689-1701. 

Noel, R.N., Atibalentja, N., and L. L. Domier. 2005. Emended description of Pasteuria 

nishizawae. Int. J. Systematics and Evolut. Microbiol. 55: 1681-1685. 

Sayre, R. M. and M. P. Starr. 1985. Pasteuriapenetrans (ex Thorne, 1940) nom: rev., comb. n, 

sp. m., a mycelial and endospore-forming bacterium parasitic in plant-parasitic nematodes. 

Proceedings of the Helrninothological Society of Washington 52: 149-165. 

Sayre, R. M., W. P. Wergin, J. M. Schmidt, and M.P. Starr. 1991. Pasteuria nishizawae sp. 

nov., a mycelial and endospore-forming bacterium parasitic on cyst nematodes of genera 

Heterodera and Globodera. Res. Microbiol. 14: 55 1-564. 

Sturhan, D. 1988. New host and geographical records of nematode-parasitic bacteria of the 

Pasteuria penetrans group. Nematologica 34: 350-356.