United States Prevention, Pesticides EPA712-C-96-339
Environmental Protection and Toxic Substances February 1996
Agency (7101)
&EPA Microbial Pesticide
Test Guidelines
OPPTS 885.5300
Expression in a
Freshwater Environment
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INTRODUCTION
This guideline is one of a series of test guidelines that have been
developed by the Office of Prevention, Pesticides and Toxic Substances,
United States Environmental Protection Agency for use in the testing of
pesticides and toxic substances, and the development of test data that must
be submitted to the Agency for review under Federal regulations.
The Office of Prevention, Pesticides and Toxic Substances (OPPTS)
has developed this guideline through a process of harmonization that
blended the testing guidance and requirements that existed in the Office
of Pollution Prevention and Toxics (OPPT) and appeared in Title 40,
Chapter I, Subchapter R of the Code of Federal Regulations (CFR), the
Office of Pesticide Programs (OPP) which appeared in publications of the
National Technical Information Service (NTIS) and the guidelines pub-
lished by the Organization for Economic Cooperation and Development
(OECD).
The purpose of harmonizing these guidelines into a single set of
OPPTS guidelines is to minimize variations among the testing procedures
that must be performed to meet the data requirements of the U. S. Environ-
mental Protection Agency under the Toxic Substances Control Act (15
U.S.C. 2601) and the Federal Insecticide, Fungicide and Rodenticide Act
(7U.S.C. I36,etseq.).
Final Guideline Release: This guideline is available from the U.S.
Government Printing Office, Washington, DC 20402 on The Federal Bul-
letin Board. By modem dial 202-512-1387, telnet and ftp:
fedbbs.access.gpo.gov (IP 162.140.64.19), internet: http://
fedbbs.access.gpo.gov, or call 202-512-0132 for disks or paper copies.
This guideline is also available electronically in ASCII and PDF (portable
document format) from the EPA Public Access Gopher (gopher.epa.gov)
under the heading "Environmental Test Methods and Guidelines."
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OPPTS 885.5300 Expression in a freshwater environment.
(a) Scope—(1) Applicability. This guideline is intended to meet test-
ing requirements of the Federal Insecticide, Fungicide, and Rodenticide
Act (FIFRA) (7 U.S.C. 136, et seq.\
(2) Background. The source material used in developing this har-
monized OPPTS test guideline is OPP guideline 155A-11.
(b) General. This guideline is to be used with OPPTS 885.5000.
(1) Data an the expression of a microbial pest control agent (MPCA)
in a freshwater environment are required by 40 CFR 158.740 to support
the registration of each end-use product intended for outdoor application
on fresh water and each manufacturing-use product that legally may be
used to formulate such an end-use product when toxic or pathogenic ef-
fects are observed in any of the following Tier I tests:
(i) Avian oral pathogenicity/toxicity test: Tier I. (OPPTS 885.4050)
(ii) Avian respiratory pathogenicity test (OPPTS 885.4100).
(iii) Wild mammal toxicity and pathogenicity test (OPPTS 885.4150)
(iv) Freshwater fish toxicity and pathogenicity testing (OPPTS
885.4200).
(v) Freshwater aquatic invertebrate toxicity and pathogenicity tests
(OPPTS 885.4240).
(vi) Plant studies—aquatic (OPPTS 885.4300).
(2) See 40 CFR 158.50 and 158.740 to determine whether these data
must be submitted.
(c) Test standards. (1) Tests shall be conducted in a simulated aquat-
ic environment (e.g. aquarium with bottom sediment) to determine whether
the MPCA is able to survive, persist, and replicate in a freshwater environ-
ment consisting of fresh water and bottom sediment representative of the
proposed use site. The following parameters should be varied to determine
their effect on the survival and growth of the MPCA population:
(i) Temperature.
(ii) pH.
(iii) Nutrients.
(iv) Sunlight.
(v) Oxygen content.
(vi) Hardness.
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(vii) Turbulence.
(2) The values selected for each parameter listed should be selected
to approximate the conditions expected at the intended use site.
(3) Specialized laboratory studies designed to determine the MPCA's
growth requirements (e.g., temperature, pH, sunlight, and oxygen) may
supplement the study described in this guideline. Specialized laboratory
studies may demonstrate that the MPCA will be unable to survive and
persist in a freshwater environment. In such instances, the Agency will
consider studies an individual basis to meet the intent of testing in lieu
of the study described in this guideline.
(d) Test substance. A typical end-use product or the technical grade
of the active ingredient shall be tested.
(e) Test duration. Data to establish a population decline curve should
be collected at intervals until two half-life determinations have been made
or until data establish that the microbial agent population is able to main-
tain itself in a freshwater environment at or above the level present imme-
diately after test initiation.
(f) Reporting and evaluation of data. The reporting and evaluation
provisions are the same as those set forth in OPPTS 885.5000.
(g) Tier progression. If results of this study and use patterns informa-
tion indicate that the MPCA is likely to enter and is able to persist in
a freshwater environment so that the susceptible nontarget organisms tested
in Tier I are likely to be exposed, the appropriate testing in Tier III
(OPPTS 885.4550 through OPPTS 885.4850) is required as specified in
40 CFR 158.740.
(h) References. The following references contain useful for develop-
ing acceptable protocols:
(1) Anthony, D.W. et al. Field tests with Nosema Algerae Vavra and
Undeen (Microsporida, Nosematidae) against Anopheles albinamus Wiede-
mann in Panama. Miscellaneous Publications of the Entomological Society
of America 11:17-28 (1978).
(2) Brand, R.J. et al. Methods for assessing field persistence of Bacil-
lus thuringiensis spores. Journal of Invertebrate Pathology 25:199-208
(1975).
(3) EPA. Impact of the use of microorganisms on the aquatic environ-
ment. EPA publication 660-3-75-001. Technical Publications Office, En-
vironmental Protection Agency, National Environmental Research Center,
Corvallis, OR 97330 (1975).
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(4) Hostetter, D.L. et al. Persistence of formulations of Bacillus
thuringiensis spores and crystals on eastern red cedar foliage in Missouri
Journal of the Kansas Entomological Society 48:189-193 (1975).
(5) Ignoffo, C.N. et al. Stability of Bacillus thuringiensis and
Baculovirus heliothis on soybean foliage. Environmental Entomology
3:117-119(1974).
(6) Kaya, H.K. Persistence of spores of Pleistophora schuber
(Onidospora: Microsporida) in the field and their application in microbial
control. Journal of Invertebrate Pathology 26:329-332 (1975).
(7) Pinnock, D.E. et al. The field persistence of Bacillus thuringiensis
spores. Journal of Invertebrate Pathology 18:405-411 (1971).
(8) Sinclair, J.L. and M. Alexander. Role of resistance to starvation
in bacterial survival in sewage and lake water. Applied Environmental
Microbiology 48:410-415 (1984).
(9) Young, S.Y. Pre- and posttreatment assessment of virus levels.
Selected papers from EPA-USDA Working Symposium. M. Summers, R.
Engler, L. Falcon, and P. Vail (eds.) American Society of Microbiology
(1975).
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