United States
Environmental Protection
Prevention, Pesticides
and Toxic Substances
April 1996
&EPA Ecological Effects Test
OPPTS 850.2400
Wild mammal acute

Public Draft"

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
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
(7 U.S.C. 136, etseq.).
Public Draft Access Information: This draft guideline is part of a
series of related harmonized guidelines that need to be considered as a
unit. For copies: These guidelines are available electronically from the
EPA Public Access Gopher (gopher.epa.gov) under the heading "Environ-
mental Test Methods and Guidelines" or in paper by contacting the OPP
Public Docket at (703) 305-5805 or by e-mail:
To Submit Comments: Interested persons are invited to submit com-
ments. By mail: Public Docket and Freedom of Information Section, Office
of Pesticide Programs, Field Operations Division (7506C), Environmental
Protection Agency, 401 M St. SW., Washington, DC 20460. In person:
bring to: Rm. 1132, Crystal Mall #2, 1921 Jefferson Davis Highway, Ar-
lington, VA. Comments may also be submitted electronically by sending
electronic mail (e-mail) to: guidelines@epamail.epa.gov.
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, or call 202-512-0135 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

OPPTS 850.2400 Wild mammal acute toxicity.
(a)	Scope—(1) Applicability. This guideline is intended to meet test-
ing requirements of both the Federal Insecticide, Fungicide, and
Rodenticide Act (FIFRA) (7 U.S.C. 136, et seq.) and the Toxic Substances
Control Act (TSCA) (15 U.S.C. 2601).
(2) Background. The source material used in developing this har-
monized OPPTS test guideline is OPP 71-3 Wild Mammal Toxicity Test
(Pesticide Assessment Guidelines, Subdivision E—Hazard Evaluation;
Wildlife and Aquatic Organisms) EPA report 540/09-82-024, 1982.
(b)	When required—(1) Data on the toxicity of a pesticide to wild
mammals are required by 40 CFR 158.145 on a case-by-case basis to sup-
port the registration of an end-use product intended for outdoor applica-
tion. The toxicity data required by 40 CFR 158.135 for evaluating hazards
to humans and domestic animals are normally adequate to indicate hazard
to wild mammals. Under certain conditions, however, these data are not
sufficient to assess the potential hazard to wild mammals which are likely
to be exposed to an end-use product. Examples of circumstances when
data on the toxicity of a pesticide to wild mammals may be required by
40 CFR 158.145 include, but are not limited to, the following:
(1)	When data (e.g. data required by 40 CFR 158.135) indicate that
there is considerable variation in the sensitivity of different mammalian
species to the toxic effects of a pesticide, and when there is evidence that
wild mammals are heavily exposed to pesticides; and
(ii) When pesticides with bactericidal properties will be applied to
the forage of wild ruminants, and toxicological data do not include infor-
mation on possible interference with rumen fermentation in domestic (or
wild) ruminants.
(2)	See 40 CFR 158.50, "Formulators' exemption," to determine
whether these data must be submitted. Section II-A of this Subdivision
provides an additional discussion on this subject.
(c)	Test standards. Data sufficient to satisfy the requirements in 40
CFR 158.145 should be derived from tests which comply with the general
test standards in § 70-3 and all of the following standards:
(1) Test substance—(i) Generally, data should be derived from test-
ing conducted with the technical grade of each active ingredient in the
(ii) In special circumstances, data from testing conducted with the
applicant's end-use formulation or a typical end-use product may be re-
quired by 40 CFR 158.75(b) to support the registration of an end-use prod-
uct. Examples of such circumstances include, but are not limited to:

(A)	When effects of the product on the rumen fermentation proces
need to by determined, or
(B)	When secondary toxicity studies need to be performed (e.g. prod-
uct is introduced into prey species which is fed to predator species).
(2)	Species. Testing should be performed on a mammalian species
representative of those found in the area(s) likely to be affected by the
proposed use pattern(s). Test animals may be pen-reared or wild captured,
but should be phenotypically indistinguishable from wild mammals. In no
case should endangered or threatened animals be used for testing.
(3)	Toxicity determination. When the animals are large or the spe-
cies is relatively scarce, a study which determines only the approximate
maximum tolerated dosage for the test species may be acceptable. In all
other cases, the acute oral LD50, dietary LC50, or dietary no-effect level
should be determined, following consultation between the Agency and the
(d)	Reporting and evaluation of data. In addition to the information
provided in § 70-4, test reports should contain the following information:
(1)	Age of each animal and how determined;
(2)	Mean body weight for each test and control group at initiation
and termination of test;
(3)	Number of animals of each sex of animal tested;
(4)	Total food consumption for each test and control group;
(5)	Test diet;
(6)	Dose schedule;
(7)	Mortality;
(8)	Number and circumstances of accidental deaths or injuries;
(9)	LD50 (in mg/kg) or LC50 (in ppm) with 95 percent confidence
limits, or estimated maximum tolerated dose;
(10)	Specified methods used to calculate LD50 or LC50; and
(11)	Slope of the dose-response line.
(e)	Acceptable protocols. Because the Agency intends that toxicity
tests on mammals, other than those required by 40 CFR 158.135, be con-
ducted only to access specific situations, no single test methodology can
provide adequate guidance for all cases. In addition, there are no widely
accepted protocols that include husbandry practices appropriate to a diver-
sity of mammals. Therefore, the test methodologies should be determined

on a case-by-case basis. Appropriate tests methods should be developed
by the registrant in consultation with the Agency. The following are of-
fered for guidance.
(1)	Dietary LC50 and no-effect level tests. Methods for dietary tests
may be adapted from the subchronic oral dosing studies for mammals in
OPPTS 870.3100 and/or from the avian dietary LC50 study in OPPTS
(2)	Toxicity studies for large and relatively scarce mammals. An
example of an acceptable protocol for toxicity studies with mammals that
are large, relatively scarce, or otherwise difficult to obtain is provided as
a modification of a protocol that appears in an unpublished draft report
to EPA from the American Institute of Biological Sciences (AIBS), titled
Analysis of Specialized Pesticide Problems, Volume VI, Wildlife Toxi-
cology Study, pages 4 to 9. This report is dated October, 1974 and was
funded under EPA Contract No. 68-01-2457.