United States	Prevention, Pesticides	EPA712-C-96-122
Environmental Protection	and Toxic Substances	April 1996
Agency	(7101)
&EPA Ecological Effects Test
OPPTS 850.1500
Fish life cycle toxicity

"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 850.1500 Fish life cycle 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 72-5 Life-Cycle Tests of Fish (Pes-
ticide Assessment Guidelines, Subdivision E—Hazard Evaluation; Wildlife
and Aquatic Organisms) EPA report 540/09-82-024, 1982.
(b)	When required. (1) Data obtained from a life-cycle test of fish
are required by 40 CFR 158.145 to support the registration of an end-
use product intended to be applied directly to water or expected to trans-
port to water from the intended use site, and when any of the following
conditions apply:
(1)	If the estimated environmental concentration is equal to or greater
than one-tenth of the no-effect level in the fish early life-stage or inverte-
brate life-cycle test; or
(ii) If studies of other organisms indicate the reproductive physiology
of fish may be affected.
(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 the following test standards:
(1)	Test substance. Data shall be derived from testing conducted with
the technical grade of each active ingredient in the product.
(2)	Duration of tests. Fish should be cultured in the presence of the
test substance from one stage of the life cycle to at least the same stage
of the next generation (e.g. egg to egg).
(3)	Species. Testing should be performed on a freshwater fish (e.g.
fathead minnow). An estuarine species (e.g. sheepshead minnow) may be
used if the pesticide is expected to enter the estuarine environment.
(4)	Concentration analysis. The concentration of the test substance
in the water should be determined at the start of the study and periodically
throughout the study to verify concentrations.
(d)	Reporting and evaluation of data. In addition to the basic infor-
mation provided in § 70-4, the test report should contain the following
information (where appropriate):

(1)	Reproductive effects;
(2)	Detailed records of spawning, egg numbers, fertility, and fecun-
(3)	No-effect level, and mortality data;
(4)	Statistical evaluation of effects;
(5)	Locomotion, behavioral, physiological, and pathological effects;
(6)	Definition of the criteria used to determine effects;
(7)	Summary of general observation of signs of intoxication or other
(8)	Stage of life cycle in which organisms were tested;
(9)	Duration of the test; and
(10)	Concentration analysis.
(e) Acceptable protocol—(1) Freshwater fish life-cycle test. An ex-
ample of an acceptable protocol is found in the following reference:
National Water Quality Laboratory Committee on Aquatic Bioassays.
1971. Recommended bioassay procedure for fathead minnow Pimephales
promelas (Rafinesque) chronic tests. (Revised January, 1972). Pp. 15-24
in Biological Field and Laboratory Methods. U. S. Environmental Protec-
tion Agency, Office of Res. and Dev. EPA-670/4-73-001.
(2) Estuarine fish life-cycle test. Examples of acceptable protocols
are found in the following references:
(i) Schimmel, S.C., and D.J. Hansen. 1974. Sheepshead minnow
Cyprinodon variegatus: an estuarine fish suitable for chronic (entire
lifecycle) bioassays. Proc. 28th Ann. Cong. S.E. Assoc. Game-Fish Comm.
Pp. 392-398.
(11)	Hansen, D.J., P.R. Parrish, S.C. Schimmel, and L.R. Goodman.
1978. Life-cycle toxicity test using sheepshead minnows (Cyprinodon
variegatus). Pp. 109-116 in Bioassay Procedures for the Ocean Disposal
Permit Program. U.S. Environmental Protection Agency, Office of Re-
search and Development. EPA-600/9-78-010.