United States
Environmental Protection
Prevention, Pesticides
and Toxic Substances
EPA 712-C-96-155
April 1996
&EPA Ecological Effects Test
OPPTS 850.4300
Terrestrial Plants Field
Study, Tier III

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 850.4300 Terrestrial plants field study, Tier III.
(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 124-1 Terrestrial Field Testing
(Pesticide Assessment Guidelines, Subdivision J—Hazard Evaluation;
Nontarget Plants) EPA report 540/09-82-020, 1982.
(3)	Test objective. This guideline should be used in conjunction with
OPPTS guideline 850.4000, Background—Nontarget plant testing, which
provides general information and overall guidance for the nontarget plants
test guidelines.
(i)	General. (A) Terrestrial field testing studies are designed to pro-
vide phytotoxicity data on a pesticide. These phytotoxicity data are needed
to evaluate the level of pesticide exposure to nontarget terrestrial plants
and to assess the impact of pesticides on endangered and threatened plants
as noted under the Endangered Species Act. Where a phytotoxic effect
is noted in one or more plants, additional terrestrial field testing studies
may be required. These data are required by 40 CFR 158.150 to support
the registration of any pesticide intended for outdoor use under the Federal
Insecticide, Fungicide and Rodenticide Act (FIFRA), as amended.
(B) Pesticides with outdoor use patterns that do not readily release
the pesticide to the environment do not have to be evaluated using this
phytotoxicity test. These use patterns include tree injection, subsurface soil
applications, recapture systems, wick applications, and swimming pool
uses. If any of these use patterns do readily expose nontarget plants to
the pesticide, as through vapors, the pesticide phytotoxicity potential may
need to be evaluated.
(ii)	Objective of the terrestrial field testing Tier III test. (A) The
objective of the Tier III terrestrial field testing study is to determine if
a pesticide exerts a detrimental effect to plants during critical stages in
their development. The test is performed on species from a cross-section
of the nontarget terrestrial plant population. This is a multiple dose test
designed to evaluate the phytotoxic effects of the pesticide over a wide
range of anticipated pesticide quantities as may be found in the environ-
(B) The Tier III terrestrial nontarget plant phytotoxicity field studies
are only required if greater than 25 percent adverse effects on plant growth
for terrestrial plants are expected to occur when the product is used as
directed by the label (the estimated environmental concentration exceeds
the EC25 terrestrial value.

(C) The Tier III tests are expected to provide information on det-
rimental effects to plants during critical stages of development. The typical
end-use product (TEP) is used to assess effects on a broader range of
nontarget plant species in a number of geographical areas.
(b) Test standards. In addition to the general test standards set forth
in OPPTS 850.4000, the test standards for this guideline are the same as
those listed in OPPTS 850.4100, with the following modifications:
(1)	Test substance. The test substance shouldl be the end-use product
or a representative end-use product from the same major formulation cat-
egory for that general use pattern. Examples of major formulation cat-
egories are: Wettable powders, emulsifiable concentrates, and granulars.
(If the manufacturing-use product is usually formulated into end-use prod-
ucts comprising two or more major formulation categories, a separate
study must be performed with a typical end-use product for each category).
The test substance used should contain the highest percentage of active
ingredient (AI) and/or be the most widely used (total pounds AI).
(2)	Application levels. The dosages tested should be the same as
those employed in the Tier III test, OPPTS 850.4225.
(3)	Species, (i) Representatives of the following plant groups are to
be tested, subject to the limitations of paragraph (b)(2)(iii) of this guide-
(A)	Dicotyledonae (dicots), representatives of three families.
(B)	Monocotyledonae (monocots), representatives of three families.
(C)	Vascular Cryptogamae (ferns and allies), representatives of two
(D)	Bryophyta (mosses) or Hepatophyta (liverworts), one representa-
tive (for wetland use patterns only).
(E)	Gymnospermae (conifers), one representative.
(ii)	Plant species used for testing Tiers I and II can be used to satisfy
the monocot or dicot test plant requirements of this guideline.
(iii)	If any of the plant groups are not likely to be exposed to the
pesticide under normal conditions of use, testing of such groups is not
required. Justification for elimination of a test species or group should
be included in the test report.
(iv)	Additional plant species may be required if the general selectivity
of the pesticide cannot be readily identified.
(4)	Test conditions. Plants are to be grown under field-use conditions
similar to those of the natural habitat of the plants in nature.

(5)	Duration. The test duration should be of sufficient length to as-
sess multiple applications directed by the label. Observations should con-
tinue for the entire life cycle of the test plants with observations every
2 to 4 weeks.
(6)	Season of application. The test substance is to be applied over
a period of time or season according to the proposed label instructions.
(7)	Test locations. The pesticide should be tested in those geographic
locations where it is expected to be used, as based on proposed label use
sites. Where important species diversity and physiographic differences
occur within a region of intended application, regional testing may be inad-
equate, and testing at a more specific region or biome level may be re-
quired. United States regional areas of potential testing include: North-
eastern temperate deciduous, Southeastern temperate deciduous, Northern
grassland (prairie), Southern grassland (prairie), Northwestern (and Alas-
kan) conifer forest and high desert, Southwestern chaparral Mediterranean
and low desert, and Hawaiian and Caribbean tropical regions.
(c)	Reporting. In addition to the information required in OPPTS
850.4000 and 850.4100, the test report should include the test conditions
employed (including the soil and environmental conditions) and the deter-
mination of the 50 percent detrimental effect level.
(d)	Data reporting. (1) The registrant's report on terrestrial field test-
ing studies should include all information necessary to provide:
(1)	A complete and accurate description of the field treatments and
(ii)	Sampling data and phytotoxicity rating.
(iii)	Data on storage of the plant material, if so performed.
(iv)	Results of any chemical analysis of the plant material.
(v)	Reporting of the data, rating system, and statistical analysis.
(vi)	Quality control measures/precautions taken to ensure the fidelity
of the operations.
(2)	Each laboratory/greenhouse/small field plot terrestrial field testing
report should include the following information:
(i) General information. (A) Cooperator or researcher (name and
address), test location (county and state; country, if outside of the United
States), and date of study.
(B) Name (and signature), title, organization, address, and telephone
number of the persons responsible for planning/supervising/monitoring
and, for field plot studies, applying the pesticide.

(C)	Trial identification number.
(D)	Quality assurance indicating: Control measures/precautions fol-
lowed to ensure the fidelity of the phytotoxicity determinations, record-
keeping procedures and availability of logbooks, skill of the laboratory
personnel, status of the field and supporting laboratory equipment; degree
of adherence to good laboratory practices; and degree of adherence to good
agricultural practices in maintaining healthy plants.
(E)	Other information the registrant considers appropriate and relevant
to provide a complete and thorough description of the test procedures and
(ii)	Test substance (pesticide). (A) Identification of the test pesticide
AI including chemical name, common name (ANSI, BSI, ISO, WSSA),
and company developmental/experimental name.
(B)	AI percentage in the end-use product or representative end-use
product from the same major formulation category for that general use
(C)	Dose rates in terms of AI per area of land or of leaf (if leaf-
area-index is provided).
(D)	Dose rates in terms of less than the maximum label rate with
dosages in a geometrical progression of no more than twofold and with
subtoxic (
(D) Identification of the date of planting, date of pesticide application,
and date of phytotoxicity rating or harvest.
(iv)	Site of the test. (A) Site description of the terrestrial field testing
study such as a grassland, forested area, fallow field, tilled field.
(B)	Location of the test sites that represent the general regional areas
of potential usage: Northeastern temperate deciduous, Southeastern tem-
perate deciduous, Northern grassland (cool prairie), Southern grassland
(warm prairie), Northwestern (and Alaskan) conifer forest and high desert,
Southwestern chaparral Mediterranean and low desert, and Hawaiian and
Caribbean semitropical and tropical regions.
(C)	Climatological data during the test (records of applicable condi-
tions for the type of site, i.e., temperature, thermoperiod, rainfall or water-
ing regime, light regime—intensity and quality, relative humidity, wind
(D)	Field lay-out (for field plots), e.g., size and number of control
and experimental plots;.number of plants per plot/unit area.
(E)	Population density of seeds or plants.
(F)	Cultural practices such as cultivation and irrigation.
(G)	Substrate characteristics of the sites (name/designation of soil
type and its physical and chemical properties, including pH and percent
organic matter, presence and depth of fragipan or shallow bedrock, etc.).
(v)	Results. (A) Phytotoxicity rating (including a description of the
rating system) for each plant or group of plants (population) in the test.
(B)	Weight, height or other growth parameters that may have been
measured to ascertain toxic effects of the pesticide upon the plants.
(C)	Statistical analysis of the results including environmental or effec-
tive concentration (EC) values.
(vi)	Evaluation. Determination as to whether additional phytotoxicity
testing will be necessary to characterize the phytotoxic nature of the chem-
(e) References. The following references should be consulted for ad-
ditional background material on this test guideline.
(1)	Little, T.M. and F.J. Hills. Agricultural Experimentation - Design
and Analysis. Wiley, NY (1978).
(2)	Truelove, B., ed. Research Methods in Weed Science. Southern
Weed Science Society., Auburn, AL. Auburn Printing (1977).

(3)	Ratch, H. and J.S. Fletcher. Plant Reproduction and/or Life Cycle
Testing. In EPA Publ. EPA/600/9-91/041, Plant Tier Testing: A Workshop
to Evaluate Nontarget Plant Testing in Subdivision J Pesticide Guidelines,
Nov. 29-Dec. 1, 1990, ERL, Corvallis, OR. pp. 80-89 (1991).
(4)	Pfleeger, T. Impact of Airborne Pesticides on Natural Plant Com-
munities. In EPA Publ. EPA/600/9-91/041, Plant Tier Testing: A Work-
shop to Evaluate Nontarget Plant Testing in Subdivision J Pesticide Guide-
lines, Nov. 29-Dec. 1, 1990, ERL, Corvallis, OR. pp. 108-123 (1991).