Attachment 4-3
Guidance for Developing Ecological
Soil Screening Levels (Eco-SSLs)
Eco-SSL Standard Operating Procedure (SOP) #4: Wildlife Toxicity
Reference Value Literature Review, Data Extraction and Coding
OSWER Directive 92857-55
Issued November 2003
Revised June 2005
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Attachment 4-3
Standard Operating Procedure (SOP) #4: Wildlife Toxicity
Reference Value Literature Review, Data Extraction and Coding
for
Ecological Soil Screening Levels (Eco-SSLs)
OSWER Directive 92857-55
June 2005
Soil Scraoninn LĞwl*
Prepared for USEPA Region 8 by
Syracuse Research Corporation
Denver, Colorado 80123
maintained by
Computer Sciences Corporation
Duluth, Minnesota 55804
Contract 68-W01-032, Task Order No. 24
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TABLE OF CONTENTS
1.0 INTRODUCTION U.
1.1 Purpose 1-1
1.2 Wildlife TRY Database 1-2
2.0 REVIEW OF LITERATURE AND REJECTION CRITERIA 2J.
3.0 WILDLIFE TRY DATABASE WEBSITE 3J.
3.1 Location and Log-On 3-1
3.2 Navigation 3-1
4.0 CODING GUIDELINES AND DATA ENTRY 4J.
4.1 Article Information 4-1
4.2 Study Information 4-4
4.3 Exposure Information 4-5
4.4 Endpoint Information 4-24
4.5 Data Evaluation Score 4-55
5.0 QUALITY ASSURANCE AND QA REPORTS 5J.
6.0 DOWNLOADS 6J.
APPENDIX A A- 1
APPENDIXB B- 1
REFERENCES R- 1
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LIST OF TABLES
Table 1. Literature Rejection Categories 2-1
Table 2. Contaminants of Concern 4-2
Table 3. Percentages of Metal 4-6
Table 4a. Order, Family, and Common Name for Avian Test Species 4-10
Table 4b. Order, Family, and Common Name for Mammalian Test Species 4-11
Table 5. Organism Source Code 4-12
Table 6. Control Type Code Descriptions 4-12
Table 7. Nutrition Requirements 4-13
Table 8. Concentration Units and Conversions to Dose 4-17
Table 9. Dose Units and Conversion to mg/kg BW/day 4-17
Table 10. Method of Contaminant Analysis Code Descriptions 4-19
Table 1 la. Application Frequency Code Descriptions 4-20
Table 1 Ib. Exposure Type Code Descriptions 4-21
Table 12. Test Location Code Descriptions 4-21
Table 13. Standard Study Guidelines and Reporting Parameters 4-23
Table 14. Exposure Duration and Age Units 4-25
Table 15. Lifestage Code Descriptions 4-26
Table 16. Effect Group Descriptions 4-27
Table 17. Effect Groups, Types and Measures 4-28
Table 17. Effect Groups, Types and Measures 4-29
Table 17. Effect Groups, Types and Measures 4-32
Table 17. Effect Groups, Types and Measures 4-34
Table 17. Effect Groups, Types and Measures 4-35
Table 17. Effect Groups, Types and Measures 4-36
Table 17. Effect Groups, Types and Measures 4-37
Table 18. Response Sites and Codes 4-37
Table 19. Body Weight Units and Conversions 4-46
Table 20. Default Body Weights 4-47
Table 21. Units and Conversions 4-53
Table 22. Summary of Data Evaluation Scoring System 4-56
Table 23. Default Weaning, Puberty, and Lifespan 4-62
LIST OF FIGURES
Figure 4-1. Decision tree for selecting hypothesis-testing procedures 4-44
11
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1.0 INTRODUCTION
1.1 Purpose
The United States Environmental Protection Agency (USEPA) Office of Emergency and
Remedial Response (OERR) with a multi-stakeholder workgroup has developed risk-based soil
screening levels (Eco-SSLs). Eco-SSLs are concentrations of contaminants in soils that are
protective of ecological receptors that commonly come into contact with soil or ingest biota that
live in or on soil. Eco-SSLs are derived separately for four groups of ecological receptors:
mammals, birds, plants, and soil invertebrates. As such, these values are presumed to provide
adequate protection of terrestrial ecosystems.
The Eco-SSLs should be used in the baseline ERA process to identify the contaminants that need
to be evaluated further in the characterization of exposure, effects and risk characterization. The
Eco-SSLs should be used during Step 2 of the Superfund ERA process, the screening-level risk
calculation. This step normally is completed at a time when limited soil concentration data are
available, and other site-specific data (e.g., contaminant bioavailability information, area use
factors) are not available. It is expected that the Eco-SSLs will be used to screen the site soil
data to identify those contaminants that are not of potential ecological concern and do not need
to be considered in the subsequent baseline ERA.
Plant and soil biota Eco-SSLs are developed from available plant, soil invertebrate and microbial
toxicity data. The mammal and bird Eco-SSLs are the result of back-calculations from a Hazard
Quotient (HQ) of 1.0. The HQ is equal to the dose (associated with the contaminant
concentration in soil) divided by a toxicity reference value (TRV). Generic food chain models
are used to estimate the relationship between the concentration of the contaminant in soil and the
dose for the receptor (mg per kg body weight per day). The TRV represents a numerical
estimate of a no adverse effect level (dose) for the respective contaminant.
The methods for deriving the oral TRVs needed for calculation of Eco-SSLs for mammals and
birds are contained within four standard operating procedures (SOPs):
Eco-SSL SOP #3 Literature Search and Retrieval (Attachment 4-2)
Eco-SSL SOP #4 Literature Review, Data Extraction and Coding (Attachment 4-3)
Eco-SSL SOP #5 Data Evaluation (Attachment 4-4)
Eco-SSL SOP #6 Derivation of the Oral TRV (Attachment 4-5)
This document serves as SOP #4 which is Attachment 4-4 of the Eco-SSL guidance document.
It describes the procedures used for review and extraction of data from toxicological studies
identified as a result of SOP #3 (Attachment 4-2) and also serves as a user's manual for the web-
Guidance for Developing Eco-SSLs Attachment 4-3 1-1 June 2005
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based data entry system used to guide the data extraction process. The extracted data are
evaluated (scored) for their usefulness in establishing an oral TRV according to procedures
provided in SOP #5 (Attachment 4-4). The extracted and scored data are then used to derive
TRVs for mammals and birds, according to the procedures outlined in SOP #6 (Attachment 4-5).
1.2 Wildlife TRV Database
The Wildlife TRV database is a tool designed to facilitate efficient and accurate data extraction
from reviewed toxicological studies. Importing the data directly into an electronic database
facilitates the necessary sorting, searching and presentation of the data for the purposes of TRV
derivation. The original database was designed using Microsoft Access and included a series of
data entry forms. It was envisioned that each of the parties responsible for data entry would
receive a copy of the Access database on disk. After all toxicity studies had been entered and
coded by individual parties, each remote database would then be transferred and merged into a
master Access database., The use of the Access-based data entry system was reevaluated as a
result of changes in the data entry process and the addition of USEPA regional users. Several
issues were identified, including: 1) how to update future changes to the database after the initial
distribution, 2) how to effectively merge and incorporate all remote databases into the master
database, 3) how to distribute the completed master database to all interested parties after the
data entry process has been completed, and 4) how to distribute the database for review by
external parties.
A web-based data entry system was proposed to resolve these issues. The web-based data entry
system allows for remote access from any computer with Internet capabilities. Entry to the site
is password-protected and limited to only those individuals responsible for data entry. All
information entered is sent directly to a master database, avoiding quality assurance problems
associated with merging multiple sources into one database. This system provides immediate
access to entered data. Any changes to the data entry process or scoring are immediately
reflected on the website. The website also allows users to view summaries of entered
information in a format designed for quality assurance (QA) review. Information in the system
can be downloaded into Microsoft Access format.
The results of the Eco-SSL coding effort are transferred to EPA, Mid-Continent Ecology
Division, Duluth, MN for incorporation into the ECOTOX database. The coding guidelines used
here for the Wildlife TRV effort follow the same basic structure as those used by EPA for
TERRETOX. There are, however, some necessary additions and exclusions from the
TERRETOX coding system. The TRV database is focused on identifying the no observed
adverse effect level (NOAEL) and lowest observed adverse effect level (LOAEL) doses in
relevant toxicological studies. Experimental data for toxicological endpoints are not entered into
the TRV database.
Guidance for Developing Eco-SSLs Attachment 4-3 1-2 June 2005
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2.0 REVIEW OF LITERATURE AND REJECTION CRITERIA
At this point in the Wildlife TRY derivation process, hard copies of literature identified as a
result of SOP #5 are available to the User. A unique reference number is assigned to each article
identified as a result of the literature search process. This reference number serves as a key to
the full citation which is recorded in a commercially available bibliographic software program
(ProCite or Reference Manager, ISI Researchsoft). The hard copies of the literature are housed
at the EPA Region 8 offices in Denver, Colorado and at the EPA Mid-Continent Ecology
Division in Duluth, MN.
The bibliographic reference file contains information on the article title, authors, journal or
report title, date, volume, issue, page numbers, abstract, keywords, and article retrieval status. A
unique Eco-SSL/TRV identification number provides the link between the data entered on the
website and the article information identified in the literature search and recorded in the
bibliographic reference file. This number is located in the upper-right corner of the article on a
small white label. Additional information provided on this label includes the first author of the
article and the identity of the contaminant of concern.
Example label:
Contractor
Auth: Smith
Eco-SSL TRV:45
Cobalt
A preliminary review is conducted on each article to determine whether the study contains data
suitable for Wildlife TRV derivation. Table 1 provides a category listing of the types of
publications and studies that are not included in the effort. These categories are referred to as
rejection categories or criteria.
Table 1. Literature Rejection Categories
Rejection Criteria
ABSTRACT
(Abstract)
ACUTE STUDIES
(Acu)
AIR POLLUTION
(Air P)
ALTERED RECEPTOR
(Alt)
AQUATIC STUDIES
(Aquatic)
ANATOMICAL STUDIES
(Anat)
Description
Abstracts of journal publications or conference
presentations
Single oral dose or exposure duration of three days or less.
Studies describing the results for air pollution studies.
Studies that describe the effects of the contaminant on
surgically-altered or chemically-modified receptors (e.g.,
right nephrectomy, left renal artery ligature, hormone
implant, etc.).
Studies that investigate toxicity in aquatic organisms
Studies of anatomy. Instance where the contaminant is
used in physical studies (e.g., silver nitrate staining for
histology).
Basis
Reference type
Exposure
Exposure
Experimental design
Experimental design
Experimental design
Guidance for Developing Eco-SSLs Attachment 4-3 2-1
June 2005
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Table 1. Literature Rejection Categories
Rejection Criteria
BACTERIA
(Bact)
BIOACCUMULATION
SURVEY
(Bio Ace)
BIOLOGICAL TOXICANT
(BioX)
BIOMARKER
(Biom)
CARCINOGENICITY
STUDIES
(Carcin)
CHEMICAL METHODS
(Chem Meth)
CONFERENCE
PROCEEDINGS
(CP)
DEAD
(Dead)
DISSERTATIONS
(Diss)
DRUG
(Drug)
DUPLICATE DATA
(Dup)
ECOLOGICAL
INTERACTIONS
(Ecol)
EFFLUENT
(Effl)
CHEMICAL
FATE/METABOLISM
(Fate)
FOREIGN LANGUAGE
(FL)
FOOD STUDIES
(Food)
FUNGUS
(Fungus)
Description
Studies on bacteria or susceptibility to bacterial infection
Studies reporting the measurement of the concentration of
the contaminant in tissues.
Studies of biological toxicants, including venoms, fungal
toxins, Bacillus thuringiensis, other plant, animal, or
microbial extracts or toxins.
Studies reporting results for a biomarker having no
reported association with an adverse effect and an exposure
dose (or concentration).
Studies that report data only for carcinogenic endpoints
such as tumor induction. Papers that report systemic
toxicity data are retained for coding of appropriate
endpoints.
Studies reporting methods for determination of
contaminants, purification of chemicals, etc. Studies
describing the preparation and analysis of the contaminant
in the tissues of the receptor.
Studies reported in conference and symposium
proceedings.
Studies reporting results for dead organisms. Studies.
reporting field mortalities with necropsy data where it is
not possible to establish the dose to the organism.
Dissertations are excluded. However, dissertations should
be flagged for possible future use.
Studies reporting results for testing of drug and therapeutic
effects and side-effects. Therapeutic drugs include vitamins
and minerals. Studies of some minerals may be included if
there is potential for adverse effects.
Studies reporting results that are duplicated in a separate
publication. The publication with the earlier year is used.
Studies of ecological processes that do not investigate
effects of contaminant exposure (e.g., studies of "silver"
fox natural history; studies on ferrets identified in iron
search).
Studies reporting effects of effluent, sewage, or polluted
runoff.
Studies reporting what happens to the contaminant, rather
than what happens to the organism. Studies describing the
intermediary metabolism of the contaminant (e.g.,
radioactive tracer studies) without description of adverse
effects.
Studies in languages other than English
Food science studies conducted to improve production of
food for human consumption.
Studies on fungus
Basis
Toxicant type
Experimental design
Toxicant type
Endpoint
Endpoint
Experimental design
Data source
Exposure
Data source
Endpoint
Experimental design
Exposure
Effect
Foreign Language
Effect
Receptor
Guidance for Developing Eco-SSLs Attachment 4-3 2-2
June 2005
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Table 1. Literature Rejection Categories
Rejection Criteria
GENE
(Gene)
HUMAN HEALTH
(HHE)
IMMUNOLOGY
(IMM)
INVERTEBRATE
(Invert)
IN VITRO
(In Vit)
LEAD SHOT
(Lead shot)
METHODS
(Meth)
MINERAL REQUIREMENTS
(Mineral)
MIXTURE
(Mix)
MODELING
(Model)
NO CONTAMINANT OF
CONCERN
(No COC)
NO CONTROL
(No Control)
NO DATA
(No Data)
NO DOSE or CONC
(No Dose)
NO DURATION
(NoDur)
NO EFFECT
(No Efct)
Description
Studies of genotoxicity (chromosomal aberrations and
mutagenicity).
Studies with human subjects.
Studies on the effects of contaminants on immuno logical
endpoints.
Studies that investigate the effects of contaminants on
terrestrial invertebrates are excluded.
In vitro studies, including exposure of cell cultures, excised
tissues and/or excised organs.
Studies administering lead shot as the exposure form.
These studies are labeled separately for possible later
retrieval and review.
Studies reporting methods or methods development
without usable toxicity test results for specific endpoints.
Studies examining the minerals required for better
production of animals for human consumption, unless there
is potential for adverse effects.
Studies that report data for combinations of single
toxicants (e.g. cadmium and copper) are excluded.
Exposure in a field setting from contaminated natural soils
or waste application to soil may be coded as Field Survey.
Studies reporting the use of existing data for modeling,
i.e., no new organism toxicity data are reported. Studies
which extrapolate effects based on known relationships
between parameters and adverse effects.
Studies that do not examine the toxicity of Eco-SSL
contaminants of concern
Studies which lack a control or which have a control that is
classified as invalid for derivation of TRVs.
Studies for which results are stated in text but no data is
provided. Also refers to studies with insufficient data
where results are reported for only one organism per
exposure concentration or dose. Also refers to studies
where no data is provided but the text reports statistical
comparison results and p values.
Studies with no usable dose or concentration reported.
These are usually identified after examination of full paper.
This includes studies which examine effects after exposure
to contaminant ceases. This also includes studies where
offspring are exposed in utero and/or during lactation and
then after weaning to similar concentrations (or doses) as
their parents. Dose cannot be determined. In some cases,
where exposure was during gestation and effects are
measured after cessation of exposure (after), data are
retained to record reproductive latent effects.
Studies with no exposure duration. These are usually
identified after examination of full paper.
Studies with no relevant effect evaluated in a biological
test species or data not reported for effect discussed.
Basis
Endpoint
Receptor
Endpoint
Receptor
Effect
Exposure
Experimental design
Effect
Exposure
Modeling/Existing
data
Exposure
Experimental design
Experimental design
Exposure
Exposure
Effect
Guidance for Developing Eco-SSLs Attachment 4-3 2-3
June 2005
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Table 1. Literature Rejection Categories
Rejection Criteria
NO ORAL
(No Oral)
NO ORGANISM
(No Org)
NOT AVAILABLE
(Not Avail)
NOT PRIMARY
(Not Prim)
NO TOXICANT
(No Tox)
NUTRIENT DEFICIENCY
(Nut def)
NUTRITION
(Nut)
OTHER AMBIENT
CONDITIONS
(OAC)
OIL
(Oil)
ORGANIC METAL
(Org Met)
LEAD BEHAVIOR OR HIGH
DOSE MODELS
(Pb Behav)
PHYSIOLOGY STUDIES
(Phys)
PLANT
(Plant)
PRIMATE
(Prim)
Description
Studies using non-oral routes of contaminant
administration including intraperitoneal injection, other
injection, inhalation, and dermal exposures.
Studies that do not examine or test a viable organism (also
see in vitro rejection category).
Papers that could not be located. Citation from electronic
searches may be incorrect or the source is not readily
available.
Papers that are not the original compilation and/or
publication of the experimental data.
No toxicant used. Publications often report responses to
changes in water or soil chemistry variables, e.g., pH or
temperature. Such publications are not included.
Studies of the effects of nutrient deficiencies. Nutritional
deficient diet is identified by the author. If reviewer is
uncertain then the administrator should be consulted.
Effects associated with added nutrients are coded.
Studies examining the best or minimum level of a chemical
in the diet for improvement of health or maintenance of
animals in captivity.
Studies which examine other ambient conditions: pH,
salinity, DO, UV, radiation, etc.
Studies which examine the effects of oil and petroleum
products.
Studies which examine the effects of organic metals. This
includes tetraethyl lead, triethyl lead, chromium
picolinate, phenylarsonic acid, roxarsone, 3-nitro-4-
phenylarsonic acid,, zinc phosphide,
monomethylarsonic acid (MMA), dimethylarsinic acid
(DMA), trimethylarsine oxide (TMAO), or arsenobetaine
(AsBe) and other organo metallic fungicides. Metal
acetates and methionines are not rejected and are
evaluated.
There are a high number of studies in the literature that
expose rats or mice to high concentrations of lead in
drinking water (0.1, 1 to 2% solutions) and then observe
behavior in offspring, and/or pathology changes in the
brain of the exposed dam and/or the progeny. Only a
representative subset of these studies were coded.
Behavior studies examining complex behavior (learned
tasks) were also not coded.
Physiology studies where adverse effects are not associated
with exposure to contaminants of concern.
Studies of terrestrial plants are excluded.
Primate studies are excluded.
Basis
Exposure
Receptor
Data source
Data source
Exposure
Exposure
Exposure
Toxicant
Toxicant
Exposure
Exposure and endpoint
Effects
Receptor
Receptor
Guidance for Developing Eco-SSLs Attachment 4-3 2-4
June 2005
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Table 1. Literature Rejection Categories
Rejection Criteria
PUBLAS
(Publ as)
QSAR
(QSAR)
REGULATIONS
(Reg)
REVIEW
(Rev)
SEDIMENT CONC
(Sed)
SLUDGE
SOIL CONC
(Soil)
STRESSOR
(QAC)
SURVEY
(Surv)
REPTILE OR AMPHIBIAN
(Hop)
UNRELATED
(Unrel)
WATER QUALITY STUDY
(Wqual)
YEAST
(Yeast)
Description
The author states that the information in this report has
been published in another source. Data are recorded from
only one source. The secondary citation is noted as Publ
As.
Derivation of Quantitative Structure- Activity Relationships
is a form of modeling. QSAR publications are rejected if
raw toxicity data are not reported or if the toxicity data are
published elsewhere as original data.
Regulations and related publications that are not a primary
source of data.
Studies in which the data reported in the article are not
primary data from research conducted by the author. The
publication is a compilation of data published elsewhere.
These publications are reviewed manually to identify other
relevant literature.
Studies in which the only exposure concentration/dose
reported is for the level of a toxicant in sediment.
Studies on the effects of ingestion of soils amended with
sewage sludge
Studies in which the only exposure concentration/dose
reported is for the level of a toxicant in soil.
Studies examining the interaction of a stressor (e.g.,
radiation, heat, etc.) and the contaminant, where the effect
of the contaminant alone cannot be isolated.
Studies reporting the toxicity of a contaminant in the field
over a period of time. Often neither a duration nor an
exposure concentration is reported.
Studies on reptiles and amphibians. These papers flagged
for possible later review.
Studies that are unrelated to contaminant exposure and
response and/or the receptor groups of interest.
Studies of water quality
Studies of yeast
Basis
Data source
Model
Data source
Data source
Exposure
Exposure
Exposure
Exposure
Exposure
Receptor
Relevance
Relevance
Receptor
If a retrieved article is rejected after review, the user records the reason for rejection in the
bibliographic reference file and the article is not considered further in the process. The results of
the literature review and the application of rejection criteria are described for each contaminant
of concern.. SOP #6 (Attachment 4-5) describes the process for deriving the Wildlife TRV.
Guidance for Developing Eco-SSLs Attachment 4-3 2-5
June 2005
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3.0 WILDLIFE TRV DATABASE WEBSITE
3.1 Location and Log-On
The Wildlife TRV Database is located in Duluth, MN and maintained by the USEPA Mid-
Continent Division. For quality assurance purposes, the Eco-SSL Wildlife TRV Database is
accessible only to authorized users. It is important that users not give their log on information to
others.
The Wildlife TRV Database website is accessed from the Explorer or Netscape browsers by
typing http://trv.ecodev.com/ in the address bar. The TRV Homepage will appear and display a
prompt to log on. The User clicks on "log" to access the Log On page. The User enters a
username and password as directed and clicks "Log On" to access the database.
3.2 Navigation
The TRV Database contains a sidebar for navigation among the database functions. Navigation
choices include Home. Log Out Admin. Data Entry, and Password. Most features needed by
TRV Database users are located under the Data Entry feature. A brief description of each
function is provided below.
Home
Clicking on the Home option will return the User to the log on, which prompts the User for
username and password.
Logout
The Logout option is used to exit the Wildlife TRV Database.
Admin
The Admin feature allows users with administrative authorization (Administrative Users) to
permanently delete articles or alter database features such as data entry codes or programming
code for calculations.
Data Entry
The Data Entry option displays the main menu for data entry navigation. Six options are
available for this feature:
1. Add Article
2. Article List
3. A Targeted Article List
4. QA'd Article List
Guidance for Developing Eco-SSLs Attachment 4-3 3-1 June 2005
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5. Reports
6. Download database
A brief description of each option is provided below.
Add Article: This option allows the User to add information from a new article. This
function is discussed in more detail in Section 4.
Article List: This option displays all articles currently in the TRV system. The User may
View/Edit Articles, Add Phases to an article, Delete Articles, access the
Exposures/Endpoint edit screens, or access the QA report (a printable version of the
information entered for the article), by clicking on the appropriate column. Listed
articles may be changed by any user if they have not been locked by the QA approval
process (see Section 6). When an article has been approved following a QA review, the
Add Phase, Delete Article and Exposures edit options are disabled. Article Approval
information is then displayed on the QA report, along with any QA comments.
A Targeted Article List: This option allows the User to create a list of articles for a
specific SSL#, chemical, or author. The retrieved list displays the selected articles in the
same manner, and offers the same QA functionality, as the full Article List.
QA'd Article List: This option displays a list of articles that have received QA approval.
The display information includes Record #, Chemical, Author, Reviewer, Date reviewed,
Approver, Date Approved, an Edit Article function, and QA report display. Articles that
have passed QA are locked for editing by users without Administrative User status.
Locking means that the Edit Article feature in this list does not allow reviewers to change
the QA status of an article. However, a reviewer who disagrees with a QA revision may
provide an explanation in the comment box.
If an approved article is unlocked by an Administrative User (i.e., the Article Approved
field is changed to "No" or "Not Checked"), the article disappears from the QA'd Article
List, and all editing functionality is restored.
Reports: Three types of reports are available under this function:
QA Modifications: This report displays a list of articles that have been quality
assured, but not necessarily approved. The display includes contaminant of
concern, SSL (Record) #, Ecoref # (i.e., the number used to identify the article in
the ECOTOX database), Author, Reviewer, Review date, Approver, QA date,
Last change date, and whether or not the article has been approved (Apr?). This
report allows reviewers could keep track of quality assurance revisions and to
maintain consistency of coding.
Guidance for Developing Eco-SSLs Attachment 4-3 3-2 June 2005
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Article Counts: This report provides information on all contaminants of concern,
the total articles reviewed for each chemical, which articles were reviewed by
Duluth Offsite, how many articles were approved, and the number of articles with
at least one score of greater than 65.
Article Information: Information on contaminant of concern, SSL#, Ecoref#,
Author, Reviewer, Review date, High Score, and approval status (Apr?) is
provided for each chemical. This report function provides a "sort by" window,
which allows the User to sort by each parameter.
Password
The password option allows individual users to change their password.
Guidance for Developing Eco-SSLs Attachment 4-3 3-3 June 2005
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4.0 CODING GUIDELINES AND DATA ENTRY
Entry of study-specific toxicological information into the TRY Database is facilitated by data
entry screens which prompt the User for required information. This interface helps ensure that
relevant data from each study is completely and consistently entered. Required information is
divided into five categories with a corresponding data entry screen for each: Article Information,
Study Information, Exposure Information, Endpoint Information, and Score Information. A
navigation bar, which summarizes the specific article, phase, and endpoint which is currently
being scored, is provided at the top of each data entry screen to identify the User's location
throughout the data entry process.
The data entry process is initiated by clicking on the Data Entry option located in the navigation
bar. The User selects Complete Entry to begin entering information from a selected article or
report. Once data entry has begun for a specific article or report, continue to enter information
until all endpoints have been scored. This "start-to-finish" process ensures fewer errors resulting
from incomplete entries. In addition, there is a time limit for data entry. The User is
automatically logged out if no data entry activity is registered for a period of one hour.
4.1 Article Information
Record Number
The Record Number is a unique number assigned to an article identified in the literature search.
The Record Number provides the link between the data entered on the website and the article
information in the bibliographic reference file (e.g., ProCite or Reference Manager). This
number is located on a small white label placed in the upper-right corner of the article. The User
enters the number in the numeric field provided for the Record Number (eg.: 45 in the example
below).
Example label: Contractor ID Publication first author
Contractor Auth: Smith
Eco-SSL TRV:45 Cobalt
Reference ID# Contaminant of Concern
Contaminant of Concern (COO
The contaminant form tested in the reviewed study is entered at the "Exposure Information"
screen. To ensure quality and consistency, a pull down list is provided for all contaminants
which are to be reviewed for the Eco-SSL effort. This list is presented in Table 2. The User
Guidance for Developing Eco-SSLs Attachment 4-3 4-1 June 2005
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selects the contaminant from the pull down list for entry into the database record. If results for
several contaminants of concern (COCs) are available in a single article, separate results are
entered for each COC.
Special cases for COC data entry are DDT and its metabolites and the polycyclic aromatic
hydrocarbons (PAHs). Data for DDT metabolites (DDE, ODD, DDA, and DDMU) are entered
as chemical forms of Total DDT. If an article contains experimental data for multiple forms,
data for the parent compound and/or individual metabolites are entered as separate phases.
The PAHs of concern are listed individually in Table 2. The specific approach to evaluation of
PAH toxicity and derivation of PAH TRVs is currently under review by EPA. The procedures
for entering data on this class of contaminants will be added to this SOP in a future update.
Table 2. Contaminants of Concern
Contaminant
Code
Contaminant Name
Chlorinated Organics
Did
PCB
DDT
ODD*
DDE*
DDA*
DDMU*
PCP
Dieldrin
Total PCBs
Total DDT - DDT
Total DDT - ODD
Total DDT - DDE
Total DDT - DDA
Total DDT - DDMU
PCP (Pentachlorophenol)
Other Organics
RDX
TNT
RDX (Hexahydro-l,3,5-trinitro-l,3,5-
triazine)
TNT (Trinitrotoluene)
Metals
Al
Ba
Sb
As
Be
Cd
Cr
Co
Cu
Fe
Pb
Mn
Ni
Se
Ag
V
Zn
Aluminum
Barium
Antimony
Arsenic
Beryllium
Cadmium
Chromium
Cobalt
Copper
Iron
Lead
Manganese
Nickel
Selenium
Silver
Vanadium
Zinc
Contaminant
Code
Contaminant Name
Polycyclic Aromatic Hydrocarbons (PAHs)**
Dmg
Dma
Ace
Acy
Ani
Ant
Baa
Bap
Bkf
Bghip
Bbf
Chr
Dbaha
Dbaep
Dbf
Fla
Fl
Ind
Nap
Phe
Pyr
3,3'-Dimethylbenzidine
7, 1 2-Dimethy Ibenz(a)- anthracene
Acenaphthene
Acenaphthylene
Aniline
Anthracene
Benz(a)anthracene
Benzo(a)pyrene
Benzo(k)fluoranthene
Benzo(g,h,I)perylene
Benzo(b)fluoranthene
Chrysene
Dibenz(a,h)anthracene
Dibenzo(a,e)pyrene
Dibenzofuran
Fluoranthene
Fluorene
Indeno( 1 ,2,3-cd)pyrene
Naphthalene
Phenanthrene
Pyrene
* Considered a chemical form of DDT and is coded as a separate phase under the COC of Total DDT
* * The specific approach to grouping of PAHs and derivation of TRVs is under consideration by USEPA and has
not yet been finalized
Guidance for Developing Eco-SSLs Attachment 4-3 4-2
June 2005
-------
Author Key
The Author Key is a text field designed to provide a citation for the article entered. This citation
is used to verify the record number and is incorporated into the navigation bar at the top of each
page. Author information is entered in the same way the article would be cited in a scientific
document, with the author's last name(s) separated by a comma and the year. If there is one
author, the citation appears as "Smith, 1997"; if there are two authors, the citation appears as
"Smith and Jones, 1997"; if there are three or more authors, the citation appears as "Smith et al.,
1997". The first or middle name initials are not used in the Author Key.
Primary Source
The toxicity data used for the Eco-SSL Wildlife TRVs are taken from primary sources only. A
primary source is the original compilation and/or publication of the experimental data.
Secondary sources are defined as studies where the data reported is not from research conducted
by the author and/or the publication is a compilation of data published elsewhere. Secondary
sources are coded as reviews ("Rev"; see Table 1) and are rejected for use (i.e., toxicological
data from these sources are not entered into the TRY Database). However, secondary sources
are examined to identify other relevant literature. This process is referred to as a manual review.
The User selects "Yes" or "No" for primary source by checking the appropriate box. If "No" is
selected, the information entered to this point is saved and the program exits to the "Data Entry"
screen.
Results Reported for Exposure to a Single Contaminant
The Wildlife TRY database compiles data from experiments which examine exposure to a single
contaminant. Studies that report results for concurrent exposure to multiple contaminants are
rejected for use as a mixture ("mix"; see Table 1). In cases where exposures are for
contaminants in sediment, soil or food, the exposure should be for a single contaminant in the
environmental matrix. The sediment, soil or food cannot contain other contaminants in excess of
nutritional requirements. The User selects "Yes" or "No" for exposure to a single contaminant
by checking the appropriate box. If "No" is selected, the information entered to this point is
saved and the program exits to the "Data Entry" screen.
When the "Article Information" screen is completed, the User verifies that all data entered are
correct and then clicks on "Next" at the bottom of the screen to continue. The User should not
use the browser back arrow to return to a previous data entry screen to correct errors, because
use of this key results in a deletion of information.
Guidance for Developing Eco-SSLs Attachment 4-3 4-3 June 2005
-------
4.2 Study Information
Study Phase determination
The study phase is the basic unit of experimental design for the Wildlife TRY Database.
Multiple study phases are present if the study reports different results for any of the following
parameters: test organism (different species /strain or lifestage), chemical form (e.g. DDE and
p,p'DDT, Cadmium chloride and Cadmium sulfate, etc.), range of chemical doses or
concentrations, test location, exposure type, control type, total number of doses, application
frequency, or route of exposure. The User enters a description of each phase, including the basis
for selection and differences in parameters, in the text box (remarks field) provided. The
following is an example of data that would be coded as separate phases:
Example: Phase 1 - oral exposure to Cadmium chloride in food to rats for 10 weeks
Phase 2 - oral exposure to Cadmium chloride in food to mice for 10 weeks
The following cases would not be coded as multiple phases: 1) Experiments that perform interim
measurements at intermediate time points in addition to terminal measurements; 2) Studies on
metals where the dose is expressed as the metal (note: this is the only instance in which
contaminant form does not trigger a new phase); 3) Results for male and female exposure groups
within the same experiment. Results for males and females are not coded as separate phases
because this could potentially create replicate NOAEL and LOAEL values. The User should
select and enter the results for the most sensitive gender. Results for the opposite gender should
be recorded in the NOAEL/LOAEL comment fields. In cases, where the male and female
organisms have different exposure concentrations or doses, then they should be entered as
separate phases.
In some cases it may be appropriate to code male and female results separately where more
accurate exposure doses result. This is the case if authors report body weights specific to each of
sexes or report respective doses for each of the sexes in mg/kg bw/day. It is more accurate to use
the reported body weights for each sex or to use the reported doses. In these instances, the user
should enter only the most conservative results across the sexes and not all endpoints for all
sexes. A note should be made in the NOAEL/LOAEL comment field concerning the results for
the sex not entered.
If multiple phases of a study report the same NOAEL and LOAEL concentrations (or doses) for
the same endpoint and test species, the User enters results for only one of the Phases. The phase
entered should be the one that will give the highest total score. Replicate tests are generally
considered one phase. Typically, the results for the shortest exposure duration that reports the
most conservative results (lowest NOAEL or LOAEL) should be entered (see detailed comments
on identification of the NOAEL and LOAEL below). If the author reports different doses for
different exposure durations, the reviewer enters the data for each exposure duration as a
separate phase. For gestational exposure studies that report separate results for different
Guidance for Developing Eco-SSLs Attachment 4-3 4-4 June 2005
-------
gestational days of exposure, the reviewer can enter the results as separate Phases. The rationale
for data entry decisions is recorded in the comments field.
When the "Study Information" screen is completed, the User verifies that all data entered are
correct and then clicks on "Next" at the bottom of the screen to continue. The newly entered
data are recorded in the database at this time. The User should not use the browser back arrow
to return to a previous data entry screen to correct errors, as use of this key results in deletion of
information.
When data entry for an individual phase is completed, the database will ask the User if another
phase is required. At the end of data entry, the database will automatically calculate the number
of phases for each article. This value is included in the QA report.
4.3 Exposure Information
Phase Number
The phase number is automatically generated by the application and corresponds to the phases
briefly described in the "Study Information" section. The User should verify that the phase
number is correct. If there are any discrepancies, the User should record the specific information
and contact a database administrator.
Contaminant Form
The form of contaminant used in the exposure is recorded by the User in the text box provided.
The form can be entered as a name or as a contaminant formula (eg.: Cadmium Chloride or
CdCl2). If a specific contaminant form is not provided in the article, the User enters the general
name for the contaminant (e.g., cadmium). Organic forms of metals are not used for derivation
of Wildlife TRVs and should not be coded (with the exception of acetate forms which readily
dissociate in solution).
Administered Amount of a Contaminant (% Molecular Weight)
Toxicological studies administer metals using compounds which contain various amounts of the
metal by weight. Some studies report concentrations (or doses) as units of metal per amount of
exposure medium (water or diet) (e.g., mg of Co per kg of diet), while others report
concentrations (or doses) based on the compound used (e.g., mg of cobalt chloride per kg of
diet). For example, if the administered compound is cadmium chloride, then only 61.32 percent
by weight was delivered as cadmium (based on the molecular weight (MW) for cadmium
chloride (CdCl2) of 183.32 g/mol and the MW for cadmium of 112.41 g/mol). A dose of
cadmium chloride of 5 mg/kg is therefore equal to 3.1 mg of cadmium/kg (i.e., 5 mg/kg *
61.32% = 3.1 mg/kg). Table 3 provides a list of contaminant forms and the respective
percentages of metal by weight. The User enters the listed percent in the numeric field provided.
If the exposure is reported as pure contaminant, the User enters the number 100.
Guidance for Developing Eco-SSLs Attachment 4-3 4-5 June 2005
-------
It is important to examine and enter data for metals accurately, as failure to do so may introduce
errors into the calculation of the NOAEL and LOAEL. If, for example, the authors give the
chemical form as lead acetate, but report the chemical concentration as 50 mg Pb/kg, then it
should be assumed that the concentration is based on lead, not lead acetate, and the percentage
of MW would be 100. However, if the chemical concentration had been 50 mg/kg, the
percentage of MW would be 54.61. In another case, if the paper using a chemical form reports
the concentration or dose based on a portion of the form, e.g., reports the concentration based on
selenite for sodium selenite, the chemical should be reported as the portion of the form (selenite),
and the percentage of MW would be the amount of selenium in selenite.
For organic chemicals, the study should be reviewed for information on the purity of the tested
material. The purity of an organic chemical (including DDT and metabolites) is assumed to be
100%, unless the paper specifically reports material of lesser purity was used (e. g. 80%
technical grade DDT). If a lesser purity is reported the user should enter the appropriate amount.
Table 3. Percentages of Metal and Occurrence in Soil
Contaminant
Aluminum
Aluminum
Aluminum
Aluminum
Aluminum
Aluminum
Aluminum
Aluminum
Aluminum
Aluminum
Aluminum
Aluminum
Antimony
Antimony
Antimony
Antimony
Antimony
Antimony
Antimony
Antimony
Antimony
Arsenic
Arsenic
Arsenic
Barium
Compound
Aluminum chloride (A1C13)
Aluminum fluoride (A1F3)
Aluminum nitrate (A1N309)
Aluminum potassium sulfate (A1KO8S2)
Aluminum sulfate (A1(SO4)3)
Aluminum sulfate hydrate (A1(SO4)3H20
Aluminum nitrate nonahydrate (A1H18N3O18)
Aluminum chloride hexahydrate (A1C13H12O6)
Aluminum trihydrate (A1H3O3)
Aluminum sulfate octahydrate (A12H36O30S3)
Aluminum fluoride dihydrate (A1F3H6O3)
Aluminum sulfate hexadecahydrate (A12H28O26S3,
Potassium antimonate (pyro) (K4O7Sb2)
Potassium antimonate (pyro) tetrathydrate
(K2H2Sb207).
Potassium antimonate hydrate (KSbO3)
Antimony potassium tartrate (Sb2H4K2O12)
Antimony trichloride (SbCl3)
Antimony trifluoride (SbF3)
Antimony trioxide (Sb2O3)
Antimony bisulfide (Sb2S3)
Potassium hexahydroxoantimonate (H6KO6Sb)
Sodium arsenate (NaAsO4)
Sodium arsenite (NaAsO2)
Sodium arsenate (generic form) (AsH2NaO4)
Barium carbonate (Ba CO3)
CAS#
7446-70-0
7784-18-1
13473-90-0
10043-67-1
10043-01-3
57292-32-7
7784-27-2
7784-13-6
21645-51-2
7784-31-8
15098-87-0
16828-11-8
29638-69-5
10090-54-7
10090-54-7
11071-15-1
10025-91-9
7783-56-4
1309-64-4
1345-04-6
12208-13-8
13464-38-5
7784-46-5
7631-89-2
513-77-9
%of
MWas
Metal
20.23
32.13
12.67
10.45
15.77
14.98
7.19
11.18
34.59
8.10
19.55
9.08
47.05
46.31
46.31
39.67
53.38
68.11
83.53
71.69
46.32
36.04
56.5
45.71
69.59
Could be
Found in Soil
Environment
No
Yes
Yes
No
Yes
Yes
Yes
No
Yes
Yes
Yes
Yes
No
No
No
No
Yes
No
Yes
Yes
No
Yes
Yes
Yes
Yes
Reference
2
3
2,3
2,3
2
3
3
3
2
2
2
1
1
2
Guidance for Developing Eco-SSLs Attachment 4-3
4-6
June 2005
-------
Table 3. Percentages of Metal and Occurrence in Soil
Contaminant
Barium
Barium
Barium
Barium
Barium
Barium
Barium
Beryllium
Beryllium
Beryllium
Beryllium
Beryllium
Beryllium
Beryllium
Beryllium
Cadmium
Cadmium
Cadmium
Cadmium
Cadmium
Cadmium
Cadmium
Cadmium
Chromium
Chromium
Chromium
Chromium
Chromium
Chromium
Chromium
Chromium
Chromium
Chromium
Chromium
Chromium
Chromium
Chromium
Cobalt
Cobalt
Cobalt
Cobalt
Cobalt
Compound
Barium acetate (Ba (C2H8O2)2
Barium chloride dihydrate (BaCl2H402)
Barium sulfate (BaO4S)
Barium nitrate (BaN2O6)
Barium chloride (BaCl2)
Barite (barium sulfate) (BaO4S)
Barium sulfide (BaS)
Beryllium chloride (BeCl2)
Beryllium fluoride (BeF2)
Beryllium hydroxide (BeH2O2)
Beryllium nitrate trihydrate (Be(NO3)2-3H2O)
Beryllium nitrate (Be(NO3)2)
Beryllium silicate (Be2O4Si)
Beryllium sulfate (BeO4S)
Beryllium sulfate tetrahydrate (BeH8O8S)
Cadmium acetate (C4H6CdO4)
Cadmium bromide (CdBr2)
Cadmium chloride (CdCl2)
Cadmium iodide (CdI2)
Cadmium nitrate (CdN2O6)
Cadmium sulfate (CdSO4)
Cadmium chloride hydrate (Cd2Cl4H10O5)
Cadmium sulfate hydrate (CdH16O12S)
Chromium (Cr)
Chromic acid (VI) (CrH2O4)
Sodium chromate (VI) (CrNa2O4)
Chromium fluoride (III) (CrF3)
Chromium chloride (CrCl3)
Chromium potassium sulfate (III) (CrKO8S2)
Sodium dichromate (VI) (Cr2Na2O7)
Chromium (III) nitrate (CrN3O9)
Chromate (CrO4)
Chromium sulfate pentahydrate (III) (Cr2O12S3)
Hexavalent chromium ion (Cr^+6)
Chromium nitrate nonahydrate (CrH18N3O18)
Potassium chromate (CrK2O4)
Potassium dichromate (Cr2K2O7)
Cobalt acetate (CoO4C4H6)
Cobalt chloride (CoCl2)
Cobalt chloride hexahydrate (CoCl2+6H20)
Cobalt nitrate Co(NO3)2
Cobalt sulfate (CoSO4)
CAS#
543-80-6
10326-27-9
7727-43-7
10022-31-8
10361-37-2
13462-86-7
21109-95-5
7787-47-5
7787-49-7
13327-32-7
7787-55-5
13597-99-4
15191-85-2
13510-49-1
7787-56-6
543-90-8
7789-42-6
10108-64-2
7790-80-9
10325-94-7
10124-36-4
7790-78-5
7790-84-3
7440-47-3
7738-94-5
7775-11-3
7788-97-8
10025-73-7
10141-00-1
10588-01-9
13548-38-4
11104-59-9
15244-38-9
18540-29-9
7789-02-8
7789-00-6
7778-50-9
71-48-7
7646-79-9
7791-13-1
10141-05-6
10124-43-3
%of
MWas
Metal
53.77
56.22
58.84
52.55
65.95
58.84
81.07
11.27
19.17
20.94
4.82
6.77
16.37
8.58
5.09
48.77
41.29
61.32
30.69
47.55
53.92
49.23
31.88
100
44.06
32.10
47.71
32.83
18.36
39.70
21.85
44.83
26.52
100
13.00
26.78
35.35
33.29
45.39
24.9
32.22
38.02
Could be
Found in Soil
Environment
No
Yes
Yes
Yes
Yes
Yes
No
Yes
No
Yes
Yes
Yes
Yes
Yes
Yes
No
No
Yes
No
Yes
Yes
Yes
Yes
Yes
No
Yes
No
Yes
Yes
Yes
Yes
Yes
Yes
Yes
Yes
Yes
No
Yes
Yes
Yes
Yes
Reference
3
2
3
3
2
2
3
3
3
3
3
1
1
1
1
1
1
3
1
1
3
3
3
3
2
Guidance for Developing Eco-SSLs Attachment 4-3
4-7
June 2005
-------
Table 3. Percentages of Metal and Occurrence in Soil
Contaminant
Cobalt
Cobalt
Copper
Copper
Copper
Copper
Copper
Copper
Copper
Copper
Copper
Copper
Copper
Copper
Iron
Iron
Iron
Iron
Iron
Iron
Iron
Iron
Iron
Iron
Iron
Lead
Lead
Lead
Lead
Lead
Lead
Lead
Manganese
Manganese
Manganese
Nickel
Nickel
Nickel
Nickel
Nickel
Nickel
Selenium
Compound
Cobalt sulfate heptahydrate (CoSO4x7H20)
Cobalt (II) formate (CoO4C2H2)
Copper chloride (CuCl2)
Copper (II) sulfate (CuSO4)
Copper (I) acetate (C2H3CuO2)
Copper oxychloride (Cu2Cl(OH)3)
Copper acetate (CuO4C4H6)
Cupric acetate (CuO4C4H6)
Cupric nitrate (Cu(NO3)2
Cupric chloride (CuCl2)
Cuprous chloride (CuCl)
Cupric perchlorate hexahydrate (Cl2CuH12O14)
Cupric nitrate hemipentahydrate (Cu2H10N4O19)
Copper chloride dihydrate (Cl2CuH4O2)
Ferric chloride (Cl3Fe)
Ferrous chloride (Cl2Fe)
Sulfonic acid, iron salt (Fe2O12S3)
Ferric hydroxide (H3FeO3)
Ferrous sulfide (FeS)
Ferrous sulfate (FeO4S)
Ferric sulfate (Fe2O12S3)
Ferrous hydroxide (Fe2H3O3)
Ferric sulfate hydrate (Fe2O12S3)
Iron trichloride (FeCl3)
Iron (II) dichloride tetrahydrate (FeCl2H8O4)
Lead acetate (C4H6O4Pb)
Lead carbonate (PbCO3)
Lead chloride (PbCl2)
Lead nitrate (Pb(NO3)2
Lead oxide (PbO)
Lead powder (Pb)
Lead sulfate (PbSO4)
Manganese (II) chloride (Cl2Mn)
Manganese (II) nitrate (MnN2O6)
Manganese (II) nitrate hydrate (H2MnN2O7)
Nickel chloride hexahydrate (Cl2H12NiO6)
Nickelous chloride (Cl2Ni)
Nickelous nitrate (N2NiO6)
Nickel sulfate hexahydrate (H12NiO10S)
Nickelous acetate tetrahydrate (C4H6NiO4)
Nickel (II) chloride hydrate (H12N2NiO12)
Selenium dioxide (O2Se)
CAS#
10026-24-1
544-18-3
1344-67-8
7758-98-7
598-54-9
1332-65-6
4180-12-5
142-71-2
3251-23-8
7447-39-4
7758-89-6
13770-18-8
19004-19-4
10125-13-0
7705-08-0
7758-94-3
10124-49-9
1309-33-7
1317-37-9
7720-78-7
10028-22-5
18624-44-7
10028-22-5
7705-08-0
13478-10-9
301-04-2
598-63-0
7758-95-4
10099-74-8
1317-36-8
7439-92-1
7446-14-2
7773-01-5
10377-66-9
15710-66-4
7791-20-0
7718-54-9
13138-45-9
10101-97-0
373-02-4
13478-00-7
7446-08-4
%of
MWas
Metal
21.91
39.55
47.27
39.81
51.84
59.51
51.84
34.99
33.88
47.27
64.19
17.15
27.32
37.28
34.43
44.06
27.93
52.26
63.53
36.77
27.93
52.26
27.93
34.43
28.09
54.61
77.55
74.50
62.56
92.83
100
68.32
43.66
30.70
27.89
24.69
45.29
32.12
22.33
33.20
20.18
71.16
Could be
Found in Soil
Environment
Yes
No
Yes
Yes
No
No
No
No
Yes
Yes
Yes
No
Yes
Yes
Yes
Yes
No
Yes
Yes
No
Yes
Yes
Yes
Yes
Yes
Yes
Yes
Yes
Yes
Yes
Yes
Yes
Yes
No
Yes
No
Reference
2
1
1
1
1
1
2
2
2
1
2
2
1
4
Guidance for Developing Eco-SSLs Attachment 4-3
4-8
June 2005
-------
Table 3. Percentages of Metal and Occurrence in Soil
Contaminant
Selenium
Selenium
Selenium
Selenium
Selenium
Selenium
Selenium
Selenium
Selenium
Selenium
Vanadium
Vanadium
Vanadium
Vanadium
Vanadium
Vanadium
Zinc
Zinc
Zinc
Zinc
Zinc
Zinc
Zinc
Zinc
Zinc
Zinc
Zinc
Compound
Potassium selenate (K2O4Se)
Potassium selenite (K2O3Se)
Hydrogen selenide (H2Se)
Selenious acid (H2O3Se)
Sodium selenate (Na2O4Se)
Sodium selenite (Na2O3Se)
Sodium selenide (Na2Se)
Selenium sulfide (S2Se)
Selenocystine (C6H12N2O4Se2)
Selenomethionine (environmental form)
(C5HuN02Se)
Sodium Orthovanadate
Vanadium (III) chloride (C13V)
Vanadyl trichloride (C13OV)
Vanadic acid, Ammonium salt (H4NO3V)
Sodium vanadate (NaVO3)
Vanadic acid, Trisodium salt (Na3O4V)
Zinc chloride (Cl2Zn)
Zinc nitrate (N2O6Zn)
Zinc sulfate (Zn SO4)
Zinc acetate (C4H6O4Zn)
Zinc peroxide (O2Zn)
Zinc phosphide (Zn3P2)
Zinc sulfate heptahydrate (H14OuSZn)
Zinc bromide (Zn Br2)
Zinc iodide (Zn I2)
Zinc nitrate hexahydrate (H12N2O12Zn)
Zinc acetate dihydrate (C4H10O6Zn)
CAS#
7790-59-2
10431-47-7
7783-07-5
7783-00-8
13410-01-0
10102-18-8
1313-85-5
7488-56-4
1464-43-3
1464-42-2
13721-39-6
7718-98-1
7727-18-6
7803-55-6
13718-26-8
13721-39-6
7646-85-7
7779-88-6
7733-02-0
557-34-6
1314-22-3
1314-84-7
7446-20-0
7699-45-8
10139-47-6
10196-18-6
5970-45-6
%of
MWas
Metal
35.71
38.49
97.51
61.22
41.79
45.66
63.20
55.19
47.27
40.26
21.27
32.38
29.39
43.55
41.78
26.70
47.98
34.52
40.50
35.64
67.14
76.00
22.74
29.04
20.49
21.98
29.79
Could be
Found in Soil
Environment
Yes
Yes
Yes
Yes
Yes
Yes
Yes
No
Yes
Yes
Yes
Yes
Yes
Yes
Yes
Yes
Yes
Yes
Yes
No
No
No
Yes
No
No
Yes
No
Reference
4
4
4
4
4
4
4
2
4
1
1
1
1
1
1
1
1
'Alloway (1990)
2Merck Index
3Bodeketal. (1988)
4Shamberger(1983)
Species Common Name/Laboratory Strain
The common name or laboratory strain of the test organism is entered in the text box provided.
Common name examples include: mouse, rat, dog, chicken, etc. Experiments testing different
strains if the same species should be coded as different phases.
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4-9
June 2005
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Genus and Species
The scientific name (genus and species) of the test organism is entered in the text box provided.
If the genus and species are not reported in the article, use the Species Lookup Table next to the
Species text box to find the correct scientific name.
Table 4a. Order, Family, and Common Name for Avian Test Species
Common Name
Bobwhite, northern
Chicken
Chicken
Chicken
Cormorant, double-
Cowbird, brown-headed
Dove, Ringed turtle-
Dove, rock
Duck, dabbling
Duck, American black
Duck, mallard
Duck, wood
Eagle, bald
Finch, striated
Finch, zebra
Goose, swan
Guineafowl, helmeted
Hawk, red-tailed
Heron, black-crowned
House sparrow
Kestrel, American
Owl, common bam
Owl eastern screech
Pheasant, ring-necked
Pigeon
Quail
Quail, Japanese
Robin, American
Shrike, loggerhead
Starling, European
Titmouse, tufted
Turkey
Order
Galliformes
Galliformes
Galliformes
Galliformes
Ciconiiformes
Passeriformes
Columbiformes
Columbiformes
Anseriformes
Anseriformes
Anseriformes
Anseriformes
Ciconiiformes
Passeriformes
Passeriformes
Anseriformes
Galliformes
Ciconiiformes
Ciconiiformes
Passeriformes
Ciconiiformes
Strigiformes
Strigiformes
Galliformes
Columbiformes
Galliformes
Galliformes
Passeriformes
Passeriformes
Passeriformes
Passeriformes
Galliformes
Family
Odontophoridae
Phasianidae
Phasianidae
Phasianidae
Phalacrocoracidae
Fringillidae
Columbidae
Columbidae
Anatidae
Anatidae
Anatidae
Anatidae
Accipitridae
Passeridae
Estrildidae
Anatidae
Numididae
Accipitridae
Ardeidae
Passeridae
Falconidae
Tytonidae
Strigidae
Phasianidae
Columbidae
Phasianidae
Phasianidae
Muscicapidae
Laniidae
Sturnidae
Paridae
Phasianidae
Genus
Colinus
Gallus
Gallus
Gallus
Phalacrocorax
Molothrus
Streptopelia
Columba
Anas
Anas
Anas
Aix
Haliaeetus
Lonchura
Poephila
Anser
Numida
Buteo
Nycticorax
Passer
Falco
Tyto
Otus
Phasianus
Columba
Coturnix
Coturnix
Turdus
Lanius
Sturnus
Parus
Meleagris
Species
virginianus
gallus
domesticus
sp.
auritus
ater
risoria
livia
sp.
rubripes
platyrhynchos
sponsa
leucocephalus
striata
guttata
cygnoides
meleagris
jamaicensis
nycticorax
domesticus
sparverius
alba
asio
colchicus
sp.
coturnix
japonica
migratorius
ludovicianus
vulgaris
bicolor
gallopavo
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June 2005
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Table 4b. Order, Family, and Common Name for Mammalian Test Species
Common Name
Bank vole
Bat, little brown
Blesbok
Cattle
Dog
Goat
Guinea pig
Hamster, golden
Mink
Mouse
Mouse, white-footed
Mouse, house
Mouse, oldfield
Pig
Pig, domestic/feral
Pronghorn
Rabbit, European
Rat, black
Rat
Rat, cotton
Rat, Norway
Sheep, Ball
Sheep
Shrew, short-tailed
Shrew, common
Vole, short-tailed
Vole, meadow
Wapiti
Whitetail deer
Order
Rodentia
Chiroptera
Artiodactyla
Artiodactyla
Carnivora
Artiodactyla
Rodentia
Rodentia
Carnivora
Rodentia
Rodentia
Rodentia
Rodentia
Artiodactyla
Artiodactyla
Artiodactyla
Lagomorpha
Rodentia
Rodentia
Rodentia
Rodentia
Artiodactyla
Artiodactyla
Insectivora
Insectivora
Rodentia
Rodentia
Artiodactyla
Artiodactyla
Family
Muridae
Vespertilionidae
Bovidae
Bovidae
Canidae
Bovidae
Caviidae
Muridae
Mustelidae
Muridae
Muridae
Muridae
Muridae
Suidae
Suidae
Antilocapridae
Leporidae
Muridae
Muridae
Muridae
Muridae
Bovidae
Bovidae
Soricidae
Soricidae
Muridae
Muridae
Cervidae
Cervidae
Genus
Clethrionomys
Myotis
Damaliscus
Bos
Canis
Capra
Cavia
Mesocricetus
Mustela
Mus
Peromyscus
Mus
Peromyscus
Sus
Sus
Antilocapra
Oryctolagus
Rattus
Rattus
Sigmodon
Rattus
Ovis
Ovis
Blarina
Sorex
Microtus
Microtus
Cervus
Odocoileus
Species
glareolus
lucifugus
pygargus
taurus
familiaris
hircus
porcellus
auratus
vison
sp.
leucopus
musculus
polionotus
sp.
scrofa
americana
cuniculus
rattus
sp.
hispidus
norvegicus
dalli
aries
brevicauda
araneus
agrestis
pennsylvanicus
elaphus
virginianus
Organism Source
The source of the test organism is selected from the pull down list. A detailed description of each
organism source is available under the description link to the right of the pull down list. The list of
available organism sources is provided in Table 5. The User should use the code for COM to
denote if laboratory species was obtained from an outside source. Confusion occurs when a
Guidance for Developing Eco-SSLs Attachment 4-3 4-11
June 2005
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laboratory rat strain is from a commercial source. In this case the code COM should be used. If
there is no mention of source for laboratory strains the User should assign the LAB code.
Table 5. Organism Source Code
Code
CBC
COM
DOM
GAM
GOV
LAB
NR
WLD
Organism Source Description
Captive Breeding Colony
Commercial Source
Domestic Strain
Game Farm Strain
Government Agency Source
Laboratory Strain
Not Reported
Wild Strain
Control Type
The effects of contaminant exposure are evaluated by comparing groups of exposed organisms to
one or more groups of untreated organisms. These untreated organisms are designated as the
controls. The User selects the type of test control(s) used in the study from the Control Type pull
down list. Detailed descriptions of each available control type are available under the description
link to the right of the pull down list. The list of available control types is provided in Table 6. If
the study reports multiple controls, select "M" for Multiple and briefly describe the control types
in the comments text box provided. If the authors report that the control had a background level of
the contaminant, this should be entered in the control comment field. Studies which use control
types coded as historical B, H, K, P, Z and NR lack an acceptable control group and are not used
for the derivation of Wildlife TRVs. Studies with gavage exposures must have a vehicle or sham
control group to have a valid control. Gavage studies without a valid control should be rejected as
"no control".
Table 6. Control Type Code Descriptions
Code
B
C
H
Validity of
Control
Invalid
Valid
Invalid
Description
Baseline or Background Control: parameters of actual or representative test species
measured either before or after administration of test contaminant, though not as part of the
same test scenario.
Concurrent Control: controls are run simultaneously with the contaminant exposure, e.g. in
the laboratory where a contaminant free test chamber is used or in field studies where the
control data are obtained upstream from the exposure data; also includes field tests where the
controls are run in a separate system, i.e., pond A and pond B or field A and field B.
Historical Control: applicable to natural field system testing, data collected prior to
exposure often during an independent long-term survey of the area; see also B - Baseline
Control. In laboratory studies, the term historical control refers to compilation of data for a
specific endpoint in multiple experiments over time.
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June 2005
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Table 6. Control Type Code Descriptions
Code
K
M
P
V
Z
NR
Validity of
Control
Invalid
Valid, if one
control type is
valid.
Invalid
Valid
Invalid
Invalid
Description
Insufficient Information: Data for control are presented, but without accompanying
methodology to identify procedures used
Multiple: multiple controls were reported, e.g. historic and concurrent
Positive control: employs a test substance known to give a positive response for a
endpoint in the test organism.
specific
Carrier, vehicle, or solvent control: test organisms are concurrently exposed to the carrier
or solvent (the "vehicle") used to administer the contaminant
No Control: no controls were used in the study
Not Reported: there is no information about presence or absence of controls provided in the
publication
For some elements it may be necessary to establish if a control diet is provided or potentially
toxic exposures are administered. Table 7 provides a list of the elements that are considered to be
essential elements and nutrient requirements for domestic species. These ranges of essential
nutrient requirements should be used to establish if exposures are nutritionally deficient,
sufficient or potentially toxic (higher than nutrient requirements).
Table 7. Nutrition Requirements
Element
Copper
Amount
(mg/kg diet dw)
3.6 to 16.8
4.4 to 20.8
4
8
4 to 5
6
8 to 25
5
6
4
6
5 to 8
8 to 10
4.3tol7.2
7.0 to 21
5.8 to 28.4
4 to 5
6
8
Species
Cattle (juvenile)
Cattle (pregnant or lactating)
Chicken (growing) (8 to 18 w)
Chicken (juvenile) (0 to 8 w)
Chicken, immature (egg laying)
Guinea pig
Horse (foal)
Japanese quail (starting, growing and breeding)
Mouse
Pig (growing)
Pig (juvenile)
Rat
Ruminants
Sheep (juvenile)
Sheep (pregnant ewe)
Sheep (lactating ewe)
Swine and poultry
Turkey (0 to 8 wk and egg laying)
Turkey (8 to 24 wk and breeding)
Reference
Underwood and Suttle (1999)
Underwood and Suttle (1999)
Underwood and Suttle (1999)
NRC(1994)
NRC(1994)
NRC(1995)
Underwood and Suttle (1999)
NRC(1994)
NRC(1995)
Underwood and Suttle (1999)
Underwood and Suttle (1999)
NRC(1995)
NAS (1980)
Underwood and Suttle (1999)
Underwood and Suttle (1999)
Underwood and Suttle (1999)
NAS (1980)
NRC(1994)
NRC(1994)
Guidance for Developing Eco-SSLs Attachment 4-3 4-13
June 2005
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Table 7. Nutrition Requirements
Element
Cobalt
Iron
Manganese
Nickel
Selenium
Amount
(mg/kg diet dw)
0.04 to 0.06
0.10
0.11
0.05 to 0.08
0.11
50
40
30 to 60
56 to 80
38 to 56
80
120
80
60
40 to 80
30 to 50
25 to 40
80
60
50
10 to 25
28 to 60
17 to 25
60
33
30
55
4.5
50
40
60
10
70
60
10
4
8.5
10
50
Ito 16
30
60
55
O.50
0.05 to 0.3
0.036 to 0.062
0.044 to 0.070
0.20 to 0.28
0.10 to 0.15
0.05 to 0.08
0.15
Species
Cattle, deer, and goat
Ruminants
Ruminants (cattle, sheep, goat)
Sheep
Sheep (growing)
Cattle
Cattle (calf)
Cattle (juvenile)
Chicken, immature (egg laying)
Chicken, mature (egg laying)
Chicken , broiler (0 to 8 wk)
Japanese quail (starting, growing and breeding)
Pig (adult female)
Pig (up to 20 kg body weight)
Poultry (first 4 weeks of life)
Sheep
Sheep (juvenile)
Turkey (0 to 4 wk)
Turkey (4 to 16 wk and egg laying)
Turkey (16 to 24 wk and breeding)
Cattle
Chicken, immature (egg laying)
Chicken, mature (egg laying)
Chicken (0 to 8 wk)
Chicken (breeding)
Chicken (growing)
Chicken
Dog
Duck (0 to 2 wk)
Guinea pig
Japanese quail (starting, growing and breeding)
Mouse
Pheasant (ring-necked) (0 to 8 wk)
Pheasant (ring-necked) (9 to 17 wk and breeding)
Pig (female)
Pig (growing) and adult
Rabbit
Rat
Rat
Sheep and goat
Sheep
Turkey (0 to 24 wk; breeding and laying)
Turkey
Ruminants
Ruminants
Cattle (beef)
Cattle (dairy)
Chicken and turkey
Chicken, immature (egg laying)
Chicken, mature (egg laying)
Chicken (0 to 8 wk)
Reference
Underwood and Suttle (1999)
NAS (1980)
McDowell (1985)
Underwood and Suttle (1999)
Underwood and Suttle (1999)
McDowell (1985)
NAS (1980)
Underwood and Suttle (1999)
NRC(1994)
NRC (1994)
NRC (1994)
NRC (1994)
Underwood and Suttle (1999)
Underwood and Suttle (1999)
Underwood and Suttle (1999)
McDowell (1985)
Underwood and Suttle (1999)
NRC (1994)
NRC (1994)
NRC (1994
Underwood and Suttle (1999)
NRC (1994)
NRC (1994)
NRC (1994)
Underwood and Suttle (1999)
Underwood and Suttle (1999)
NAS (1980)
NAS (1980)
NRC (1994)
NRC (1995)
NRC (1994)
NRC (1995)
NRC (1994)
NRC (1994)
Underwood and Suttle (1999)
Underwood and Suttle (1999)
NAS (1980)
NRC (1995)
NAS (1980)
Underwood and Suttle (1999)
NAS (1980)
NRC (1994)
NAS (1980)
McDowell (1985)
McDowell (1985)
Underwood and Suttle (1999)
Underwood and Suttle (1999)
Underwood and Suttle (1999)
NRC (1994)
NRC (1994)
NRC (1994)
Guidance for Developing Eco-SSLs Attachment 4-3 4-14
June 2005
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Table 7. Nutrition Requirements
Element
Selenium
Vanadium
Zinc
Amount
(mg/kg diet dw)
0.20
0.10
0.150
0.20
0.150
0.1 50 to 0.400
0.028 to 0.043
0.031 to 0.055
0.10
0.05
0.20
0.05 to 0.5
0.1
20 to 50
9 to 14
33 to 40
29 to 44
65
35
40
40 to 100
60
20
25
50
10
60
12 to 25
18 to 33
17
70
65
50
40
Species
Duck (0 to 2 wk)
Horse
Guinea pig
Japanese quail (starting, growing and breeding)
Mouse
Rat
Sheep (female)
Sheep (juvenile)
Pig (growing 10 to 1 10 kg)
Pig (lactating)
Turkey (0 to 24 wk; breeding and laying)
Chicken (juvenile) and rat
Rats
Cattle
Cattle (calf)
Chicken, immature (egg laying)
Chicken , mature (egg laying)
Chicken (breeding)
Chicken (juvenile growing)
Chicken (0 to 8 wk)
Domestic animals and fowl
Duck (0 to 2 wk)
Guinea pig
Japanese Quail (starting and growing)
Japanese Quail (breeding)
Mouse
Pheasant (ring-necked) (0 to 17 wk and breeding)
Rat
Sheep (juvenile)
Sheep (juvenile)
Turkey (0 to 4 wk)
Turkey (4 to 8 wk and egg laying)
Turkey (8 to 12 wk)
Turkey (12 to 24 wk and breeders)
Reference
NRC(1994)
Underwood and Suttle (1999)
NRC(1995)
NRC (1994)
NRC(1995)
NRC (1995)
Underwood and Suttle (1999)
Underwood and Suttle (1999)
Underwood and Suttle (1999)
Underwood and Suttle (1999)
NRC (1994)
McDowell (1985)
NAS (1980)
McDowell (1985)
McDowell (1985)
NRC (1994)
NRC (1994)
Underwood and Suttle (1999)
Underwood and Suttle (1999)
NRC (1994)
NAS (1980)
NRC (1994)
NRC (1995)
NRC (1994)
NRC (1994)
NRC (1995)
NRC (1994)
NRC (1995)
Underwood and Suttle (1999)
Underwood and Suttle (1999)
NRC (1994)
NRC (1994)
NRC (1994)
NRC (1994)
Number of Concentrations or Doses Tested
The total number of different concentrations or doses administered to the test organism is entered
for the specific phase in the numeric field provided. The total number of concentrations (or
doses) includes the control(s). For example, for a study which has four exposure groups of 5, 10,
20, 50 mg/kg and a valid control, the number 5 would be entered. A study must have at least two
concentrations or doses (i.e., a valid control and one exposure group) to be used for derivation of
Wildlife TRVs.
If a diet deficiency dose is reported for the chemical of concern (e.g. 0/0.2/1/2 mg/kg Selenium
and 0.2 level is identified as the normal diet level), then code the normal diet (e.g., 0.2 mg/kg) as
the control value (e.g., 0/1/2 mg/kg Se) and note the normal diet level (e.g., 0.2 mg/kg Se) in the
Guidance for Developing Eco-SSLs Attachment 4-3 4-15
June 2005
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control comments as the background level. The User should not include the diet deficient level in
the number of total doses.
Test Concentrations or Test Doses with Units
The test concentration is the amount of contaminant to which the test organism is exposed per
unit of exposure medium (water, diet or other medium). The test dose is the amount of
contaminant administered to the test organism per unit of body weight in a specified period of
time. Doses are preferred over concentrations for the purpose of establishing a wildlife TRV, but
they are not reported in many toxicological studies.
If both dry and wet weight concentrations or doses are provided in the study, use the dry weight
values, as this is the more conservative measurement. Tests in which doses varied over the
exposure period after the initial dose level are coded only if the result is reported before a change
in dose.
Information from gavage and capsule studies needs to be carefully examined to determine
whether the contaminant was administered on a mg/kg body weight or mg/organism basis. In
either case, the administered amount is a dose, and should not be entered as a concentration.
In cases where nursing offspring are exposed via the mother's milk, the concentration of
contaminant in milk must be reported in order for the results to be coded as a juvenile lifestage
effect. In most developmental and reproductive studies, exposure concentrations or doses are
reported only for the mother and are not specifically estimated or reported for the offspring.
If only exposure concentrations are reported in the study, the User should not calculate the
respective dose. The application is designed to calculate the dose automatically based on the
reported concentrations and User-supplied body weight and ingestion rate parameters. The User
should enter either the reported exposure concentrations OR doses, but not both, in the field. The
concentrations or doses are separated by a forward-slash in the text box provided. The control(s)
should be included as the first in the series (eg.: 0 / 5 /10/207 50), and should always be zero. If
a background level of contaminant is reported for the control, that information should be entered
in the control comment field.
There is an exception to the rule to allow the system to calculate the dose. For studies that report
different ingestion and body weights for each of the exposure levels, it is more accurate to
calculate the doses for each exposure level and enter these in the system.
A separate data entry field allows the User to select the appropriate units for concentration (or
dose) from the pull down list. The list of available concentration units and conversions to dose is
shown in Table 7. The list of available dose units and conversion to standard units of mg/kg/day
is shown in Table 8. A detailed description of the available units is provided under the description
link to the right of the pull down list. The application assumes that doses which do not specify a
unit of time (e.g., g/org or g/kg BW) are in or have been converted to units of per day prior to
entry.
Guidance for Developing Eco-SSLs Attachment 4-3 4-16 June 2005
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Table 8. Concentration Units and Conversions to Dose
Concentration Fields
% in diet
g/g
g/kg
g/kg/d
g/L
mg/g
mg/kg
mg/kg/d
mg/1
mg/ml
ng/g
ng/kg
ng/1
ng/mg
ppb
ppm
ug/g
ug/kg
ug/1
ug/mg
mg%
g%
percent in diet
grams per g
grams per kilogram
grams per kilogram per day
grams per liter
milligrams per gram
milligrams per kilogram
milligrams per kilogram per
day
milligrams per liter
milligrams per milliliter
nanograms per gram
nanograms per kilogram
nanograms per liter
nanograms per milligram
parts per billion
parts per million
micrograms per gram
micrograms per kilogram
micrograms per liter
micrograms per milligram
milligram percent by weight in
diet or drinking water (i.e.,
mg/100 g diet or mg/1 00 mL
drinking water)
gram percent by weight in diet
or drinking water (i.e., g/lOOg
diet or g/lOOmL drinking
water)
Conversion to
Concentration (C) as
mg/kg or mg/L
multiply by 10000
multiply by 1,000,000
multiply by 1,000
multiply by 1,000
multiply by 1,000
multiply by 1,000
multiply by 1
multiply by 1
multiply by 1
multiply by 1000
multiply by 0.001
multiply by 0.000001
multiply by 0.000001
multiply by 1
multiply by 0.001
multiply by 1
multiply by 1
multiply by 0.001
multiply by 0.001
multiply by 1000
multiply by 10
multiply by 10,000
Conversion to Dose as mg/kg BW/day
Multiply C by the IR (kg/day) and divide by BW in kg
Multiply C by IR (kg/day) and divide by BW in kg
Multiply C by IR (kg/day) and divide by BW in kg
Multiply C by IR (kg/day) and divide by BW in kg
Multiply C by IR (L/day) and divide by BW in kg
Multiply C by IR (kg/day) and divide by BW in kg
Multiply C IR (kg/day) and divide by BW in kg
Multiply C by IR (kg/day) and divide by BW in kg
Multiply C by IR (L/day) and divide by BW in kg
Multiply C by IR (L/day) and divide by BW in kg
Multiply C by IR (kg/day) and divide by BW in kg
Multiply C by IR (kg/day) and divide by BW in kg
Multiply C by IR (L/day) and divide by BW in kg
Multiply C by IR (kg/day) and divide by BW in kg
Multiply C by IR (kg/day) and divide by BW in kg
Multiply C IR (kg/day) and divide by BW in kg
Multiply C by IR (kg/day) and divide by BW in kg
Multiply C by IR (kg/day) and divide by BW in kg
Multiply C by IR (L/day) and divide by BW in kg
Multiply C by IR (kg/day) and divide by BW in kg
Multiply C by IR (L or kg/day) and divide by BW in kg
Multiply C by IR (L or kg/day) and divide by BW in kg
Table 9. Dose Units and Conversion to mg/kg BW/day
Dose Fields*
g/d
g/gBW
g/kgBW
g/kg BW /d
g/org
g/org/d
kg/d
kg/org
kg/org/d
mg/d
mg/g BW
grams per day
grams per gram body weight
grams per kilogram body weight
grams per kilogram body weight per day
grams per organism
grams per organism per day
kilograms per day
kilograms per organism
kilograms per organism per day
milligrams per day
milligrams per gram body weight
Conversion to mg/kg BW/day
multiply by 1,000 then divide by BW in kg
multiply by 1,000,000
multiply by 1,000
multiply by 1,000
multiply by 1,000, divide by BW in kg
multiply by 1,000 then divide by BW in kg
multiply by 1,000,000 and divide by BW in kg
multiply by 1,000,000, divide by BW in kg
multiply by 1,000,000 and divide by BW in kg
divide by BW in kg
multiply by 1000
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June 2005
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Table 9. Dose Units and Conversion to mg/kg BW/day
Dose Fields*
mg/g BW/d
mg/kg BW
mg/kg BW/d
mg/org
mg/org/d
ng/g bw
ng/g bw/d
ng/kg BW
ng/kg BW/d
ng/org
ug/org
ug/org/d
ug/kg BW
ug/kg BW/d
milligrams per gram body weight per day
milligrams per kilogram body weight
milligrams per kilogram body weight per day
milligrams per organism
milligrams per organism per day
nanograms per gram body weight
nanograms per gram body weight per day
nanograms per kilogram body weight
nanograms per kilogram body weight per day
nanograms per organism
micrograms per organism
micrograms per organism per day
micrograms per kilogram body weight
micrograms per kilogram body weight per day
Conversion to mg/kg BW/day
multiply by 1000
multiply by 1
multiply by 1
divide by BW in kg
divide by BW in kg
multiply by 0.001
multiply by 0.001
multiply by 0.000001
multiply by 0.000001
multiply by 0.000001, divide by BW in kg
multiply by 0.001, divide by BW in kg
multiply by 0.001, divide by BW in kg
multiply by 0.001
multiply by 0.001
*The application assumes that doses which do not specify a unit of time (e.g., g/org or g/kg B W) are in or have been
converted to units of per day prior to entry
In cases where the dose is not reported on a per day basis, the User must manually adjust the
NOAEL and/or LOAEL for the duration between administered doses. For example, this
conversion would be required in a study that administered the test material every other day or
only on five out of seven days per week. An example of adjustment for duration is provided in
the text box to the right. In cases where the reported concentration or dose units are not provided
in the pull down list, the User must convert the reported results to one of the units available for
selection.
Are Absorbed Doses Reported?
An absorbed dose is defined as the amount
of the exposure dose which is absorbed into
the bloodstream. For example, if 80 percent
of an exposure dose of 10 mg/kg BW/day is
absorbed, the absorbed dose is 8 mg/kg
BW/day. Absorbed doses are most often
reported in toxicokinetic studies. They are
not typically reported in the type of toxicity
studies reviewed for inclusion in the TRV
database.
The User selects "Yes" or "No" by checking
the appropriate box. If "Yes" is selected,
the User enters a brief description of how
the absorbed doses were measured and
reported in the text box provided. For
example, the absorbed dose is estimated in
Example for Conversion to Duration-Adjusted
Concentration or Dose Units
A study reports a NOAEL dose administered as 10 ug
per animal every two days. The User needs to convert
this dose to any set of units that can be entered into the
application. The User chooses to convert the dose to
mg per day by multiplying the dose by a conversion
factor for ug to mg of 0.001 and dividing by 2 to
achieve an administered concentration of 0.005 mg per
day. The User can now enter this result and select the
mg per day units from the dose fields. The User
should enter the conversions in detail in the comment
field provided.
Guidance for Developing Eco-SSLs Attachment 4-3 4-18
June 2005
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some studies as the difference between administered dose or intake of the contaminant and the
amount of contaminant or metabolite(s) excreted in feces and/or urine. When blood residue
levels are reported, the User should select "No" for the Absorbed Doses check box, but enter the
code "BL-RSDE" in the Absorbed Doses comments field.
Method of Contaminant Analysis
This field enables the User to indicate whether the tested concentrations (or doses) were
quantified or if nominal (target) values are reported. The User selects the method of contaminant
analysis from the pull down list provided. A detailed description of each method of analysis is
available under the "Description" link to the right of the pull down list. The list of available
contaminant analysis methods is shown in Table 10. The User selects "M" if the test
concentrations (or doses) are quantified and the results are reported. The User selects "U" if
nominal (target) values are reported or the method of contaminant analysis is not clear from the
information provided in the study. To complete data entry for this field, the User needs to read
the text of the paper carefully to determine whether exposure concentrations in the diet or
drinking water are verified by contaminant analysis. Some studies that verify or measure the
concentration or doses administered indicate in the text of the paper that analysis was performed,
but do not report data for the measured dose levels. The method of contaminant analysis in this
case is coded as unmeasured but verified (UX).
Table 10. Method of Contaminant Analysis Code Descriptions
Code
Measured (M)
Unmeasured (U)
Unmeasured but
Verified (UX)
Method of Contaminant Analysis Description
Exposure and/or observation concentrations or doses are verified by analytical methods and are
reported as quantitative measured values.
Exposure and/or observation concentrations or doses are clearly identified as nominal values; or
when the author does not report any information about whether the concentrations were measured
or nominal, i.e. unmeasured is used as a default value when there is no information provided about
the contaminant concentrations.
Unmeasured exposure concentrations or doses are clearly identified as nominal values.
The author reports that contaminant analysis was done to verify the nominal values, but
analvtical results are not ret>orted in the article.
Measured Concentrations/Measured Doses with Units
The measured concentration is the amount of the contaminant in the exposure medium as
determined by analysis. The measured dose is the amount of contaminant in the exposure vehicle
as determined by analysis, expressed per unit of organism (amount of contaminant per unit body
weight or per organism), and administered in a specified period of time. Doses are preferred over
concentrations for derivation of Wildlife TRVs. If only concentrations are reported in the study,
the User does not calculate the respective dose. The application is designed to calculate the dose
automatically. The User enters either the measured concentrations OR doses (not both) for each
of the treatment groups separated by a forward-slash in the text box provided. The control(s) are
included first in the series, (eg.: 0 / 4.8 /10.2 /18.9 / 51.1). The User next selects the appropriate
units associated with the measured concentrations or doses reported in the study from the pull
Guidance for Developing Eco-SSLs Attachment 4-3 4-19 June 2005
-------
down list. A detailed description of available units is provided under the description link to the
right of the pull down list. The list of available units is shown in Table 8.
If both measured and nominal doses or concentrations are reported, the User enters the measured
data only.
Application Frequency
The frequency of the exposure application is selected from the pull down list. For exposures in
which there are "X" applications per a given time period, the User enters the number of
applications in the numerical field provided. A detailed description of the selections available for
application frequency is available under the description link to the right of the pull down list. The
list of available application frequency selections is shown in Table 1 la.
The User should note that this feature is NOT used to automatically calculate duration-adjusted
doses (e.g. for doses administered every other day or five days per week). Duration-adjusted dose
conversion must be performed manually by the User as described above.
Table lla. Application Frequency Code Descriptions
ADL
CON
DLY
EOD
X
Xperh
Xperd
Xperw
X per mo
NR
Ad libitum; without limit or restraint
Continual; non-pulsed
Daily; dosing regime not specified
Every other day
Dosed x time(s) per study period; e.g. 1 time = IX
X times per hour
X times per day
X times per week
X times per month
Not Reported
Exposure Type
The exposure type represents the method by which the contaminant is administered to the test
organism. Studies reporting results for oral exposures (diet, gavage, capsule and drinking water)
are used exclusively for the purpose of establishing the Wildlife TRVs. Studies reporting an
exposure type other than oral should have been excluded earlier in the process by application of
the Literature Rejection Criteria described in Section 2. If the User discovers a study reporting
results for non-oral exposures at this point of the data entry process, the information entered to
this point is saved and the program exits to the "Data Entry" screen.
The following are special cases that might be encountered in coding exposure type and route. If
the publication only indicates "diet" as an exposure, the User should choose "FD" (contaminant
incorporated into the food) for the exposure route. If the exposure route changes within a test, the
data are entered as a separate phase. Dosing of progeny via the mother's milk in reproductive
Guidance for Developing Eco-SSLs Attachment 4-3 4-20
June 2005
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and/or developmental studies is considered a food (FD) route, not a drinking water route. The
User should note that the concentration of the contaminant of concern in mother's milk must be
reported in order to code data for juvenile animals exposed via lactation.
Table lib. Exposure Type Code Descriptions
Code
FD
DR
CH
GV
OR
Description
contaminant incorporated into the food
contaminant incorporated into the drinking water
choice of treated or untreated food or drinking water
gavage
other oral (e.g. capsule)
Route of Exposure
The route of exposure is directly related to the "Exposure Type" as described in Table 1 Ib.
Because the Wildlife TRVs are based on data from oral exposure studies, only codes specific to
oral exposures are available in the pull down list.
Test Location
The User selects the appropriate location or setting in which the experiment is reported to be
conducted from the pull down list. The list of test locations and definitions is provided in Table
12. The listed options are based on Rand (1995). If the test location is not specified, the User is
instructed to select "NR" for Not Reported.
Table 12. Test Location Code Descriptions
FieldA*
FieldN*
FieldU*
Lab*
NR*
Field, Artificial - a simulated or artificial field study is conducted in "an artificially bounded
system that is a simplification of a specific ecosystem", e.g., aviaries, pens, enclosures
Field, Natural- a natural field study is one "in which both the test system [...] and exposure
to the stressor are "naturally" derived"; e.g., sprayed agricultural field or orchard plots, field
surveys
Field - Unable to determine whether natural or artificial setting
Laboratory indoor setting
Not Reported - unable to determine if laboratory or field
* Rand (1995)
Experimental Design
The purpose of the experimental design field is to capture information on the overall purpose and
design of the study and to record important information that is not entered elsewhere. The User
enters a statement on the objective of the experiment and a brief description of the experimental
design in the text box provided. Care should also be taken to enter information on reproductive
Guidance for Developing Eco-SSLs Attachment 4-3 4-21
June 2005
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stages in this section. The experimental design should include, but is not limited to, information
specific to the number of experiments conducted, dosing design, types of endpoints recorded,
exposure durations (including the full duration of the experiment and interim time points), and
statistical methods used to analyze the results. In studies with multiple phases this information
may overlap information entered in the remarks field. In this case, enter "see remarks" for
information that would be otherwise duplicated. An example of an experimental design entry is
provided in the text box . Six main points to capture in this section: (1) Purpose of study; (2)
Brief description of exposure (species, initial age, chemical, and duration of exposure); (3) time
intervals when food consumption was monitored and dosing design; (4) time intervals when
effect measurements were taken; (5) brief summary of effect groups that were measured during
the study; (6) statistical analysis
If test methods are not reported in the current paper, but are reported in another paper being used
for TRV, the User should enter the SSL-ID number for the paper containing the full description of
test methods.
Test Conditions
A checklist of representative USEPA (2002) and ASTM (2001a-d) standard guidelines and
reporting parameters for toxicological studies is provided as Table 13. Comparable toxicology
study guidelines are also published by the U.S. Food and Drug Administration (USFDA, 2002)
and the Organisation for Economic Development and Cooperation (OECD, 2002). These
guidelines provide detailed descriptions of acceptable methodology for specific types of toxicity
tests and are designed to generate consistently acceptable data for regulatory purposes. Studies
conducted by the National Toxicology Program follow procedures comparable to those described
in the USEPA guidelines, and should be considered guideline studies for purposes of deriving
Wildlife TRVs..
For derivation of Wildlife TRVs, the
objective in reviewing studies for
guideline compliance is to determine
whether the study authors used acceptable
general methodology and animal
husbandry procedures. A comparison of
reporting parameters for 16 standard
toxicological test protocols in Table 13
indicates that descriptions of procurement,
animal care, characterization of the
material tested are common to many
standard guidelines. Therefore, reporting
of information on these parameters in a
study under evaluation for derivation of a
TRV is considered a good indication that
an acceptable approach to testing was
used. The User evaluates the test
Experimental Design - Example Text
Purpose of study was to assess the effect of cadmium
on energy metabolism and tissue metal concentrations.
Juvenile mallard drakes were exposed to cadmium in
feed for 41 to 43 days. Food consumption was
monitored for 3 day periods at biweekly intervals.
Body weights were determined on days 0, 21, and 42.
Terminal endpoints included selected organ weights;
concentration of cadmium, copper, and zinc in liver
and kidney; biochemical measurements in liver, blodd,
kidney, blood, and urine. One-way ANOVA was used
to test for significant effects. Duncan's multiple range
test was used to test for significant effects in pair-wise
comparisons among variables significantly affected by
treatment.
Guidance for Developing Eco-SSLs Attachment 4-3 4-22
June 2005
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conditions reported in the study by comparison to the parameters in Table 13. The User then
chooses the appropriate description from the pull down list based on the test conditions and
parameters reported in the study. If the test conditions are not reported in the current paper, but
are reported in a separate paper being used for TRV, the User records the SSL-ID number for the
source of the test methods in the Experimental Design comment field.
Experiment Sample Size
The experiment sample size should be summarized within this field. The user should denote the
number of experimental animals per exposure dose or concentration as well as the controls. Any
reduction in animal numbers during the experiment for reasons other than contaminant exposure
should be denoted (losses from disease, handling, etc.,).
Table 13. Standard Study Guidelines and Reporting Parameters
Test Conditions
Source of Test Animals
Health of Test Animals
Age of Test Animals
Acclimation procedures
Assignment of animals to
housing
Description of basal diet
(including source,
diluents and
supplements)
Nutrient content of diet
Water
Description of housing
conditions (including
size, type, material)
Temperature
Photoperiod
Lighting intensity
Humidity
Frequency, duration and
methods of observation
General description of
facilities
Description of test
substance (including
CAS number, purity,
source, solvent or carrier,
if used.)
Test Protocols
Avian
Dietary
OPPTS
850.2200
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
Avian
Reproduction
OPPTS
850.2300*
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
90 day Oral
Study in Rats
ASTME
1372-95
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
Chronic
Oral Study
in Rats
ASTME
1619-95
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
Sub acute
Dietary
with
Avian
Species
ASTME
857-87
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
Reproductive
Studies with
Avian Species
ASTME
1062-86
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
Developmental
Toxicity in
Rats and
Rabbits
ASTM E 1483-
92
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
Reproduction
and Fertility
Study in Rats
ASTME
1062-86*
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
* These test guidelines have recently been withdrawn without designation of replacement guidelines. Information on replacement
guidelines will be included in future updates of this SOP as it becomes available.
Guidance for Developing Eco-SSLs Attachment 4-3 4-23
June 2005
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The "Exposure Information" screen is now complete. The User now verifies that all data entered
are correct and click on "Next" at the bottom of the screen to continue. The User should not use
the browser back arrow to return to a previous data entry screen to correct errors. Using the back
arrow results in deletion of information.
4.4 Endpoint Information
Exposure Duration and Units
The exposure duration for a specific endpoint
is entered in the numeric field provided. For
example, if contaminant exposure lasts for ten
weeks, the User enters the number 10. For
studies that report dosages (or concentrations)
that are varied during the period of exposure,
the User evaluates each unique dosage
duration as a separate endpoint. The units
associated with the exposure duration are
selected from the pull down list provided.
The list of available units is shown in Table
14. Studies of three days or less are
considered acute and are not coded. Results
reported for recovery periods (i.e., after
exposure to the contaminant ceases) longer
than one day post-exposure are not coded. In
the comment field the User should also record
the exposure durations at which
measurements of the endpoint was made (e.g.
1, 14 and 28 days).
The general rules for determining the
exposure duration for each LOAEL/NOAEL
are as follows:
Coding Gestational and Lactation Exposure
Studies
Gestational Exposures
Enter results for gestational exposures as
Phases for the mother not the progeny
Effects to the progeny are coded as
reproductive endpoints (REP)
Enter the exposure duration as the time
chemical was administered during gestation.
If exact time is not reported estimate based
on gestation of test animal.
Lactation Exposures
Enter results for lactation exposures as
Phases separate for the mother and the
progeny
For the Phase for the mother effects to the
progeny are coded as reproductive endpoints
(REP)
Enter the exposure duration as the time
chemical was administered during lactation.
If exact time is not reported estimate based
on lactation time for the test animal.
If it is possible to quantify exposures for
progeny during lactation (i.e. the
concentration of contaminant is measured in
the mother's milk) then enter exposure
duration as the time during lactation that the
progeny was exposed.
1. The exposure duration coded should be for
the first occurrence of an adverse effect.
If no clear dose response exists at the
shortest exposure duration, the User
should move to the next exposure duration and reassess the dose-response relationship.
2.. The first (shortest) exposure duration reporting an adverse should be coded, as long as a clear
dose response relationship is evident in the data collected at this time point. The User should
record in the exposure duration comment field the exposure durations at which further (later)
measurements are reported.
Guidance for Developing Eco-SSLs Attachment 4-3 4-24
June 2005
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3. If none of the exposure durations are associated with an adverse effect, the longest exposure
duration is selected.
Table 14. Exposure Duration and Age Units
s
mi
h
d
w
mo
yr
If
NR
second
minute
hour
day
week
month
year
lifetime
Not Reported
Age with Units
The User enters the age of the test organism at the beginning of the study in the numeric field
provided. For example, if the study reports that two-week-old ducklings are exposed at the start
of the study, the User enters the number 2. The appropriate units are selected from the pull down
list provided. The list of available age units is shown in Table 14. For designation between adult,
juvenile, sexually mature, sexually immature, the user should assign the terms juvenile, adult and
sexually mature according to the guidelines in Table 20. If the author identifies the test
organisms as sexually mature or sexually immature this should be preferentially assigned.
Sex
The User selects the sex of the test organism from the pull down list provided. "M" is selected
for male organisms. "F" is selected for female organisms. If organisms of both sexes are tested,
"BH" is selected for "Both Male and Female". "NR" is selected for Not Reported if the sex of the
test organisms is not specified.
Lifestage
The lifestage of the test organism is selected from the pull down list provided. The list of
available lifestages is shown as Table 15. If the lifestage of the test organism is not reported then
it can be estimated based on the reported body weight of the organism using the default body
weight table. If the lifestage is not evident from the study, then "NR" is selected for Not
Reported. The lifestage in which the result is reported is used when an organism is dosed through
multiple lifestages. For example, if a female is exposed before mating, through gestation, and
into lactation, the lactation (LC) lifestage is used.
The User should note that the pull down list for lifestage includes larvae (LV), nauplii (NU) and
pupa (PU) lifestages for terrestrial insects. These are included for possible future applications and
do not apply to the coding process for Wildlife TRVs.
Guidance for Developing Eco-SSLs Attachment 4-3 4-25
June 2005
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Is This a Critical Lifestage?
A lifestage is defined as "critical" if it is critical to the survival and reproduction of the species.
These lifestages may or may not be more sensitive to contaminant exposure. Exposures during
these critical lifestages are preferred in the derivation of wildlife TRVs. Table 15 identifies the
lifestages from the pull down list considered to be "critical". The User selects "Yes" or "No" by
checking the appropriate box. If the lifestage is not specified, the User should check "NR" for
Not Reported. There may be some cases where the User must use professional judgement to
classify certain exposures as critical; these cases should be documented by comments. Critical
exposure periods include those during lactation and gestation. When reproductive effects are
reported for adult males or females, the lifestage is identified as critical.
Determination of whether a lifestage in avian studies is critical is done in the following way. The
lifestage code LB is used for birds that are producing eggs during the study period. The lifestage
LB is applied only to REP endpoints. This is by definition a critical lifestage that applies to all
measured REP endpoints. The AD lifestage code is assigned to adult birds that are not laying
eggs during the study period. The User should note that the relevant lifestage code for birds that
stop producing eggs as a result of treatment is LB.
Table 15. Lifestage Code Descriptions
Code
AD
EG
EM
IM
JV
LB
MA
NR
SA
SI
SM
YO
YY
GE
LC
Lifestage
adult
egg
embryo
immature
juvenile; includes yearling,
fledgling, hatchling, weanling
egg laying bird
mature
not reported
subadult
sexually immature
sexually mature
young
young of year
Gestational Exposures
Lactation
Critical
(Yes or No)
No
Yes
Yes
Yes
Yes
Yes
No
No
No
No
No
Yes
Yes
Yes
Yes
Effect Group
Exposure of test organisms to contaminants can result in both positive and adverse effects. The
reviewer should code only potentially adverse effects. Possible adverse effects that may be
reported in toxicological studies are divided into nine Effect Groups. These groups were
developed as part of the coding system devised for ECOTOX by EPA Duluth. The nine Effect
Groups are accumulation (ACC), behavior (BEH), biochemistry (BIO), growth (GRO), mortality
(MOR), pathology (PTH), physiology (PHY), population (POP), and reproduction (REP). A brief
Guidance for Developing Eco-SSLs Attachment 4-3 4-26
June 2005
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description of each effect group is provided under the description link to the right of the pull
down list. The list of available Effect Groups is shown and described in Table 16.
The User selects the appropriate effect group from the pull down list provided. The User should
consult both Tables 16 and 17, which provide the Effect Types and Measures that are specific to
the Effect Groups to identify the appropriate Effect Group for the endpoint described in the study
under review.
Table 16. Effect Group Descriptions
Code
Description
ACC
Accumulation: a general term describing the process (bioaccumulation) by which contaminants are taken into
and stored in plants or animals; bioaccumulation occurs when the rate of contaminant uptake exceeds the rate
of elimination of the same contaminant; therefore accumulation measurements include uptake (UPTK) and
elimination (ELIM) rates as well as actual tissue concentrations (RSDE); accumulation endpoints include the
asymptotic threshold concentration (ATCN), bioconcentration factor (BCF) and bioaccumulation factor (BAF).
BEH
Behavior: a general term characterizing overt activity of an organism represented by three effect groups -
avoidance (AVO), general behavior (BEH), and feeding behavior (FDB). Examples of behavioral
measurements include stimulus avoidance (STIM), feeding changes (FDNG), general reproductive success (
RSUC), and general activity levels (ACTV).
BIO
Biochemical: measurement of biotransformation or metabolism of chemical compounds, modes of toxic
action, and biochemical responses in animals including three effect groups - chemical (CHM), enzyme (ENZ)
and hormone (FIRM) effects. Examples of biochemical measurements include chemical parameters such as
cell (CCHG) or amino acid (AMAC) changes, enzyme parameters such as transferase, oxidase or hydrolase
reactions, and measurements of hormone response levels. NOTE: Biochemical responses associated with
heavy metal accumulation (e.g., zinc content) should not be coded. Responses that are indicative of a target
tissue response (e.g., calcium content in bone) should be coded.
GRO
Growth: a broad category which encompasses measures of weight and length and includes effects on
development (DVP), growth (GRO) and morphology (MPH). Morphology: measurements and endpoints
which address the structure (bones) and form (organ/tissue development) of an organism, at any stage of its life
history.
MOR
Mortality: measurements and endpoints where the cause of death is by direct action of the contaminant; e.g.
an endpoint such as the LD50 estimates the lethal dose to 50% of the exposed population whereas
measurements count the actual number dead or the percentage reduction within a population as a result of the
exposure.
PTH
Pathology: measurements and endpoints regarding the causes, nature and effects of diseases and other
abnormalities; the four effect groups include histology (EDS), immunotoxicity
(IMM), intoxication (ITX), and gross wasting effects (GRS). Immunotoxicity, parasites and tumor effects are
not coded for TRVs for the Eco-SSLs.
POP
Population: measurements and endpoints regarding a group of organisms or plants of the same species
occupying the same area at a given time. Measurements include abundance, biomass, size and age class
structures.
PHY
Physiology: measurements and endpoints regarding changes and activity in cells and tissues of plants or
animals.
REP
Reproduction: measurements and endpoints to track the effect of toxicants on the reproductive cycle.
Effect Type
The available Effect Types are provided in a pull down list. The listed Effect Types are specific
to the Effect Group previously selected by the User. The appropriate Effect Type for the endpoint
is selected from the pull down list provided. The available selections are listed in Table 17.
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June 2005
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Effect Measure
The effect measure is a variable used to determine organism response to contaminant exposure.
The available Effect Measures in the pull down list correspond to the previously selected Effect
Type. The User selects the Effect Measure from the pull down list. The measurement code
"XXXX" denotes a temporary code that needs to be validated and assigned an acronym. The list
of available selections is provided in Table 17. To avoid repetitive entries of NOAEL and
LOAEL values and to make the coding process more efficient, the User is instructed to record
only one result per Effect Group. The most conservative result (lowest NOAEL or LOAEL)
should be recorded. In cases where there are biochemical, behavior, or pathology changes
reported in offspring then two reproduction endpoints can be coded one reporting these effects in
the progeny and a second reporting effects for the parent(s).
Table 17. Effect Groups, Types and Measures
BEHAVIOR EFFECT GROUP (BEH)
(AVO) Avoidance Effect Type Measurement Code and Definition
CHEM
FOOD
STIM
WATR
contaminant avoidance
food avoidance
stimulus avoidance
water avoidance
(BEH) General Behavior Effect Type Measurement Code and Definition
ACTP
ACTV
AGGT
AMBU
EQUL
GBHV
LOCO
DPLY
DRMT
FRZG
GBHV
GPRD
INST
NMVM
NVOC
RRSP
RSPT
VCLF
accuracy of learned behavior
activity, general
aggression
ambulatory /circadian rhythm
balance and equilibrium
behavioral changes
distance
displaying behavior
dormant, adverse condition
freezing behavior
general behavior
general production
induced sleeping time
number of movements
number of vocalizations
righting response
response time to stimulus
visual cliff
(FOB) Feeding Behavior Effect Type Measurement Code and Definition
BGNB
CAIN
FCNS
FDNG
begging behavior
caloric intake
food consumption
feeding behavior
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June 2005
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Table 17. Effect Groups, Types and Measures
FEFF
FSTR
FTIM
GFDB
WCON
feeding efficiency
food storage
feeding time
general feeding behavior
water consumption
BIOCHEMISTRY EFFECT GROUP (BIO)
(CHM) Biochemical Effect Type Measurement Code and Definition
5HAA
AABA
ALAN
ALBM
ALBE
AMAC
AMMO
AMNH
ANBC
ARGI
ASCA
ASHC
ASIS
ASPA
ATPT
B2MG
BASO
BIOT
BUNT
CALC
CAPH
CCHG
CHLN
CHLR
CHOL
CREA
DISC
DTBL
EOSN
ERTH
ESAA
FECO
FEPR
FFTA
GBCM
GLCN
GLTH
GLUC
GLYC
5-hydroxyindole acetic acid
alpha-aminobutyric acid
alanine
albumins
albumen energy
amino acids, general term
ammonia
p-amino hippurate
aniline binding capability
arginine
ascorbic acid
ash content
amyloidosis
apartate
adenosine triphosphate
beta2-microglobulin
basophil
biotin content
blood urea nitrogen
calcium
calcium/phosphorus ratio
cell changes
choline
chloride
cholesterol
creatinine
dethylsuccinate hdyrolysis
direct bih'rubin (conjugated)
eosinophil
erythoroblasts
amino acids, essential
iron content (do not code)
free erythrocyte protoporphyrins
fatty acids, free
general biochemical
glycine
glutathione
glucose
glycogen
Guidance for Developing Eco-SSLs Attachment 4-3 4-29
June 2005
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Table 17. Effect Groups, Types and Measures
GMIN
HEME
HEMT
glutamine
heme content
general hematology
(CHM) Biochemical Effect Type Measurement Code and Definition
HIST
HMCT
HMGL
IBIL
ILEU
LACT
LCTA
LEUC
LEUK
LIPD
LMPH
LPSA
LYSI
MCHC
MCPR
MCPV
METH
MONO
NADP
NEAA
NEFA
NEUT
ORNI
OSRS
PCLV
PHPH
PHEN
PPHT
PHSP
PCON
PHST
PORP
KCON
PRTO
PYRV
RGSH
NPSH
RBCE
RBVL
RETI
SERI
NACO
SOMC
histidine
hematocrit (anemia)
hemoglobin
indirect bilirubin (free)
isoleucine
lactate
lactic acid
leucine
leukocytes
lipid
lymphocyte
lipid soluble antioxidants
lysine
mean corpuscular hemoglobin
microsomal proteins
mean corpuscular volume
methionine
monocyte
nicatinamide-adenine dinucleotide phosphate, reduced
amino acides, nonessential
fatty acids, nonesterified
neutrophil
ornithine
osmotic resistance/RBC
packed cell volume
pH
phenyalanine
phosphate
phosphatide phosphorus
phosphorus
phospholipid content, total
porphyrin
potassium
protoporhyrin
pyruvate
reduced gluthione
nonprotein sulfhydryl
red blood cell
relative blood volume (volume/lOOg body weight)
reticulocytes
serine
sodium
somatomedin C
Guidance for Developing Eco-SSLs Attachment 4-3 4-30
June 2005
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Table 17. Effect Groups, Types and Measures
SPLO
splenocytes
SRTN
serotonin
TEAM
tetraethylammonium
(CHM) Biochemical Effect Type Measurement Code and Definition
TFAA
amino acids, total free
THBA
thiobarbituric acid
THIA
thiamine
THRE
threonine
THRM
thrombocytes
TLBL
bih'rubin, total
PRTL
protein, total
TRIE
tributyrin
TRIG
triglycerides
TYRO
tyrosine
TRYP
tryptophan
TTAA
amino acids, total
TWBC
white blood cell count, total
UBWB
white blood cell, undifferentiated blasts
UREA
urea
URIC
uric acid
VALI
valine
VTD3
vitamin D3
VTMA
vitamin A
ZNPR
zinc protoporphyrin
(ENZ) Enzyme Effect Type Measurement Code and Definition
20HB
2-OH biphenyl hydroxylase
40HB
4-OH biphenyl hydroxylase
450R
Cyt P450 reductase
DHYD
NADPH dehydrogenase
AATT
alanine aminotransferase
ACHE
acetylcholinesterase
ACPH
acid phosphatase
AEPX
aldrin epoxidase
AHHD
aryl hydrocarbon hydrolase
AHDX
aniline hydroxylase
ALAD
(delta) -aminolevulinic acid dehydrogenase
ALAS
(gamma) y-ALA synthetase
ALDO
aldolase
ALPH
alkaline phosphatase
ANAE
alpha naphthyl acetat esterase
APND
aminopyin n-demethylase
ATRP
alanine transpeptidase
ASAT
aspartate aminotransferase
BAPH
benzo(a)pyrene hydroxylase
BCHE
buterylcholinesterase
BCOD
butoxycoumurin O-dealkylase
BHXA
benzpyrene hydroxylase
Guidance for Developing Eco-SSLs Attachment 4-3 4-31
June 2005
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Table 17. Effect Groups, Types and Measures
BPND
BROD
CAAH
benzphetamine-n-demethylase
benzylresorufin O-deethylase
carbonic anhydrase
(ENZ) Enzyme Effect Type Measurement Code and Definition
CACA
CATP
CYB5
CCOX
CEST\
CRKI
NCCR
EPHY
ECOD
EROD
ESTE
FDPA
GENZ
G6PD
GGTR
GLAD
GLPX
GLRE
GLTR
GOTR
GPTR
GSTR
HXBH
LADH
LDMD
MADH
MCOD
MG6P
MAOA
MGAT
NKAT
ORCT
P450
PBES
PBHD
PCOD
PNAD
PROD
SBDH
choline acetyltransferase
calcium ATPase
cytochrome B-5
cytochrome C-oxidase
chloinesterase
creatine kinase
NADPH cytochrome C reductase
epoxide hydrase
ethoxycoumurin O-deethylase
7-ethoxyresorufin O-deethylase
esterase
fructos-diphosphate aldolase
general enzyme
glucose-6-phosphate dehydrogenase
(gamma) Y-glutamyltransferase
glutamic acid dehydrogenase
gluathione peroxidase
gluthione reductase
glucouronyl transferase
glutamic -oxaloacetic transaminase
glutamic pyruvic transaminase
glutathione S-transferase
hexobarbital hydroxylase
lactate dehydrogenase
lactate dehydrogenase/malic dehydrogenase ratio
malic dehydrogenase
methoxycoumarin O-dealkylase
microsomal glucose 6-phosphatase
mono amino oxidase
magnesium ATPase
sodium potassium ATPase
ornithine carbamoyl transferase
cytochrome P450 proteins
phenyl benzoate esterase
pentobarbital hydroxylase
propoxycoumarin O-dealkylase
p-nitroanisole demethylase
pentylresorufin O-deethylase
sorbitol dehydrogenase
(ENZ) Enzyme Effect Type Measurement Code and Definition
SCDH
SCOT
SGPT
succinate dehydrogenase
serum glutamate oxalo aetate transaminase
serum glutamic pyruvic transaminase
Guidance for Developing Eco-SSLs Attachment 4-3 4-32
June 2005
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Table 17. Effect Groups, Types and Measures
THTR
TRIE
thiotransferase
triacetin esterase
(HRM) Hormone Effect Type Measurement Code and Definition
ANDR
CORT
CRTS
DOPA
EPIN
ESDL
ESTR
FOSH
GHRM
GTHH
GHRM
LUTH
NORE
PROS
TSTR
THYR
TRII
androgen
corticosterone
cortisol
dopamine
epinephirine
17-beta estradiol
estrogen
follicle stimulating hormone
general hormone
growth hormone
hormone, changes in
luteinizing hormone
norephinephrine
progesterone
testosterone
thyroxine
tridothyronine
Growth Effect Group (GRO)
(DVP) Development Effect Type Measurement Code and Definition
EMDV
FLDG
GDPV
LRGN
WEAN
embryo development
fledged/female or /brood
general development
limb regeneration
weaned
(GRO) Growth Effect Type Measurement Code and Definition
BODL
BDWT
GGRO
body length changes - non-adult organisms only (see PTH GRS for adults)
body weight changes - non-adult organisms only (see PTH GRS for adults)
general growth
(MPH) Morphology Effect Type Measurement Code and Definition
cose
CRLT
FRLT
GMPH
HULT
MOSC
MUSC
OVLT
RULT
SOSC
caudal ossification center
crown-rump length
feather length
general morphological changes
humerus length
metacarpal ossification center
muscle changes
oviduct length
radius-ulna length
sternal ossification center
Guidance for Developing Eco-SSLs Attachment 4-3 4-33
June 2005
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Table 17. Effect Groups, Types and Measures
(MPH) Morphology Effect Type Measurement Code and Definition
SRIB
TRLT
TELT
TTLT
supernumerary ribs
tarsus length
testis length
tibiotarsus length
MORTALITY (MOR) EFFECT GROUP
(MOR) Mortality Effect Type Measurement Code and Definition
GMOR
LFSP
MDTH
MORT
SURV
TDTH
TKNO
general mortality
lifespan
mean time of death
mortality
survival
time to death
knockdown
PATHOLOGY (PTH) EFFECT GROUP
(GRS) Gross Wasting Effect Type Measurement Code and Definition
BODL
BDWT
body length changes - adult organisms only (see GRO GRO for non-adults (jv, ma, sm, etc.)
body weight changes - adult organisms only (see GRO GRO for non-adults (jv, ma, sm, etc.)
(ORW) Organ Weight Effect Type Measurement Code and Definition
ORWT
SMIX
organ weight changes
organ weight in relationship to body weight
(HIS) Histology Effect Type Measurement Code and Definition
ARTS
CTYP
EDMA
ENCP
GHIS
GLBM
GLSN
HEMR
HYPL
IIBD
NCRO
NPHR
TFLR
USTR
arteriosclerosis
percent cell type
edema
encephalopathy
histological changes, general
glomerular basement membrane
gross lesions
hemorrhage
hyperplasia
intranuclear inclusion bodies
necrosis
nephrosis
tissue fluorescence in UV light
ultrastructural changes
(ITX) Intoxication Effect Type Measurement Code and Definition
ANOR
ATAX
CONV
GITX
IMBL
INCO
GITX
PARL
TINT
TREM
anorexia
ataxia
convulsions
general intoxication
immobile
incoordination
intoxication, general
paralysis
time to signs of intoxication
tremors
Guidance for Developing Eco-SSLs Attachment 4-3 4-34
June 2005
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Table 17. Effect Groups, Types and Measures
POPULATION (POP) EFFECT GROUP
(POP) Population Effect Type Measurement Code and Definition
PBMS
DVRS
GPOP
INDX
ABND
NCHG
RCPR
SEXR
TRAP
biomass or weight for total population
diversity and evenness
general population effect
index to population size, count, number
number of animals/population and population density (N/area)
population change (change in N/change in time)
recapture ratio
sex ratio
trappability
PHYSIOLOGY (PHY) EFFECT GROUP
(PHY) Physiology Effect Type Measurement Code and Definition
ADPO
BLPR
BTMP
CRDY
DIFD
BDVL
EECG
EXCR
FDCV
GPHY
GLFR
HTRT
HYDR
NRGM
META
RPRT
OSMO
PRIN
IRRI
THRG
PROT
oxidative phosphorylation
blood pressure
body temperature
crop dysfunction
digestibility of food
blood volume
electroencephalogram
excretion rate
food conversion efficiency
general physiology
glomular filtration rate
heart rate
hydration
metabolizable energy
metabolic rate
respiratory rate
serum/plasma osmolality
PR intervals
skin irritation
thermoregulation
Prothrombin time
REPRODUCTION (REP) EFFECT GROUP
(REP) Reproduction Effect Type Measurement Code and Definition
ABNM
BEFF
BNDG
COUR
CYNG
DEYO
EGPN
PERT
GREP
GIDX
abnormal
breeding efficiency
pair bonding nesting behavior
courtship behavior
care of young, nest attentiveness
death of young
eggs per nest
fertility
general reproduction
gestation index
Guidance for Developing Eco-SSLs Attachment 4-3
4-35
June 2005
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Table 17. Effect Groups, Types and Measures
(REP) Reproduction Effect Type Measurement Code and Definition
GSTT
HTCH
LACG
NANT
NCLU
NDAY
NINC
NOPN
NSNT
NSTI
NSTS
NTSZ
NUNT
OBRD
ODVP
OEGP
OHIS
ORWT
OTHR
OVRT
PBEH
PIPD
PLBR
PRFM
PROG
PRWT
PVOP
RBEH
SBRD
RHIS
RPRD
RSEM
RSUC
SPCL
SPCV
TEDG
TERA
TEWT
TPRD
gestation time
hatch
lactating
nests abandoned
corpus lutea, number of
number of days between eggs laid
number of nests incubated
number of organisms per nest
successful nests
nest initiation
number of active nests
nest size
unsuccessful nests
open brood
offspring development
onset of egg production
offspring histology
reproductive organ weight
other; use this to code any effects (pathology, biochemistry, etc.) reported in offspring of
contaminant-exposed mothers
ovulation rate
progeny behavior
pipped
pairs with litter or brood
pregnant females in a population
progeny counts/numbers
progeny weight (TBWT, LTWT)
premature or delayed vaginal opening
reproductive behavior changes
sealed brood
reproductive organ histology
reproductive capacity
resorbed embryos
reproductive success (general)
sperm cell counts
sperm cell viability
testes degeneration
teratogenic measurements
testes weight
total production
(EGG) EGG Effect Type Measurement Code and Definition
ALWT
CRAK
EGVL
EGWT
EQUA
albumen weight
cracked eggs
egg volume
egg weight
egg quality
Guidance for Developing Eco-SSLs Attachment 4-3
4-36
June 2005
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Table 17. Effect Groups, Types and Measures
ESIN
eggshell index
(EGG) EGG Effect Type Measurement Code and Definition
ESLT
ESQU
ESTH
ESWD
ESWT
FTEG
GEGG
SHLL
SHSZ
SFYK
YOLK
YKWT
eggshell length
eggshell quality
eggshell thicknes
eggshell width
eggshell weight
fertile egg
general egg effects
percent shell
shell size
soft yolk
yolk, percent
yolk weight
Response Site
The response site is the specific location (e.g., organ or tissue) where an effect is observed. The
response site for mortality (MOR) or behavioral (BEH) effects is not applicable and whole
organism should be selected. The response site that is specific to the endpoint is selected from
the pull down list. The list of available selections is shown in Table 18. If the response site is not
reported, "NR" is selected for Not Reported.
Table 18. Response Sites and Codes
Code
AG
AD
AR
AS
AL
AT
AF
AP
BI
BL
BV
BO
BM
BR
BT
BU
CA
CH
CL
CG
CO
CR
Response Site
Accessory Gland
Adipose Tissue
Adrenal Gland
Air Sac
Albumen (egg white)
Alimentary Tract
Amniotic Fluid
Appendages
Bile
Blood
Blood Vessel
Bone
Bone Marrow
Brain
Breast
Bursa
Cartilage
Cord, spinal
Claw
Cloacal gland
Collagen
Crop
Code
MK
MT
MU
MB
MO
NG
NE
NK
NR
OL
0V
OD
PS
PE
PI
PC
PL
PG
PR
PY
RC
RT
Response Site
Milk, lactating females
Multiple Tissue/Organs
Muscle
Muscle+Bone
Mucous
Nasal Gland
Nervous Tissue
Neck
Not Reported
Olfactory
Ovaries
Oviduct
Pancreas
Penis
Pituitary Gland
Placenta
Plasma
Prostate Gland
Proventriculus
Progeny
Rectum
Reproductive Tissue
Guidance for Developing Eco-SSLs Attachment 4-3 4-37
June 2005
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Table 18. Response Sites and Codes
Code
DG
DT
EG
EU
EM
EN
ER
ES
EY
FE
FM
FO
FT
GB
GT
GZ
GO
HA
HD
HE
HM
HG
HY
IN
KI
LD
LG
LI
LU
MM
MS
MC
Response Site
Digestive Gland
Digestive Tract
Egg
Egg Cuticle
Embryo
Entrails
Erythrocyte
Esophagus
Eye
Feathers
Femur
Foot
Fetus
Gall Bladder
Gastrointestinal Tract
Gizzard
Gonads
Hair
Head
Heart
Humerus
Harderian Gland
Hypothalamus
Intestinal Tract
Kidney
Lipid, Fat
Leg
Liver
Lungs
Mammary Tissue
Mesenteric Lymph Node
Micro some
Code
SV
SE
SR
SN
SK
SG
SL
SM
SP
SH
ST
SX
TA
TE
TG
TH
TB
TI
TS
TY
UB
UR
UT
VA
VD
VE
VI
WI
WO
YO
XX
Response Site
Seminal Vesicle
Sensory Organs
Serum
Skeleton
Skin, Epidermis
Shell Gland
Shell
Sperm
Spleen
Stomach
Soft Tissue
Submaxillary Gland
Tail
Testes
Thigh muscle
Teeth
Tibia
Tissue
Thymus
Thyroid
Urinary Bladder
Urine
Uterus
Vagina
Vas Deferens
Vertebra
Viscera
Wings
Whole Organism
Yolk
Temporary code, requires validation
Endpoint Comments
The endpoint comment field allows the User to enter specific notes concerning the selected
endpoint. In cases where multiple effect types are reported, the User records only one
measurement and response site. The User enters the rationale for selection of a particular
endpoint for data entry in the comments field. The comment field is also used to record other
effect types, measurements, and/or response sites in the same effect group that are not chosen for
coding. Abbreviations are used as much as possible in the comment field.
Identify the NOAEL and or LOAEL
The User is required to review each toxicological study and to identify No Observed Adverse
Effect Level (NOAEL) and/or Lowest Observed Adverse Effect Level (LOAEL) values. The
NOAEL is defined as the highest concentration (or dose) used in a study for which there is no
Guidance for Developing Eco-SSLs Attachment 4-3 4-3 8
June 2005
-------
statistically significant adverse effect observed in the test organism. The LOAEL is defined as
the lowest concentration (or dose) at which a statistically significant adverse effect is observed in
contaminant-exposed organisms when compared to controls. The identification of the NOAEL
and LOAEL is the most critical step in the data entry process, as these values will be ultimately
be used to derive the Wildlife TRVs.
It is important to note that the NOAEL and LOAEL are endpoint specific. For example, the
selected LOAEL for a growth endpoint may be 5.7 mg/kg BW/day whereas the LOAEL for a
pathological endpoint may be 2.3 mg/kg BW/day.
Publications or documents which report studies of interest to the regulatory community may
identify both a NOAEL and a LOAEL, only a NOAEL, or only a LOAEL. In cases where values
are identified by the study author, this fact should be noted in the NOAEL/LOAEL comments
field. The User should note NOAEL and LOAEL values are often reported only for the most
sensitive endpoint in the study. If NOAEL and/or LOAEL values are identified by the study
authors, these are considered to be the default values for entry into the TRY database. However,
NOAEL and LOAEL values identified by the study authors should be carefully reviewed by the
User for consistency with TRV coding guidelines.
Many publications, particularly those reporting basic toxicological research, do not identify
NOAEL or LOAEL values. In these cases, the User must determine whether there are sufficient
data available to determine NOAEL and/or LOAEL values. Four general cases are possible for
data adequacy:
The data have been analyzed using appropriate statistical methods
The data have been analyzed, but the statistical methods are not appropriate
No statistical analysis was performed, but sufficient data are available to perform an
independent analysis
No statistical analysis was reported and insufficient data are available to perform an
independent analysis
The process of identifying a NOAEL and/or LOAEL begins by determining whether statistical
analysis of the data was performed. This may be obvious from presentation of data in summary
tables. However, the User should be prepared to carefully examine the text in the methods
section, text of the results section, and footnotes in data tables for information on statistical
analysis and results. In cases were no statistically significant results were observed, the only
indication that statistical analysis was performed may be a description provided in the methods
section. If an appropriate statistical analysis has been performed, the User applies the general
rules below to identify NOAEL and/or LOAEL values.
The general rules for determining each LOAEL/NOAEL and their exposure durations are as
follows:
1. The User identifies a NOAEL in the following cases where: 1) there are sporadic, statistically
significant differences, but no clear dose response (e.g., a statistically significant difference is
Guidance for Developing Eco-SSLs Attachment 4-3 4-39 June 2005
-------
reported at a low or mid dose but not at higher doses); and/or 2) there is anecdotal information to
suggest that the apparent difference is a statistical artifact rather than a real effect. Statistical
significance must be at least a p value of 0.10 or less.
2. In individual studies which report results at multiple time points, the exposure duration coded
should be the first (earliest) occurrence of an adverse effect. The concentration or dose at which
the adverse effect occurs is the LOAEL. If a clear dose-response relationship is not evident at the
shortest exposure duration, the User examines data for the subsequent exposure durations in a
stepwise manner to identify the LOAEL.
3. When selecting among multiple effect measures for the same effect type, the first (shortest)
exposure duration resulting in the lowest LOAEL should be coded, provided a clear dose
response relationship is evident.
4. If there is no adverse effect reported and/or a dose-response relationship is not evident, the
User selects the highest dose (or concentration) at the longest duration as a NOAEL value.
The User should carefully review the methods and results sections and footnotes of the data
summary tables before making a determination that statistical analysis was not conducted.
The following examples are provided to assist reviewers in properly assigning NOAEL and
LOAEL values and the corresponding exposure durations:
Example 1
DurationVDoses
1 week
2 weeks
3 weeks
10 mg/kg/d
no
sig
no
20 mg/kg/d
no
sig
no
30 mg/kg/d
no
sig
no
TRY Duration
NOAEL/LOAEL
2 week
LOAEL 10
Example 2
DurationVDoses
1 week
2 weeks
3 weeks
10 mg/kg/d
no
no
no
20 mg/kg/d
no
sig
no
30 mg/kg/d
sig
no
sig
TRY Duration
NOAEL/LOAEL:
1 week
NOAEL 20; LOAEL 30
Guidance for Developing Eco-SSLs Attachment 4-3 4-40
June 2005
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Example 3
DurationVDoses
1 week
2 weeks
3 weeks
10 mg/kg/d
no
no
no
20 mg/kg/d
sig
no
sig
30 mg/kg/d
no
sig
no
TRY Duration
NOAEL/LOAEL:
2 week
NOAEL 20 LOAEL 30
Example 4
DurationVDoses
1 week
2 weeks
3 weeks
10 mg/kg/d
sig
no
no
20 mg/kg/d
no
sig
no
30 mg/kg/d
no
no
sig
TRY Duration
NOAEL/LOAEL:
3 week
NOAEL 20 LOAEL 30
Example 5
DurationVDoses
1 week
2 weeks
3 weeks
10 mg/kg/d
no
no
sig
20 mg/kg/d
no
sig
no
30 mg/kg/d
sig
no
no
TRY Duration
NOAEL/LOAEL:
1 week
NOAEL 20 LOAEL 30
Example 6
DurationVDoses
1 week
2 weeks
10 mg/kg/d
no
no
20 mg/kg/d
no
no
30 mg/kg/d
no
no
TRY Duration
NOAEL/LOAEL:
3 week
NOAEL 30
Guidance for Developing Eco-SSLs Attachment 4-3 4-41
June 2005
-------
3 weeks
no
no
no
Example 7
Duration/Doses
1 week
2 weeks
3 weeks
10 mg/kg/d
no
no
sig
20 mg/kg/d
sig
sig
no
30 mg/kg/d
no
no
no
TRY Duration
NOAEL/LOAEL:
3 week
NOAEL 30
(No dose response)
Example 8
DurationVDoses
1 week
2 weeks
3 weeks
10 mg/kg/d
no
no
no
20 mg/kg/d
sig
sig
no
30 mg/kg/d
no
no
no
TRY Duration
NOAEL/LOAEL:
3 week
NOAEL 30
(No dose response)
Example 9
DurationVDoses
1 week
2 weeks
3 weeks
10 mg/kg/d
no
no
no
20 mg/kg/d
sig
sig
sig
30 mg/kg/d
no
no
no
TRY Duration
NOAEL/LOAEL:
3 week
NOAEL 30
(No dose response)
The User should take special care in identifying NOAEL and LOAEL values for nutrients. In
some cases, contaminants that are also nutrients may give a biphasic dose-response curve, with
the lower doses eliciting a positive effect and higher doses causing a negative response. The User
selects the NOAEL and LOAEL for the second (adverse) phase of the dose response curve in
these instances. For purposes of the Wildlife TRY effort, only data for no effects or adverse
effects are recorded.
Guidance for Developing Eco-SSLs Attachment 4-3 4-42
June 2005
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A nutrient study that examines a relevant endpoint (i.e., one with the potential for adverse effects)
is coded, even in the absence of observed adverse effects. In this case, the User selects the
highest dose as a NOAEL. Beneficial effects are not coded.
In some cases, the statistical analyses used in a study may not be appropriate or adequate for the
particular study design. Alternatively, the study authors may not have performed a statistical
analysis of the data. In either case, the User has three options. The first option is to analyze or
re-analyze the data using appropriate statistical procedures and record the results (Figure 4-1)
(Gad, 1998). The second option is to determine a NOAEL or LOAEL based on the
preponderance of the data. The third option is rejection of the study and assignment of a data
evaluation score of 0. In this case, the study result would be rejected and not used in the
derivation of wildlife TRVs. In most cases a statistical analyses of the data will be completed in
cases where the study does not use any statistics. There are expected to be very few instances
where statistical analyses is recalculated by the User because the original method was inadequate.
The statistical re-analyses or application of these three options can be completed by the user but
will more often be the responsibility of the quality assurance reviewer.
If no statistical analysis was performed in a study and insufficient data are available to perform an
independent analysis, the User should reject the study and assign a data evaluation score of zero.
In this case, the study result would be rejected and not used in the derivation of wildlife TRVs.
This action is based on a low degree of confidence in studies which do not adequately report the
details of experimental design and results.
In theory, the threshold for the particular adverse effect lies between the NOAEL and the
LOAEL. Recent publications have reviewed the weaknesses of the use of NOAELs in risk
assessments (e.g., USEPA, 1995). Some analyses of acute toxicity data have shown that
NOAELs can represent as much as a 30% or 40% difference from control (as a result of low
statistical power), while other studies have identified LOAELs that are incorrectly low as a result
of statistical artifacts. While it is hoped that NOAELs and LOAELs bracket the threshold
concentration, their determination is a function of the spacing of dietary concentration and the
statistical power of the test.
NOAEL and LOAEL Units
The units associated with the NOAEL and LOAEL are automatically assigned by the application
based upon the units previously selected when describing the exposure concentrations or doses
(see the Exposure Information section). If measured concentrations are entered, these units are
preferentially returned as the units for the NOAEL field.
Is the NOAEL or LOAEL Reported by the Author?
If the NOAEL and/or the LOAEL are identified and reported by the author, the User selects "Yes"
by checking the appropriate box. If the NOAEL and/or LOAEL are assigned by the reviewer,
based on information provided in text, tables or figures, the User selects "No" by checking the
appropriate box.
Guidance for Developing Eco-SSLs Attachment 4-3 4-43 June 2005
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Assumed Parametric Data
Continuous data such as body
weights, blood-cell counts, etc.
Visually examine the
data.
Do thev aroear
NO
Comparison of 3 or
more groups
Comparison
of two
groups
Bartlett's
homogenecity
of variance
NOT SIGNIF.
SIGNIF.
{Homogeneous}
{Heterogeneous}
VOT NORMAL (+)
Analysis of
Variance
NOT SIGNIF.
SIGNIF.
Data not
significant.
No more tests
Intracomparison only
of groups vs. controls?
Group sizes approx.
equal?
Categorical (Quantal) Data
Frequency data such as
mortalities, pathology findings,
Comparison of
two groups
Comparison of 3 or
more groups
Non-Parametric Data
Includes data such as
percentage values, ranks, etc.
NOT SIGNIF.
{Homogt n}
No (to either question) Yes (to both
Student's t-test
Dt = (Nj + N2) - 2
{Heterogen}
N, =N,
N,
Comparison of two Comparison of 3 or
groups or if variance in more groups all
one or more groups = 0 with some variation
(no variation within within group
I
I
Student's t-test
Dt = N! - 1
* If plot does not clearly demonstrate, lack of normality exact tests may be employed.
If continuous data, Kalmogorov Smirnov test.
If discontinuous data. Chi-Snuare Goodness of Fit test mav he used.
Kruskal-Wallis
Non-
Parametric
Anova
SIGNIF. NOT SIGNIF.
Figure 4-1.
Decision tree for selecting hypothesis-testing
procedures (from Gad, 1998)
Distribution
Free Multiple
Comparisons
Data not
significant.
No more
tests
NOT SIGNIF.
Guidance for Developing Eco-SSLs Attachment 4-3
4-44
March 2005
-------
NOAEL and LOAEL Comments
The NOAEL/LOAEL comment field is used to record any specific information pertaining to the
selection of NOAEL and/or LOAEL. Statistical analysis information, including p-values (when
applicable), is entered in this field. Any information regarding lack of statistical significance at
different exposure durations during the test is noted here (i.e, no significance). The specific
location of the NOAEL and LOAEL results in the paper, including references to the text page
number and table or figure numbers, is recorded by the User in this comment field.
Is Wet Weight Reported?
The Eco-SSL for wildlife is reported as a "safe concentration" in soil on a dry weight basis. The
estimation (or back calculation) from a safe dose to an associated safe soil concentration requires
the TRY to be expressed on a dry weight basis. The requires that the estimation of a dose (mg of
contaminant per kg BW of the test organism per day) from dietary exposure concentrations be
based on units per dry weight diet.
If the study reports that the dietary exposure concentrations are expressed on a wet weight basis,
then the User should select "Yes" by checking the appropriate box. If the dietary concentration
units are reported as dry weight, select "No" by checking the appropriate box. If the dietary
concentration units are not specified as wet weight or dry weight, select "NR" for Not Reported.
Also select "NR" if a drinking water, gavage or other oral study is being entered. For studies
where NR is entered, the entered results are assumed to be reported in dry weight and are not
converted by the application. This is assumed to be conservative as conversion to dry-weight
results in higher LOAEL and NOAEL dose values.
If Wet Weight is Reported, Is the Percent Moisture Reported?
If the dietary concentration level units are reported as wet weight and the percent moisture is also
reported, the User selects "Yes" by checking the appropriate box. If percent moisture is not
reported, the User selects "No" by checking the appropriate box. For drinking water studies, the
User selects "NR" by checking the appropriate box.
Percent Moisture (%)
If the percent moisture in the exposure media is reported, the User enters the percent moisture in
the numeric field provided. For example, if the percent moisture for laboratory rat chow is
reported as 3 percent, the number 3 is entered. The number 100 should be entered for drinking
water studies. If the percent moisture is not report the application assumes 5%.
Is the Body Weight Reported?
The User should review the study to determine if the test organism body weights are reported. If
body weights are reported, the User selects "Yes" by checking the appropriate box. If body
weights are not reported, the User selects "No" by checking the appropriate box. In cases where a
Guidance for Developing Eco-SSLs Attachment 4-3 4-45 June 2005
-------
body weight is reported in the citation (e.g., initial weight) but is not specific for the age or time
the endpoint measurement was made, the body weight is still considered to be "reported". In
cases where body weight data are used from a different experiment within the same citation, the
user identifies the body weight as "not reported".
Body Weight with Units
If body weight data are reported in the study, the User selects an appropriate value to be used by
the application to calculate either a NOAEL or LOAEL dose. The User should select the body
weight reported for the appropriate NOAEL or LOAEL exposure level group. The highest body
weight should be used if both NOAEL and LOAEL exposure level groups are identified. If
results are reported for both male and female organisms, the User should record the highest body
weight for the appropriate NOAEL or LOAEL exposure (body weights are not averaged). If
authors report total weight gain, that amount is used to adjust the appropriate initial weight to the
approximate body weight at the time the endpoint was evaluated. This calculation should be
documented in the body weight comments section. The body weight is entered in the numeric
field provided. The User then selects the appropriate units for the reported body weight from the
pull down list. The list of available units and conversion factors is provided in Table 19.
In cases where the study reports an initial weight, but does not report body weight or body weight
gain for interim or terminal time points in the study, the User enters the initial body weight. Use
of this data represents a conservative approach to calculation of the intake rate of the medium and
subsequent calculation of the NOAEL and/or LOAEL value.
Table 19. Body Weight Units and Conversions
Body Weight Fields
ngbw
ugbw
mgbw
gbw
kgbw
Ibbw
nanograms body weight
micrograms body weight
milligrams body weight
grams body weight
kilograms body weight
pounds body weight
Conversion to BW in kg
multiply by 0.000000000001
multiply by 0.000000001
multiply by 0.000001
multiply by 0.001
none
multiply by 0.4535924
If body weight data are not reported in the study, the User must select an appropriate default body
weight. Table 20 provides a summary of default body weight values that are organism-, sex- and
age-specific. The User selects the appropriate default body weight and enters the result in the
numeric field provided. The largest body weight corresponding to the lifestage(s) used in the
experiment should be used. Default body weight units are reported in kilograms (kg). If a default
body weight value is not available in Table 20, the User may enter an appropriate value identified
from another source. If an alternate value is entered, the User should enter the value in units of
kg and provide a description of the value and reference in the body weight comment field.
Guidance for Developing Eco-SSLs Attachment 4-3 4-46
June 2005
-------
Body Weight Comments
The User enters information specific to any of the following in the comment field provided for the
body weights:
1) A description of the body weight selected or calculated from the study for entry. The
description should include the rationale for selection, sex of the organism, any
calculations and appropriate references to study table (including dose or concentration,
exposure duration if relevant, figure and page numbers).
2) A description of any value selected from the default table and rationale for selection.
3) A description of any alternative value selected from additional sources and the appropriate
reference.
Table 20. Default Body Weights
General
Organism Type
cat
cat
cat
cat
cat
cat
cattle
chicken
chicken
chicken
chicken
chicken
chicken
chicken
chicken
dog
dog
dog
dog
dog
dog
dove
dove
Specific
Organism Type
unspecified
unspecified
unspecified
unspecified
unspecified
unspecified
unspecified
unspecified
unspecified
domestic
domestic
domestic
domestic
domestic
domestic
unspecified
unspecified
unspecified
unspecified
unspecified
unspecified
ringed turtle
ringed turtle
Sex
M
M
M
F
F
F
BH
M
F
BH
BH
BH
BH
BH
BH
M
M
M
F
F
F
B
B
Age
weaning to 90 days
90 days to 1 year
1 year or older
weaning to 90 days
90 days to 1 year
1 year or older
Adult
Older than 30 days
Older than 30 days
1 day
3 days
7 days
14 days
20 days
28 days
weaning to 90 days
90 days to 1 year
1 year or older
weaning to 90 days
90 days to 1 year
1 year or older
0 days
7 days
Lifestage
juvenile
sexually mature
adult
juvenile
sexually mature
adult
adult
adult
adult
juvenile
juvenile
juvenile
juvenile
juvenile
juvenile
juvenile
sexually mature
adult
juvenile
sexually mature
adult
juvenile
juvenile
Default
BW (kg)
1.72
3.66
4
1.49
2.96
3.1
272
1.3
1.6
0.0397
0.0543
0.084
0.328
0.564
1.042
2.4
10.8
14
1.97
10.1
14
0.010
0.057
Reference
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
Osweiler et al., 1976
USEPA, 1987
USEPA, 1987
Diazetal 1994
Franson and Custer,
1982
Southern and Baker,
1981
Southern and Baker,
1981
Donaldson and
McGowan, 1989
Diazetal., 1994
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
Carriereetal., 1986
Carriere et al., 1986;
Kendall and Scanlon,
Guidance for Developing Eco-SSLs Attachment 4-3 4-47
June 2005
-------
Table 20. Default Body Weights
General
Organism Type
dove
dove
dove
dove
duck
duck
duck
duck
duck
duck
duck
duck
duck
duck
duck
duck
duck
duck
duck
duck
duck
duck
duck
duck
eagle
eagle
gerbil
gerbil
gerbil
gerbil
gerbil
gerbil
guinea pig
guinea pig
guinea pig
guinea pig
guinea pig
guinea pig
Specific
Organism Type
ringed turtle
ringed turtle
ringed turtle
ringed turtle
mallard
mallard
mallard
mallard
white pekin
white pekin
white pekin
white pekin
white pekin
white pekin
white pekin
white pekin
white pekin
white pekin
white pekin
white pekin
white pekin
white pekin
white pekin
white pekin
bald eagle
bald eagle
unspecified
unspecified
unspecified
unspecified
unspecified
unspecified
unspecified
unspecified
unspecified
unspecified
unspecified
unspecified
Sex
B
B
B
B
F
M
JV
JV
B
B
M
F
M
F
M
F
M
F
M
F
M
F
M
F
M
F
M
M
M
F
F
F
M
M
M
F
F
F
Age
14 days
21 days
28 days
Adult
Adult
Adult
10 days
30 days
0 days
7 days
14 days
14 days
21 days
21 days
28 days
28 days
35 days
35 days
42 days
42 days
49 days
49 days
56 days
56 days
Adult
Adult
weaning to 90 days
90 days to 1 year
1 year or older
weaning to 90 days
90 days to 1 year
1 year or older
weaning to 90 days
90 days to 1 year
1 year or older
weaning to 90 days
90 days to 1 year
1 year or older
Lifestage
juvenile
juvenile
juvenile
adult
adult
adult
juvenile
juvenile
juvenile
juvenile
juvenile
juvenile
juvenile
juvenile
juvenile
juvenile
juvenile
juvenile
juvenile
juvenile
juvenile
juvenile
juvenile
juvenile
adult
adult
juvenile
sexually mature
adult
juvenile
sexually mature
adult
juvenile
sexually mature
adult
juvenile
sexually mature
adult
Default
BW (kg)
0.092
0.117
0.134
0.144
1.1
1.2
0.092
0.46
0.06
0.27
0.78
0.74
1.38
1.28
1.96
1.82
2.49
2.30
2.96
2.73
3.34
3.06
3.61
3.29
4.13
5.35
0.048
0.084
0.1
0.04
0.073
0.09
0.48
0.89
1
0.39
0.86
0.9
Reference
1981
Carriere et al, 1986;
Kendall and Scanlon,
1981
Carriere et al., 1986;
Kendall and Scanlon,
1981
Carriere etal., 1986
Carriere etal, 1986;
Kendall and Scanlon,
1981; Keith and
Mitchell, 1993
USEPA, 1993
USEPA, 1993
USEPA, 1993
USEPA, 1993
NRC, 1994
NRC, 1994
NRC, 1994
NRC, 1994
NRC, 1994
NRC, 1994
NRC, 1994
NRC, 1994
NRC, 1994
NRC, 1994
NRC, 1994
NRC, 1994
NRC, 1994
NRC, 1994
NRC, 1994
NRC, 1994
Dunning, 1993
Dunning, 1993
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
Guidance for Developing Eco-SSLs Attachment 4-3 4-48
June 2005
-------
Table 20. Default Body Weights
General
Organism Type
hamster
hamster
hamster
hamster
hamster
hamster
hamster
hamster
hamster
hamster
hamster
hamster
hamster
hamster
hamster
hamster
hamster
hamster
horse
horse
horse
horse
mink
mink
mink
mink
mouse
mouse
mouse
mouse
mouse
mouse
mouse
mouse
mouse
mouse
mouse
mouse
mouse
Specific
Organism Type
golden Syrian
golden Syrian
golden Syrian
golden Syrian
golden Syrian
golden Syrian
Chinese &
Djungarain
Chinese &
Djungarain
Chinese &
Djungarain
Chinese &
Djungarain
Chinese &
Djungarain
Chinese &
Djungarain
unspecified
unspecified
unspecified
unspecified
unspecified
unspecified
unspecified
unspecified
unspecified
unspecified
unspecified
unspecified
unspecified
unspecified
BAF1
BAF1
BAF1
BAF1
BAF1
BAF1
B6C3F1
B6C3F1
B6C3F1
B6C3F1
B6C3F1
B6C3F1
deer mouse
Sex
M
M
M
F
F
F
M
M
M
F
F
F
M
M
M
F
F
F
B
M
M
M
M
F
M
F
M
M
M
F
F
F
M
M
M
F
F
F
M
Age
weaning to 90 days
90 days to 1 year
1 year or older
weaning to 90 days
90 days to 1 year
1 year or older
weaning to 90 days
90 days to 1 year
1 year or older
weaning to 90 days
90 days to 1 year
1 year or older
weaning to 90 days
90 days to 1 year
1 year or older
weaning to 90 days
90 days to 1 year
1 year or older
> 2 year
2 to 5 months
5 to 12 months
12 to 24 months
weaning to 49 days
weaning to 49 days
> 1 year
> 1 year
weaning to 90 days
90 days to 1 year
1 year or older
weaning to 90 days
90 days to 1 year
1 year or older
weaning to 90 days
90 days to 1 year
1 year or older
weaning to 90 days
90 days to 1 year
1 year or older
Adult
Lifestage
juvenile
sexually mature
adult
juvenile
sexually mature
adult
juvenile
sexually mature
adult
juvenile
sexually mature
adult
juvenile
sexually mature
adult
juvenile
sexually mature
adult
adult
foal
weanling
yearling
juvenile
juvenile
adult
adult
juvenile
sexually mature
adult
juvenile
sexually mature
adult
juvenile
sexually mature
adult
juvenile
sexually mature
adult
adult
Default
BW (kg)
0.097
0.134
0.15
0.095
0.145
0.16
0.03
0.041
0.04
0.025
0.038
0.035
0.0635
0.0875
0.095
0.2425
0.5025
1.03
498.95
181.44
317.51
408.23
1.02
0.583
1.18
0.596
0.0223
0.0261
0.035
0.0204
0.0222
0.03
0.0316
0.0373
0.04
0.0246
0.0353
0.035
0.02
Reference
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
Jurgens(1993)
Jurgens(1993)
Jurgens(1993)
Jurgens(1993)
USEPA 1993
USEPA 1993
USEPA 1993
USEPA 1993
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1993
Guidance for Developing Eco-SSLs Attachment 4-3 4-49
June 2005
-------
Table 20. Default Body Weights
General
Organism Type
mouse
mouse
mouse
mouse
mouse
mouse
mouse
pheasant
pheasant
owl
owl
pigeon
pigeon
Pig
Pig
Pig
Pig
quail
quail
quail
quail
quail
quail
rabbit
rabbit
rabbit
rabbit
rabbit
rabbit
rat
rat
rat
rat
rat
rat
rat
rat
rat
rat
rat
rat
rat
rat
Specific
Organism Type
deer mouse
unspecified
unspecified
unspecified
unspecified
unspecified
unspecified
ring-necked
ring-necked
barn owl
barn owl
pigeon
pigeon
domestic
domestic
miniature
miniature
Japanese
Japanese
bobwhite
bobwhite
bobwhite
bobwhite
unspecified
unspecified
unspecified
unspecified
unspecified
unspecified
Fischer 344
Fischer 344
Fischer 344
Fischer 344
Fischer 344
Fischer 344
Long-Evans
Long-Evans
Long-Evans
Long-Evans
Long-Evans
Long-Evans
Osborne-Mendel
Osborne-Mendel
Sex
F
M
M
M
F
F
F
F
M
M
F
F
M
M
F
M
F
F
M
F
M
JV
JV
M
M
M
F
F
F
M
M
M
F
F
F
M
M
M
F
F
F
M
M
Age
Adult
weaning to 90 days
90 days to 1 year
1 year or older
weaning to 90 days
90 days to 1 year
1 year or older
Adult
Adult
Adult
Adult
Adult
Adult
Adult
Adult
Adult
Adult
Adult
Adult
Adult
Adult
10 days
30 days
weaning to 90 days
90 days to 1 year
1 year or older
weaning to 90 days
90 days to 1 year
1 year or older
weaning to 90 days
90 days to 1 year
1 year or older
weaning to 90 days
90 days to 1 year
1 year or older
weaning to 90 days
90 days to 1 year
1 year or older
weaning to 90 days
90 days to 1 year
1 year or older
weaning to 90 days
90 days to 1 year
Lifestage
adult
juvenile
sexually mature
adult
juvenile
sexually mature
adult
adult
adult
adult
adult
adult
adult
adult
adult
adult
adult
adult
adult
adult
adult
juvenile
juvenile
juvenile
sexually mature
adult
juvenile
sexually mature
adult
juvenile
sexually mature
adult
juvenile
sexually mature
adult
juvenile
sexually mature
adult
juvenile
sexually mature
adult
juvenile
sexually mature
Default
BW (kg)
0.019
0.02695
0.0317
0.0375
0.0225
0.02875
0.0325
0.95
1.3
0.479
0.568
0.340
0.369
225
225
72.5
72.5
0.1
0.09
0.17
0.16
0.012
0.04
2.86
3.76
4
3.1
3.93
4.1
0.18
0.38
0.4
0.124
0.229
0.25
0.248
0.472
0.5
0.179
0.344
0.35
0.263
0.514
Reference
USEPA, 1993
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
Dunning, 1993
Dunning, 1993
Dunning, 1993
Dunning, 1993
Dunning, 1993
Dunning, 1993
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
Dunning, 1993
Dunning, 1993
USEPA, 1993
USEPA, 1993
USEPA, 1993
USEPA, 1993
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
Guidance for Developing Eco-SSLs Attachment 4-3 4-50
June 2005
-------
Table 20. Default Body Weights
General
Organism Type
rat
rat
rat
rat
rat
rat
rat
rat
rat
rat
rat
rat
rat
rat
rat
rat
rat
rat
rat
rat
rat
rat
sheep
sheep
sheep
sheep
sheep
sheep
sheep
sheep
shrew
shrew
sparrow
Specific
Organism Type
Osborne-Mendel
Osborne-Mendel
Osborne-Mendel
Osborne-Mendel
Sprague-Dawley
Sprague-Dawley
Sprague-Dawley
Sprague-Dawley
Sprague-Dawley
Sprague-Dawley
Wistar
Wistar
Wistar
Wistar
Wistar
Wistar
unspecified
unspecified
unspecified
unspecified
unspecified
unspecified
Old Norse
Old Norse
Dala
Dala
Chun forest
Chun forest
Chun forest
domestic
short-tailed
short-tailed
white-throated
Sex
M
F
F
F
M
M
M
F
F
F
M
M
M
F
F
F
M
M
M
F
F
F
M
F
M
F
BH
M
F
B
M
F
B
Age
1 year or older
weaning to 90 days
90 days to 1 year
1 year or older
weaning to 90 days
90 days to 1 year
1 year or older
weaning to 90 days
90 days to 1 year
1 year or older
weaning to 90 days
90 days to 1 year
1 year or older
weaning to 90 days
90 days to 1 year
1 year or older
weaning to 90 days
90 days to 1 year
1 year or older
weaning to 90 days
90 days to 1 year
1 year or older
Adult
Adult
Adult
Adult
Juvenile
Adult
Adult
1 week
Adult
Adult
Adult
Lifestage
adult
juvenile
sexually mature
adult
juvenile
sexually mature
adult
juvenile
sexually mature
adult
juvenile
sexually mature
adult
juvenile
sexually mature
adult
juvenile
sexually mature
adult
juvenile
sexually mature
adult
adult
adult
adult
adult
Juvenile
Adult
Adult
juvenile
adult
adult
adult
Default
BW (kg)
0.55
0.201
0.389
0.4
0.267
0.523
0.6
0.204
0.338
0.35
0.217
0.462
0.5
0.156
0.297
0.32
0.235
0.4702
0.51
0.2024
0.3846
0.4
43
32
115
80
24
91.5
73
46.81
0.017
0.017
0.0259
Reference
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
www. ansi. okstate . edu/br
eeds/sheep/
www. ansi. okstate . edu/br
eeds/sheep/
Geertman,2001
Geertman, 2001
www. ansi. okstate . edu/br
eeds/sheep/ based on
data for related strain
(Kerry Hill)
www. ansi. okstate . edu/br
eeds/sheep/ based on
data for related strain
(mean of Kerry Hill and
Shropshire)
www. ansi. okstate . edu/br
eeds/sheep/ based on
data for related strain
(mean of Kerry Hill and
Shropshire)
Steinheim et al., 2002
USEPA, 1993
USEPA, 1993
Dunning, 1993
Guidance for Developing Eco-SSLs Attachment 4-3 4-51
June 2005
-------
Table 20. Default Body Weights
General
Organism Type
starling
starling
turkey
turkey
turkey
turkey
turkey
turkey
turkey
turkey
turkey
turkey
turkey
turkey
turkey
turkey
turkey
turkey
turkey
turkey
vole
vole
vole
finch
Specific
Organism Type
starling
starling
domestic
domestic
domestic
domestic
domestic
domestic
domestic
domestic
domestic
domestic
domestic
domestic
domestic
domestic
domestic
domestic
domestic
domestic
prairie vole
meadow vole
meadow vole
zebra finch
Sex
M
F
M
F
M
F
M
F
M
F
M
F
M
F
M
F
M
F
M
F
BH
M
F
B
Age
Adult
Adult
20 w
20 w
25 w
25 w
30 w
30 w
35 w
35 w
40 w
40 w
45 w
45 w
50 w
50 w
55 w
55 w
60 w
60 w
Adult
Adult
Adult
Adult
Lifestage
adult
adult
juvenile
juvenile
juvenile
juvenile
juvenile
juvenile
juvenile
juvenile
juvenile
juvenile
juvenile
juvenile
juvenile
juvenile
adult
adult
adult
adult
adult
adult
adult
adult
Default
BW (kg)
0.0847
0.0799
14.3
8.4
16.4
9.8
19.1
11.1
20.7
11.1
21.8
10.8
22.5
10.5
23.2
10.5
23.9
10.5
24.5
10.6
0.042
0.043
0.039
12
Reference
Dunning, 1993
Dunning, 1993
NRC, 1994
NRC, 1994
NRC, 1994
NRC, 1994
NRC, 1994
NRC, 1994
NRC, 1994
NRC, 1994
NRC, 1994
NRC, 1994
NRC, 1994
NRC, 1994
NRC, 1994
NRC, 1994
NRC, 1994
NRC, 1994
NRC, 1994
NRC, 1994
USEPA, 1993
USEPA, 1993
USEPA, 1993
Dunning, 1993
Is the Intake Rate Reported?
The Intake Rate fields refer to the intake rate of the exposure medium (diet or drinking water), not
the contaminant. The intake rate is used to convert contaminant concentration data to a
contaminant dose. If intake rates are reported, the User selects "Yes" by checking the appropriate
box. If intake rates are not reported, the User selects "No" by checking the appropriate box. In
gavage or other oral exposures (capsule), the User selects "No" by checking the appropriate box,
but overrides the dose quantification score to reflect that intake rate was "reported."
Intake Rate with Units
If the intake rate is reported in the study, the User selects the appropriate value to be used by the
application to calculate either a NOAEL or LOAEL dose. If the study reports doses, the intake
rate does not need to be entered. If the intake rate for a treatment group is variable over the course
of the experiment, enter the average rate and note this in the comments field. The User should
select the intake rate reported for the appropriate NOAEL or LOAEL exposure level group. The
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highest intake rate should be used if both NOAEL and LOAEL exposure level groups are
identified. The intake rate is entered in the numeric field provided. The application assumes that
the intake rate entered (for dietary studies) is dry weight-based. If the User gathers information
from the study that reports otherwise, then the User should convert the intake rate to a dry weight
basis and report in detail the necessary conversion in the Intake Rate Comment Field. Next the
User selects the appropriate units associated with the intake rate from the pull down list. The list of
intake rate units is provided in Table 21.
If the intake rate is not reported. The User does not enter an intake rate, the application calculates
the intake rate automatically using allometric equations based on the body weight, specific class
and exposure route for the test organism. The intake rate is calculated and reported in the Score
Information Screen in units of kg bw per day or L per day (see Appendix A).
Table 21. Units and Conversions for Intake Rate of Medium
Intake Rate Fields
kg/d (or L/d)
kg/kg BW/day
or
L/kg BW/day
kg/org/d or
L/org/d
g/d or ml/day
g/kg BW/d or
ml/kg BW/d
g/org/d or
ml/org/d
mg/d or ul/d
mg/kg BW/d or
ul/kg BW/d
mg/org/d or
ul/org/d
ug/d
ug/kg bw/d
ug/org/d
ng/d
ng/kg bw/d
ng/org/d
kilograms or liters per day
kilograms or liters per kilogram BW per
day
kilograms or liters per organism per day
grams per day
grams per kilogram BW per day
grams per organism per day
milligrams per day
milligrams per kilogram BW per day
milligrams per organism per day
micrograms per day
micrograms per kilogram BW per day
micrograms per organism per day
nanograms per day
nanograms per kilogram BW per day
nanograms per organism per day
Conversion to kg/day or L/day
multiply by 1
multiply by BW in kg
multiply by 1
multiply by 0.001
multiply by 0.001 then multiply by BW in kg
multiply by 0.001
multiply by 0.000001
multiply by 0.000001 then multiply by BW in kg
multiply by 0.000001
multiply by 0.000000001
multiply by 0.000000001 then multiply by BW in
kg
multiply by 0.000000001
multiply by 0.000000000001
multiply by 0.000000000001 thenmultiply by BW
in kg
multiply by 0.000000000001
Intake Rate Comments
In the comment field provided for the body weights, the User enters information specific to any of
the following:
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A description of the intake rate selected or calculated from the study for entry. The
description should include the rationale for selection, any calculations and
appropriate references to study table, figure and page numbers.
A description of any value selected from the default table and rationale.
A description of any alternative value selected from additional sources and the
appropriate reference.
Endpoint Sample Size
The user enters information in this field on the number of experimental animals for the specific
endpoint entered for each exposure concentration or dose. Any specific reasons for loss of
experimental animals not associated with exposure to the contaminant should be noted.
Results for the NOAEL
These fields allow the User to enter information concerning the experimental results for the
NOAEL exposure (dose) level. The User enters information here in instances where ONLY A
NOAEL is reported and no LOAEL is reported. In these instances, it is important to evaluate the
power of a study design to detect an adverse effect, if it were present. A detailed description of the
power calculation is provided as Appendix B. Statistical power is calculated using the number of
test organisms, the mean endpoint response for control and treated organisms, and the standard
deviations of the means as inputs. If the distribution of values in the control group and the
exposed group are both approximately normal and the variance is similar for both, the power of the
study can be estimated from the information entered below. The numeric fields provided for
power calculation data entry cannot be blank. If any of the required input data are missing (i.e.,
because they are not reported for the endpoint), the study power is not calculated and the
application reports "not calculated".
The User should note that data from figures are not used to calculate power.
Number of Exposed Organisms. The User enters the total number of organisms exposed
in the numeric field provided. If the total number of exposed organisms is not reported, the
User leaves the numeric field blank. A blank field is evaluated as null and power is not
calculated.
Number of Control Organisms. The User enters the total number of control organisms
exposed at the NOAEL dose in the numeric field provided. If the total number of control
organisms is not reported, the field is left blank. The blank field is evaluated as null and
power is not calculated by the application.
Mean of Endpoint in Exposed Organisms. The User enters the mean of the NOAEL
result for the exposed organisms in the numeric field provided. If a mean value is not
Guidance for Developing Eco-SSLs Attachment 4-3 4-54 June 2005
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reported, -99 is entered in the field. This value is evaluated as null and power is not
calculated.
Mean of Endpoint in Control Organisms. The User enters the mean for the control
group in the numeric field provided. If a mean value is not reported, -99 is entered in the
field. This value is evaluated by the system as null and power is not calculated.
Standard Deviation of Endpoint in Exposed Organisms. The standard deviation is
required for calculation of statistical power. The database permits entry of either the
standard deviation or the standard error of the mean for the endpoint of interest. The User
selects the appropriate data type and enters the value in the numeric field provided. If a
standard error is entered, the database automatically estimates the standard deviation as the
product of the standard error multiplied by the square root of the sample size n. If neither a
standard deviation nor standard error value is provided, -99 is entered in the field. The
field is evaluated as null and power is not calculated.
Standard Deviation of Endpoint in Control Organisms
The standard deviation is required for calculation of statistical power. The database
permits entry of either the standard deviation or the standard error of the control mean for
the endpoint of interest. The User selects the appropriate data type and enters the value in
the numeric field provided. If a standard error is entered, the database automatically
estimates the standard deviation as the product of the standard error multiplied by the
square root of the sample size n. If neither a standard deviation nor standard error value is
provided, -99 is entered in the field. The field is evaluated as null and power is not
calculated.
Confidence Alpha. This is the statistical significance level chosen to declare a treatment
response as significantly different different from the control response. The default value
of alpha is 0.05. However, the system allows the User to select other statistical
significance levels from the pull-down list provided. The User should seek approval from
an Administrator before using alpha values other than the default of 0.05.
At this point in the data entry process, the "Endpoint Information" screen is now complete. The
User verifies that all entered data are correct and clicks on the "Next" button at the bottom of the
screen to continue. The User should not use the browser back arrow to return to a previous data
entry screen to correct errors, as deletion of data results.
4.5 Data Evaluation Score
For the convenience of the User, the Data Evaluation screen provides a summary of the
information required to determine a data evaluation score for each endpoint entered. This
summary is provided at the top of the Score Information screen. The Data Evaluation Scoring
system is described in SOP #7 (Attachment 4-5).
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For this summary screen, the data previously entered for body weight, intake rate, and the NOAEL
and/or LOAEL are converted to the appropriate units for calculation of a final NOAEL and/or
LOAEL value expressed as mg/kg-day. Each of these conversions are described in detail below:
Body Weight. The application automatically converts body weights and units entered by
the User to units of kilograms. The equations used for the conversion of body weight data
are presented in Table 19.
Intake Rate. The application converts the exposure medium intake rate entered by the
user to units of kilograms of food or liters of drinking water per organism per day. The
equations that are used for this conversion are presented in Table 21. If the intake rate is
assigned by the application, based on the default allometric equations for food and water
ingestion, no conversion is required as the equations estimate intake rates in the appropriate
units.
Conversion to Dose. The application converts the entered NOAEL and/or LOAEL
concentration or dose values to the appropriate units of mg of contaminant per kg BW per
day. The equations used for these conversions are provided in Table 9. If the NOAEL
and/or LOAEL concentrations are expressed on a wet weight basis in the study, the
application makes the appropriate conversion to dry weight based on the moisture content
entered by the User.
The final data evaluation score assigned to the NOAEL and/or LOAEL is based on the addition of
individual scores for ten study attributes. These ten attribute scores are described in the following
subsection and are summarized in Table 22. For each attribute, a score is assigned ranging from 0
(no merit for derivation of a TRV) to 10 (extremely valuable and relevant for derivation of a
TRV). It is important to note that a low score does not imply that the study is poor, only that it is
not optimal for deriving a TRV.
To determine the final data evaluation score, the User selects the appropriate score from the pull
down list provided for each of the study attributes. The application defaults to the appropriate
score based on the information entered. The User can, however, alter the default scores under
special circumstances. If any of the individual attribute scores are equal to 0 the total score is
equal to 0 and the study is not used for the derivation of Wildlife TRVs.
Table 22. Summary of Data Evaluation Scoring System
Attribute
1 . Data source
"Source Score"
2. Contaminant
Form
"Chemical Form
Score"
Description
Primary
Secondary
Contaminant form is known and is the same or similar to the of medium of concern
Contaminant form is irrelevant to absorption or biological activity
Contaminant form is known and is different from that found in the medium of concern
Contaminant form is not reported (this includes situations when the contaminant is just listed
as "Lead" or "Selenium")
Score
10
0
10
10
5
4
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Table 22. Summary of Data Evaluation Scoring System
Attribute
Description
Score
3. Test Substrate
"Test Substrate
Score"
Test substance concentrations reported as actual measured values (M), verified nominal (UX)
and/or doses administered by gavage
Test substance concentrations reported as nominal values (U)
Test substance concentrations not reported
10
5
0
4. Dose
Quantification
"Dose
Quantification
Score"
Administered doses reported as mg/kg-BW (includes gavage doses reported in these units)
Administered doses need to be calculated and intake rates and body weights provided
Administered doses need to be calculated and only one value (intake or body weight)
provided (if study is gavage or other capsule, intake is "provided")
Administered doses need to be calculated based on estimated intake rates and body weights
Administered doses cannot be calculated from the information provided
10
7
6
5
0
5. Dose Range
"Dose Range
Score"
Both a NOAEL and a LOAEL are identified; values are within a factor of 3
Both a NOAEL and a LOAEL are identified; values are within a factor of 10
Both a NOAEL and a LOAEL are identified; values are not within a factor of 10
Only a NOAEL or a LOAEL is identified
Study lacks a suitable control group
10
8
6
4
0
6. Dose Route
"Dose Route
Score"
Chemical incorporated into food (including mother's milk)
Other oral (gavage, capsule)
Chemical incorporated into drinking water
Not dietary, other oral, or drinking water or not reported or choice of treated and non
treated food or water
10
7. Endpoint
"Endpoint Score"
Reported endpoint is a reproductive or population effect (REP) (POP)
Reported endpoint is lethality (chronic or subchronic exposures (MOR)
Reported endpoint is reduction in growth (GRO)
Reported endpoint is sublethal change in organ function, behavior or neurological function
(BEH, PHY, PTH)
Reported endpoint is a biomarker of exposure with unknown relationship to fitness (BIO)
10
9
8. Exposure
Duration
"Exposure
Duration Score"
Exposure duration encompasses multiple lifestages of test species
Exposure duration is at least 0.1 times the expected life span of the test species or occurs
during a critical life phase
Exposure duration is shorter than 0.1 times the expected life span of the test species and
multiple doses or concentrations are administered
Exposure duration is shorter than 0.1 times the expected life span of the test species and
only a single dose or concentration is administered.
Exposure duration is acute or not reported
10
10
6
3
9. Statistical
Power
"Power Score"
At least 90% chance of seeing a difference that is biologically significant
NOAEL and LOAEL available or LOAEL only available
At least 75% chance of seeing a difference that is biologically significant
At least 50% chance of seeing a difference that is biologically significant
Less than a 50% chance of detecting a difference that is biologically significant
Only NOAEL available; insufficient data reported to determine statistical power of study
10
10
8
6
3
1
10. Test
Conditions
"Test Parameter
Score"
Follows a standard guideline and reports all test parameters
Does not follow a standard guideline, but does report all test parameters
Follows a standard guideline but does not report test parameters
Does not follow a standard guideline and reports some, but not all of the test parameters
Does not report any test parameters
10
10
7
4
2
1. Data Source Score
All studies considered for TRV derivation are from primary sources. Secondary sources of data
are not used to derive Wildlife TRVs. The application automatically assigns a Source score based
on the Primary Source entry. If the "No" box is selected, the application exits completely from the
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June 2005
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program. Since the User has progressed to this point of the data entry process, the application
assumes that the study is a primary source and a score of 10 is assigned.
2. Contaminant Form Score
The wildlife TRVs are expressed in units of ingested dose (mg/kg BW/day or mg/L/day).
Expression as units of ingested dose implicitly assumes that absorption of the contaminant from
the test medium is the same as for the site medium. This assumption may be reasonable when the
two media are the same (e.g., both water or both similar food items), but may not be true if the two
media are different (e.g., test medium = water, site medium = soil). To account for the potential
difference in absorption between different media, it is necessary to convert both the ingested dose
and the TRV to units of absorbed dose:
Site Dose (absorbed) = Site Dose (ingested) * Absorption fraction from site medium
TRV(absorbed dose) = TRV(ingested dose)* Absorption fraction in test medium
Some contaminants are better absorbed and more biologically active than others. The best known
examples are differences between inorganic and organic mercury, and inorganic and organic
arsenic. Studies reporting oral absorption fraction from the test medium are preferred to those
where the absorption fraction is unknown. In the absence of oral absorption data, the assumption
of equal absorption of the contaminant from the test and site medium is reasonable when the form
of the contaminant is the same in the test medium versus the site medium. Therefore, studies
which use the same chemical form of a contaminant in the exposure medium as that typically
found on a waste site are preferred.
The User assigns a Contaminant Form score based upon the similarity of the contaminant form
used in the study to contaminant forms found in environmental media. A summary of common
contaminant forms found in environmental media is provided in Table 3. If the contaminant form
used in the study is the same or similar to that in environmental media, a score of 10 is selected by
the User. If the contaminant form is not relevant to absorption or biological activity, a score of 10
is selected. If the contaminant form is different from that in environmental media, a score of 5 is
selected. If the contaminant form is not reported (NR), a score of 4 is selected by the User.
3. Test Substrate Score
Studies that report contaminant exposure concentrations or doses in the diet or drinking water
confirmed by analytical measurement - "measured"- are preferred compared to those that do not
measure or verify the exposure doses or concentrations.
The application automatically assigns a Test Substance score based on the value the User entered
under "Method of Contaminant Analysis". If the method of contaminant analysis is measured (M)
or unmeasured but analytically verified (UX), a score of 10 is assigned. If unmeasured (U) is
entered, a score of 5 is assigned. If calculated (C) is entered, a score of 1 is assigned. Gavage
studies are considered measured, and are scored as a 10.
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4. Dose Quantification Score
Some toxicological studies report contaminant exposures in terms of dose (mg of contaminant per
unit of body weight), but some only report the concentration of the contaminant in the exposure
medium (food or drinking water). In these cases, it is necessary to convert the concentrations to a
dose using an intake rate (food or water) and a body weight. Studies that report results as doses
are preferred over those that report concentrations and the application automatically assigns these
studies a Dose Quantification Score of 10. Studies that report exposure on a concentration basis
are scored in the following manner according to preference:
If both body weight and intake rates are reported at any point of the exposure for
the test organisms in the study (the User is prompted to enter this information
earlier in the data entry process), the study endpoint receives a score of 7. The
application automatically uses the body weight and intake rate values entered
previously to convert the exposure concentrations to doses.
If only one value (intake rate or body weight) is provided for the test organisms, a
score of 6 is assigned.
If the study does not report either body weights or intake rates for the test organism,
the application assigns a score of 5. Doses are automatically calculated based on
the default body weight and intake rate values previously entered by the User.
If the administered doses cannot be calculated from the information provided, a
score of 0 is assigned by the User from the pull down menu.
If the study uses an exposure method of where the administered amount is reported as a dose in
amount of chemical per unit of body weight, the User selects the dose quantification score from
the provided pull down list as follows:
If the amount administered is reported for any exposure route in units of mg/kg
body weight, a score of 10 is assigned.
If the amount administered is in units of mg/organism, it must be divided by body
weight to convert to dose units of mg/kg/day. If the body weight is reported in the
study, a score of 7 is assigned.
If the body weight is not reported and the value needs to be estimated based on a
default, a score of 5 is assigned.
5. Dose Range Score
The TRV represents a threshold on the dose-response curve between the absence and presence of
the adverse effect of concern. Establishing this threshold involves identification of two values
from the toxicological study: a no observed adverse effect level (NOAEL) and a lowest observed
Guidance for Developing Eco-SSLs Attachment 4-3 4-59 June 2005
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adverse effect level (LOAEL). The NOAEL is defined as the highest administered dose that does
not cause a significant adverse effect. The LOAEL is defined as the lowest administered dose that
causes a significant adverse effect. Experimentally, the threshold value is estimated by assuming
it lies between the NOAEL and the LOAEL. Therefore, a study which identifies both a NOAEL
and a LOAEL is more valuable than a study that identifies only a NOAEL or LOAEL. Studies
which use a larger number and/or more closely spaced doses may provide a more refined estimate
of the NOAEL and LOAEL and are preferred.
The application automatically assigns a Dose Range score based upon the NOAEL and/or LOAEL
values entered previously by the User. This assignment appears in the pull down menu on the
score sheet. The User, however, may assign a different score from among the choices provided.
If both a NOAEL and a LOAEL are identified and the values are within a factor of 3, a score of 10
is assigned. If both a NOAEL and a LOAEL are identified and the values are within a factor of 10,
a score of 8 is assigned. If both a NOAEL and a LOAEL are identified, but the values are not
within a factor of 10, a score of 6 is assigned. If only a NOAEL or a LOAEL is identified, a score
of 4 is assigned. If the study lacks a suitable control group, a score of 0 is assigned by the User.
Unsuitable control groups include: Historical (H), No Methodology (K), and Positive (P). If the
control type is not reported (NR), a score of 0 is assigned.
6. Dose Route Score
The Eco-SSLs reflect the concentrations of contaminants in soil protective of oral exposure via
ingestion of soil or food items. Therefore, toxicological studies that use oral exposure (food,
water, gavage, or capsule) are considered to be relevant compared to studies that use other non-
oral methods of administration (inhalation, dermal, and injection routes). Studies that report
results for non-oral exposures are not used to establish TRVs and should be labeled as "non oral"
using the literature rejection criteria discussed in Section 2.0.
Dietary studies are preferred to oral exposure via gavage or capsule because they represent the
closest approximation of the intake route under natural conditions. Gavage and capsule studies are
less desirable because they do not generally reflect natural feeding behaviors and the vehicle used
to deliver the gavage dose can alter the kinetics of absorption. Drinking water is the least desirable
among the acceptable routes of administration because the Wildlife TRVs are derived for use in
assessments conducted in terrestrial rather than aquatic environments.
The application automatically assigns a Dose Route score based upon the Exposure Type and
Route of Exposure information previously entered by the User. If the Route of Exposure is via
food (FD), a score of 10 is assigned. If the route of exposure is via other oral routes (OR) or
gavage (GV), a score of 8 is assigned. If the route of exposure is via drinking water (DW), a score
of 5 is assigned. If the route of exposure is a choice between contaminated and non contaminated
media (CH), a score of 0 is assigned. If the route of exposure is not reported (NR), a score of 0 is
assigned.
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7. Endpoint Score
In most ecological risk assessments (ERAs), assessment endpoints focus on the effects of long
term exposures of contaminants on population sustainability. The specific toxicological endpoints
used as measurements of population sustainability in ERAs are site-specific. For the purposes of
identification and derivation of a TRY for calculation of an Eco-SSL, the endpoints are predefined.
The following endpoints are selected in order of preference for derivation of TRVs.
Studies measuring reproductive endpoints are considered the most appropriate and
are preferred. Reproductive endpoints are assigned a score of 10. Within the
coding system, this includes any endpoint within the reproduction (REP) effect
group (Table 17).
Studies measuring mortality or survival (chronic) as an endpoint are also considered
appropriate but are less preferable to reproductive endpoints. These study
endpoints are assigned a score of 9. Within the coding system, this includes any
endpoint within the mortality (MOR) effect group (Table 17).
Studies measuring growth are also considered appropriate for establishing TRVs.
These study endpoints are assigned a score of 8. Within the coding system, this
includes any endpoint within the (GRO) effect group (Table 17).
Studies measuring organ function, behavior or neurological function are considered
less useful in establishing TRVs. This applies to endpoints within the pathology
(PTH), behavior (BEH) or physiology (PHY) effect groups in the TRV coding
system. These study endpoints are assigned a score of 4. The User may elect to
score such studies lower if it is decided that the effect does not have an adverse
effect on organism "fitness" or health (Table 17).
Studies measuring biochemical effects or changes that are either hormonal,
chemical or enzymatic in nature are considered the least useful in establishing
TRVs. These study endpoints are assigned a score of 1. This evaluation includes
any endpoint in the biochemical (BIO) effect group of the Wildlife TRV coding
system. The User may elect to score such study measures higher if it is decided that
the measure can be related to organism "fitness" or health. Biomarkers of exposure
should always be scored as a 1.
8. Exposure Duration Score
The usefulness of a study result for derivation of a TRV is partially dependent on the duration of
the exposure. Chronic and multiple generation exposures are preferred to subchronic or acute
exposures. Chronic exposures are generally more representative of the type of exposure which
may occur at a contaminated site.
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The User assigns an Exposure Duration score based upon the duration of the study exposure and
the life span of the test organism. A summary of typical laboratory test organism life spans is
provided in Table 23. Data for representative species of wildlife are also included. In some cases,
a range of values has been presented for the longevity of wildlife species. The User should
preferentially select the average life span, if given, and secondarily select the minimum value from
that range if the exposure occurs also at a critical lifestage.
To assess if the exposure duration is representative of the expected lifespan, the User multiplies
the test organism lifespan by 0.1 and compares the result to information in Table 23. For example,
if the test organism is a gerbil with an assumed lifespan of 2.5 years (2.5 years * 0.1 = 0.25 years
or 12 weeks), an exposure duration of 9 weeks is less than 0.1 times the expected lifespan. If the
duration of the study exposure encompasses multiple generations of the test organism, a score of
10 is selected. If the duration of exposure is at least 0.1 times the expected lifespan of the test
organism or occurs during a critical lifestage, a score of 10 is selected. If the duration of exposure
is less that 0.1 times the expected lifespan of the test organism and multiple dose or concentration
groups are included in the study, a score of 6 is selected. If the duration of exposure is less that 0.1
times the expected lifespan and only a single dose or concentration group is used in addition to
controls, a score of 3 is assigned. If the exposure duration is acute (a single oral dose), a score of 0
is selected.
Table 23. Default Weaning, Puberty, and Lifespan Values for TRV Species
Group
Laboratory
Rodents
Other
Laboratory
Mammals
Other Tested
Animals
Species
Mice
Rats
Guinea Pigs
Hamsters
Gerbils
Cats
Dogs, Beagles
Rabbits, New Zealand
Vole, meadow
Shrew, short-tailed
Pig
Mink
Pheasant
Weaning (days)
21
21
14
21
21
49
42
56
21
25-30
NR
56
7-12a
Puberty (days)
50
56
70
60
70
240
240
190
>21 (male)
>42 (female)
>65 (male, lab)
>83 (male)
<365 (female)
150
300
330
Lifespan
(years)
2*
2*
6
2.5
3
15
15
6
0.1-0.25
(mean)
Use 0.25
0.37-0.38
(lab)
Use 1.04
<1.7
27
7 (mean)
10- 11 (max)
Use 9
<3
Reference
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
USEPA, 1987
Golley, 1962
Johnson and Johnson,
1982
Beer and Mcleod, 1961
Blus, 1971
Pearson, 1944
French, 1984
Dapson, 1968
Enders(1952)
Ewer (1973)
Giudice and Ratti, 2001
Guidance for Developing Eco-SSLs Attachment 4-3 4-62
June 2005
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Table 23. Default Weaning, Puberty, and Lifespan Values for TRV Species
Group
Other Tested
Animals
Species
Mallard
Chicken
Dove, mourning
Quail, bobwhite
Quail, Japanese
Weaning (days)
52-60
56
NA
15
14
NR
Puberty (days)
330
NA
80( males)
90 (females)
330
NR
Lifespan
(years)
23
24
31 (max)
1.5 (mean)
Use 1.5
6.5 (max)
3
Reference
Clappetal, 1982
Bellrose, 1976
Loekmoen et al., 1990
Krapu and Doty, 1979
USEPA, 1987
Clappetal. 1983
White etal, 1987
Mirarchi and Basket,
1994
Rosene, 1969
Porter and Terril, 1986
* Substantial strain variability
NA = Not Applicable
NR = Not Reported
a Pheasant chicks can first take flight at 7 to 1 2 days; broodomg by hen is important for the first two weeks after hatching. Broods remain intact
with hen for an indefinite period.
9. Power Score
A NOAEL is defined as the highest dose that does not cause a significant effect in the selected
endpoint when compared to the control. However, the ability to detect an effect (i.e., the reliability
of the NOAEL) depends on a number of factors. The most important are:
1) the variability of the measurement endpoint in both the control and the dosed groups
2) the number of animals in each group
That is, as variability in the measurement endpoint goes up and the number of experimental
animals goes down, the ability to detect an effect becomes very poor, and a dose which actually
causes an effect may be incorrectly identified as a NOAEL.
Statistical power is a measure of the ability to detect an effect. There are a number of standard
statistical procedures available for calculating the power of a study to detect an effect which can be
used to evaluate the reliability of NOAEL values. The statistical power test used for the
toxicological Data Evaluation process for establishing Wildlife TRVs is described in Appendix B.
If both a NOAEL and a LOAEL are reported or if only a LOAEL is reported, the power
calculation is not used and a score of 10 is assigned by the application. In cases where only a
NOAEL is reported, scores are assigned as follows. If the calculated power is greater than or
equal to 90 percent, a score of 10 is assigned. If the calculated power is greater than or equal to 75
percent, a score of 8 is assigned. If the calculated power is greater than or equal to 50 percent, a
score of 6 is assigned. If the calculated power is less than 50 percent, a score of 3 is assigned. If
Guidance for Developing Eco-SSLs Attachment 4-3 4-63
June 2005
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the power cannot be calculated because one or more of the required fields is null, a score of 1 is
assigned. In cases where the endpoint reported is mortality and the result is 0, the User should
assign a score of 10 as power cannot be calculated.
10. Test Condition Score
The User is prompted earlier in the data entry process to identify if the study follows a standard
guideline for toxicity testing and if not how many of the parameters the study reports. The
standard guidelines and test parameters are provided in Table 13. If the study follows a standard
guideline and reports all measurement parameters, then a score of 10 is assigned. If the study does
not follow standard guidelines but reports all parameters, a score of 10 is also assigned. If the
study follows a standard guideline but does not report all test parameters, then a score of 7 is
assigned. If the study does not follow a standard guideline, but reports some but not all of the test
parameters, then a score of 4 is assigned. If the study does not report any parameters, a score of 2
is assigned.
Final Total Score
The "Score Information" screen is now complete. The User verifies that all data entered are
correct and clicks on the "Calculate Score" button at the bottom of the screen to calculate the final
total score. The User should not use the browser back arrow to return to a previous data entry
screen to correct errors, this action results in a duplication of information.
The total score is based upon the evaluation of each of the ten attribute scores identified above.
The total score is calculated for a specific endpoint by taking the sum of all ten study attribute
scores (a "perfect" study is given a score of 100). However, if any one study attribute is given a
score of 0, the final score is also be set to equal 0. This ensures minimum standards for study
results that are used to derive wildlife TRVs. Studies without appropriate controls, of acute
exposure duration, without reported test substance concentrations, and non-oral exposures are
excluded from the TRY derivation process.
Several scoring examples are provided below:
Lowest Possible Total Score (all attribute scores are the minimum score without defaulting
toO:
Study Attribute
Source Score:
Dose Route Score:
Test Substrate Score:
Contaminant Form Score:
Dose Quantification Score:
Score
10
5
5
4
5
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Endpoint Score:
Dose Range Score:
Power Score:
Exposure Duration Score:
Test Parameter Score:
Total Score
1
4
1
3
2
40
Case Where Individual Attribute Score = 0
Study Attribute
Source Score:
Dose Route Score:
Test Substrate Score:
Contaminant Form Score:
Dose Quantification Score:
Endpoint Score:
Dose Range Score:
Power Score:
Exposure Duration Score:
Test Parameter Score:
Total Score
Score
10
5
1
4
0
1
4
1
3
2
0
Final Score set to zero, due to Dose Quantification Score
Highest Possible Total Score available (all attribute scores are the maximum score):
Study Attribute
Source Score:
Dose Route Score:
Test Substrate Score:
Contaminant Form Score:
Dose Quantification Score:
Endpoint Score:
Dose Range Score:
Power Score:
Exposure Duration Score:
Score
10
10
10
10
10
10
10
10
10
Guidance for Developing Eco-SSLs Attachment 4-3 4-65
June 2005
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Test Parameter Score:
Total Score
10
100
At this point of the data entry process, the User completes data entry and scoring for the selected
endpoint and clicks on "Finish this Endpoint" to proceed. The User should not use the browser
back arrow to return to a previous data entry screen to correct errors as this would result in a
duplication of information.
If there is another endpoint associated with the selected phase (the selected phase is provided in
the navigation bar at the top of the screen), the User selects "Yes" when prompted for another
endpoint and begins entry of that endpoint at the Endpoint Information screen. If there are no
other endpoints associated with the selected phase, then the User selects "No".
When the data entry process is completed, the endpoint scores are used to derive TRVs as
described in Attachments 4-5 and 4-6. Papers with a total score of 66 or higher are included in the
data set used for derivation of Wildlife TRVs. Papers with a score of 65 or less are not used for
the derivation of TRVs.
Guidance for Developing Eco-SSLs Attachment 4-3 4-66
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5.0 QUALITY ASSURANCE AND QA REPORTS
Entered data are reviewed and approved for accuracy and completeness prior to use in derivation
of Wildlife TRVs. The database provides a QA report option for manual review. Coded data are
compiled on a printable screen form. The coded data are cross-checked against the source
publication for accuracy and against the TRY coding guidelines for consistency by an
administrative user. Any errors noted are corrected in the database prior to QA approval. The
database program automatically records the date that changes were made to the TRY database as a
result of QA review, but does not track specific edits.
To perform an QA review, an Administrative User accesses the QA report for a given article. If
errors are found on the report, the User uses the editing functions available in the TRY database to
make corrections (see Section 4 for more information regarding data entry and edits). When the
correct data has been entered, the Administrative User chooses View/Edit Articles to approve the
article by choosing the "Yes" radio button located beside the text "Article Approved?". A
comment must be entered at this time, to signal to the system that the article has been approved.
This feature protects against errant clicks that may accidently change the status of the Article
Approved? radio button.
Guidance for Developing Eco-SSLs Attachment 4-3 5-1 June 2005
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6.0 DOWNLOADS
Data contained in the TRV database can be downloaded from the TRV database to a User's
computer in Microsoft Access format. To perform a download, select the "Download" function
from the Data Entry menu of the TRV database.
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APPENDIX A
ALLOMETRIC EQUATIONS FOR DEFAULT INTAKE RATES
Food Ingestion Rates
Where food ingestion rates are not reported in the individual respective toxicological studies, the
food ingestion rates are estimated using the allometric equations of Nagy (1987). Nagy (1987)
derived equations to estimate dry-weight-based food ingestion rates for mammals and birds based
on body mass. Food ingestion rates are derived using the following equations:
For mammals:
IRfood = 0.0687x5 ^F
(i)
where:
0.0687
BW
0.822 :
For birds:
Ingestion rate of food, wet weight basis (Kg/day);
Mathematical constant derived by Nagy (1987);
Body weight of the ROI (Kg); and
Mathematical constant derived by Nagy (1987).
= 0.0582x5 W
°'651
(2)
where:
0.0582 =
BW =
0.651 =
Ingestion rate of food, wet weight basis (Kg/day);
Mathematical constant derived by Nagy (1987);
Body weight of the ROI (Kg);
Mathematical constant derived by Nagy (1987); and
Water Ingestion Rates
If the water ingestion rate for the test species is not reported in the respective toxicological study
under review then the water ingestion rate for the test species is estimated used an allometric
equation. For avian species, Calder and Braun (1983) developed an equation for estimation of
drinking water ingestion (IRwater) based on the body weight of the bird where:
0.67
(3)
Guidance for Developing Eco-SSLs Attachment 4-3 A- 1
June 2005
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where:
IRwater = Ingestion rate of water, (L/day);
0.059 = Mathematical constant derived by Calder and Braun (1983);
BW = Body weight of the test species (kg); and
0.67 = Mathematical constant derived by Calder and Braun (1983).
Calder and Braun (1983) also developed an allometric equation for drinking water ingestion by
mammals.
(4)
where:
IRwater = Ingesti on rate of water, (L/day);
0.099 = Mathematical constant derived by Calder and Braun (1983);
BW = Body weight of the test species (kg); and
0.90 = Mathematical constant derived by Calder and Braun (1983).
Guidance for Developing Eco-SSLs Attachment 4-3 A-2 June 2005
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APPENDIX B
STATISTICAL POWER TEST
(Source: Rosner 1995. Fundamentals of Biostatistics)
The NOAEL is normally defined at the highest exposure level in a study that did not cause a
statistically significant difference in mean response from the control group. The test for statistical
significance that is most often used to compare the control group and an exposed group is the one-
sided t-test assuming equal variance:
7H - 777
t =
s
Jl/n
v i
s1 +(n2-
, Ğ<> - 2
where:
m = mean response of control group (nij) or test group (m2)
s = standard deviation of control group (Sj) or test group (s2)
sp = pooled standard deviation
n = number of animals in control group (nx) or test group (n2)
Power is the ability of a particular experimental study to detect a statistically significant difference
between the control group and the exposed group, if the true difference in the means is some
specified value (). The method used to calculate power for the difference in the means of two
normal distributions with equal variance in a one-tailed test is as follows:
Power = 0|Zfl) = 01 , Z,
P! I _ /1 / .. ,-I I .. l-a
where:
= Standard normal distribution function
Assumed difference between the means of the exposed and control groups
(i.e., the difference that is of concern to you as a biologically significant
effect). Choosing the value of *to use is this calculation is subjective. For
the purposes of evaluating toxicological studies as candidates for derivation
of TRVs, a default value of 20% of control is used as ? This is based on
the assumption that most experimental studies cannot detect smaller changes
with acceptable power, and that changes of 20% or less will often not result
in population level impacts, at least for many endpoints.
Guidance for Developing Eco-SSLs Attachment 4-3 B-l June 2005
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= Statistical significance level used to declare an effect different from control.
The default value of *is 0.05.
If it is not convenient to calculate the standard normal distribution function of Z..exactly, the
approximate power of a study can be determined using the following table of critical values:
Zb (Critical)
-0.674
0.000
0.674
0.842
1.282
1.645
Power
25%
50%
75%
80%
90%
95%
For example, if the calculated value of Z..is 0.93, then the power of the study to detect a difference
of size *at the 0.05% confidence level is greater than 80% but less than 90%. A similar approach
has been used in the Wildlife TRY database to evaluate the results of the power calculation and
assign a power score.
Guidance for Developing Eco-SSLs Attachment 4-3 B - 2
June 2005
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