# E C O T O X ECOTOXicology Database System TERRETOX Coding Guidelines

Prepared for

U.S. Environmental Protection Agency Office of Research and Development National Health and Environmental Effects Research Laboratory Mid-Continent Ecology Division (MED)
Duluth, Minnesota

By

Computer Sciences Corporation
Duluth, Minnesota 55802

June 2009

### TERRETOX Coding Guidelines

MED Ecotoxicology Database SOPs June 2009 i C. TERRETOX CODING GUIDELINES ......................................................................... 1 Method for Prioritizing Studies for Inclusion in the ECOTOX Database .......................... 2

1. Quality Assurance Parameters ............................................................................ 2 Reference Number (#), Author, Year ........................................................... 2 Total Tests ................................................................................................... 2 Reviewer/Review Date ................................................................................. 3 QA Date/Name ............................................................................................. 3 Test Number (TID) ...................................................................................... 3 2. Test Chemical Parameters .................................................................................. 4 Test/CAS Number/Chemical Name/ ............................................................ 6 Chemical Grade ........................................................................................... 8 Chemical Purity ............................................................................................ 8 Chemical Formulation .................................................................................. 8 Chemical Comment ..................................................................................... 8 Radiolabel .................................................................................................... 8 Chemical Abstracts Services Registry Number (CAS NUMBER) ................ 8 3. Test Information ................................................................................................... 9

A. Test Organism Information Parameters ......................................................... 9 Species Number/ Scientific/Common Name ................................................ 9 Organism Source ....................................................................................... 10 Organism Lifestage/Age ............................................................................ 10 Initial Weight (WTINT) ................................................................................ 10 Organism Comment (Org Characteristics) ................................................. 10 B. Exposure Information Parameters ............................................................... 11 Test Location ............................................................................................. 11 Exposure Type ........................................................................................... 12 Test Method (TMETH) ............................................................................... 12 Study Type (STYPE) .................................................................................. 12 Control Type .............................................................................................. 13 Number of Doses ....................................................................................... 13 Concentrations Tested (DOSES) ............................................................... 13 Application Frequency ................................................................................ 14 Exposure and Study Durations .................................................................. 14 Exposure Duration ..................................................................................... 14 Study Duration ........................................................................................... 15 Habitat Comment (HABITAT CODE)[Field parameter only] ....................... 15 Geographic Location (LOCATION)[Field parameter only] .......................... 15 Geographic Code (ST/PR/COUNTRY )[Field parameter only] .................... 15 Latitude/Longitude (LAT/LONG) [Field parameter only] ............................. 16 Application Season (AP SEAS) [Field parameter only] ............................... 16 Application Date (AP DATE)[Field parameter only] .................................... 16 Application Rate (AP RATE) ...................................................................... 16 Media Type ................................................................................................ 16 C. Media Information Parameters ..................................................................... 17

### TERRETOX Coding Guidelines

MED Ecotoxicology Database SOPs June 2009 ii Soil Type .................................................................................................... 17 Soil Texture (Sand, Silt and Clay) .............................................................. 17 Media pH .................................................................................................... 17 Temperature (Temp) .................................................................................. 17 Media Organic Matter Type and Units ....................................................... 17 Media Moisture .......................................................................................... 18 Media Cation Exchange Capacity (CEC) ................................................... 18 Soil Concentration (CONC) Measured / Dry-Wet Weight ........................... 18 Soil Concentration (CONC) Basis of Measurement: Dry/Wet Weight ........ 18 Media Comment ......................................................................................... 18 4. Exposure Information ......................................................................................... 19 Dose ID and Dose Number (No.) ............................................................... 19 Sample Number (N) ................................................................................... 20 Gender (Sex) ............................................................................................. 20 Dose .......................................................................................................... 20 Dose Statistical Method (SM) ..................................................................... 21 Dose Value ................................................................................................ 21 Dose Unit ................................................................................................... 21 Concentration Type (CONC TYPE) ........................................................... 21 Concentration Types and Definitions ......................................................... 21 Ionic Fraction ............................................................................................. 22 Chemical Analysis Method (M/U) ............................................................... 22 Remark Number/Remarks (RN) ................................................................. 23 General Comment ...................................................................................... 23 5. Results Information ............................................................................................ 23 Dose ID & Dose No .................................................................................... 29 Sample Number (SMP#) and / Units .......................................................... 29 Observation Time Duration (OBSRV DUR) ................................................ 29 Effect .......................................................................................................... 31 Effect Measurement (MEASMENT) ........................................................... 32 Endpoint (ENDPT) ..................................................................................... 32 Companion Endpoint(s) (COMPEP) .......................................................... 33 Result Set (R) ............................................................................................ 34 Significance (STAT) ................................................................................... 34 Level .......................................................................................................... 35 Paper/Reviewer Assigned Data (P or R) .................................................... 36 Trend (TREND) .......................................................................................... 38 Effect % (EFCT %) ..................................................................................... 39 Combining Effect Percent .......................................................................... 39 Response Site (SITE) ................................................................................ 39 Observed Response Value/Unit: ................................................................ 40 Weight at time of result (WTAT) ................................................................. 40 Intake Rate (INTAKE) ................................................................................ 41 Steady State (STST) .................................................................................. 41 Dry or Wet Weight (DW%) ......................................................................... 41 Percent Lipid (%LPD) ................................................................................ 41 Rating (RANK) ........................................................................................... 41

### TERRETOX Coding Guidelines

MED Ecotoxicology Database SOPs June 2009 iii Remark Number/Remarks (RN/REMARKS) .............................................. 42 General Coding Information ............................................................................. 42
TERRETOX References .................................................................................. 44

### TERRETOX Coding Guidelines

MED Ecotoxicology Database SOPs June 2009 1 

# C. TERRETOX CODING GUIDELINES

The TERRETOX Coding Guidelines presented below follow the format of the TERRETOX coding sheets located in Section 6. Supporting Materials . The TERRETOX Coding Sheet is divided into four sections: Quality Assurance Parameters, Test Information, Exposure Information, and Results Information. The field name associated with each test parameter, as pr esented on the TERRETOX Coding Sheet, is a topic heading. Below each heading is a detailed description of how to code data related to the specific test parameters. With few exceptions, reviewers should code the information as presented by the authors. Decisi ons regarding which information to code and how to represent the information in the database are based on the terrestri al plant and wildlife toxicity testing methods presented in ASTM and US EPA publications, as well as the scientific literature and toxicology textbooks. Test method publications used for the Terrestrial Plant and Wildlife Toxicity Effects (TERRETOX) database are listed in Section 6. S upporting Materials. Questions related to this document should be directed to the EPA Database Coordinator and the Lead Reviewer on the ADP Support Contract.

Note: Inclusion of publications into the TERRETOX database is determined by the test media used. If a terrestrial organism is exposed in an aqueous media, the paper will be plac ed into the AQUIRE database. Terrestrial nematodes are an example of an organism that may be coded either in TERRETO X or AQUIRE. If the test is conducted in soil media, then the data are coded in TERRETOX; if the nematode is isolated from soils and exposed using aquatic test methods, then the data are coded in AQUIRE. The exception to this rule is for hydroponic studies. Hydroponics is a terrestrial test method and is coded as such.

Note: Each publication included into the TERRETOX database must meet the five minimal criteria for acceptance (i.e. chemical, species, concentration, duration and effect). If the paper is missing one or more of these criteria ECOTOX does not search other sources to obtain the missing data piece( s). Sources such as author communications and referencing another work to obtain one of the five criteria is allowed for s pecific risk assessment/criteria projects (e.g. EcoSSL or CAD) and must be specified by the EPA Database Coordinator.

Note: Only quantitative data are encoded into TERRETOX dose re sponse encoding. If data are represented qualitatively (i.e., no numeric response values), the qualitative effects w ould be encoded as a single record by ranging all durations and concentrations.

Note: Lead shot papers are not encoded into TERRETOX. Authors often report the number of pellets fed to the animals, but not the actual dose of lead or concentration of lead/pellet. There are also instances in which the lead pellet is actually a mixture of two metals (i.e. Lead and tin).

Note: If a paper reports that the organisms used for measur ements were dead and it is unknown how long the organisms have been dead this data is not encoded, but enter A dead organisms @ in Other Effects. An author may measure the growth or weight of a dead organism. The weights of dead animals can be biased, especially for juveniles, depending on the time between death and measurement. Also, temperature may effect the measurement after death (e.g. higher brooder temperatures may cause rapid desiccation which would result in weight changes).

Note: Studies involving carbon dioxide (CO2) or ozone (03) as the toxicant are not coded into the TERRETOX database.

Note: If the author provides data from another publication, the reviewer should search the ECOTOX reference database (Reference Manager - ECOREF, INPROCESS, OUTPROCESS) to determine if this reference has been previously identified as applicable to ECOTOX. If the paper has not been identified, the reviewer should have the paper ordered by the literature acquisition staff. If the cited data is from a pub lication which is within the ECOREF file, the ECOREF number should be noted on the original paper. Data should only be c oded from the original publication to avoid duplicate data within the database.

### TERRETOX Coding Guidelines

MED Ecotoxicology Database SOPs June 2009 2 Method for Prioritizing Studies for Inclusion in the ECOTOX Database

$ For published Criteria Documents (Final and Draft Final), all data that appear in the tables will be encoded into the ECOTOX dat abase, without exception.

$ For the TRV project, all papers identified appropriate for use in deriving the TRV (Scores >65), and having MOR, REP, or GRO data will be coded into ECOTOX, without exception.

$ Data maintenance reported by users or EPA, will be addressed in a timely manner, without exception.

$ For criteria chemicals, where the Office of Water has only published the reference list, for all OPP-ESA papers, and for all general ECOTOX c oding, the following papers will be prioritized for coding:

$ Study must be published as either a journal article, thesis or numbered government report. Conference Proceedings, extended abstracts, and unpublished materials will be marked as such and removed from the coding queue.

$ Studies must be published in English. Non-English publications will be marked as such and removed from the coding queue.

$ Papers must report the use of a valid contro l. Studies that do not discuss the use of controls, or where the authors specifically state that a control was not used (ECOTOX codes NR and Z) will be marked as lacking a control ( A No control @ is placed in the Notes field (field 6) in Reference Manager) and will be removed from the coding queue.

$ The authors must report either a calc ulated endpoint, as defined by the ECOTOX coding guidelines, or presents statistically analyzed results. Papers presenting only qualitative data will be marked as such ( A No endpoint, No stats @ is placed in the Notes field (field 6) in Reference Manager) and removed from the coding queue.

$ Endpoint and/or statistical results must be su mmarized in either the text or tables. If results are only presented graphically or in figures, and specific concentrations (not ranged or approximate values) associated with the endpoint/statistical results are not presented, the paper is marked as having only graphed/figure data ( A Graphed data only @ is placed in the Notes field (field 6) in Reference Manager) , and removed from the coding queue.


## 1. Quality Assurance Parameters

Reference Number (#), Author, Year
Reference number is the unique number that identifies a particular publication. This number, automatically assigned by the dat a entry program, provides the link between data entered and the original publication. On the coding sheet, enter the reference number located in the upper right-hand corner of the hard copy of the publication, the last name of the firs t author, and the publication year in the data field REFERENCE #, AUTHOR, YEAR.

Total Tests
Total tests encoded for a publication are recorded by the reviewer in the TOTAL TESTS data field. The total test number equals the total number of results records coded in the RESULTS

### TERRETOX Coding Guidelines

MED Ecotoxicology Database SOPs June 2009 3 INFORMATION section of the coding sheet for eac h publication. Total tests are counted after the data abstraction process has been completed.

Reviewer/Review Date
The person conducting the data abstraction enters his/ her last name in the REVIEWER data field. The date on which the publication was reviewed is entered in the REVIEW DATE field using the format of month/day/year.

QA Date/Name
Following data coding and prior to data entry, an EC OTOX staff member conducts a screening check of each coding sheet to ensure completeness and accuracy of data transcription. The person conducting this quality assurance screening check enter s the date of the QA check in the QA DATE field, using the format of month/day/year, and their initials in the NAME data field.
Test Number (TID)
Each unique test is coded on a separate data sheet and assigned a unique test number (TID) by the reviewer. A unique test design may be characterized by a new test chemical, test species, test location, or exposure type. Additionally, there ar e experimental design (ED ES) parameters that will influence a test scenario sufficiently to warrant an independent Test TID. Such parameters include tests conducted at different test temperatures or conducted during different seasons. Some examples are found in Tables 1 & 2.
Table 1: A study is conducted with 2 different chemicals and the exposure for 2 species is started at 3 different lifestages. The Test IDs would be 

### :

TEST ID Unique Test Design

1 Benzene, Worm, Cocoon
2 Benzene, Worm, Juvenile
3 Benzene, Worm, Adult
4 Benzene, Bird, E gg 5 Benzene, Bird, Juvenile
6 Benzene, Bird, Adult
7 Toluene, Worm, Cocoon
8 Toluene, Worm, Juvenile
9 Toluene, Worm, Adult
10 Toluene, Bird, E gg 11 Toluene, Bird, Juvenile
12 Toluene, Bird, Adult

Table 2: A study is conducted with 1 chemical and the exposure for 1 species is conducted at 3 different temperatures. The Test IDs would be:

TEST ID Unique Test Design Exposure Info Remark

1 Benzene, Worm, 15C EDES/Conducted at 15C//

2 Benzene, Worm, 20C EDES/Conducted at 20C//

3 Benzene, Worm, 25C EDES/Conducted at 25C//

### TERRETOX Coding Guidelines

MED Ecotoxicology Database SOPs June 2009 4 If appropriate, include information about the Experiment al Design parameters in the REMARKS data field for Species Information, Ex posure Information or Soil Information as well as in the REMARKS data field for each independent Observed Response value reported.



## 2. Test Chemical Parameters
ECOTOX is catalogued by the toxicant tested using the Chemical Abstracts Service (CAS) registry number. If a CAS registry number is not availabl e through standard sources the toxicity data cannot be included in ECOTOX. Additional toxicants not included in ECOTOX are water/soil chemistry effects (e.g., pH), complex effluents, and chemical mixtures. Chemical mixtures may be interpreted broadly. For example, a pesticide formulation is a mixture of two active ingredients, each ingredient may hav e a separate CAS number. If the formulation has a single unique CAS number, the chemical reported for AQUIRE is the formulation. If two different active ingredients are mixed together in a tank t hen applied as a toxicant, the data is not coded as this is a mixture. If the exposure is based on two metal compounds but the effect is based on one ion, e.g., copper sulfate and copper chloride and Cu is the toxicant, code copper as the test chemical and report the two compounds in chemical CHARACTERISTICS



### . If herbicides/fungicides are applied as standard weed /fungus control to both control and treatment groups prior to the application of the toxicant, the toxicant effects are coded and the application of the herbicide/fungicide is noted in the Experimental Desi gn data field. If the chemicals are applied at the same time, this IS considered a mixture. Formulated or technical grade chemicals that report other chemicals as impurit ies are not considered mixtures. Species exposed to more than one chemical dur ing an exposure period are not necessarily considered mixtures. In some cases, the first chemic al applied is used to treat a deficiency, disease, parasite, etc. to ensure more healthy test organism s or may be used to induce a biological response prior to a chemical exposure. When such a treatment is described by the authors, encode the beneficial or inducing treatment as an experimental design comment and encode the toxic effects of the other chemical(s) for ECOTOX. For example, the authors state that A Grasses in all experimental sites were treated in the seedling stage of growth with bromoxynil for the control of broad-leafed weeds. No other herbicides were applied before the treatment s with dicamba or propham @ . Encode A EDES/Pretreated with bromoxynil in seedling stage for control of broad-leafed weeds// @ in the Exposure Information comment area. Encode the dicamba and propham toxicity data for ECOTOX. A similar example for lifestage studies occurs when the parental generation is exposed to one chemical prior to the gestational or post-gestational exposure of another chemical to the offspring produced, the parental data is c oded until the second chemical expos ure and the offspring data is coded only for the second chemical exposure. For the offspring records, it is noted in Experimental Design that the parents were exposed to a chemical prior to the gestational or post-gestational exposure.
Another differentiation of a mixt ure is drawn when nutrients are added to an experimental set up. If the author states that a nutrient is added at a level that is needed for growth, ECOTOX does not consider this as a mixture. If, however, the author adds a series of nutrients and toxicants to test

### TERRETOX Coding Guidelines

MED Ecotoxicology Database SOPs June 2009 5 interactive effects, ECOTOX considers this a mixt ure. The following example illustrates how nutrients and toxicants are coded.
A. Effect of copper on plant growth B. Effect of copper and nutrient on plant growth

Nutrient Copper 20 ug/l 40 ug/l Nutrient Copper 0 ug/l 10 ug/l 20 ug/l 30 ug/l 40 ug/l

0 ug/l 100% 120% 0 ug/l 100% 100% 100% 110% 120%

10 ug/l 100% 90% 10 ug/l 100% 100% 100% 100% 90%

20 ug/l 80% 80% 20 ug/l 95% 90% 80% 85% 80%

30 ug/l 60% 50% 30 ug/l 60% 60% 60% 65% 50%

40 ug/l 30% 20% 40 ug/l 20% 15% 30% 35% 30%



### A. Author states that nutrients ar e added for growth. All results coded. Each nutrient level is coded as a separate test and the nutrient level is noted in an experimental design set-up (EDES) comment.

B. Author does not state that nutrient is added for growth. Two tests are coded, one for the nutrient tested alone and a second result for the copper te sted alone. The shaded area is not coded. Mixture is noted in general remarks.

If the author does not document the basal level val ue, a determination may be possible for deficient, basal and toxic dose levels. The suggested guideline for making this determination would be interpreting dose response results for mortality, growth and reproduction to determine the deficient, basal (or basal range) and toxic levels. This coul d be accomplished by interpreting the trends for these effects. The deficient results would be excluded and basal level are coded as the control. If multiple basal values are report ed, the most optimal dose for mort ality, growth, reproduction would be considered the control value. If the toxicant added does not produce a toxicity test result (i.e., only deficient and/or basal levels), then the paper would be rejected as nutrient study.

For in situ exposures where the exposure is by default an exposure to a chemical mixture; code residue measurements or endpoints (BCF) only. No other effects or endpoints are strictly attributable to a single chemical in the same way as a residue concentration. Data for chemicals in the mixture with reported media concentrations and residue effects should be coded. If the author reports that a precip itate was formed during the addition of the chemical or during the exposure period, code A precipitate formed @ the EXP DESIGN field.

A standardized identification number and name for each chemical is recorded in the database to ensure quality and consistency. Toxicants, carri ers and positive control chemicals reported in

### TERRETOX Coding Guidelines

MED Ecotoxicology Database SOPs June 2009 6 ECOTOX are assigned a Chemical Abstract Services (CAS) Registry number and are referred to by the Collective Index (CI) standard nomenclature . The CAS number and CI name are stored in a chemical card file and an online index file (Eco Chem). EcoChem is available for screening CAS numbers and chemical names used in ECOTOX . Chemical name synonyms are stored electronically, but are also availabl e from the chemical card file.

Test/CAS Number/Chemical Name/
Record the test, carrier and/or positive control chemic al name as it is reported in the publication. The test chemical, as presented by the author, is reported on line number one (TEST). The CAS number is assigned by locating the chemical name in the EC OTOX chemical card file or in the ECOCHEM database. If the chemical name is not in the chemical card file, record a > no = in the CAS number field and the coding sheet will be referred to ECOTOX staff for CAS number verification following completion of the coding and screening quality assurance checks. In certain instances the author may modify the test chemical by neutralizing or by adding a salt. The chemical that is introduced into the organism is what should be coded as the Test Chemical.
Example: Author reports purchasing hydrogen selenite CAS#7783008

which is neutralized with NaOH Author reports administering sodium selenite CAS#10102188 to organism.

Code: Test Sodium selenite

CAS Number: 10102188 

### To assign a CAS Number from a chemical name:

Check EcoChem for the chemical name.

$ If one CAS Number is found - assign CAS Number

$ If more than one CAS Number is retrieved, check the molecular formula o If molecular formula is presented, match the molecular formula to a formula in EcoChem for the given name. [Note: Check the abstr act, methods, footnotes, table footnotes, and the Acknowledgments section for this information] o If molecular formula is not presented, do one of the following: o If a generic CAS number is available for the chemical, assign the CAS number. o If a generic CAS number is not available, check to see if the metal has a common valence (see following table), e.g. copper chloride (the +2 valence is most common), assign the CAS Number fo r the most common valence for that chemical name. o For metal compounds that do not have a more common valence (see following table) and a generic CAS number is not available for the chemical name, assign the parent metal form, e.g. Arsenic if ar senate is the chemical reported by the authors.

o For metal or nonmetal chemicals that do not fit these rules, refer the chemical to the Chemical verification sta ff for further investigation.

### TERRETOX Coding Guidelines

MED Ecotoxicology Database SOPs June 2009 7 Elements and Common Valences
(from Periodic Table of the Elements, Sargent-Welch, 1980 cat. # S-18806)

Element Symbol Common Valence Element Symbol Common Valence Actinium Ac 3 Molybdenum Mo 6 Aluminum Al 3 Neodymium Nd 3 Americium Am 3 Neptunium Np 5 Antimony Sb 3 Nickel Ni 2 Arsenic As 3 Niobium Nb 5 Astatine At -1 Nitrogen N 3 Barium Ba 2 Osmium Os 4 Berkelium Bk 3 Oxygen O -2 Beryllium Be 2 Palladium Pd 2 Bismuth Bi 3 Phosphorus P 5 Boron Be 3 Platinum Pt 4 Bromine Br -1 Plutonium Pu 4 Cadmium Cd 2 Polonium Po 4 Calcium Ca 2 Potassium K 1 Californium Cf 3 Pr aseodymium Pr 3,4 Carbon C 4 Promethium Pm 3 Cerium Ce 3 Protactinium Pa 5 Cesium Cs 1 Radium Ra 2 Chlorine Cl -1 Rhenium Re 7 Chromium Cr 3 Rhodium Rh 3 Cobalt Co 2 Rubidium Rb 1 Copper Cu 2 Ruthenium Ru 3,4 Curium Cm 3 Samarium Sm 3 Dysprosium Dy 3 Scandium Sc 3 Erbium Er 3 Selenium Se -4 Europium Eu 3 Silicon Si 4 Fluorine F -1 Silver Ag 1 Francium Fr 1 Sodium Na 1 Gadolinium Gd 3 Strontium Sr 2 Gallium Ga 3 Sulfur S 6 Germanium Ge 4 Tantalum Ta 5 Gold Au 3 Technetium Tc 7 Hafnium Hf 4 Tellurium Te -4 Holmium Ho 3 Terbium Tb 3 Hydrogen H 1 Thallium Ti 1 Indium In 3 Thorium Th 4 Iodine I -1 Thulium Tm 3 Iridium Ir 4 Tin Sn 4 Iron Fe 3 Titanium Ti 4 Lanthanum La 3 Tungsten W 6 Lead Pb 2 Uranium U 6 Lithium Li 1 Vanadium V 5 Lutetium Lu 3 Ytterbium Yb 3 Magnesium Mg 2 Yttrium Y 3 Manganese Mn 2 Zinc Zn 2 Mercury Hg 2 Zirconium Zr 4



### For the remaining chemical information lines, record the chemical name as reported by the author regarding any carriers, solvents or positive controls used for the test. If neither a carrier/solvent nor a positive control was used, report as 'NR'. If a carri er/solvent or positive control was used, circle

### TERRETOX Coding Guidelines

MED Ecotoxicology Database SOPs June 2009 8 A Carrier @ or A Positive Control @ as applicable. Frequently used carrier /solvent CAS numbers are listed in Appendix A. The CAS numbers for positive control chemicals are assigned by locating the chemical name in the ECOTOX chemical on-line or card file. If the chemical name is not in the chemical card file, record a > no = in the CAS number field and the coding sheet will be referred to ECOTOX staff for CAS number verification following completion of the coding and screening quality assurance checks.

Note: Water should not be coded as a solvent. A solvent is defined as an agent (other than water) in which the test chemical is mixed to make it miscible with dilution water befor e distribution to test chambers. Solvents or carriers are used in toxicity tests where the concentrations of the test chemic al are extremely low and a very small amount of test material must be added to the test chambers. (Rand, 1995)

Note: Exposure and observation data for carrier and positive cont rol chemicals are reported in the Exposure and Results sections. Refer to these sections for specific instructions.



### Chemical Grade
Record the chemical grade information for each chemical reported in the GRADE data field (refer to Appendix B for the applicable codes).
Chemical Purity
Record the numeric percentage information about the purit y or active ingredient of the chemical in the PURITY data field (e.g., if the author reports 97% pur ity, 97 would be entered into this data field. PU for purity would be entered into the FORMULAT ION data field (see CHEMICAL FORMULATION).

Chemical Formulation

Record the chemical formulation code for each chem ical reported in the FORMULATION data field (refer to Appendix C for the applicable codes).

The chemical formulation FORM is only coded when explicitly identified by the authors as the form (e.g., commercial) or when presented in a standard format (e.g., 25EC).

Chemical Comment
Supplemental information about the test chemical is entered into the CHEMICAL COMMENT field. If a mixture of labeled and unlabeled chemical is used, remark A labeled and unlabeled @ in this field. Record additional relevant chemical information su ch as trade names, common names, or isomers in this field.

Radiolabel
If a radiolabeled chemical is tested, record the isotope in the RADIOLABEL field (see Appendix D for codes). When the specific isotope is not reported or when multiple is otopes are reported, the field is marked with an asterisk (*). In REMARKS, note eit her RADIO/no isotope reported// or RADIO/isotope xx and isotope yy//. When both radiolabeled and unlabeled te st chemicals are used in a test, report the radiolabeled isotope and code A labeled and unlabeled @ in the CHEMICAL COMMENT field.

Note: Any REMARKS made for fields in this section will be recorded according to the instructions set forth in Test Information. 

### Chemical Abstracts Services Registry Number (CAS NUMBER)



### The CAS Number of the toxicant is recorded in the CAS NUMBER 

### field. A standardized identification number and name for each chemical recorded in the database is used for consistency. Toxicants included in the ECOTOX database are assigned a CAS registry number and are referred to by the

### TERRETOX Coding Guidelines

MED Ecotoxicology Database SOPs June 2009 9 Ninth Collective Index (CI) standard nomenclature. The CAS number and CI name are stored in a chemical card file and in an online index file (ECOCHEM) which is available electronically for screening CAS numbers and chemical names used in EC OTOX. If a hydrated form of a chemical is used in the paper, record the hydrated fo rm as reported by the author in the TEST 

### field. However, record the CAS Number for the non-hy drated form of the chemical in the CAS NUMBER 

### field.



## 3. Test Information
This section is used to report general information describing the test scenario. If any of the following information changes, a new Test ID is assigned and a new coding sheet is required. Specifically, the Test Information section describes the test organism , the test location and exposure type, information about the type of controls used, the total number of doses, and the application frequency. Refer to Table 3 for coding examples.
A. Test Organism Information Parameters

Species Number/ Scientific/Common Name
The test organism is identified by the current scient ific name as verified in the taxonomic literature. Enter the species name, as presented by the author in the SPECIES SCIENTIFIC/COMMON NAME field. Each unique test organism is assigned a s pecies number which is stored in the CRITTERS database. Locate the number for the species in t he CRITTERS database and enter it in the SPECIES NUMBER field. If the species is not in the CRI TTERS database enter 'no' in the SPECIES NUMBER field, and the coding sheet will be referred to EC OTOX staff for species verification following completion of the coding and screening quality assu rance checks. For each species number, the verified name, taxonomic hierarchy, nomenclature hist ory, and verification sources are kept on file for quality assurance documentation. Generally, when coding effects in ECOTOX, the data are reported for each individual species. Field studies may report results for a target community (e.g., beneficial and non-beneficial insects) or for an entire enclosed ecosystem (e.g. system-level primary productivity or respiration). Assign a community to the most specific taxonomic level possible (e. g. A earthworms @ would be classified as

A Oligochaeta, @ A weedy plant species @ would be classified as A Magnoliophyta. @ ). If you are not sure about the classification of a community, enter > no = in the SPECIES NUMBER field, and it will be sent to ECOTOX staff for verification. Decisions regarding the inclusion of species in TERRETOX are based on published terrestrial ecotoxicology standard methods and procedures doc umentation (eg., Menzer et al 1994; US EPA testing series; ASTM testing series). The focus for TERRETOX is to collect publications with data for soil invertebrate and microbial species, plant spec ies (agricultural and native), wildlife avian species (e.g. mallard, pheasant or bobwhite), wild mammali an species (e.g., meadow vole, deer mouse or mink), terrestrial lifestages of amphibians and reptiles, and beneficial invertebrate species (e.g., honey bee, leafcutter bee or alkali bee). If data for ot her species including laboratory, domestic or non-beneficial organisms are r eported in a publication, data for a ll test species are coded for entry into TERRETOX. Publications focusing primarily, or solely, on laboratory, domestic or non-beneficial organisms are not actively acquired or coded at this time.

### TERRETOX Coding Guidelines

MED Ecotoxicology Database SOPs June 2009 10 Organism Source
Report the source of the test organism in t he ORGANISM SOURCE data field (see Appendix E for codes). The source explicitly includes the strain of the organism, e.g. labor atory strain versus wild strain.
Organism Lifestage/Age

The LIFESTAGE/AGE data field reco rds the specific lifestage and/or age for each test organism at beginning of exposure, as reported in the paper (s ee Appendix F for lifestage codes and Appendix I for time units associated with the age of the organis m). Record the lifestage information in the first box and age information in the second box on the coding sheet. Record as 'NR' if the information is not reported in the publication.
Initial Weight (WTINT)
Report the weight and weight unit for each test organi sm at beginning of exposure, as reported in the paper as a Remark in the following format: WTINT/mean(min-max) unit//.
Organism Comment (Org Characteristics)
Report any general information provided about the test organism. Characteristics may include information such as specific strain name, cultivar , variety, length, developmental stage, hybrids or taxonomic groupings used to describe the organism being tested.

Note : Information regarding the sex of the test organism is coded in the Sex field, see Exposure Information. The sex of the organism is often directly linked to the exposure and subsequent response observations; for example, specific reproductive responses are unique to males or females.

Note: When reporting a cultivar, include > cv. = before the name of the cultivar. Include > var. = for variety in a similar manner.

### TERRETOX Coding Guidelines

MED Ecotoxicology Database SOPs June 2009 11 Table 3. Test Information Coding Sheet Example

S PECIES S CIENTIFIC /C OMMON N AME

__ Aphis sp.__________________ O RGANISM I NFORMATION

S PECIES N UMBER 5519

O RGANISM S OURCE WLD

L IFESTAGE /A GE NR 1-2 d

C HARACTERISTICS A. mellifera and A. ligustica

T EST L OCATION LAB E XPOSURE I NFORMATION

E XPOSURE T YPE FD plants sprayed outdoors in evening; moved to lab next day; bees exposed to plants in lab

E XPOSURE D URATION 6 D

S TUDY D URATION 2 WK

C ONTROL T YPE B

N UMBER OF D OSES 3

A PPLICATION F REQUENCY ADL

M EDIA TYPE NAT S OIL I NFORMATION

S OIL T YPE Pedozioic Clay -silt

S OIL T EXTURE % SA 79 SI 15 CL 6

M EDIA P H 5.6

M EDIA O RGANIC M ATTER 5 %

M EDIA M OISTURE (%) 31

M EDIA CEC NR

S OIL C ONC M EASURED )/

DRY - WET WEIGHT M DRY



### B. Exposure Information Parameters

Test Location
Report the location or setting in which the exper iment was conducted in the TEST LOCATION data field (see Appendix H). For example, a natural field study (FieldN) is an experiment conducted outdoors in a natural setting. The test organisms are sampled in the wild, e.g. population counts. Outdoor studies conducted in a simu lated environment are coded as an ar tificial field study (FieldA). Artificial field studies include organisms isolated fr om their natural environment via an enclosure of some type, e.g. cages or fencing. If the publicat ion does not provide enough information to distinguish

### TERRETOX Coding Guidelines

MED Ecotoxicology Database SOPs June 2009 12 between FieldA and FieldN, then use the code FieldU to indicate that the field test type is unknown. Laboratory tests (LAB) are conducted indoors under controlled laboratory conditions. If the location or setting cannot be identified as laboratory or field from the publication, code as Not Reported (NR).
Exposure Type
For the TERRETOX database, the term > exposure = is used to refer to the mechanism by which the toxicant was applied. Organisms are typically exposed to toxicants through diet, injection, topical or environmental routes. On occasion, an exposure may be through multiple routes (e.g., such as topical and oral). Some exposures could be coded a variety of ways. For example, exposure as an aerial spray to a field plot could be coded either as a spray applicat ion or as exposure through multiple routes, eg. topical (through skin) and diet (from consumption of exposed vegetation) for animals, or topical (through leaves) and environmental (root uptake) for plants. Within the TERRETOX database, this instance is coded as a spray application. Multip le exposure route coding is applicable when the organism is exposed through two independent applications, for example, a contaminated diet and topical application for animals or contaminated soil and leaf spray for plants. In this scenario, > MU = would be entered into the EXPOSURE TYPE data field and a remark (TYPE/ = FD = and TP// or TYPE/ = PR = and > FS = //) would be noted in the Exposure Info comments section. When coding, report the specific ex posure type, e. g., for an intercutaneous injection, code as IC (intercutaneous) not I (injection). For an environment al exposure, code as HS (hand spray) not V (environmental). If an exposure type is not reported, code as Not Reported (NR). Refer to Appendix J for exposure type codes. TERRETOX does not include in vitro assays [i.e. an ex perimental trial, involving biological matter, which is exposed to a toxicant in an artificial apparatus rather than within a living organism] in the database. Studies in which the living organism is ex posed as a whole, but an effect on an internal process is examined outside the body after the ex posure, are coded (e.g. enzyme functions). The database contains some studies using excised organs and cell cultures from plants, however these types of studies are not actively coded at this time. Future coding of these studies is under discussion. TERRETOX does not include ‘inhalation’ studies or intratracheal instillation studies (placing chemical directly into lungs). These types of studi es are rejected by using the keyword inhale.
Test Method (TMETH)
Report the name of the testing method used for conducting the toxicity experiment (e.g. ASTM (American Society for Testing and Materials) or OECD (Organization for Economic Cooperation and Development) in the following format: TMETH/te xt//. See ECOTOX Appendix II for the codes and definitions.
Study Type (STYPE)
Report the type of test set up for the toxicity study as reported by the aut hor (e.g. ACUTE, CHRONIC, ELS (Early Life Stage), FLC (Full Life Cycle), Generational) in the following format: STYPE/text//. See ECOTOX Appendix HH for codes and definitions.

### TERRETOX Coding Guidelines

MED Ecotoxicology Database SOPs June 2009 13 Control Type
Effects of toxicant exposure are evaluated by comparing the exposed organisms to untreated organisms - the controls. All toxicity tests shoul d include a concurrent control where the test conditions are identical except for the absence of t he toxicant. Some toxicity tests will also include a control for other test conditions such as the use of a solvent, feeding or a cclimation requirements, historical or pre-exposure conditions and for es tablishing reproducibility by use of a reference toxicant. (Doull et.al. 1980) 

### Report the type of test control(s) used in the st udy (Appendix M) by recording the applicable code in the CONTROL TYPE field. If more than one type of cont rol is used in the study, e.g., a dilution water and carrier control, code > M = for multiple controls. Often compar isons are made that do not meet the criteria for a control; these types of comparisons include starvation studies and acclimation periods. Report the studies that complement the toxicity test, e.g. a starvation study used in a feeding behavior or avoidance test, as a comment in the RE MARKS data field in the Test Information section of the coding sheet. Sometimes a paper will report a tabl e of baseline or historical control values. Do not code these values unless there is a direct co rrelation to a measurement or endpoint; code only control values which complement response values. When data for the control are repor ted only in graphical format, interpret the data as accurately as possible and remark that the control data were obtained from a graph in the results information section. Data points derived from a graph are most typically represented as an approximation of the data point, a range around the specific data point or as a range for all of the represented values.

If a control is identified for the test but no ex posure or results data are reported, record the appropriate control type code in the Control Type fiel d. No data will be coded in Exposure or Result Information fields.

Number of Doses
Report the total number of exposure doses, including the controls, for each independent test design in the NUMBER OF DOSES data field. If number of ex posures is not reported, e.g. in a publication reporting only calculated endpoints such as LD50s, code the field as 'NR'. Do not include endpoint or ranged doses or number of replicates in the total number of doses.
Concentrations Tested (DOSES)
Report the concentrations and the units used for the toxicity test in the following format: DOSES/value/value/value unit//. If a concentrati on range is reported use the following format DOSES/min-max unit// Occasionally an author will report a concentration as a % or fraction of an LD50 value; e.g., either the sublethal concentration used was 10% of the 96-h LD 50" or 1/10, 1/15 and 1/20 of the LD50". Such concentrations may be recalculated and used as t he concentration tested if the original LD50 concentration is provided in the publication. Flag the recalculation in the paper so that the calculation may be QAed and document the recalculation in the margin or on a blank page of the publication. Put a comma after the concentration unit and remark "Recalculated" (e.g. DOSES/3/6/15 mg/kg, recalculated//)Occasionally an author will report a conc entration as a % or fraction of an LD50 value; e.g., either the sublethal concentration used was 10% of the 96-h LD50" or 1/10, 1/15 and 1/20 of the LD50". Such concentrations may be recalculated and us ed as the concentration tested if the original LD50 concentration is provided in the publication. Flag the recalculation in the paper so that the calculation may be QAed and document the recalculat ion in the margin or on a blank page of the

### TERRETOX Coding Guidelines

MED Ecotoxicology Database SOPs June 2009 14 publication. Put a comma after the concentration uni t and remark "Recalculated" (e.g. DOSES/3/6/15 mg/kg, recalculated//)
Application Frequency
Report the frequency of the dose application in t he APPLICATION FREQUENCY data field. Refer to Appendix K for application frequency codes and units.
Exposure and Study Durations
A toxicity test may range in duration from a pre-tr eatment period through the ac tual toxicant exposure and conclude with observations of the organisms post- exposure. Duration information is coded using the units reported in the publication (see Appendix I for valid units). Refer to Table 4 for a coding example. Exposure and study durati ons are reported with the Test Information. Observation Duration is reported with the Results Information.

Table 4. Example 17-day experimental period with 2-day pre-treatment, 5-day exposure, and 10-day observation. Note: Pre-treatment days are not included in the study duration.

DURATION OF EXPERIMENT

Pre-Trt Exposure Observation

Calendar

Days 1 2 3 4 5 6 7 8 9 10 11* 12 13 14 15 16 17

Test Periods 1 2 1 2 3 4 5 1 2 3 4 5 6 7 8 9 10

Reported

Days (Study Duration) -2 -1 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15

Note: In test scenarios where incubation times are reported, i.e. enzyme fixation assays, be careful to report the toxicant exposure time not the assay incubation time. 

### Exposure Duration
The exposure duration is a mandatory field for incl usion in the TERRETOX database. In cases where the observation time is the only dur ation reported, it is assumed that the exposure duration is equivalent to the observation time. If the exposure durat ion is not reported, the paper is rejected. The period of time recorded in the EXPOSURE DURATION data field is the time of actual exposure to the chemical. For example, in Table 4 the exposure duration equals 5D. In some cases a biological time is used, such as an exposure time reported as A until hatch @

, A growing

season @ or A after the nth egg has been laid @ . Use the code from Appendix I that best describes the

### TERRETOX Coding Guidelines

MED Ecotoxicology Database SOPs June 2009 15 author's words in duration units data field. > NA = will be coded as the duration mean value for all biological time durations (e.g. EXPOSURE MEAN : NA, EXPOSURE UNIT: HT; corresponds to an until hatch duration), unless a numeric value is substituted (e.g. EXPOSURE MEAN: 2, EXPOSURE DOSE: HV; corresponds to a 2 harvests duration). However, references to time such as A observed at end of the study period @ are not coded; such papers are reject ed as having no exposure duration. For injection, diet, topical and environmental exposur es where the actual exposure is dependent on biological and environmental conditions, the exposure time is recorded as equivalent to the study time. This assumption is made to ensure consistency in data representation; it is not necessarily a true reflection of the exposure time. When exposure duration values are extrapolated from a graph the values can be approximated even when the mark does not appear on the graph. For exampl e, if the value on the graph is in the middle of two marks namely 10 and 14 days, the EXPOSURE DURATION 

### can be coded as A 12 d @ with a remark that it is from the graph: ETIME/from graph//
Study Duration
The study duration is the total time of the study excluding pre-treatment times. In the example in Table 4, the study duration is equal to 15D (5D ex posure plus a 10D observation). In cases where the observation time is the only duration reported, it is assumed that the study duration is equivalent to the observation time. The study duration will be report ed as 'NR' if no observation or study time is reported.

Note: for most field studies the exposure and study duration ar e identical because it is difficult to determine when the exposure ends. It is difficult to know when the application has completely dissipated in the environment. For lab studies the exposure and study duration may be different. This difference will be seen when there is a recovery period from exposure duration. For lab studies when the treatment is some type of injection or diet (intraperitoneally or by gavage), study duration and exposure duration are the same.



### When study duration values are extrapolated from a graph the values can be approximated even when the mark does not appear on the graph. For exampl e, if the value on the graph is in the middle of two marks namely 10 and 14 days, the STUDY DURATION 

### can be coded as A 12 d @ with a remark that it is from the graph: STIME/from graph//
Habitat Comment (HABITAT CODE)[Field parameter only]
A one-letter code based on the ecoregions derived from Bailey and UC Berkley (see ECOTOX Appendix X) is used to describe the habitat (e.g., D esert or G rassland) is coded as a remark HABCODE/code//. The descriptor field is used to reco rd the author's description of the environment, e.g. mountain, agricultural. This information is coded as a remark in the format HAB/text//. If the author does not provide any information about the habi tat, both fields are coded as NR (not reported).
Geographic Location (LOCATION)[Field parameter only]
Water body, city, county or relevant site info rmation is coded. (see ECOTOX Appendix AA for field location abbreviations.) . This information is coded as a remark in the format LOC/text//
Geographic Code (ST/PR/COUNT RY)[Field parameter only]
Code the state, province or country name of the test site along with the Geo code as a remark in the format GEO/text, code//. If the test site is not reported, an "NR" is coded. (ECOTOX Appendix BB contains a listing of country, region, pr ovince names and associated Geo code.)

### TERRETOX Coding Guidelines

MED Ecotoxicology Database SOPs June 2009 16 Latitude/Longitude (LAT/LONG) 

### [Field parameter only]
If reported by the author the lati tude and longitude are recorded as a remark in the format LAT/text// or LONG/text//. The "~" sign replaces the " E " sign in data entry. If the Lat/Long are reported in decimal degrees write out the units; see exampl e below. If not reported, NR is recorded. An example of a latitude/longitude loca tion (MED, Duluth, MN) is listed below:

Latitude: 46~50'51" N Longitude: 92~11'12" W 

### An example of a ranged latitude/l ongitude location is listed below:

Latitude: 52~30'-53~30' N Longitude: 107~30'-106~30' W 

### An example of a latitude/longitude repor ted in decimal degrees is listed below: Latitude: N36.8 decimal degrees Longitude: E115.7 decimal degrees



### Application Season (AP SEAS) [Field parameter only]
Is coded as a remark in the format AP SEAS/code//, ONLY if no application date is given by the author but the author does specify a season. This fiel d contains the season of initial application of the chemical. A list of application seasons with dates and AQUIRE codes is presented below:

Code Season WI Jan-March SP April-June SU July-Sept AU Oct-Dec NR Not Reported



### Application Date (AP DATE) 

### [Field parameter only]
The application date is the time of initial exposur e. Code the application date as a remark in the format AP DATE/date//. The format is mm-dd-yyyy, e.g. 12-01-1993. If one or more parts of the date is not reported, code the letter of the date that is missing (e.g . 12-dd-1993, mm-dd-1993, 06-15-yyyy). If more than one initial date is reported (e.g. more than one field exposed), record the dates with a comma as a separator. If one field is exposed multiple times, only report the first application date and note #x in frequency. If the application dat e is not reported, NR is recorded.
Application Rate (AP RATE)
Report the application rate value and the units that the author reports as a remark in the format AP RATE/value unit//. If an application rate is not report ed by the author, record as NR. If an exposure concentration is not reported by author, the application rate must be reported. (See ECOTOX Appendix N for application units)
Media Type
Report the type of exposure media, (e.g., natural or artificial soil, aqueous (hydroponic), filter paper), in the EXPOSURE MEDIA data field using codes pr esented in Appendix L. Report as 'NR' if you cannot determine the exposure media from the paper. If an aqueous exposure is conducted in pore water from a specific soil, report the soil parameters in the soil charac teristics fields (pH, CEC, OM, etc.). If the bottom of the experimental cham ber is covered with sand and then topped with filter

### TERRETOX Coding Guidelines

MED Ecotoxicology Database SOPs June 2009 17 paper, an Experimental Design (EDES) remark should be made that sand was used in the chamber, but MEDIA should be FLT.

C. Media Information Parameters

Soil Type
Report the scientific name of the natur al soil or commercial name of the artificial soil used in the study in the SOIL TYPE data field. If the scientific nam e is not included report the type of soil using the author's terminology, e.g., forest soil, sandy loam soil, arboreal coniferous soil.
Soil Texture (Sand, Silt and Clay)
Report the texture of the soil as stated by t he authors in the SOIL TEXTURE data field using percentages of sand (SA), silt (SI) or clay (CL).

Note: Clay may be reported as bentonite, kaolinite or montmorillonite. 

### Media pH
Report the pH of the test media used in the MEDIA pH data field. If the pH of the treated media is not presented, but the pH value is stated for the unt reated or acclimation media, code the untreated media pH and add an asterisk to the end of the val ue. If the author specifies a measurement method for the pH value (e.g., that the pH value is meas ured by pHKCl or pHCaCl2), code the pH value and identify the measurement method in the REMARKS field. If the aut hors report that a standard (see Attachment G for list of standard soils) or commercially available artificial soil is used, but do not present pH, use the pH reported in the standard test method referenced by the author. If pH is reported for the untreated or acclim ation media, code this pH value in the same way as outlined previously and denote with an asterisk. If the pH of a s pecific soil type is not given in the publication, a search of the USDA/NRCS National Cooperative So il Survey (USA) online site, at the following web address: http://www.statlab.iastate. edu/cgi-bin/osd/osdname.cgi or the United States Department of Agriculture = s Natural Resources Conservation Service National Soil Survey Center site at the following web address: http://vmhost.cdp.state.ne.us:96 can be conducted for the specific soil series. If the pH is found, range the pH values for all so il depths in the pH data field and remark in the comments section pH/from USDA web source//. Attach a printout of the pH information from this site to the publication.
Temperature (Temp)
Report the temperature of the te st media used in the following fo rmat: TEMP/mean(min -max) unit// If the temperature of the tr eated media is not presented, but the temperature value is stated for the untreated or acclimation media, code the untreated media temperat ure and add an asterisk to the end of the value. When temperatures are reported for incubation cham bers, these temperatures are acceptable for reporting as test temperatures. Do not code temperatures noted as A room temperature @

. Media Organic Matter Type and Units
Report information about the test media organic matter as presented by the author. Use the measurement value, organic matter type, and units reported by the author. Refer to Appendix FF for organic matter type codes and units. If carbon and/or nitrogen content of the soils are reported, record these values in the Soil Information Re marks section; organic matter content may be estimated from these values. If the authors report that a standard ( see Attachment G for list of

### TERRETOX Coding Guidelines

MED Ecotoxicology Database SOPs June 2009 18 standard soils) or commercially available artificial soil is used, but do not present organic matter content, use the organic matter content reported in the standard test method referenced by the author. If organic matter is reported for the untreated or acclimation media, code this organic matter value in the same way as outlined previously and denote with an asterisk. If the organic matter of a specific soil type is not given in the publication, a search of the USDA/NRCS National Cooperative Soil Survey (USA) online site, at the following web address: http://www.statlab.iastate.edu/cgibin/osd/osdname.cgi or the United States D epartment of Agriculture = s Natural Resources Conservation Service National Soil Survey C enter site at the following web address: http://vmhost.cdp.state.ne.us:96 can be conducted for the specific soil series. If the organic matter is found, range the organic matter values for all soil depths in the OM data field and remark in the comments section OM/from USDA web source. Attach a printout of the organic matter information from this site to the publication.

Media Moisture
Report percentage of moisture in the test media in the MEDIA MOISTURE data field. If moisture is reported for the untreated or acclim ation media, code this moisture percentage and denote it with an asterisk.
Media Cation Exchange Capacity (CEC)
Report cation exchange capacity and units (Refer to Appendix FF for organic matter type units) of the test media in the MEDIA CEC data field. If the ca tion exchange capacity is reported for the untreated or acclimation media, code this value and denote with an asterisk.
Soil Concentration (CONC) Measured / Dry-Wet Weight
If soil was the exposure media, use the first data field to report if the toxicant concentration was measured in the soil. If measured, code as 'M' in the SOIL CONC MEASURED data field. If not measured or no information is provided, code as 'U' or 'NR' respectively in the SOIL CONC MEASURED data field.

For instances where some treatment levels are measured and some are unmeasured/nominal, denote the SOIL CONC MEASURED as > X, = signifying that a mixtur e of measured and unmeasured values was reported in the publication, but ECOTOX reports the nominal concentrations.
Soil Concentration (CONC) Basis of Measurement: Dry/Wet Weight
Record whether soil concentration was reported based on dry or wet weight in the DRY - WET WEIGHT data field.

Media Comment
Test Information REMARKS sections are used to include additional information necessary for interpreting any of the specific test information fiel ds as well as for providing information concerning the test in general. When additional information is necessary for a given field write 'FIELD NAME/remark text//' (refer to Appendix EE for applicable field name abbreviations). For general information that is not associated with a specific fiel d, label the Remark as Other Effects (OEF). The Experimental Design (EDES) notation is used to identify information that differentiates between exposure scenarios but does not dire ctly implement changes in the data fields. Information that may make a significant change in test design includes varying exposure substrates or seasonal exposure scenarios.

### TERRETOX Coding Guidelines

MED Ecotoxicology Database SOPs June 2009 19 

## 4. Exposure Information

This section is used to record the exposure paramet ers for each specific test. A specific test is identified by the Test ID (TID) number as previously described. If the paper is a benchmark study that has been approved by EPA for dose-response coding, within each specific test, information is recorded for every treatment level including test c ontrols, positive controls, carrier controls, and toxicant exposures. Such information includes the sample number and sex, the exposure dose, whether the dose is reported in ionic form, the chemical analysis method, and any pertinent remarks (see Table 5). If it is not approved for dose-respons e coding ,within each test, two exposure records could be coded, a ranged > R = and/or endpoint = E = exposure. These exposures report the range of exposure concentrations.
Dose ID and Dose Number (No.)
Each treatment in a test is assigned a Dose ID and a Dose Number. Controls are always reported first and identified by the appropriate letter code fr om Appendix M in the DOSE ID field. Exposure doses are identified by the letter 'D'. A link is created to calculated endpoints that are dependent on multiple exposure doses by coding a line identified by the letter 'E' (the linkage 'E' is not used for BAF, TKNO or LTxx/Etxx data. These ar e linked to a specific dose). Information taken from a graph (responses that DO NOT have endpoints or statis tics) may be coded using a ranged dose 'R' that encompasses all of the exposure concentrations ex cluding the control(s). If more than one type of control is used in the study, e.g. a dilution water and ca rrier control, code two lines for control, ('C' for the dilution water control and 'V' for the carrier cont rol) in the DOSE ID field. If more than one control of a specific type is used, number each control in t he set as a replicate, e.g. 1V,. 2V If a control or treatment is identified for the test but no exposure data are repor ted, there will not be any data to code for Exposure Information or Result Informati on. See Tables 3, 4 and 5 for coding examples. When replicates are used and the results are reported separately for each replicate, code a separate line for each replicate. When the publication notes that replicat es were run but the author only reports the results as the mean of the replicate values, do not code individual lines for the replicates but instead note this information in General Remarks, ie. A x replicates; => see also the Observed Response Value section in Results Informa tion for additional instructions. When dose data are reported only in a graphical format, interpret the data as accurately as possible and remark that the data were obtained from a gr aph. Data points derived from a graph are most typically represented as an approxim ation of the data point, a range around the specific data point or as a range for all of the represented values.

Note: For the example in Table 5 , the Number of Doses reported in Test Information would be six (6) to represent the two control levels and four treatment levels; all doses tested are recorded in this field regardless of whether responses are reported. Endpoint (E) and range (R) "doses" and replicate concentrations DO NOT get counted in the total number of doses.

Table 5. Exposure Information

D OSE N O D OSE
ID N S EX DOSE SM V ALUE U NIT ION M/U RN

1 C 10 F 0 - - ppm - U 1

2 C A A A - - A - A NR

3 V A A 1 - - ug/l - M NR

### TERRETOX Coding Guidelines

MED Ecotoxicology Database SOPs June 2009 20 D OSE N O D OSE
ID N S EX DOSE SM V ALUE U NIT ION M/U RN

4 D A A 3 SE 0.01 ppm Cu A 2

5 D A A A A 0.15 A A A A

6 D A A 9 A 0.02 A A A A

7 D A A A A 0.15 A A A A

8 E NR A NR NR NR NR A A NR

G ENERAL R EMARKS 1. CONTR/Control for first generation only//
2. DOSE/Conc reported as flower residues//

DATA CONTINUES ON NEXT PAGE

Note: On occasion, when coding data for the Exposure Information section, the number of test exposures and/or replicates will exceed the allotted coding space. If this should occur, c ontinue coding on a second sheet. Note at the bottom of the exposure information section on the first page that the data continues on a second page.

Note: When concentrations are not reported for soil and pore water doses, but endpoints are reported, code exposure information as > E-dose# = NR = , then code two separate endpoints for the soil and pore water endpoints in the results section. Add a remark RVALUE/soil conc// or RVALUE/pore water conc// respectively. 

### Sample Number (N)
Sample number, denoted by an 'N' on the coding sheet, reflects the sample size at each exposure dose, i.e., the number of test organisms per treatment. Code as 'NR' if not reported.
Gender (Sex)
This field identifies the sex of the organism (male ( M), female (F) or both (B 

### ) at each exposure level. The importance of this field becomes apparent where organisms of both sexes are exposed at a given treatment level, but the observations are conducted on either the male or female. In this instance, the SEX field would be coded as B in Exposure Information, with individu al results reported for M and F in Results Information in the Sample Number Unit fiel d. See Results Information and Table 5 for coding examples. Code 'NR' 

### if not reported.

Dose
Report the exposure dose as reported in the public ation. Report the approximation (~), minus (-), greater than (>), or less than (<) symbols used by the author(s) to describe the exposure dose. The mean and/or range is coded in the DOSE data field and the unit in the UNIT field, see below. If the range values are confidence interval (CI), confidenc e limits (CL) or fiducial interval (FI) code the abbreviation in the SM data field. See t he coding example presented in Table 5 . For instances where some treatment (dose) leve ls are measured and some are unmeasured/nominal, and all unmeasured/nominal concentrations ar e reported, report t he unmeasured/nominal concentrations for each treatment level so that the range of concentrations is consistent and monotonical. Denote all of the concentration analysis methods as > X, = signifying that a mixture of measured and unmeasured va lues was reported.

### TERRETOX Coding Guidelines

MED Ecotoxicology Database SOPs June 2009 21 When coding dose response data and the author r eports both dose/concentration and application rate, the dose/concentration is reported instead of t he application rate. Report the application rate in the Exposure Remarks section in the following format AP Rate/XXXX//.

Note: If chemical concentrations (especially metals) are repor ted in terms of Total, Exchangeable, Water Soluble and Pore Water concentrations, Total is the concentration selected for entry into the dose data field. The other concentrations are reported as remarks.

Note: If a background concentration is reported for the chemical being applied, report the background value in the control dose in the DOSE field. 

### Occasionally an author will report a concentration as a % or fraction of an LD50 value; e.g., either the sublethal concentration used was 10% of the 96-h LD 50" or 1/10, 1/15 and 1/20 of the LD50". Such concentrations may be recalculated and used as the effect concentration if the original LD50 concentration is provided in the publication. Flag the recalculation in the paper so that the calculation may be QAd and document the recalculation in the margin or on a blank page of the publication. In the Remark Number/Remarks (RN) make a remark of "Recalculated"

. 

### Dose Statistical Method (SM)
Report the method used to determine the range around the Dose in the SM data field, if reported by the author(s). Use standard codes for the methods, i. e., standard deviation (SD), standard error (SE), confidence interval (CI), confidence limits (CL) or fi ducial interval (FI) or range (R). If the interval around a value is not identified in the paper as SD, SE, CI, CL, FI or R, then code as not reported (NR).

Dose Value
Report the numeric value of t he standard deviation or standard error around the Dose in the VALUE data field, as reported by the author(s).

Dose Unit
Report the measurement unit that corresponds to Dose in the UNIT data field (see Appendix N for standard units). If the author reports that the c oncentration is based on active ingredient or acid equivalent choose the unit preceded by AI or ae from the dropdown menu (e.g. AI mg/kg or ae mg/kg).
Concentration Type (CONC TYPE)

The three forms of toxicants evaluated in ECOTOX are organic compounds, metals and inorganic nonmetals. Each form can be identified as a concentrati on type code using the single letter abbreviation. Organic compounds are defined by the pesticidal te rms, formulation (F) and active ingredient (A). Publications that do not specify t he compound by the definition criteria for active ingredients are by default coded in the formulation (F) category. Metals are defined by the concentration types, tota l (T), dissolved (D), and labile/free (L); while ammonia or hydrogen sulfide compounds may have to tal concentrations (T) and/or un-ionized (U) concentrations. Organometals are coded as total (T) concentrations. Concentration Type is also linked to the Chemical Analysis Method ( METHOD ) 

### field.(see discussion below on Active Ingredient).
Concentration Types and Definitions
Organic:

### TERRETOX Coding Guidelines

MED Ecotoxicology Database SOPs June 2009 22 FORMULATION ( F ): Way in which basic pesticide (toxicant) is pr epared for practical use (Ware, 1978). Generally reserved for commercial preparation prior to actual us e and does not include the final dilution (Insect-Pest Management and Control, 1971) (e.g.; Baythroid, 2,4-D). Also included in this category are organic compounds with no pesticidal activity (e.g.; PCB, dioxin). ACTIVE INGREDIENT ( A ): Chemical substance in a product that is responsible for the pesticidal (toxic) effect (Ware, 1978). Reported as "A" when the author refers to t he concentration as active ingredient, active principle , active substance (A.S.) , acid equivalent or various grades of reagent s (ie., Analytical, Reagent or Technical). When coding, a value in the publication may be reported as A AI kg/ha @ , "AE kg/ha", or A kg AI/ha @ ; in AQUIRE this type of value is reported as > A = = for CONC TYPE , with units as kg/ha. For example, 100 kg AI/ha is reported as A = 100 kg/ha.

Note: Information reported in the PURITY field does not necessarily determine whether concentration is A or F. In addition to the description above, using A A @ as the concentration type occurs in situations such as the following: 1) Author states concentration of pesticide as A AI @

. 2) Author states %AI ( PURITY ) and reports measured concentration. 3) Author states measured concentration of a pesticide. 

### Metal/Organometals:
TOTAL ( T ): The concentration of metals determined on an unfilter ed sample after vigorous digestion, or the sum of the concentrations of metals in both dissolved and suspended fractions ( APHA et.al. 1992). Heavy metals and single elements (e.g. Na, Cl, Br) are coded as T. DISSOLVED ( D ): Those constituents of an unacidified sample that pass through a 0.45 um membrane filter (e.g. soluble metal) (APHA et.al. 1992). LABILE ( L ): The labile or free ion metal concentration dete rmined by various analytical methods. When coding, the specific labile forms or complexes are not differentiated. 

### Inorganic non-metals:
Concentrations of ammonia and hydrogen sulfide are reported in the literature in either the total or unionized form. Code the form as specified by the author. Ammonia may be reported as a variety of different forms, eg., NH 3 , NH 4 + , NH 3 -N, NH 4 OH, or NH 4 Cl. ( US EPA 1979) The author must state whether the form is T otal or U nionized; T is the default for ammonia and hydrogen sulfide papers that do not state whether total or unionized concentrations are reported. TOTAL ( T ): The dissociated, charged form of nitrogen or hydrogen related chemicals. This can take on numerous forms, e.g.; ammonium (NH + 4 ), nitrite (NO 2 ), etc. (Rand and Petrocelli, 1985).T is the default for publications that do not state whether Total or Unionized concentrations are reported. UN-IONIZED ( U ): The undissociated, uncharged form of ammonia or hydrogen sulfide. The ammonia molecule, NH 3 , is the unionized form. (In aqueous solution, ammonia assumes an equilibrium between NH 3 and NH + 4 .) The NH 3 is the toxic entity of the ammonia compound (Rand and Petrocelli, 1985).



### Ionic Fraction
For ionizing substances (e.g., metals, ammonia) , report the dose as the ion if the concentration presented by the authors is report ed as based on the ionic form of the compound (e.g., organotin as Sn ). Code the appropriate ionic symbol in the ION data field (see Appendix O for ion codes). If concentration is based on the total compound, code 'NR' in this field. For non-ionizing substances, code 'NR' in this field.
Chemical Analysis Method (M/U)

The M/U data field identifies whet her nominal or quantified exposure dose values were reported by the author(s). For the specific exposure level, r eport whether toxicant and/or carrier concentration was measured (M) or calculated/nominal/ unmeasured (U) (see Appendix P for codes and

### TERRETOX Coding Guidelines

MED Ecotoxicology Database SOPs June 2009 23 definitions). When it is not cl ear whether reported concentrations are measured, calculated or unmeasured, record as Not Reported (NR). For instances where some treatment ( dose) levels are measured and some are unmeasured/nominal, and all unmeas ured/nominal concentrations are reported, report the unmeasured/nominal concentrations for each treatment level so that the range of concentrations is consistent and monotonical. Denote all of the concentration analysis methods as > X, = signifying that a mixture of measured and unmeas ured values was reported. Measured concentration data are selected over nominal concentration results, except when the presentation of the results is more complete wit h the nominal concentration data vs the measured data (i.e., lumping records vs. separate test results).
Remark Number/Remarks (RN)
When there are remarks for a specific test, the REMARKS field as well as the Remark Number RN (remarks number) data field, will be coded. Remarks ar e identified by the coding field abbreviation listed in Appendix EE. The Remark Number (RN) field is used to link the remarks associated with each specific test. Each unique Remark is assigned a Remark Number, and only one Remark Number is used per result entry. Use an independent unique Remark Number for each section of the database, i.e., do not carry over Remarks or Rema rk Numbers from the Exposure section to the Results section. Refer to Tables 3 and 5 for coding examples.

General Comment
General information about the exposure such as any specific methodology or techniques used is recorded in the REMARKS data field with the Other E ffect (OEF) identifier. General information about the test may include names of other chemicals that were tested but were not coded for TERRETOX, results are not provided, effects that have been reported but are not lin ked to a dose, effects that are reported but are not applicable to TERRETOX (e.g. in vi tro studies, selectivity ratios, acute to chronic ratios), or effect modifiers such as c hanges in soil pH, temperature or humidity.



## 5. Results Information

This section is used to record observed effect s for each control and dose level reported for the specific test. The Dose ID and Dose No. is carri ed forward from Exposure Information. Information specific to the observed response includes the sample number and sample unit, exposure duration, descriptors of the effect observed, the respons e site, and a quantitative measure of the response. Refer to Table 6 for specific fields included in this section of the TERRETOX Coding Sheet. Terrestrial data will be encoded in the following manner (Dose-Response data will only be coding for benchmark studies agreed upon by EPA): 1. All author reported endpoints (from graphs, text and/or tables) will be encoded 2. NOEC/LOEC data points from statistically analyzed data at all measurements (i.e., author does not clearly identify these as NOEC/LOEC, but sig/nosig designations have been made), durations and response sites will be coded. These will be noted as "Reviewer Assigned" [NOTE:Positive/beneficial effects (e.g. increase in growth over control, decrease in injury over control, increase in abundance over control) that are statistically significant are coded as NOEL/NOECs. A remark in the EE REMARK field should state that the effect was a statistically significant increase or decrease over control, which ever is applicable.]

### TERRETOX Coding Guidelines

MED Ecotoxicology Database SOPs June 2009 24 3. NR-ZERO and NR-LETH will be coded for aquatic and terrestrial studies following procedures outlined in the curr ent AQUIRE coding guidelines 4. Data that is not statistically analyzed will be coded as a one line data marker, ranging all concentrations, durations and response sites per effect. 5. Data that is graphed and not statistically analyz ed or data that is qualitative will be coded as a one line data marker, ranging all concentrations, durations and response sites per effect. 6. If the data encoded by a special project is not encompassed by the rules listed above (e.g. data from special projects that used the 10% change to assign endpoints), the data will be coded as follows: For data reviewed by a special project prior to ECOTOX coding, not currently in the special projects database, the special project c oder will note that a 10% endpoint was coded. For data already in a Special project database, CSC will extract a file of endpoints (noted by 10% text) in the endpoint comments (TRV) or statistic field (PCB) and examine the data encoded by the special project For both of these cases, CSC will add an entry ( non-endpoint) to the database noting that a of 10% change occurred at the specific duration or concentration in the remarks field (e.g., 10% TRV). If data is reported for a Terrestrial MAMMAL or AVIAN receptor OR if the Reference has passed the Acceptability criteria for TRV or Ec oSSL data (plant, invert, bird, mammal) CSC will encode all dose response data, in addition to the above rules General Information on Results Coding:
Coding Data Similarly Presented Often data are reported as individual measurements as well as a mean or range for these values. Report individual test results only. However, if both raw data and percentages, e.g. number survived and % survival, are reported, both values are coded. An exception to this coding procedure occu rs when data reported for individual endpoints are graphed and mean/median data are explicitly reported. For example, when replicate LC50s are reported on graphs and the mean and median LC50s are reported in the text, code both the graphed and textual data (remark on data points taken from the graph, and also note mean LC50 or Median LC50 in the comments). Graphed Data Data points derived from a graph are represented in TERRETOX as an approximate value, a range around the specific data point or as a range for all of the represented values. The values taken from the graph must be ranged using the author = s X- and Y- axis values. Do not interpolate values that lie between axis values. A result remark is added to the observed response field denoting that the data were extracted from a graph. Graphed data are coding one of the following ways: 1. Calculated endpoints (e.g. LC50s, BAFs) are always coded separately. 2. Statistically analyzed graphed data that is graphed is coded as denoted in Example 1a & b. For Example 1a (coding dose response data for papers from the TRV/EcoSSL documents) , Data trends should be examined (i.e. areas where the exposure changes from not significantly different to signifi cantly different from the control values) and coded. The maximum number of result records that can be coded from statistically analyzed graphed data is 2n-1, where n is the number of concentrations including the control. If all data points for a single dose are significant, non-significant or have multiple significance (i.e. no clear response) combine all durations into a single record (see Example 1, doses 2D, 4D and 7D). If a dose presents a clear trend (e.g. non-significant for t he first three weeks and significant for the last two weeks) code two results for the dose. Combine the durations for t he first three weeks and code the Statistical Significance as

A nosig @ for the first result and combine the durations for the second two weeks and code the Statistical Significance as

A sig @ . Code records similarly if part of the dose is significant or not significant and the other part has multiple significance

### TERRETOX Coding Guidelines

MED Ecotoxicology Database SOPs June 2009 25 (See Example 1, doses 3D, 5D and 6D. For Example 1b (General ECOTOX coding) reviewer assigned endpoints should be coded at each duration, no dose response data encoded unl ess the paper is from the TRV/EcoSSL documents. 3. For data that is not statistically analyzed, two lines ar e coded, one line for the control and a second line for all doses. For the ranged doses, the exposure number would be coded as #R (# representing the next dose level) and the exposure concentrations would be ranged in the Dose data field. See Example 2 for the coding of non-analyzed data.

### TERRETOX Coding Guidelines

MED Ecotoxicology Database SOPs June 2009 26 Example 1. Statistically Analyzed Data



## Rat weight (g)

800

700 0 ppm 1 ppm 600

Terretox Results Coding for Example 1 a (For dose response coding, i.e. data from the TRV/EcoSSL documents)

OBSERVED RESPONSE VALUE/UNIT ID SMP #

UNITS OBSERV

DUR EFFECT MEAS

MENT END

PNT R STAT LEVEL P

R SITE

MEAN RANGE SM VALUE UNIT D

W

% REMARK

1C NR OR 1-5 w GRO WGHT NR -1 NR NR P W O NR ~100/ - ~700/ NR NR g NR 1. RVALUE/ FROM GRAPH//

2D NR OR 1-5 w GRO WGHT NR -1 nosig p<0.05 P W O NR ~100/ - ~700/ NR NR g NR 1. RVALUE/ FROM GRAPH//

3D NR OR 1-3 w GRO WGHT NR -1 nosig p<0.05 P W O NR >100/ - <500/ NR NR g NR 1. RVALUE/ FROM GRAPH//

3D NR OR 4-5 w GRO WGHT NR -1 mult P<0.05 P W O NR >500/ - <700/ NR NR g NR 1. RVALUE/ FROM GRAPH// 

## 0 100 200 300 400 500

1 week 2 week 3 week 4 week 5 week 5 ppm 10 ppm 50 ppm 100 ppm 500 ppm 

# *

*

* * 

# * * *

* *

* 

# * * *

### TERRETOX Coding Guidelines

MED Ecotoxicology Database SOPs June 2009 27 OBSERVED RESPONSE VALUE/UNIT ID SMP #

UNITS OBSERV

DUR EFFECT MEAS

MENT END

PNT R STAT LEVEL P

R SITE

MEAN RANGE SM VALUE UNIT D

W

% REMARK

4D NR OR 1-5 w GRO WGHT NR -1 mult P<0.05 P W O NR >0/ - <700/ NR NR g NR 1. RVALUE/ FROM GRAPH//

5D NR OR 1-3 w GRO WGHT NR -1 nosig p<0.05 P W O NR >100/ - <500/ NR NR g NR 1. RVALUE/ FROM GRAPH//

5D NR OR 4-5 w GRO WGHT NR -1 sig p<0.05 P W O NR >400/ - <~700/ NR NR g NR 1. RVALUE/F ROM GRAPH//

6D NR OR 1-3 w GRO WGHT NR -1 mult p<0.05 P W O NR >100/ - <500/ NR NR g NR 1. RVALUE/ FROM GRAPH//

6D NR OR 4-5 w GRO WGHT N -1 sig p<0.05 P W O NR ~400/ - <~500/ NR NR g NR 1. RVALUE/ FROM GRAPH//

7D NR OR 1-5 w GRO WGHT NR -1 sig p<0.05 P W O NR >0/ - <~400/ NR NR g NR 1. RVALUE/ FROM GRAPH//



### Terretox Results Coding for Example 1 b (General ECOTOX coding)

OBSERVED RESPONSE VALUE/UNIT ID SMP #

UNITS OBSERV

DUR EFFECT MEAS

MENT END

PNT R STAT LEVEL P

R SITE

MEAN RANGE SM VALUE UNIT D

W

% REMARK

8E NR OR 1 w GRO WGHT NOEL -1 nosig p<0.05 R WO NR 100/ - NR NR ppm NR 1. RVALUE/ FROM GRAPH//

8E NR OR 1 w GRO WGHT LOEL -1 sig p<0.05 R WO NR 500/ NR NR ppm NR 1. RVALUE/ FROM GRAPH//

8E NR OR 2 w GRO WGHT NR -1 MULT p<0.05 R WO NR 1-500/ NR NR ppm NR 1. RVALUE/ FROM GRAPH//

8E NR OR 3 w GRO WGHT NOEL -1 nosig p<0.05 R WO NR 100/ - NR NR ppm NR 1. RVALUE/ FROM GRAPH//

8E NR OR 3 w GRO WGHT LOEL -1 sig p<0.05 R WO NR 500/ NR NR ppm NR 1. RVALUE/ FROM GRAPH//

8E NR OR 4 w GRO WGHT NOEL -1 nosig p<0.05 R WO NR 1/ NR NR ppm NR 1. RVALUE/F ROM GRAPH//

8E NR OR 4 w GRO WGHT LOEL -1 sig p<0.05 R WO NR 5/ NR NR ppm NR 1. RVALUE/ FROM GRAPH//

8E NR OR 5 w GRO WGHT NOEL -1 nosig p<0.05 R WO NR 10/ NR NR ppm NR 1. RVALUE/ FROM GRAPH//

### TERRETOX Coding Guidelines

MED Ecotoxicology Database SOPs June 2009 28 OBSERVED RESPONSE VALUE/UNIT ID SMP #

UNITS OBSERV

DUR EFFECT MEAS

MENT END

PNT R STAT LEVEL P

R SITE

MEAN RANGE SM VALUE UNIT D

W

% REMARK

8E NR OR 5 w GRO WGHT LOEL -1 sig p<0.05 R WO NR 50/ NR NR ppm NR 1. RVALUE/ FROM GRAPH//



### Example 2. Non-analyzed Data

Rat Weight (g)

0 100 200 300 400 500 600 700 800

1 week 2 week 3 week 4 week 5 week 0 ppm 1 ppm 5 ppm 10 ppm 50 ppm 100 ppm 500 ppm



### Terretox Results Coding for Example 2

OBSERVED RESPONSE VALUE/UNIT ID SMP #

UNITS OBSERV

DUR EFFECT MEAS

MENT END

PNT R STAT LEVEL P

R SITE

MEAN RANGE SM VALUE UNIT D

W

% REMARK

1C NR OR 1-5 w GRO WGHT NR -1 NR NR P W O NR ~100/ - ~700/ NR NR g NR 1. RVALUE/ FROM GRAPH//

8R NR OR 1-5 w GRO WGHT NR -1 NR NR P W O NR ~100/ - ~700/ NR NR g NR 1. RVALUE/ FROM GRAPH//

Species Data Data may be reported for an individual species as well as for a community or population. For example an author may report that biomass for an earthworm, Eisenia fetida, has decreased and that the invertebrate population biomass has increased. Report the measurements and endpoints as reported by the author for both the individual species and the species group.

### TERRETOX Coding Guidelines

MED Ecotoxicology Database SOPs June 2009 29 Dose ID & Dose No

This is the same Dose ID and Number as recor ded for each treatment level under the in Exposure Information. Transcribe the ID and Dose Number for each treatment level.
Sample Number (SMP#) and / Units
The sample number reflects the sample size (e .g., 10 embryos) that the observation or response value is based on at each exposure level. For endpoints based on calculations (e.g. LD50, NOEL, etc.) rather than individual dose measurements, the sample number will be coded as 'NR'. Code 'NR' if no information about the observed sample has been reported. Sample units correspond to the sample number; i.e ., the unit on which the measurement or endpoint is based (see Appendix Q for applicable codes). Code ' NR' if the sample unit is not reported.

EXAMPLE: 

### For a sample size of 190 eggs, the sample unit is eggs (EG); therefore, if the effect measurement is HTCH, and the observation response value is 90%, then 90% of 190 eggs hatched.

Note: For generational studies and measurements based on the progeny , note F1, F2, etc. in the sample units field.

Note: If a sample number is not provided, but a "unit" is, always enter the unit in the sample units field.

Note: A FieldN test scenario involves exposing plots or sample areas, in addition to specific test organisms. Usually the number of exposed organisms is unknown. The number of plots or sample areas is coded as '#/EU = (the number of experimental units) in Results Information rather than in the Exposure Information Sample Number field. See Table 6 for coding examples 

### Observation Time Duration (OBSRV DUR)
The Observation Duration reported includes exposure ti me plus any additional days up to the time at which the response to the toxicant was observed. It does not include pre-treatment time. In the example in Table 4, the observation was made on day 11; therefore, the obser vation duration time is 9D. If the observation time is not reported or unable to be explicitly determined, code as less than or equal to (<=) the exposure duration. NR s hould not be coded in this data field. Observations during the pretreatment time are r eported as negative values. For the Table 4 example, the observation time for a pretreatment sample co llected on day 2 of the pr etreatment period would be recorded as -1D. Report as > -x = any pretreatment response observations for which time is unknown. In some cases a biological time is used, such as an observation time reported as A until hatch @

,

A growing season @ or A after the nth egg has been laid @ . Use the code from Appendix I that best describes the author's words in duration units data field. > NA = will be coded as the duration mean value for all biological time durations (e.g . OBSERVATION MEAN: NA, OBSERVATION UNIT: HT; corresponds to an until hatch duration), unless a numeric value is substituted (e.g. OBSERVATION MEAN: 2, OBSERVATION DOSE: HV; corresponds to a 2 harvests duration). However, references to time such as A observed at end of the study period @ are not coded; such papers are rejected as having no observation duration.

Note: In test scenarios where incubation times are reported, e.g .. enzyme fixation assays, be careful to report the toxicant exposure time not the assay incubation time.

Note: In test scenarios that involve generational studies, the observation duration times are reported from the time the parents were exposed. For example the parents were exposed for 10 months prior to mating, and the progeny was born 2 months later, the observation duration for both the adult REP PROG effect and for the juvenile DVP ABNL is 12 months.

### TERRETOX Coding Guidelines

MED Ecotoxicology Database SOPs June 2009 30 The exposure duration would be the same for both - 10 months . The only difference between the two effects is in the sample unit. The sample unit for the adult effect w ould be 'AD' and for the juveniles it would be 'F1'. Table 6 . Results Information Coding Examples

Dose

ID

&

No. SMP

#

UNIT OBS

DUR EFCT MEAS END

PT R S

T

A

T L V L P

R S

I

T

E OBSERV.RESP

VALUE/UNIT

X Range SM Value Unit DW % %

L

P

D R

A

N

K R

N REMARKS

1C 10 F 5h MOR MDTH NR -1 N R N R N R NR 11.5 SD 7.8 d NR N R - 1 1 RVALUE/ from graph//

2 MSMT/ asymptotic level//

1C A @ A ACC RSDE A -1 " A A WO 1245 ug/g W

25 42 - 2

3V A @ A MOR MDTH A -1 " A A NR 15.8 SD 5.9 d NR N R - 1

4D A @ A A A A -1 " A A A 12.8 SD 7.6 d A A - 1

5D A @ A A A A -1 " A A A 15.6 SD 5.5 d A A - 1

5D A @ A ACC RSDE A -1 " A A WO 1459 ug/g W

33 44 - 2

6D A @ A MOR MDTH A -1 " A A NR 16.6 SD 8.0 d NR N R - 1

1C 10 EU 1-5d REP PROG NR -1 N R N R N R NR 680 (645-690) eg/d NR N R - N R

2C A A A A A -1 " A A A 983 (825-1012) eg/d A A - N R

3V A@ A A A A -1 " A A A 259 (243-272) eg/d A A - N R

4D A A A A A -1 SI G <

. 0 5 P A 246 (232-257) eg/d A A - N R

### TERRETOX Coding Guidelines

MED Ecotoxicology Database SOPs June 2009 31

Dose

ID

&

No. SMP

#

UNIT OBS

DUR EFCT MEAS END

PT R S

T

A

T L V L P

R S

I

T

E OBSERV.RESP

VALUE/UNIT

X Range SM Value Unit DW % %

L

P

D R

A

N

K R

N REMARKS

5D A A A A A -1 " A A A 255 (242-267) eg/d A A - N R

Data for REP/PROG would be continued for dose levels 3 and 5

8E NR NR 20d MOR MORT LD50 -1 N R N R P NR 9.8 (5.6 -11.2) d NR N R - 1



### When coding the endpoints LTxx or ETxx, which ar e based on the time it takes to get a XX% response, code the associated effect response time and not the total exposure time in the OBSERVED DURATION field.
Example 1: A test with earthworms is run for 10 hours, but it takes 6 hours for the "T1/2" duration which is the time to take 50% to burrow. The author reported T1/2 endpoint is similar to an ET50 endpoint, therefore a reviewer assigned ET50 is coded.

Code: Assigned Endpoint: R Endpoint: ET50 Effect: BEH Measurement: BBBH Observation Duration: 6 h

Example 2: A 4 week study is run with quail and reports mortality. The author reports the LT50 value at 10 mg/kg, 20 mg/kg and 30 mg/kg as 3.5 weeks, 2 weeks and 1 week, respectively.

Code: Assigned Endpoint: P Endpoint: LT50 Effect: MOR Meas urement: MORT Duration: 3.5 wk Concentration: 10 mg/kg Assigned Endpoint: P Endpoint: LT50 Effect: MOR Measuremen t: MORT Duration: 2 wk Concentration: 20 mg/kg Assigned Endpoint: P Endpoint: LT50 Effect: MO R Measurement: MORT Duration: 1 wk Concentration: 30 mg/kg


### When observed duration values are extrapolated from a graph the val ues can be approximated even when the mark does not appear on the graph. For exampl e, if the value on the graph is in the middle of two marks namely 10 and 14 days, the OBS DUR 

### can be coded as A 12 d @ with a remark that it is from the graph: OTIME/from graph//

Effect
Ecotoxicology is the study of the A toxic effects of natural or artificial substances on living organisms (e.g. fish, birds, plants) ... @ The effects may manifest at various levels of organization from subcellular through individual organisms to communities and ecosystems. Effects may be both positive and adverse; toxicology focuses on the adverse effe cts. Adverse effects include short-term and longterm lethality and sub-lethal effects such as changes in behavior, growth, development, reproduction, uptake and elimination, and ti ssue structure. (Rand 1995)

### TERRETOX Coding Guidelines

MED Ecotoxicology Database SOPs June 2009 32 In TERRETOX, effect groups include accumulation, behav ior, biochemistry, cellular, growth, mortality, physiology, population, reproduction and ecosystem (s ee Appendix R for definitions). Within each effect group, the observed effect must be quantified in a reproducible way (e.g. visual rating or scoring). In TERRETOX, two mechanisms are used to represent the observed effect: measurements and endpoints. Measurements include quant itative observations that de scribe and evaluate biological responses to toxicants, while endpoints are based on ca lculations derived from statistical analysis of the observations. Therefore, while measurements are direct biological observations, endpoints provide a statistical comparison of responses to toxicants. Coding criteria for each of these mechanisms is described below, directly fo llowing the General Notes section. The > General Notes = section provides guidelines for extracting e ffect/ measurement data from the publication.
Effect Measurement (MEASMENT)
Generally, A measures @ or A measurements @ are variables used to aid in the interpretation of the degree of response to a toxicant by an organism. For example, measures of behavioral effects in TERRETOX include general behavioral changes (BEH GBHV), changes in feeding activity (FDB FDNG), and stimulus avoidance (AVO STIM). Appendi x S lists the measurements currently used for each of the effects in the TERRETOX database. If mo re than one organism is measured in either an artificial (pot, cage, aquaria, etc.) or natural popul ation (field, water body, etc.),reviewing staff will code the effect and measurement as POP BMAS (population biomass). For instances where there is an individual biomass (one organism, artificial or natural), reviewing staff will code GRO BMAS (growth biomass). Endpoint/Result Set (R)/ Stat/Level/A ssigned (PR) (ENDPT/STAT/Level/P or R)
Endpoint (ENDPT)
An endpoint is a value derived from statistical analysi s or calculation of a specific measurement, or series of measurements, made during the tes t. Endpoints may be classified as measurement endpoints or assessment endpoints. Assessment endpoint s refer to environmental parameters such as population, community or ecosystem measurement s, e.g., growth rates or sustainable yields. Measurement endpoints refer to specific variables that are used to evaluate the assessment endpoints, such as diversity or evenness. (Hoffman et.al. 1995; US EPA 1996) 

### The ECOTOX databases utilize assessment and measurement endpoints which quantitatively represent the response(s) of a given individual, popul ation, or community to the effects of a toxic agent. Appendix T lists and defines endpoints used in TERRETOX. For each endpoint, effect and measurement must also be coded. Refer to Table 6, Results Information, for coding examples. For some endpoints, linkage to an exposure dose, and therefore an Exposure Dose Number, is especially important. These endpoints include BAFs or time associated endpoints such as LTxx and ETxx and TKNO. However, endpoints that are not linked to a specific concentration, e.g., LDxx, are not associated with an exposure number because the observed result is based on a calculated rather than an observed dose. These endpoints are linked wit h a placeholding Dose ID and Dose Number. The linkage is noted by E in the DOSE ID dat a field and the associated DOSE NO. in the EXPOSURE information. Refer to Table 5, Re sults Information, for coding examples. In contrast, NOEC/NOELs and LOEC/LOELs are the endpoints used to represent a statistically significant range within the tested concentrations. The NOEC/NOEL is the highest tested concentration having no statistically significant adv erse effect and the LOEC/LOEL is the lowest tested concentration having a statistically significant adverse effect. (Rand 1995) 

### NOEC/NOELs and

### TERRETOX Coding Guidelines

MED Ecotoxicology Database SOPs June 2009 33 LOEC/LOELs are also linked by the place holding exposure number, as indicated in the previous paragraph. Note: Positive/beneficial effects (e.g. increase in growth over control, decrease in injury over control, increase in abundance over control) that are statistically significant are coded as NOEL/NOECs. A remark in the comments field should state that the effect was a statistically significant increase or decrease over control, which ever is applicable. For endpoints of ETXX or LTXX, code both the observation duration and the observed response value with the same values. On occasion, authors will report LD50 information in the methods section of their publication, without reporting any accompanying test procedure information. These test results are coded if enough information is provided by the authors to verify t hat the value(s) were not published elsewhere, and that the study meets all five minimal criteria for acceptance . The term "Methods LD50" is coded in the EE REMARK.
Companion Endpoint(s) (COMPEP)
If the endpoint that is being coded has a A companion endpoint(s) @ record that is also being coded, group the records by coding a numeric value in the following format: COMPEP/number//. For each group of companion endpoints within a test use the same number. Unique companion endpoint groups are defined by the same effect, measurem ent, response site and duration. For example:
Example 1: The NOEC, LOEC and MATCs are reported fo r Growth - Length, Growth - Weight and Mortality - Mortality, the Growth Length endpoints are companion endpoints group 1, the Growth Weight are companion endpoints group 2 and the Mortality Endpoints are companion endpoints group 3. The data is coded as: Endpoint: NOEC Effect: GRO Measurement: LGTH COMPEP/1// Endpoint: LOEC Effect: GRO Measurement: LGTH COMPEP/1// Endpoint: MATC Effect: GRO Measurement: LGTH COMPEP/1//

Endpoint: NOEC Effect: GRO Measurement: WGHT COMPEP/2// Endpoint: LOEC Effect: GRO Measurement: WGHT COMPEP/2// Endpoint: MATC Effect: GRO Measurement: WGHT COMPEP/2//

Endpoint: NOEC Effect: MOR Measurement: MORT COMPEP/3// Endpoint: LOEC Effect: MOR Measurement: MORT COMPEP/3// Endpoint: MATC Effect: MOR Measurement: MORT COMPEP/3// 

### If an endpoint does not have a companion endpoint code NR. The numbering of companion endpoints starts from 1 again when a new TID is used. For example:
Endrin (TID 1) Endpoint: NOEC Effect: GRO Measurement: LGTH COMPEP/1// Endpoint: LOEC Effect: GRO Measurement: LGTH COMPEP/1// Endpoint: MATC Effect: GRO Measurement: LGTH COMPEP/1//

Heptachlor: (TID 2) Endpoint: NOEC Effect: GRO Measurement: WGHT COMPEP/1// Endpoint: LOEC Effect: GRO Measurement: WGHT COMPEP/1// Endpoint: MATC Effect: GRO Measurement: WGHT COMPEP/1//

Methoxychlor (TID 3) Endpoint: NOEC Effect: MOR Measurement: MORT COMPEP/1//

### TERRETOX Coding Guidelines

MED Ecotoxicology Database SOPs June 2009 34 Endpoint: LOEC Effect: MOR Measurement: MORT COMPEP/1// Endpoint: MATC Effect: MOR Measurement: MORT COMPEP/1// 

### Result Set (R)
This field is used to link effects and endpoints toget her for data output display. The default entry for this field is -1. Enter positive integers for each specific effect/endpoint linkage. For example, a mortality data table and an LD50 as well as a body we ight growth table and an EC50 are reported. Code all of the mortality data t able and the LD50 with a '1' in the R data field and code '2' in the R data field for the growth and EC50 data. If there are endpoints without data or vice versa, code '-1' in the R data field. See Table 6, for examples.
Significance (STAT)
The significance or STAT data field is coded with SIG or NOSIG if the author has presented statistical analysis of the test result as compared to the contro ls. As a general rule, if statistics are presented in the publication, assume that the exposure treatments were compared to the control. Statistical tests that measure differences betw een treatments are not coded. See Tables 7 and 8 for coding examples. When statistical comparisons are pr esented for multiple controls (e.g., statistics in relation to a standard and solvent control), both sets of results are coded. In these instances, note the specific type of control used in the statisti cal analysis in the Remarks section.
The STAT data field is coded as "NR" for reco rds having an endpoint of LCxx, ECxx, LTxx, BAF, ETxx, ICxx, LDxx. For NOEC/NOEL, LOEC/LOEL and statistically analyzed effects results without endpoints, code the significance as reported by the aut hor(s), or 'NR' if statistical results are not presented in the publication. If the author states that there is a 'statistically significant' increase or decrease in an observed effect (whether or not they report the st atistical method used) but does not report a level of statistical significance level or identify a method of statistical analysis, code 'SIG' or 'NOSIG' and 'NR' in LEVEL field. If the author states there is a significant in crease or decrease in an observed effect but does not say it is "statistically significant," code 'NR' in the STAT data field.

Note: For concurrent control results, there is no STAT or LEVEL defined. Statistical significance is compared to the control values.

Note: Coding statistics from least square differences (LSD). If a paper presents data in the following format, determine the statistical significance by the following calculation.

Dose Response

Control 1 2.4

Dose 2 4.6

Dose 3 4.8

Dose 4 6.7 LSD(0.05) 1.1

Subtract or add the LSD value (1.1) to the control value to get the lowest value that is significantly different. In this example, anything above 3.5 or below 1.3 is a significant value at the p=0.05 level. Therefore, all doses are significantly different from the control because they are all greater than 3.5 (Pers. Comm., R. Regal, UMD Statistics Dept., 1999).

### TERRETOX Coding Guidelines

MED Ecotoxicology Database SOPs June 2009 35 Level
The level of significance (e.g. test statistic) is coded when the author has reported statistical analysis in the test result. Terminology for significance le vel may be presented as: p =; p< or alpha value; χ 2



### . The terminology is equivalent and is generally in t he range of 0.001 to 0.10. See Tables 7 & 8 for coding examples.

The LEVEL field is coded as "NA" for records having an endpoint of MATC, LCxx, ECxx, LTxx, BCF, ETxx, ICxx, LDxx, LETC, BCFD, NR-LETH, and NR-ZE RO. However, when the confidence level is other than 95%, the level is coded as reported.

. If an author reports two levels of statistical significance (e.g. 0.05 and 0.01, 5% and 1%) for the same data point and the significance is different at the levels, the level at which the data is first statistically significant is coded. For example: Premature Molting for 15 days is analyzed at 5% and 1%

Concentration (ng/larva) % premature molting 5% , 1% * Code

Control 0 d , C

1 15.38 c, C

30 66.67 b, B

60 92.59 a, AB

100 90.00 a, AB

200 96.67 a, A LOEL at 5% significance at 1 ng/larva

* Same characters in a column indicate no significant difference 

### If an author reports two levels of statistical significance (e.g. 0.05 and 0.01, 5% and 1%) for the same data point and the significance is the same at both le vels, the most sensitive level (e.g. 0.01) at which the data is first statistically significant is coded. For example: Mortality for 9 days is analyzed at p=0.05 and p=0.01

Concentration (ng/larva) % Mortality P=0.05 , p=0.01 * Code

Control 6.3 b , B

25 6.4 b, B

50 24.6 a, A

100 20.2 a, A

150 32.2 a, A NOEL at p=0.01 at 25 ng/larva, LOEL at p=0.01 at 50 ng/larva

* Same characters in a column indicate no significant difference
If an author reports p values but the level at which the data ar e considered significant is not reported by the author, use the conservative significance level of P<=0.05 as a benchmar k. The ENDPOINT ASSIGNED is coded as R and a remark of “SIG ASSIGNED” is made on SIGNIFICANCE.

### TERRETOX Coding Guidelines

MED Ecotoxicology Database SOPs June 2009 36 Example: The paper reports gene expression of 9 genes at one concentration and gives p values but does not report the level that is considered significant. Using a benchmark leve l of significance of p<=0.05 the following endpoints are coded:

mRNA Name P VALUE ENDPOINT ASSIGNED ENDPOINT SIGNIF REMARKS

BHMT 0.57 R NOEL ANOSIG/ SIGNIF/SIG ASSIGNED//

ETIF 0.04 R LOEL ASIG/ SIGNIF/SIG ASSIGNED//

UCE 0.01 R LOEL ASIG/ SIGNIF/SIG ASSIGNED//

Tubulin 0.21 R NOEL ANOSIG/ SIGNIF/SIG ASSIGNED//
Trypsin 0.07 R NOEL ANOSIG/ SIGNIF/SIG ASSIGNED//

UBI 0.14 R NOEL ANOSIG/ SIGNIF/SIG ASSIGNED//

SAMS 0.32 R NOEL ANOSIG/ SIGNIF/SIG ASSIGNED//
Hcy a 0.72 R NOEL ANOSIG/ SIGNIF/SIG ASSIGNED//
Hcy b 0.31 R NOEL ANOSIG/ SIGNIF/SIG ASSIGNED//



### Paper/Reviewer Assigned Data (P or R)
The PR data field is used to identify the source of the effect or endpoint information. If the effect or endpoint was reported by the author in the publication a 'P' is coded in t he PR data field; if the effect or endpoint was assigned by the reviewer, an 'R' is coded. See Tables 7 & 8 for coding examples. Endpoints calculated by the author must be specific ally identified, i.e., LD50, LT50 or NOEL/NOEC (see Appendix T for endpoint codes and definitions). Reviewers will follow these guidelines in assigning endpoints:

1. B Y DEFINITION : If the author does not actually state that the value is an LD50 but states that A concentration x is the dose estimated to be lethal to 50% of the test organisms @ , the reviewer should code this as an LD50 endpoint because the author defines the LD50. Such a designation is accompanied by noting > R = in the PR data field. 2. When the author provides text which identifies a value as the A highest tested concentration having no statistically significant adverse effect @ , the reviewer should code this as a NOEL/NOEC ; the A lowest tested concentration having a statistically significant adverse effect @ is coded as a LOEL/LOEC . In both cases, the PR field will be coded as 'R' to reflect reviewer assignment of an endpoint. Because LOEL/NOEL va lues are assigned under very specific experimental and statistical conditions, TERRETOX reviewers will be assigni ng complementary NOELs or LOELs only when the author assigns either a LOEL or NOEL. 3. When the author provides statistical information, which designates concentrations as significantly different from the control, the reviewer will code this information as SIG or NOS IG. The reviewer will also report the level of significance in the LEVEL data p-value in the p-value field. 4. If the author presents biologically significant data which does not include statistical analysis because all of the animals died, the reviewer should not interpret this data as a non-monotonical response. Code the appropriate biologically significant endpoints as NOEC/LOEC values. For example:
Example 1

WHOLE BODY GROWTH
CONTROL 139 G 10 UG/ML 138 G 20 UG/ML 136 G 30 UG/ML 100 G* 40 UG/ML 95 G* 50 UG/ML ALL DEAD

### TERRETOX Coding Guidelines

MED Ecotoxicology Database SOPs June 2009 37 * = SIGNIFICANT AT 0.05 CODE: EFFECT: GRO (GROWTH) ENDPOINTS: NOEC @ 20 UG/ML MEASURE: WGHT (WEIGHT) LOEC @ 30 UG/ML
Example 2

% DEFORMITY
CONTROL 0% 10 UG/ML 0% 20 UG/ML 0% 30 UG/ML 0% 40 UG/ML 100% 50 UG/ML ALL DEAD

CODE: EFFECT: DVP (DEVELOPMENT) ENDPOINTS: NOEC @ 30 UG/ML MEASURE: DFRM (DEFORMATION) LOEC @ 40 UG/ML

Table 7. Coding Statistical and Endpoint Data Directly from a Table.

A The data from our experiments is shown in the following table. @

# Conc ug/g Survival % Stat sig p<0.05 Calc Endpt

C1 0 97 NR NR

D2 7 97 NOSIG NR

D3 15 75 NOSIG NOEL

D4 30 26 SIG LOEL

D5 50 0 SIG NR

In this example, the raw data table is coded as follows ( only for TRV/EcoSSL data as dose response, otherwise only the NOEL/LOEL data is encoded):

DOSE EFFECT MEASMENT ENDPT R STAT LEVEL P R

C1 MOR SURV NR 1 NR NR P

D2 MOR SURV NR 1 NOSIG p<0.05 P

D3 MOR SURV NR 1 NOSIG p<0.05 P

D4 MOR SURV NR 1 SIG p<0.05 P

D5 MOR SURV NR 1 SIG p<0.05 P

E6 MOR SURV NOEL 1 NOSIG p<0.05 P

E6 MOR SURV LOEL 1 SIG p<0.05 P

Table 8 . Coding Statistical Data Directly from a Table with a Reviewer Assigned Endpoint.

A The data from our experiments, in Table Z, shows that the concentration that had no observable effect on mortality was 7 ug/g. @

Table Z: Mortality of Eisenia fetida to copper

### TERRETOX Coding Guidelines

MED Ecotoxicology Database SOPs June 2009 38 # Conc ug/g Survival %

C1 0 97

D2 7 97

D3 15 75

D4 30 26

D5 50 0

In this example, the table is coded as follows (only for TRV/EcoSSL data as dose response, otherwise only the NOEL/LOEL data is encoded:

DOSE EFFECT MEASMENT ENDPT R STAT LEVEL P R

C1 MOR SURV NR 1 NR NR P

D2 MOR SURV NR 1 NR NR P

D3 MOR SURV NR 1 NR NR P

D4 MOR SURV NR 1 NR NR P

D5 MOR SURV NR 1 NR NR P

E6 MOR SURV NOEL 1 NOSIG NR R

E6 MOR SURV LOEL 1 SIG NR R

Standard methods recommend that when determining a NOEL/L OEL, at least three exposure concentrations be used (Menzer 1994 at 1406);. If the author uses only one exposure concentration A ND assigns a NOEL/LOEL or SIG/NOSIG result, a Remark noting A only conc tested @ will be coded. 

### Trend (TREND)
The observed or measured response trend as com pared to the control is coded when reported or graphically displayed. Report the trend as a remark in the format TREND/code//. When assigning a trend to a record, it should reflec t the measurement which may or may not reflect the effect. For example, when authors report a decreas e in survival; the effect is reported as MOR, and the trend is associated with the measur ement; i.e., decrease in survival.
Example: EFCT: MOR TREND: Dec MEASURE: SURV 

### The trend for BCF, LCxx, LTxx is coded as "inc". The trend for ECxx, NOEC, LOEC , and MATC will be either "inc", "dec", A chg @ or NR depending on the results of the test. In instances when a trend is non-monotonical code A chg @ . The trend is noted as a two or three letter code:

CODE : TREND :
INC increase
DEC decrease NEF no observed effect; e.g., when coding NR-ZERO the trend is NEF CHG no clear trend, results are variable (e.g. any combination of above trends listed) NR no trend reported or if no control response is reported then the trend is not able to be identified

Example: When a clear response, or lack thereof, is observ ed within an effect, it is coded as either INC, DEC, or NEF. The measurement used to evaluate the effect is reported in the MEASURE field, for example:

EFCT: GRO TREND: INC MEASURE: LGTH

### TERRETOX Coding Guidelines

MED Ecotoxicology Database SOPs June 2009 39 When measurements do not report quant ifiable data, data are combined into one record. If these data report multiple trends, code CHG in the TREND 

### field and report the individual trends in RESULT_REMARK 

### field as in the following example:
Example: EFCT: HIS TREND: CHG MEASURE: GHIS MSMT/EDMA,DISO,EHYP// 

### Effect % (EFCT %)

The EFCT% 

### is coded when the effect is reported as a percent change, e.g. percent of the total population or percent increase or decrease. If the author reports the number dead (i.e., A 5 of 20") do not recalculate as a percent.
Example 1: A 80% mortality @ EFCT: MOR TREND: INC EFCT %: 80 MEASURE: MORT
Example 2: A 25% survival @ EFCT: MOR TREND: DEC EFCT%: 25 MEASURE: SURV
Example 3: A 5 of 20 died @ EFCT: MOR TREND: INC EFCT%: NR MEASURE: MORT
Example 4: A 45% inc EFCT: ENZ TREND: INC EFCT%: 45 Measure: ATPA ATPase activity @ 

### If the percent effect is coded from a graph, code the percent values using a qualifier, ie. <, >, or ~, using only the graphical intervals reported on the gr aph. Place a slash in the field and code EFCT%/ from graph// in the RESULT REMARK 

### field. If the percent effect is graphed and is not clear enough to extrapolate, code A graphed @ in EFCT % 

### field. If the effect percent is not reported, the field is coded as NR. If the percent effect is presented as A xx% of the control @ , place a "/" in the EFCT % 

### field and code: EFCT %/xx% of control// in the REMARK 

### field.
Example: GRO WGHT Dec 20-30% of the control

EFCT: GRO MEASURE: WGHT EFCT%: / TREND: DEC Remarks: Efct%/20-30% of control// 

### For NR-ZERO and NR-LETH endpoints, the EFCT % 

### field should be coded as 0% and 100%, MORT respectively. When no stats and effect % are presented at the hi ghest concentration, the concentrations should be ranged and the EFCT % 

### should be presented as < the effect % from the paper.
Combining Effect Percent

When data for an effect are combined because a stat istical analysis was not applied and/or a clear dose response was not observed, and several percent effect values are presented, there are two different ways to report data. If the author reports the effect m easurement on a single parameter, the effect percent is reported as a range. Example 1: 30-75% mortality EFCT: MOR MEASURE: MOR TREND: INC EFCT%: 30-75 Example 2: 20-30% dec O2 consumption EFCT : PHY MEASURE: OXYG TREND: DEC EFCT%: 20-30 

### Response Site (SITE)
The specific site at which an effect measurement wa s observed is coded in the SITE data field, e.g. for residues (RSDE) recorded in the A liver, @ enter 'LI' in the SITE data field (see Appendix U for applicable codes). Response site is valid entry fo r GRO, AEG, CEL, PHY, DVP, GEN, REP, HIS,

### TERRETOX Coding Guidelines

MED Ecotoxicology Database SOPs June 2009 40 ENZ, BCM, HRM, INJ, MPH and ACC effect groups (see Appendix S for effect group and measurement codes). If a response site is not repor ted or not applicable, e.g. mortality, behavioral effects, code the site as Not Reported (NR). If data are presented without statisti cal analysis in a graph or figure, results for each measurement are combined by response site.
Observed Response Value/Unit:
Mean, Range, Statistical Method (SM), Value, Unit (Mean, Range, SM, Value, Unit) 

### Enter the greater than (>), less than (<), minus (-) or approximation (~)symbols, if reported, as used by the author(s) to describe the response value pr eceding the MEAN or RANGE data field entries. Report the mean or single observed response value, as reported in the publicat ion, in the MEAN data field. When individual response values are repor ted along with a sum of all values, report each individual response value as well as the sum/total value on separate result lines. Report the range or confidence (or fiducial) intervals (or limits) which are defined as CL (confidence limit), CI (confidence interval), FI (fiducial interval), or FL (fiducial limit) of the response value in the RANGE data field. The type of dat a stored in the RANGE data field will be identified in the SM data field (e.g., Data reported as a range (with a mean) or confidence interval (with an endpoint) will be specifically identified in the SM field. It is also assumed that the confidence interval is calculated at 95% and is noted in the LEVEL data field.

When the measurement unit includes a standard deviati on (SD) or standard error (SE), specifically identify these types of ranges in the SM data field. Report the numeric value of the standard deviation or standard error in the VALUE data field. Report the measurement unit which corresponds to the MEAN and/or RANGE entry in the UNIT data field (see Appendix N for standard units).

Refer to Table 6 in Results Information for coded examples. Table 6a. provides a standard deviation example, Table 6b. provides an example of a range, and Table 6c. provides a confidence interval example. Occasionally an author will report a concentration as a % or fraction of an LD50 value; e.g., either the sublethal concentration used was 10% of the 96-h LD 50" or 1/10, 1/15 and 1/20 of the LD50". Such concentrations may be recalculated and used as the effect concentration if the original LD50 concentration is provided in the publication. Flag the recalculation in the paper so that the calculation may be QAd and document the recalculation in the margin or on a blank page of the publication. Put a slash next to the concentration value and note in the remarks area: RVALUE/Recalculated//.
Weight at time of result (WTAT)
The body weight of the organism at the time of the observed effect response is reported. If multiple body weights are given throughout the test, report the we ight at the time the response is reported. If a range of weight is given for the organisms, report the range of weights. R eport the treatment group weight, if weights are given for both a treatment group and a control group. The data is coded as follows: WTAT/mean(min-max) unit//.

### TERRETOX Coding Guidelines

MED Ecotoxicology Database SOPs June 2009 41 Intake Rate (INTAKE)
Report the intake rate of the exposur e media (not the contaminant) at t he time of the result (or closest rate to that time) for diet studies (through food or drinking water) as reported by the author in the following format: INTAKE/value unit//.
Steady State (STST)

Record whether the residue/bioconcentration/bioaccumu lation data at the time of the result is at steady state or equilibrium using the following format: STST/YES// or STST/NO//. If the author does not report in the text that the data is at steady state or equilibrium but represents the data graphically, this information can be determi ned by the graphed data. If the residue or BCF data is presented in a graph and the data points flatten out and continue to flatten during the exposure phase, the point at which it flattens out is the steady state or equilibrium point. For Example:



# BCF values

12 h 0 10 20 30 40 50 60 70 80 90 100

24 h 48 h 72 h 96 h 120 h
10 ug/l

The steady state or equilibrium is reached at 48 hours. STST is coded as YES for the BCF values at 48, 72, 96 and 120 hours. If the author does not report this information code: STST/NR//.
Dry or Wet Weight (DW%)
Record whether the residue/bioconcentration/bioaccumulation or growth data ar e reported on a dry or wet weight basis in the DW% data field. If percent moisture is reported, record the percentage value also, e.g. W75%.
Percent Lipid (%LPD)
If percent lipid information is provided in the publicati on, record as a % value in the %LPD data field. If the data are not reported in the publication, code as 'NR'.
Rating (RANK)
Following evaluation by EcoSSL (Ecological Soil Sc reening Levels) Task Group reviewers, this field will be marked for each test result for each publicati on to indicate the Evaluation Criteria Score and

### TERRETOX Coding Guidelines

MED Ecotoxicology Database SOPs June 2009 42 selected benchmark value ranking for determining the Ec oSSL value. Prior to this, leave this data field blank.
Remark Number/Remarks (RN/REMARKS)
When there are remarks for a specific test, the RE MARKS field as well as the Remark Number (RN) field, will be used. The Remark Number field is used to link the remarks associated with each specific test result. Each unique Remark is assigned a Remarks Number and only one Remark Number is used per result entry. Use a unique Remark Number for each section of the database, i.e., do not carry over Remarks or Remark Numbers from t he Exposure Information to the Results Information sections. Remarks are preceded by the Remarks Number and identified by the field abbreviation listed in Appendix EE. Refer to Table 6 for coding examples.
General Coding Information
Q. What is encoded from a publication? A. All quantitative data are encoded from the publication. Each data poi nt from tables, text and graphs is coded. Graphical data may be coded as ranges (1 resu lt for the control and 1 result for all of the doses), unless statistical analysis is performed. Graphed data is reported by using <, > or ~ values. These values must be the values noted by the axis marks from the graph. If duplicate results are reported in text and tabular format, note in the ma rgin of the paper that the text information was coded from Table N. Non-quantitative data are noted in t he general remarks section.

Q. Are abstractors allowed to interpret results from publications? A. All information from a paper is abstracted us ing the author's terminology and numeric values. Exceptions to this include the expansion of exponential numbers and when the author's "words" match the standard definition effects and endpoints. If an endpoint is "interpreted" by an abstractor, it is noted by an 'R' in the ASSIGNED P/R data field.

Q. How and why are comments made? A. In general, comments are used to better define or capture the researcher's intent. THESE ARE USED SPARINGLY. Comments are linked to coded fields by an identifier in the appropriate comments field (Organism, Exposure, Soil, General or Results informa tion/remarks data fields). For example, a RESPONSE VALUE comment of median LC50 is located in Result remarks data field as Rvalue/median LC50//. Some comments are not link ed to a specific data field (i.e. exposure temperature or in vitro studies). These comment s are also noted in the appropriate comments field (i.e. exposure temperature in Exposure information and in vitro in the general remarks data fields).

Q. Is anything written in the original paper by the data abstractors? A. Abstractors should note any co mments about abstraction in the margins or on the tables/graphs of the original paper. This would include the Test ID Number for each unique test design, reason for data not being coded, LD50s outside of confidence interv als, errors between text and tables, or other anomalies.

### TERRETOX Coding Guidelines

MED Ecotoxicology Database SOPs June 2009 43 Q. What happens if an endpoint is outside the confidence interval/limit or text and tabular or text and abstract data points differ? A. The abstractor encodes only the endpoint value and notes that the range was not coded in the original publication. Textual information is used ov er all other data, unless the value is noted in another section of the paper. Then, the most frequent value is encoded.

### TERRETOX Coding Guidelines

MED Ecotoxicology Database SOPs June 2009 44 6. Supporting Materials

TERRETOX References
American Society for Testing and Materials. 1996. E 555-95 Standard Practice for Determining Acute Oral LD50 for Testing Vertebrate Control Agents. In: Annual Book of Standards, Section 11. Water and Environmental Technology, Vol. 11.05 Biological Effects and Environmental Fate; Biotechnology; Pesticides:109-110. [replaces E 555-75] American Society for Testing and Materials. 1996. E 857-87 Standard Practice for Conducting Subacute Dietary Toxicity Tests with Avian Species. In: Annual Book of Standards, Section 11. Water and Environmental Technology, Vol. 11.05 Biological Effects and Environmental Fate; Biotechnology; Pesticides:278-282. [replaces E 857-81] American Society for Testing and Materials. 1996. E 1062-86 Standard Practice for Conducting Reproductive Studies with Avian Species. In: A nnual Book of Standards, Section 11. Water and Environmental Technology, Vol. 11.05 Biological Effects and Environmental Fate; Biotechnology; Pesticides:418-428. Casarett, L.J., J.Doull, C.D.Klassen, and M.O.Amdur. 1986. Casarett and Doull = s Toxicology. The Basic Science of Poisons. Third Edition, Macmill an Publishing Co., Inc., NY [classic toxicology textbook] Hill, B.H. 1997. Aquatic plant communities for impact monitoring and assessment. Chapter Ten IN: W.Wang, J.W.Gorsuch, and J.S.Hughes (Eds), Plants for Environmental Studies, CRC Lewis Publishers: 277-305. [excellent overview] Hoffman, D.J. 1995. Wildlife Toxicity Testing. In: D.J.Hoffman, B.A.Rattner, G.A.Burton,Jr., and J.Cairns,Jr. (Eds), Handbook of Ecotoxicology. CRC Press, Inc., Boca Raton, FL:47-69. [good overview of wildlife toxicity testing methods] Hoffman,D.J., B.A.Rattner, G.A.Bur ton,Jr., and J.Cairns,Jr. (Eds) 1995. Handbook of Ecotoxicology. CRC Press, Inc., Boca Raton, FL [general refe rence with chapters on ecot oxicological effects, specific contaminant sources and effects, case histories, predictive ecotoxicology and risk assessment] Lewis, M.A. and W. Wang. 1997. Water quality and aquatic plants. Chapter Six IN: W.Wang, J.W.Gorsuch, and J.S.Hughes (Eds), Plants for En vironmental Studies, CRC Lewis Publishers: 142- 175. [excellent overview]
Menzer, R.E., M.A. Lewis and A.Fairbrother. 1994. Me thods in Environmental Toxicology. Chapter 38 In: W. Hayes (Ed), Principles and Methods of To xicology, Third Edition, Raven Press, NY:1391- 1418. [excellent overview of aquatic and terrestrial (plant and animal) methods] Merriam Webster (No Date). Merriam-Webster = s Collegiate Dictionary. [On-line dictionary]. Available: http://www.m-w.com/dictionary.htm [1999, 2000]. Morris, W. (Ed) (1979). The American Herit age Dictionary of the English Language (New College ed.). Boston: Houghton Mifflin Company. Parker, S.P. (Ed) (1994). McGraw-Hill Dicti onary of Scientific and Technical Terms (5 th 

### ed.). Ney York: McGraw-Hill, Inc. Schaeffer, D.J. 1991. A Toxicological Perspective on Ecosystem Characteristics to Track Sustainable Development. Ecotoxicol. Environ. Saf. 22:225-239. [pro vides a general overview of toxicological and ecosystem endpoints]

### TERRETOX Coding Guidelines

MED Ecotoxicology Database SOPs June 2009 45 US Environmental Protection Agency. 1988. Guidance Document for Conducting Terrestrial Field Studies. EPA-540/09-88-109, Office of Pe sticide Programs, Washington, DC US Environmental Protection Agency. 1986. Haza rd Evaluation Division, Standard Evaluation Procedure, Avian Reproduction Tes t. EPA-540/9-86-139, Office of Pest icide Programs, Washington, DC [general info, no specific requirements provided] US Environmental Protection Agency. 1986. Haza rd Evaluation Division, Standard Evaluation Procedure, Field Testing for Pollinators. EPA- 540/9-86-140, Office of Pesticides Programs, Washington, DC US Environmental Protection Agency. 1985. Haza rd Evaluation Division, Standard Evaluation Procedure, Honey Bee - Acute Contact LD50. EPA- 540/9-85-002, Office of Pesticides Programs, Washington, DC US Environmental Protection Agency. 1985. Ha zard Evaluation Division, Standard Evaluation Procedure, Honey Bee - Toxicity of Residues on Foliage. EPA-540/9-85-003, Office of Pesticides Programs, Washington, DC US Environmental Protection Agency. 1985. Haza rd Evaluation Division, Standard Evaluation Procedure, Wild Mammal Toxicity Test. EPA- 540/9-85-004, Office of Pesticide Programs, Washington, DC US Environmental Protection Agency. 1985. Haza rd Evaluation Division, Standard Evaluation Procedure, Avian Single-Dose Oral LD50. EPA- 540/9-85-007, Office of Pesticide Programs, Washington, DC US Environmental Protection Agency. 1985. Haza rd Evaluation Division, Standard Evaluation Procedure, Avian Dietary LC50 Test. EPA-540/9-85-008, Office of Pesticide Programs, Washington, DC

U.S. Environmental Pr otection Agency. 2009. MED Ecotoxicology Database Standard Operating Procedures (prepared by Computer Sciences Corporation), Mid-Continent Ecology Division, Duluth, MN US Environmental Protection Agen cy. 1982. Pesticide Assessment Guidelines Subdivision L Hazard Evaluation: Nontarget Insects. EPA-540/9-82-019, Office of Pesticides and Toxic Substances, Washington, DC US Environmental Protection Agen cy. 1982. Pesticide Assessment Gui delines Subdivision E, Hazard Evaluation: Wildlife and Aquatic Organisms. EPA-540/9-82-024, Office of Pesticide Programs, Washington, DC [overview of EPA standard methods for terrestrial and aquatic, laboratory and field, pesticide exposures] US Environmental Protection Agency. 1996. Pr oposed Guidelines for Ecological Risk Assessment, Risk Assessment Forum, EPA/630/R-95/002B, US EPA, Washington, DC [on order from US EPA 7/97) Wang,W., J.W.Gorsuch, and J.S.Hughes (Eds). 1997. Plants for Environmental Studies, CRC Lewis Publishers:563 pp. [excellent overview] W.B. Saunders Company (1994). Dorland = s Illustrated Medical Dictionary (28 th 

### ed.). Philadelphia: Harcourt Brace & Company.TERRETOX Coding GuidelinesTERRETOX LAB CODING SHEET (February ##, 2007)CHEMICALGRADEPURITYFORMULATIONCHARACTERISTICSRADIO LABTID_____________CAS NUMBER1. TEST2. Positive control/carrier3. Positive control/carrierREF #, AUTHOR, YEARTOTAL TESTSREVIEWERDATE //QA Date / /NameSpecies Scientific/Common NameRemarksDOSENO.DOSEIDNSEXDOSESMVALUEUNITIONM/URNSPECIES NUMBERORGANISMINFOORGANISM SOURCELIFESTAGE/AGEORG CHARACTERISTICSTEST LOCATIONEXPOSURE INFOEXPOSURE TYPECONTROL TYPENUMBER OF DOSESAPPL FREQUENCYEXPOSURE DURATIONSTUDY DURATIONGENERALREMARKSEXPOSURE MEDIASOIL INFOSOIL TYPESOIL TEXTURESA_______%SI_____%CL____________%WTINT/GEO/MEDIACECDOSES/LAT/MEDIA PHSTYPE/AP DATE/MEDIA ORGANIC MATTERHAB/AP SEAS/MEDIA MOISTURELOC/AP RATE/SOIL CONC MEASUREDDRY-WETWEIGHTMED Ecotoxicology Database SOPs June 2009 46TERRETOX Coding GuidelinesMED Ecotoxicology Database SOPs June 2009 47Reference Number__________________CAS#_________TID______________Observed Response Value/UnitD%RLOC#DOSENO/IDSMP #UNITSOBSERVDUREFFECTMEASMENTENDPNTRSTATLEVELPRSITEMEANRANGESMVALUEUNITSW%LPDTNGRNREMARKSWTAT/CONCTYPETREND/EFCT%STST/INTAKE/COMPEP/TEMP/WTAT/CONCTYPETREND/EFCT%STST/INTAKE/COMPEP/TEMP/WTAT/CONCTYPETREND/EFCT%STST/INTAKE/COMPEP/TEMP/WTAT/CONCTYPETREND/EFCT%STST/INTAKE/COMPEP/TEMP/WTAT/CONCTYPETREND/EFCT%STST/INTAKE/COMPEP/TEMP/WTAT/CONCTYPETREND/EFCT%STST/INTAKE/COMPEP/TEMP/TERRATOX CODING SHEET