United States
       Environmental Protection
       Agency
Office of Water (4305)
EPA-823-K-09-001
  August 2009
&EPA   AQUATOX (RELEASE 3)

        MODELING ENVIRONMENTAL FATE
          AND ECOLOGICAL EFFECTS IN
             AQUATIC ECOSYSTEMS
            VOLUME 1: USER'S MANUAL

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AQUATOX (RELEASE 3)

MODELING ENVIRONMENTAL FATE
  AND ECOLOGICAL EFFECTS IN
    AQUATIC ECOSYSTEMS
   VOLUME 1: USER'S MANUAL
       Jonathan S. Clough
           AUGUST 2009
   U.S. ENVIRONMENTAL PROTECTION AGENCY
         OFFICE OF WATER
    OFFICE OF SCIENCE AND TECHNOLOGY
        WASHINGTON DC 20460

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Disclaimer

This document describes the interface of the aquatic ecosystem model AQUATOX, Release 3.
Anticipated users of this document include persons who are interested in using the model,
including but not limited to researchers and regulators. The model described in this document is
not required, and the document does not change any legal requirements or impose legally
binding requirements on EPA, states, tribes or the regulated community. This document has
been approved for publication by the Office of Science and Technology, Office of Water, U.S.
Environmental Protection Agency. Mention of trade names, commercial products or
organizations does not imply endorsement or recommendation for use.
Acknowledgements

This model has been developed and documented by Dr. Richard A. Park of Eco Modeling and by
Jonathan S. Clough of Warren Pinnacle Consulting, Inc. under subcontract to Eco Modeling.  The
work was funded with Federal funds from the U.S. Environmental Protection Agency, Office of
Science and Technology under contract number 68-C-01-0037 to AQUA TERRA Consultants,
Anthony Donigian, Work Assignment Manager.  Integration of Interspecies Correlation Estimation
(Web-ICE) was made possible due to the work of US. EPA Office of Research and Development
Gulf Breeze, the University of Missouri-Columbia, and the US Geological Survey.

The assistance, advice, and comments of the EPA work assignment manager, Marjorie Coombs
Wellman of the Exposure Assessment Branch, Office of Science and Technology have been of
great value in developing this model and preparing this report. Further technical and financial
support from David A. Mauriello, Rufus Morison, and Donald Rodier of the Office of Pollution
Prevention and Toxics is gratefully acknowledged. Marietta Echeverria, Office of Pesticide
Program, contributed to the integrity of the model through her careful analysis and comparison
with EXAMS.  Release 2 of this model underwent independent peer review by Donald DeAngelis,
Robert Pastorok, and Frieda Taub; and Release 3 underwent peer review by Marty Matlock,
Damian Preziosi, and Frieda Taub. Their diligence is greatly appreciated.

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AQUATOX (RELEASE 3) VOLUME 1: USER'S MANUAL
Table of Contents:


  Getting Started	1
    AQUATOX: A Brief Overview	1
    Installation Considerations	2
    Loading a Study	2
    The Main Window	3
    Saving a Study	5

  Working With Existing Simulations	5
    What is in an "APS" file?	5
    Libraries and Parameters in a Simulation	6
    Exploring State Variables	6
    Site Parameters	25
    Exploring Model Setup	29
    Running the Simulation	35
    Viewing Graphical Output	35
    Viewing Table  Output	45
    Viewing Uncertainty Output	45
    Viewing Sensitivity Output	47
    Exporting Results	48
    Clearing Results	48
    Export to Release 2.2/3.0 Databases	49
    "Taking Parameters" from Another Study	49

  Other Interface Components	49
    Using the Toolbar	49
    Editing the Toolbar	50
    The AQUATOX Wizard	50
    Exporting Parameters as Text	55
    Aquatic-Dependent Vertebrates	56
    Batch Mode	57

  Setting Up Simulation, Single Segment	58
    Data Requirements	58
    Site Types	59
    Starting with a  Surrogate Simulation	60
    Starting from Scratch	61
    Water Volume  Modeling Options	61
    Adding a State Variable	61
    Using Sediment Bed Models and Data Requirements	62
    Model Calibration	66
    Model Validation	68

  Working with a Linked Simulation	68
    Viewing Linkages	69
    Examining  Linked Output	70
    Examining  a Single Segment	70
    Exporting All Graphs to Word (Linked)	71
    Tributary Input Segments	71

  Setting Up a  Linked Simulation	72
    Additional Data Requirements for Linked Mode	72

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  Using a Template Segment	73
  Excel Template Import Capability	73
  Adding a New Segment	76
  Linking Segments Together	77
  Water Balance Considerations	78
  Stratification in a Linked System	78

Tutorials	79
  Simple Tutorial	79
  Stream Tutorial	85

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AQUATOX (RELEASE 3)  USER'S MANUAL



Getting Started

AQUATOX:  A Brief Overview

AQUATOX is a simulation model for aquatic systems. AQUATOX predicts the fate of various
pollutants, such as nutrients and organic chemicals, and their effects on the ecosystem, including
fish, invertebrates, and aquatic plants. AQUATOX is a valuable tool for ecologists, biologists,
water quality modelers, and anyone involved in performing ecological risk assessments for
aquatic ecosystems.

AQUATOX simulates the transfer of biomass, energy and  chemicals from one compartment of
the ecosystem to another. It does this by simultaneously computing each of the most important
chemical or biological  processes for each day of the simulation period; therefore it is known as a
process-based  or mechanistic model. AQUATOX can predict not only the environmental fate of
chemicals in aquatic ecosystems, but also their direct and  indirect effects on the resident
organisms. Therefore  it has the potential to establish causal  links between chemical water quality
and biological response and aquatic life uses.

AQUATOX is the only general ecological risk model that represents the combined environmental
fate and effects of conventional pollutants, such as nutrients and sediments, and toxic chemicals
in aquatic ecosystems. It considers several trophic levels,  including attached and planktonic algae
and submerged aquatic vegetation,  invertebrates, and forage, bottom-feeding, and game fish; it
also represents associated organic toxicants.  It has been  implemented for streams, ponds, lakes,
estuaries, reservoirs, and experimental enclosures.

The fate portion of the model, which is applicable especially to organic toxicants, includes:
partitioning among organisms, suspended and sedimented detritus, suspended and sedimented
inorganic sediments, and water; volatilization; hydrolysis; photolysis; ionization; and microbial
degradation. The effects portion of the model includes: acute toxicity to the various organisms
modeled; and indirect  effects such as release of grazing and predation pressure, increase in
detritus and recycling of nutrients from killed organisms, dissolved oxygen sag due to increased
decomposition, and loss of food base for animals.

AQUATOX is the latest in a long series of models, starting with the aquatic ecosystem model
CLEAN (Park et al., 1974) and subsequently improved in consultation with numerous researchers
at various European hydrobiological laboratories, resulting in the CLEANER series (Park et al.,
1975, 1979, 1980; Park, 1978; Scavia and Park, 1976) and LAKETRACE (Collins and Park,
1989). The MACROPHYTE model, developed for the U.S. Army Corps of Engineers (Collins et
al., 1985), provided  additional capability for representing submersed aquatic vegetation. Another
series started with the toxic fate model PEST, developed to complement CLEANER (Park et al.,
1980, 1982), and continued with the TOXTRACE model (Park, 1984) and the spreadsheet
equilibrium fugacity PART model. AQUATOX combined algorithms from these models with an
ecotoxicological construct borrowed from the FGETS model  (Suarez and Barber, 1992); and
additional code was written as required for a truly integrative fate and effects model (Park, 1990,
1993).  In the late 1990s, AQUATOX was restructured and linked to Microsoft Windows interfaces
to provide even greater flexibility, capacity for additional compartments, and user friendliness.

   •   AQUATOX  Release 1  was produced in 2002 and was the first EPA release to run under
       Windows.
   •   AQUATOX  Release 2 was completed in 2003 and included more state variables and
       multi-age-class fish along with a refined user-interface.
   •   AQUATOX  Release 2.1 was completed in 2005 and included additional chemical
       modeling options and variable stoichiometry among  numerous other refinements.

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AQUATOX (RELEASE 3)  USER'S MANUAL
       AQUATOX Release 2.2 was completed in 2006 and included updated simulations and
       parameter databases along with minor interface enhancements.
       AQUATOX Release 3 was completed in 2009 and includes linked segments, simulations
       of estuaries, dramatically improved output capabilities, and many other model
       improvements.
Installation Considerations

To install the AQUATOX Model, an "Installshieldฎ" is utilized.

Through its install program, AQUATOX will produce the following folder structure:

    •   DATABASE:  holds libraries of parameters, chemical toxicity databases, and
       interspecies regression models for toxicity (see ICE).
    •   OUTPUT:  holds files produced by AQUATOX to produce tabular data and graphs. Also
       a default location for exporting results.
    •   PROGRAM: holds the executable itself and helper files
    •   STUDIES: holds the simulations included with the Installshield.  New simulations may be
       saved in this location or anywhere else that is desirable.  A "Tutorial" directory is also
       included here with files required for the tutorials in this document. This directory also
       includes some ancillary files such as a parameter import template, descriptions of files,
       etc.

Be sure that you have read-write access to each of these directories wherever you are installing
AQUATOX.

If you install AQUATOX in the same location as a previous installation, you will no longer be able
to access the previous version and any modifications you made to the databases or default study
files (*.APS) may be lost.

The Borland Database Engine is also installed as part of the installation of AQUATOX.

Some additional notes:

    •   AQUATOX must be initially installed by an Administrator.
    •   If AQUATOX will exclusively be run using an "administrator" account, no additional
       changes need to be made.
    •   If AQUATOX will be run by a user with a "standard" account, full read and write privileges
       must be given to that user for the directory in which AQUATOX has been installed.
    •   An alternative to adding read and write privileges: is to install AQUATOX in a directory
       where the user already has read and write privileges. This  is the best choice for
       managed desktop systems.

For more information see the AQUATOX Release 3 Installation Guide.

Loading a Study

The APS file is the basic unit in AQUATOX; it contains site data, loadings, and parameter values
used in a simulation; and it may contain results from a prior simulation.   Click on File in the menu
bar to get the pull-down file menu, and click on Open. You will then be given a choice of
AQUATOX study files to load.

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AQUATOX (RELEASE 3)  USER'S MANUAL
.Open
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AQUATOX (RELEASE 3)  USER'S MANUAL
The Chemical button brings up the loadings screen for the organic toxicant, if modeled. Double
clicking on the state variable named "Dissolved org. toxicant" that will appear at the top of the
list of state and driving variables (if a chemical is included in the simulation) has the same effect.

The Site button loads the site characteristic screen .

Setup allows the user to set the dates of the simulation, and to specify various options such as
the control setup, uncertainty analysis, and saving biologic rates.

Notes provides a window for writing comments on the study.

Birds, Mink... links to a model of chemical uptake for shorebirds or any other animals that feed
exclusively on aquatic prey.

The Sed Layers button brings up parameters and initial conditions for the multi-layer sediment
model or the sediment diagenesis model if they are included in the simulation.

Perturbed starts the simulation with changed conditions, such as with a toxicant.

Control starts a simulation without the stressor; the user can use Control Setup to specify what
is changed and what is held  constant, or parameters may be changed between running the
control and  perturbed simulations.

Output presents the results  as a series of charts and graphs .

The output can be exported as database, spreadsheet, or text files by clicking on Export Results
or Export Control

The State and Driving Variables In Study show the full list of state variables within the
simulation.  Variables can be added to or deleted from this list using the buttons at the bottom of
the list. Animals, plants, and detritus within this list can have up to twenty organic chemicals
associated with them.

The Edit with Wizard button allows you to edit the current simulation with the AQUATOX
Wizard.

If you hit the Help button from the Main Screen, you will jump to this topic in the help screen.

When this screen is viewed in Linked Mode it represents a single linked segment.  In this case,
two additional buttons are available. Stratification affects the setup of vertically stratified
segments in linked mode. Morphometry allows the user to specify a time series of cross section
areas for each segment.

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AQUATOX (RELEASE 3) USER'S MANUAL
r-—— — — — — ————————————————————— -^^ 	 T
.1||, Ruin R MN,aps- - Main Window •
A QUA TOX: Study Information
EPA Release 3,0
Study Name: Rum R, 18 MM
j
Model Run Status:
Perturbed Run: No Results Attached
Control Run: No Ctrl. Results Attached
Data Operations: Program Operations:
Q||| Initial Conds,
S^r1 Chemical
Site
Ta^lP^j
^B Setup

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AQUATOX (RELEASE 3)  USER'S MANUAL
    •   List of state variables and driving variables utilized and their loadings, "underlying
       parameters," and initial conditions;
    •   Site specific and remineralization Parameters;
    •   Model setup information;
    •   Boundary condition loadings for a simulation;
    •   Results from any simulation that may have previously been run, including "rates";
    •   External data that has been imported into the simulation for plotting against results;
    •   The graph library that has been produced for that simulation;
    •   The uncertainty mode or sensitivity mode setup including distributions chosen and
       correlation information.

Not included in the APS file are uncertainty output, which is saved in a database format (*.db or
*.dbf)  and sensitivity output which is in Excel format (*.xls).  Libraries of parameters are not
included in the APS file either, other than those parameters that have been "loaded" into a
simulation and therefore  have relevance to that simulation.

Libraries and Parameters in a Simulation

There is a basic dichotomy in working with AQUATOX. You have a choice of editing database
files in the general "library" or of opening and editing a particular study. Studies are self-
contained files with all the information on a particular simulation, including initial conditions,
loadings, parameter values, first and last dates for the simulation, and simulation results.
Parameter values can be edited, but changes apply on/yto that study.  The intent is to be able to
archive a model application so that all assumptions and results are saved for future reference.
This is especially important for regulatory applications that are subject to later review. (Of course,
you also should archive the version of AQUATOX that was used.)

Parameter and site records that will  be used  repeatedly should be saved in the appropriate
library. Each  library is a  database in Paradox format with records for each organism, chemical, or
site. Generally, editing of parameters should be done in the library mode to maintain consistency
among studies.  In contrast, if a site  record is only going to be used for a single study, it may be
desirable to create it within the study. Study records can be copied into the library; so the choice
of where to edit parameters is up to the user. It is the user's responsibility, though, to synchronize
parameter values  among studies. This can be done by saving  a record to a library and then
loading that record to each study.

To create or edit a record for general use, click on  Library in the menu bar. You can then click
on the specific library from the pull-down menu.

See Also:  Types  of Libraries

Exploring State Variables

State Variables

State  variables are those ecosystem components that are being simulated. These include
organic toxicants,  nutrients and dissolved gasses,  organism and detrital compartments and their
associated toxicants, and other variables traditionally considered driving variables, such as water
inflow, temperature, pH,  light, and wind.

AQUATOX is a powerful  model because you can add or delete state variables.  It is even
possible to remove all biotic components in order to model a tank or other sterile system.  In
general, the fewer state variables, the better. In particular, unnecessary state variables slow
down  the simulation and  create additional requirements for verification. This  is especially true for

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AQUATOX (RELEASE 3)  USER'S MANUAL
streams, which tend to be more dynamic and therefore slower to simulate. Nevertheless, often it
is desirable to model a food web rather than a food chain, for example to examine the possibility
of less tolerant organisms being replaced by more tolerant organisms as environmental
perturbations occur. The choice of which state variables to model depends to a large extent on
the purpose of the modeling application and the availability of data pertaining to the state
variables.

Modifying the State Variable List

To Delete a state variable, select the variable you wish to delete from the state variables list by
clicking on it. To select multiple state variables, hold down the control key while clicking more
than one variable on the list. Then, click on the delete button and confirm the deletion.  There
are several state variables (such as nutrients) that are basic to an AQUATOX simulation, and that
therefore cannot be deleted.

To Add a state variable, click on the Add button and choose the variable you wish to add from
the dialog box that appears. Note that the names of the taxonomic groups and ecologic guilds on
the main study screen are followed by the names of the specific groups in brackets.  After clicking
the Add button, you will first be prompted to choose the taxonomic group, ecologic guild, or
chemical compartment you wish to add.  Then, you will be  prompted to load the chemical or
species specific parameters from the appropriate library.

You may also modify the state variable list using the AQUATOX Wizard.

After animal or plant state variables have been added  or removed one critical step is to view the
trophic interactions for the simulation to ensure that the resulting food web is reasonable.
AQUATOX does have default trophic interactions within each organism,  but not knowing which
organisms are going to be included in each simulation, it will usually require modification. The
best way to access trophic interactions is by selecting  "Edit Trophic Interactions" under the
"Study" menu, or by clicking on  "Trophic Matrix" which is a button found within  Animal state
variable loadings screens.

Initial Conditions and Loadings

Initial values and loadings are needed for all the state variables or compartments  simulated.
These are input on the loadings screen.  If one or more toxicants are modeled, then initial
concentrations associated with the biota can also be specified. Constant loadings for plants and
invertebrates can be considered  as "seed" values, although care should  be taken to use small
values or the loadings can dominate the simulation.  Even  periphyton and zoobenthos may be
maintained through drift from upstream, and a constant loading is appropriate.  Likewise,
macrophytes may die back in winter and sprout from rhizomes; because rhizomes are not
explicitly modeled, a small loading is the mechanism for reestablishing the population in the
simulation when environmental conditions become more favorable.

Of course, upstream loadings may be significant inputs to a reach or lake.  These may  be
represented by constant or dynamic (time-varying) loadings.  AQUATOX has a very flexible
interpolation routine to obtain daily values from irregular data points and  even time series
occurring or extending outside the simulation period. Dynamic loadings  can be entered directly
on the loadings screen, or they can be entered or obtained offline and imported into the model.
Imported data can be in a variety of formats, which are evident when the "Change"  button  is
used. Loadings can be altered by means of a multiplier (the "Multiply loading by" button). This
procedure is especially useful for analyzing various loading scenarios. It is also a way to correct
or convert data series. However, ordinarily the multipliers are set to 1 for the Control simulation,
so use for other than perturbations is discouraged.

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AQUATOX (RELEASE 3)  USER'S MANUAL
Loadings in "inflow water" are closely related to the volume of inflow water specified (or
calculated as a result of choices) in the water volume screen.  In other words, loadings in unit per
liter of water must have an associated inflow of water in order to be relevant to the simulation.

On the other hand, nonpoint-source (NPS), and point-source (PS) loadings are input in units
of grams per day and are not affected by the quantity of inflow water. AQUATOX ignores the
quantity of water that is associated with NPS and PS loadings as a model simplification; however,
this water can be directly modeled in the multi-segment model with the  use of Tributary Input
Segments.

Direct precipitation loadings are a function of the site's surface area with units of grams per meter
squared per day. This input field includes  both wet and dry precipitation.

Any of the time-series loadings may be imported, exported, or cleared using the "Change" button
found directly below their listing.

Also see this note about dynamic loadings.

Importing and Exporting Loadings

Loadings and other time series may be imported from Excel format, Tab delimited format, DBase
format, Paradox Database Format, or Comma Separated Value  (CSV) format.

To change the format, move to the List Files of Type menu and select a different format. The
required input format will be specified in the box at the upper left. For example, an Excel data file
should have the date in column A of the first worksheet and the data to be  imported in Column B.
    Excel Data

    Column A of the first sheet in
    the workbook should hold the
    date. Column B should
    hold data to be imported. (Or,
    specify' row, col, sheet below)
When you select a file, a preview will appear in the lower box. You can "update" this preview by
pressing the button above it.

For Excel only, you may specify where the data you wish to import appears.  This is helpful when
working with very large spreadsheets with multiple sets of data on them. The worksheet number
is specified first and then the start row, date column, and value columns may also be specified as
shown below.
   Sheet #  1    Start Row  1     Date Col  lA   Val Col  B
To import from Excel, Microsoft Excel 2000 or a later version of Microsoft Excel must be installed
on the computer.

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AQUATOX (RELEASE 3)  USER'S MANUAL
Some data, for which there are units conversions available, may be converted on the fly using the
conversion check-box.
Loadings may also be exported from AQUATOX by selecting the "Export" tab at the top of the
screen.  Time series that are no longer relevant may be cleared from AQUATOX by selecting the
"Clear Data" tab at the top of the screen.

Important Note about Dynamic Loadings:

Dynamic loadings are loadings that are variable over the simulation's time-period.  These
loadings are entered using a list of dates and associated loadings.

During a simulation, if the date that is being simulated appears on the input list of dates, the
loading is taken directly from the list.  If the current date in a simulation occurs between two
dates, interpolation is used to determine the correct loading value. Because of this interpolation,
if the intent is to represent a spike such as from storm runoff on a particular day, the spike loading
should be bracketed by zero ("0") loadings.

If the current date in a simulation occurs  before the first date or after the last date of the
loading time series, AQUATOX assumes that the loadings "wrap around" with an annual
cycle. Specifically, the AQUATOX algorithm will step towards the input data in one-year
increments until the derived date falls within the input time series. The model will then interpolate
the results, if necessary,  and assign the results to the date being modeled. In this  manner, if you
had two years of loadings but ran the model for eight years,  the model would repeat the second
year of loadings in an annual cycle for the last seven years of the simulation. (Note:  if a different
type of annual cycle or interpolation  is desired this can be derived outside of the AQUATOX
interface and then imported into the  model.)

Exercise caution  when modeling multiple years using loadings data from only one  or a few years.
Sporadic loadings, which would be expected in that one particular year may inappropriately be
repeated in successive years.  If you do not wish loadings to be repeated, enter values ("0" or
otherwise) for the first and last days of the simulation.

If there is only one dynamic loading point present, this is interpreted in the same manner as a
constant load.

Dissolved Org. Tox Loadings

This initial conditions and loadings screen contains a few items that are unique to dissolved
organic toxicants.

The gas-phase concentration input allows the user to enter a constant concentration of toxicant
in the air (g/m3) that affects the degree of volatilization (potentially in either direction).

The biotransformation button may be used to access the biotransformation screen which is only
relevant when multiple chemicals are modeled, and only then when parent compounds
and daughter products are both included in the mix.

The toxicity data button allows the user to specify direct effects from the toxicant to  biotic
elements in the simulation in the chemical toxicity data screen.

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AQUATOX (RELEASE 3)  USER'S MANUAL
Nutrient Loadings
The nutrient initial conditions and loadings screens include the capability to model Total N and
Total P along with all of the items contained on other loadings screens.
When initial conditions, inflows, or other loadings are entered as Total N, by selecting the
appropriate check-box (e.g. "Init. Cond. is Total N"), model inputs are located in the "Nitrate"
initial conditions and loadings screen and  loadings on the ammonia screen are grayed out as they
are not relevant.  (Total N is assumed to be 12% ammonia for inflow and nonpoint-source
loadings and 15% ammonia for point-source loadings.)
When Total N or Total P  are used as model inputs, AQUATOX calculates the dissolved content
by subtracting out loading inputs for suspended and dissolved detritus and suspended algae as
discussed in section 5.4  of the technical documentation.
Detrital Initial Conditions and Loadings
A complex loading screen is necessary for organic matter inputs in the water column. AQUATOX
simulates Organic Matter (dry weight); however, the user can input data as Organic Carbon or
Biochemical Oxygen Demand (BOD) and the model will make the necessary conversions.
(See the technical documentation for more information about how the model converts organic
carbon and BOD loadings into organic matter.)
       Suspended and Dissolved Detritus    I
        Initial Condition:
         1          mg/Ldry    Input is:
        Loadings from Inflow:     r Organic Matter
      r Use Constant Loading of     r Organic Carbon
                  mg/Ldry
      !• use Dynamic Loadings
             Date
[Loading
             6/12/2000
             6/29/2000
             7/19/2000
             8/2/2000
             8/17/2000
             9/5/2000
             9/13/2000
             9/25/2000
      2.7
      3.1
      1.7
      1.9
        3
      2.4
      2.6
          v
  A.  I Change
mg / L dry
             Multiply loading by 1
  Motes:
       nut99LB2.xls & nutrientOO,1.jcls
       Rum River obs values
                                    Percent that is Paniculate
                             Breakdown of Initial Condition:
                                     pO          percent
                             For  Inflow          j^j Loadings:
                                   (• Use Constant Percentage of
                                     p"           percent
                                   r UseTimeseries of Percentages
                                  [Loading
                                                                                 percent
                                                                          Change
              Point Sources  I \  Non-Point Sources
                  i
                                           Dissolved / Particuiate Percentages |
                                            Labile / Refractory Percentages
                                       Q.K.  MX Cancel
Organic matter initial conditions and loadings are divided into four compartments:
    •   particulate refractory detritus;
                                                 10

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AQUATOX (RELEASE 3)  USER'S MANUAL
    •   participate labile detritus;
    •   dissolved refractory detritus; and
    •   dissolved labile organic matter.

Initial conditions and loadings are parsed by specifying % Particulate and % Refractory which
can be entered as constant or time-varying percentages (0-100). Loadings of organic matter can
be constant or dynamic (time series) for concentrations in inflowing water (mg/L), and for mass
from point sources and non-point sources (g/d). Toxicants associated with detritus also can be
specified (ug/kg).

Organic matter loadings in "inflow water" are closely related to the volume of inflow water
specified (or calculated as a result of choices) in the water volume screen.   In other words,
loadings in unit per liter of water must have an associated inflow of water in order to be relevant
to a simulation.
Separate state variable input screens are provided for refractory and labile organic matter within
the sediment bed. The initial conditions are giv
Associated toxicants are given as ug/kg (ppb).
the sediment bed. The initial conditions are given as g/m2, and the loadings are given as mg/L.
See section 5.1 of the technical documentation for more information.  Also see this note about
dynamic loadings.

Temperature Data Screen

The annual mean and range in temperature from the site underlying data screen can be used, or
time series can be entered—in which case make sure that the complete time period being
simulated  is covered. If the system stratifies then temperatures must be given for both  epilimnion
and hypolimnion.

If a system may thermally stratify then hypolimnion temperature data may also be entered here.
Under the model's default behavior, thermal stratification will be assumed when vertical
temperature differences exceed three degrees; at all other times a well-mixed system is assumed
and modeled.  However, there is an additional button on this screen labeled "Stratification
Options" that allows a user to modify this behavior by specifying dates of stratification,
thermocline depth, and flow routing options.

For more information on temperature modeling, see section 3.5 of the technical documentation. If
time series are entered you may wish to see the note about dynamic loadings.

Wind Loadings Screen

(To access this screen double-click on "Wind Loading" on the state variables list in the main
window.)

Within the wind loadings screen, you can either use a default time series for loadings, a constant
wind loading, or enter dynamic loadings.

Wind  is  an important driving variable  because it determines the stability of blue-green algal
blooms, affects reaeration or oxygen  exchange, and  controls volatilization of some organic
chemicals. Wind also can affect the degree of mixing in estuaries.  Wind is usually measured at
meteorological stations at a height of 10 m and is expressed as m/s. Wind is less important for
rivers and streams.
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For the default time series, wind is computed using a complex Fourier series of sines and cosines
fora 365-day repeating period with a user-supplied mean value. The edit box for the mean
value (in m/s) appears in the upper right hand of the wind loadings screen.

For more information on wind, see section 3.6 of the technical documentation.

Light Loadings Screen

(To access this screen double-click on "Light" on the state variables list in the main window.)

When entering light data, the user has three options:  Constant, time series, or annual mean and
range may be given for light  in Langleys/day. If annual mean and range are used, these
parameters must be filled-in  in the site underlying data screen.

Also, photoperiod can be auto-calculated from latitude or entered manually.  The latter can be
useful when modeling experimental facilities.

Conversions into Langleys per day (Ly/d) are provided using the "convert" button; imported time
series may also  be converted.

See section 3.6  of the technical documentation. Also see this note about dynamic loadings.

pH Screen

(To access this screen double-click on pH on the state variables list in the main window.)

pH is important in AQUATOX for several reasons.

    •    lonization of ammonia is sensitive to pH;
    •    Nitrification rates are sensitive;
    •    Hydrolysis of organic chemicals can be  sensitive to pH;
    •    Calcite precipitation  is predicted as a function of pH;
    •    Conversion of refractory detritus to labile detritus is affected by  pH; and
    •    Decomposition of organic matter is affected;

A user may input a time series of pH values here or calculate pH values using a simple semi-
empirical formulation that requires a Mean Total Alkalinity input on this screen (see section 5.7 of
the technical documentation  for more information.)

When a time series is utilized the rules for dynamic loadings apply.

The pH  state variable cannot be deleted from an AQUATOX simulation.

Water Volume Data

(To access this screen double-click on "Light" on the state variables list in the main window.)

Considerable flexibility exists to compute or specify water volume.  Depending on the method
chosen, inflow or discharge values  may be required. The Manning's equation can be used to
compute changing volumes in a stream. The simplest procedure is to hold volume constant at
the initial condition. Volume  can also be computed dynamically using both inflow and discharge,
which are input on this screen; however, it also uses the annual evaporation rate, which  is input
on the site underlying data screen.  When  available, a  known time series can be  entered or
imported.  Flow data can  be  imported in several formats, including USGS tab-delimited; however,

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AQUATOX (RELEASE 3)  USER'S MANUAL
recent changes in the USGS format, including variable header material, makes this prone to
problems. If the data do not appear in the preview window then the flow data will have to be
converted in a spreadsheet from cfs to m3/d, and the date column and flow column then exported
as a tab-delimited or comma-separated file suitable for importing into AQUATOX.

Notes:

The initial condition water volume is determined from the inputs on this screen and not from the
site underlying data. However, a "Get Initial Cond. from Site Data" button exists to allow the
initial condition to be copied from that screen.

The Mannings Equation Method (streams only) requires discharge data.  Inflow data and site
volume are calculated using Manning's Equation.  Careful attention should be given to the
"Channel Slope" and "Manning's Coefficient" parameters entered in the "Stream Data" screen
(within the site underlying data screen.)

The Keep Constant at Initial Condition Level Method requires inflow data. Discharge is
calculated based on inflow and evaporation.

If you choose to Calculate Dynamically, volume is calculated based on inflow, outflow and
evaporation.

The Utilize Known Values Method requires a time series of known volumes and inflow data.
Outflow is calculated taking evaporation into account.

The differential equation that calculates the water volume of the system is:

                     SVolume / 6 t = Inflow - Outflow - Evaporation

The "Stratification Options" button allows a user to modify default model behavior regarding
stratification by specifying dates of stratification, thermocline depth, or flow routing options.

For more information on morphometry and volume, see section 3.1 of the technical
documentation.

Initial Conditions Summary Screen

Initial conditions can be displayed and printed for all state variables in this summary screen and
any associated toxicants (reach this screen by clicking on the Initial Conds. button on the main
screen.)

These initial conditions cannot be edited on this screen  because of the complexities of the
suspended and dissolved detritus screen.

Parameters

Parameters provide values for coefficients in the process equations. Although default values are
given, the user has great flexibility in specifying values to represent site-specific species or
groups.

There are five libraries of parameter values that may be loaded  into a simulation.

The Chemical Library contains parameters for organic chemicals that could be associated with a
simulation. Within a simulation, chemical parameters may be found by double-clicking a
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AQUATOX (RELEASE 3)  USER'S MANUAL
Dissolved org. toxicant on the state-variable list and then choosing Edit underlying data.  See
also: Chemical Data Screen

The Animal Library contains parameters for fish and invertebrates that could be associated with
a simulation. Within a simulation, animal parameters may be found by double-clicking an animal
on the state-variable list and then choosing Edit underlying data. See also: Animal Data
Screen

The Plant Library contains parameters for algae and macrophytes that could be added to a
simulation.  Within a simulation, plant parameters may be found by double-clicking a plant on the
state-variable list and then choosing Edit underlying data. See also Plant Data Screen

The Site Library contains parameters for representative sites that could be modeled by
AQUATOX. Within a simulation, the site parameters can be found by clicking the Site button and
then by clicking on the Edit Underlying Site Data button that appears. See also Site Data
Screen

The Remineralization Library contains parameters about the detritus and nutrients associated
with a site.  To find these parameters within a simulation click on the Site button and then by
clicking on the Remineralization button that appears.

When editing libraries, a GridMode button  may be pressed that toggles editing between a matrix
of all parameters in the library or a close-up look at the parameters, units, and comments for one
specific entry in the library.

To add a new library entry you may press the New button and a blank set of parameters is
created for data entry.  Alternatively, after modifying a parameter set in a simulation, those
parameter values may be saved  back into the library (either overwriting the original entry or the
name may be changed to add additional information to the library.)

When you are editing the underlying data that applies to a simulation, you can either load from or
save those parameters to the library file by clicking on the Load from Library or the Save to
Library  buttons at the top left of the screen.

Chemical  Properties and Fate Data Screen

Required chemical "underlying parameters" and units can be found on this screen. These
parameters govern chemical fate and partitioning  behavior.

Note that the organic-sediment/detritus and water partition coefficient can be calculated
dynamically or entered manually by the user. Sorption to inorganic sediments is only relevant
when the multi-layer sediment model is incorporated.

Parameters on this screen govern the chemical fate processes of

    •   ionization,
    •   volatilization,
    •   hydrolysis,
    •   photolysis,
    •   sorption, and
    •   microbial degradation.
    •   PFA Parameter Screen
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AQUATOX (RELEASE 3) USER'S MANUAL
You may also edit the toxicity data for the relevant chemical by selecting the Toxicity Data button
at the top right of the screen.

To maximize comprehension, parameters on this user interface screen are described with several
English words rather than symbolically. Appendix B of the AQUATOX Technical
documentation contains a full description of each of the parameters shown here under
"Chemical Underlying Data" as well as their manner of referral in the equations of the technical
documentation (often a shorter variable name).  In this manner, a user can use this appendix as a
reference to search the technical documentation and find all equations in which each parameter
is utilized. Advanced users can also easily find  the parameters within the AQUATOX source
code as the "internal" variable names are also listed within  Appendix B.

For more information on modeling organic chemicals, see chapter 8 of the technical
documentation.

Chemical Toxicity Data Screen

This screen is where all of the chemical toxicity  parameters are located.  To get to this screen go
to Chemical Underlying Data and select the "Toxicity Data" button. Or use the button on the
chemical's initial conditions and loadings screen

There are multiple options for entering uptake rate constant (k1), the elimination rate constant
(k2) and the bioconcentration factor (BCF) or allowing the model to calculate these parameters
(BCF=k1/k2)

Additionally, elimination rates may be estimated using the octanol water partition coefficient
(Kow).

If the user only has toxicity data for a few species, an extensive library of regressions
(Interspecies Correlation Estimation, or Web-ICE) is built into the model (Asfaw and Mayer,
2003). This may be accessed by using the button at the bottom of the screen  ("Interspecies
Toxicity Correlation Models").

By entering both LC50 and EC50 values for a species the application factor can be computed.
The user has the option of applying that same ratio to the rest of the species in the animal or
plant toxicity screen using the buttons Estimate animal LCSOs... and Estimate plant ECSOs....
Animal and Plant toxicity require different parameters so they are given separate matrices on the
screen.  The animal toxicity data appears above the plant toxicity data.

The animal toxicity parameters are as follows:

       Animal Name, must match the "toxicity data" record in the relevant Animal Data screen.
       LC50, (ug/L), external concentration of  toxicant at which 50% of population is killed
       LC50 exp. time, (h), exposure time in toxicity determination
       Elim Rate Const. (K2), (1/d), elimination rate constant
       K1  Uptake Const. (L/kg) optional uptake rate constant (dry weight)
       BCF, (L/kg) optional Bioconcentration Factor (dry weight)
       Biotrnsfm rate const, (1/d), Daily  rate  of biotransformation of this toxicant
       EC50 growth, (ug/L), external concentration of toxicant at which there is a 50% reduction
           in growth
       Growth exp. time, (h), exposure time in toxicity determination
       EC50 repro., (ug/L), external concentration of toxicant at which there  is a 50% reduction
           in reproduction
       Repro exp. time, (h), exposure time in  toxicity determination
       Ave wet wt., (g), mean wet weight of organism
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AQUATOX (RELEASE 3)  USER'S MANUAL
       Lipid frac., (g lipid/g organism), fraction of lipid in organism (wet weight)
       Drift Thresh, (|jg/L), concentration at which invertebrate drift is initiated

The plant toxicity parameters are as follows

       Plant Name, must match the "toxicity data" record in the relevant Plant Data screen.
       EC50 photo, (|jg/L), external concentration of toxicant at which there is 50% reduction in
           photosynthesis
       EC50 exp. time, (h), exposure time in toxicity determination
       EC50 dislodge, (ug/L), external concentration of toxicant at which 50% of algae is
           dislodged (not applicable for phytoplankton)
       Elim Rate Const. (K2), (1/d), elimination rate constant
       K1  Uptake Const. (L/kg) optional uptake rate constant (dry weight)
       BCF, (L/kg) optional Bioconcentration Factor (dry weight)
       Biotrnsfm. rate const., (1/d), Biotransformation Rate
       LC50, (ug/L), external concentration of toxicant at which 50% of population is killed
       LC50 exp time, (h), exposure time in toxicity determination
       Lipid frac., (g lipid/g organism), fraction of lipid in organism (wet weight)
Buttons at the bottom of the screen:

Estimate Animal or Plant K2s using Kow.  Elimination rate constants may be estimated using
the chemical's KOW data. Animal estimates  utilize the "wet weight" and "lipid frac." fields,
whereas plant estimates utilize only the fraction lipid field within the plant's toxicity record. For a
macrophyte-specific calculation to be performed, the plant name must include the substring
"macroph."

LC50 / EC50 estimation: Using the EC50 to LC50 ratio for one species, the EC50 or LC50 can
be estimated for other species within the simulation.

Interspecies Toxicity Correlation Models:  Utilize the integrated ICE regression databases.

Extensive information about the AQUATOX ecotoxicology model may be found in chapter 9 of the
technical documentation.
Using ICE for Regressions

This screen allows the user to utilize a database of regressions to populate all of the species in
their simulation with toxicity data.

Often LC50 data will only be available for one or two of the many species that a user wishes to
include in a simulation.  To alleviate this problem, a substantial database of regressions is
available (ICE) as developed by the US. EPA Office of Research and Development, the
University of Missouri-Columbia, and  the US Geological Survey (Asfaw and Mayer, 2003). The
ICE database includes over 2000 models with over 100 aquatic species as "surrogates."
Regressions may be made on the basis of species, families, or genera.  The database also
includes goodness of fit information for regressions so their suitability for a given application may
be ascertained (see statistic definitions below).

The steps that a user can take to use ICE within AQUATOX are as follows:

    •   Invoke the ICE interface from the AQUATOX "Chemical Toxicity Parameter" screen;
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AQUATOX (RELEASE 3)  USER'S MANUAL
    •   "Step 1:" Choose from the six available ICE databases (species, genus, and family by
       either scientific names or common names), by using the drop-down list provided;
    •   "Step 2:"  Choose a "surrogate species" that  matches a  species  for which there  is
       observed LCSOdata (drop-down list);
    •   "Step 3:" Choose a "predicted species" that matches a species that you wish to model
       (and for which LCSOdata is unavailable);
    •   "Step 4:"  Examine the goodness  of fit  for your model  and  evaluate whether  it  is
       appropriate for your purposes. Where there  are multiple surrogates  for the  desired
       predicted species, compare the statistics and choose the best surrogate/predicted pair.
    •   "Step 5:"Apply the model by assigning the surrogate and  predicted  species to  species
       within the chemical's  toxicity  record.  Use the drop-down  menus shown to apply the
       correlation within your toxicity database.  The changes will  be made  when you return to
       the toxicity screen from the ICE interface.
You may also visit the Web-ICE Website using the URL or the button displayed at the top of the
screen in case an updated database is available.

Graphs may  be displayed on  a  log-scale  or arithmetic  basis and the user may change the
confidence interval plotted.

Statistics provided for goodness of fit are defined below (source: Raimondo et al., 2007)

    •   Sample Size (n) - The  number of data points used to build the  model. In general, the
       higher the number of samples, the more robust the model.
    •   Intercept (a)-The Iog10 value of the predicted taxon toxicity when the Iog10 of the
       surrogate species toxicity is 0.
    •   Regression Coefficient (slope b) - The regression coefficient, represents the change in
       Iog10 value of the predicted taxon toxicity for every change in Iog10 value of the
       surrogate species toxicity.
    •   Average value of X (log IcSO) - The average of toxicity values for the surrogate species
       used in the model, log-transformed.
    •   Mean Square Error (MSE) - An unbiased estimator of the variance of the regression
       line.
    •   Standard Error of Slope (SEB)-A measure of the variability of the derived slope.
    •   Correlation Coefficient Sqr (R2) - The proportion of the data variability that is explained
       by the model. The greater the R-squared value and the closer it is to one, the more
       robust the model is in describing the relationship between the predicted and surrogate
       taxa.
    •   Probability (Pr) that slope <> 0, p-value - The significance level of the linear
       association and the probability that the linear association was a result  of random data.
       The lower the p-value, the stronger the association among the surrogate and predicted
       taxa. Model p-values of < 0.00001 are reported as 0.00000.

Raimondo, S., Vivian, D.N., Barren, M.G., 2007. Web-based Interspecies Correlation Estimation
(Web-ICE) for Acute Toxicity: User Manual. Version 1.1. EPA/600/R-07/071, U.S. Environmental
Protection Agency, Gulf Breeze, FL.

Biotransformation Screen

When multiple chemicals are modeled, the biotransformation screen indicates which chemicals
can be converted to other chemicals within the simulation.

For the chemical indicated in the title of the screen, the columns of the matrix indicate the
ecological compartments in which that chemical can break down and the rows of the matrix
represent the other chemicals within the simulation into which the chemical could be


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AQUATOX (RELEASE 3)  USER'S MANUAL
transformed.  Each cell indicates the percentage of the chemical that is biotransformed into the
daughter product (chemical indicated in the row title).  Each column must sum to 100 percent; the
"other" row autocalculates to make this possible.

The rate of biotransformation in each species is governed by the biotransformation parameter
found in the chemical toxicity record screen.

Categories in which biotransformation may be specified are:

    •   Aerobic Microb.: Aerobic microbial degradation within detritus;
    •   Anaerobic: Anaerobic microbial degradation within detritus;
    •   In Algae:  Plant compartments;
    •   Benthic Insect:  Invertebrates specified with an animal type of "Benthic Insect";
    •   Other Invert: Other invertebrates;
    •   In Fish.

In some cases, modeling a specific biotransformation pattern within a particular species will be
desirable. In this case, using the "Add Species Specific Data" button will make this possible.
You will be able to select a modeled species in the simulation and then specify the appropriate
biotransformation rate within that species.

Plant Data Screen

Several fields near the top of the screen require explanation.  If you click on the arrow to the right
of Plant type, you will be given a choice. The choice of Plant type is important because different
types have different physical or biological processes that apply to them. For instance,
phytoplankton are subject to sinking, but not periphyton, which are attached to a surface.
Conversely periphyton are limited somewhat by very slow current velocity; but not phytoplankton,
which are adapted to still water.  If "macrophytes" are chosen  the species must be distinguished
as "benthic," "rooted floating," "or "free-floating." Table 5 in the Technical Documentation helps
clarify some of the differences between plant types.

Although Plant type is important for determining which processes do or do not apply to the state
variable, the Taxonomic Type field is included as an organizing tool and does not affect model
output.

The Toxicity Record field within this screen links to the "plant name" within each chemical's
toxicity data.  In this manner, several plants could link to the same toxicity record if that is
desired. You may select a record from the list or type a new name if the desired plant name does
not appear in the pulldown list.

Phytoplankton and periphyton compartments may be linked together. When viewed in a
simulation, a plant will have an "Edit All Plant Linkages" button available at the top of the
screen.  Periphyton also have a "Periphyton Linkage" button which allows you to  edit the same
information but only for the species shown.  It is considerably  more powerful and user-friendly to
use the "Edit AN" button.

A few notes regarding the some of the most important parameters:

    •   Photosynthesis and growth rates are quite sensitive to the saturating light and P half-
       saturation coefficients along with optimum temperature and the maximum photosynthetic
       rate;
    •   The sedimentation rate should be small for streams and much larger for lakes;
    •   The Critical Force (FCrit) for scour is quite important for periphyton;


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    •   The phytoplankton mortality coefficient may be adjusted for a particular site;
    •   The exponential mortality coefficient (which increases the mortality for suboptimal
       conditions) may need to be adjusted if blooms crash too quickly or not quickly enough.
    •   Occasionally the extinction coefficient may need to be increased if algal growth is too
       strong-thai is the  principal means of negative feedback, and can vary among groups.
    •   Salinity effects are only relevant if "salinity" is included within the simulation in which case
       photosynthesis and mortality may be affected.

To maximize comprehension, parameters on this user interface screen are described with several
English words rather than  symbolically.  Appendix B of the AQUATOX Technical
documentation contains a full description of each  of the parameters shown here under "Plant
Underlying Data" as well as their manner of referral in the equations of the technical
documentation (often a shorter variable name).  In this manner, a user can use this appendix as a
reference to search the technical documentation and find all equations in which each parameter
is utilized.  Advanced users can also easily find the parameters within the AQUATOX source
code as the "internal" variable names are also listed within Appendix B.

See section 4.1 and 4.2 of the technical documentation for extensive discussion of modeling
algae and  macrophytes.

Plant Linkages

Individual periphyton to phytoplankton linkages may be edited using the "Periphyton Linkage
button." Using the "Edit AM" button allows you to edit these linkages for the entire simulation
simultaneously. All periphyton in a simulation are listed here along with drop-down boxes that
allow the user to link these to available phytoplankton compartments. For convenience sake,
multiple periphyton compartments may be linked to the same phytoplankton compartment.

Periphyton may slough or  be scoured, contributing  to suspended algae; this may be reflected in
the chlorophyll a observed in the water column.  Using this screen, periphyton can be linked to a
phytoplankton compartment  so that chlorophyll a results reflect periphyton sloughing.  One-third
of periphyton is assumed to become the linked phytoplankton and two thirds are assumed to
become suspended detritus  in a sloughing event. When not linked, AQUATOX assumes that all
sloughed periphyton becomes detritus.

Additionally, when phytoplankton undergoes sedimentation it will now be incorporated into the
linked periphyton layer if such a linkage exists.  If multiple periphyton species are linked to a
single phytoplankton species, biomass is distributed to periphyton weighted by the mass of each
periphyton compartment.  If no linkage is present, settling phytoplankton  are assumed to
contribute  to sedimented detritus.

Animal Data Screen

This screen displays all of the relevant parameters  for the animal that has been selected.

The Toxicity Record field within this screen links to the "animal name" within each chemical's
toxicity data. In this manner, several animals could link to the same toxicity record if that is
desired. You may select a record from the list or type a new name if the  desired animal name
does not appear in the pull-down list.  To edit toxicity linkages for all plants and animals in a
simulation simultaneously, click the "Edit AM" button.

As was the case in  the Plant Data Screen, the choice of Animal type is important because
different types  have different physical or biological processes that apply to them.  For instance,
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AQUATOX (RELEASE 3)  USER'S MANUAL
benthic insects are subject to emergence but other animal types are not.  Table 6 in the Technical
Documentation describes the differences between animal types.

For "Benthic Invertebrates," a benthic designation box is available. This box does not affect
model execution but has effects on the calculation of Biological Metrics (see section 4.6 of the
technical documentation.)

Although Animal type is important for determining which processes do or do not apply to the
state variable, the Taxonomic Type or Guild field is included as an organizing tool and does not
affect model output.

Sensitivity of animals to sediments comes into this  parameter screen in several places.  The
feeding effects are found in the top of the screen with the feeding parameters. Lethal effects and
percent-embeddedness effects are found towards the bottom of the screen with the other
mortality parameters.

The fraction in water column field is only relevant  to models running the multi-layer sediment
model and determines how much of the organism is exposed to the water column as opposed to
pore waters in the top layer of sediment.

Bioaccumulation Data includes the lifespan and fraction lipid. Uptake and maximum
bioaccumulation of organic chemicals are sensitive  to these parameters.

Low Oxygen Effects parameters are available for lethality and growth and reproduction
effects.  Ammonia toxicity parameters follow below that followed by Salinity effects.

AQUATOX can model two size classes for each fish species. Records for different size classes
are linked by clicking on Size-Class Links and choosing the correct record from the list given.

Fish may spawn automatically using a formulation based on the optimal temperature parameter
or specific spawning dates may be entered under spawning parameters.

For fish, consumption and  respiration rates can be directly entered into the model (top of screen)
or allometric models can be utilized (bottom of screen). Allometric models within AQUATOX
can calculate consumption and respiration rates as  functions of weight and temperature.

A few notes regarding the some of the more important parameters:

    •  Biomass predictions are sensitive to "Maximum Consumption" rate and "Endogenous
       Respiration" rates when not calculated based on weight (allometics);
    •  Intrinsic mortality rate ("mortality coefficient") is often a site-specific response and is
       therefore subject to calibration;
    •  The  optimum  temperature can have a significant effect on biomass computations;
    •  The  minimum prey for feeding affects the efficiency of foraging behavior;
    •  Allometric means the change in metabolic rate in relation to the size of the organism;
       allometric parameter values may be found in the Wisconsin Bioenergetics Model
       documentation (Hewett and Johnson, 1992; Hanson et al., 1997).

To maximize comprehension, parameters on this  user interface screen are described with English
words rather than symbolically. Appendix B of the  AQUATOX Technical documentation
contains a full description of each of the parameters shown here under "Animal Underlying
Data" as well as their manner of referral in the equations of the technical documentation (often a
shorter variable name). In this manner, a user can use this appendix as a reference to search the
technical documentation and find all equations in which each parameter is utilized. Advanced
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AQUATOX (RELEASE 3)  USER'S MANUAL
users can also easily find the parameters within the AQUATOX source code as the
"internal" variable names are also listed within Appendix B.

See section 4.3 of the technical documentation for extensive discussion of modeling animals
within AQUATOX.
Animal Linkages

Fish may be modeled as a single "stand-alone" compartment, they may be linked by size-class,
using the species data screen or an age-class fish may be modeled.

To get  a good summary of the animal linkages currently utilized by the model, these can be
viewed in Step-7 of the AQUATOX Wizard (example graphic below). This screen also provides a
convenient interface to add size-class or age-class fish.
  Below is the list offish Included in the current simulation.  Click the [Add] or
  [Remove] buttons to modify this list, or the [Next] button to move on.


                            Fish Species in Simulation:
              iShiner: large forage fish. sLngle-conigartiaiieiit fish
              LargeHiQuthBass : game fish, two size-class Qsh
              White Bass : multi-age fish. 15 age-classes
                     Add a Fish Species
Trophic Interactions

When you load an animal into an AQUATOX simulation, you are also loading a trophic interaction
matrix for that particular animal. This trophic interaction table is important because it defines
food-web relationships and assimilation efficiencies.

Within AQUATOX, there are two ways of viewing trophic interactions for a given simulation, on a
species-by-species basis or to view the trophic interactions matrix for the whole simulation. The
species-by-species manner of viewing trophic interactions is generally less useful but is
maintained so that users can edit trophic interactions for libraries as well as for specific
simulations.
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Species Specific Trophic Interactions:
 ACIUATOX—  Trophic Interactions of Single Species
                    Trophic Interactions of Tubifex tubtfex:
                            Preference (ratio)  Egestion jfrac.)
                            i
                            I
                            0.5
Leidy & Ploskey, 1980, p. C4
                          View Organisms in Current Study Only
This screen shows the preferences and egestion factors for a single species. To get to the
single-species trophic interaction screen, within the Animal Library, click on the Trophic
Interactions button that appears at the top right of the screen. To get to the trophic interaction
matrix for an animal within a simulation, click on that same button within the animal's underlying
data.

Within a simulation, you will generally only be interested in viewing an animal's trophic
interactions with organisms within that simulation. To view preferences for all available organism
compartments, clicking  the View All Data button at the bottom of the screen.

The preference ratio indicates the animal's preference to consume a particular compartment on
the list.  Each preference is a fractional preference relative to the other food items on the list, and
it need not add to one.  Each time-step, AQUATOX will normalize these fractional preferences for
the food sources that are available at that time.

The egestion fraction represents the portion of food that is not assimilated for a given animal and
food compartment combination.
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 Trophic Interaction Matrix Screen:
                                                                               """• ™

       • Show Preferences   r Show Egestion Coefficients    <~ Show Comments
                                    Roofer, Bra* white Percr Catfish
                                                                     OK	jB X Cancel
Export Preference Matrix to Excel
This screen enables the user to see all trophic interactions for all animals in a particular species
against all prey items.  Generally, this is a more useful interface for examining and entering
information about the food web.  Preferences are normalized to 100% when the screen is first
shown. After data are  entered or modified, these preferences can  be renormalized by clicking the
"renormalize" button at the top of the screen.

By clicking on the radio buttons at the top of the screen, the user may toggle between
preferences, egestion coefficients, and comments. The matrix may also be exported to Excel for
printing or sharing.

Species Data Screen

The species data screen  allows the user to represent a single species offish with two state
variables representing  two size classes of that fish, generally young of the year (YOY) and adult.
To access this screen select "Size Class Links" from the Animal's underlying data.
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AQUATOX (RELEASE 3)  USER'S MANUAL
                       LgGameFisM: [Smallmouth Bass, Lg]
                       is the same species as
                       SmGameFishl: [Smallmouth Bass, YOi
                       ** No other state variable **
                         Help
OK
Cancel
Large gamefish can only be paired with species loaded into a small gamefish compartment.
Similarly, large bottom fish can only be paired with small bottom fish and large forage fish can
only be paired with small forage fish compartments.  Any species that can be matched within the
simulation will appear on the list within this screen.

To get a good summary of the animal linkages currently utilized by the model, these can be
viewed in Step-7 of the AQUATOX Wizard. This screen also provides a convenient interface to
add size-class or age-class fish.

A similar dialog box may also be utilized to link periphyton to phytoplankton compartments, but
the Plant Linkages screen (accessible by using the "Edit AM" button  in plant underlying data) is
a preferable interface for this procedure as all plant linkages may be viewed simultaneously.

Mult! Age Fish Data Screen

AQUATOX can model one species offish as having multiple age classes with input from this
series of specialized screens.  To access this screen one must first add a multi-age class fish to
the simulation (see below) and then double-click on "Multi.Age-Class Fish " in the state variable
list.
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AQUATOX (RELEASE 3)  USER'S MANUAL
 Input Parameters for Multi-Age Fish
                                             	

         Mutti Age Fish: [Largemouth Bass ]

  General i Initial Condition  Inflow Loadings  Toxicant I.C.  Toxicant Loadings  Lipid Frac. M
         General Parameters:
         Fish Name:   | Large mouth Bass


         Sexually mature at age:  3        years
                 Edit YOY Underlying Data
      Edit Underlying Data for Fish Over One Year in Age
     YOY Trophic Interactions     Older Trophic Interactions
                                                    Help
OK
X Cancel
Tabs at the top enable the user to switch between the different screens, such as Initial
Conditions, Inflow Loadings, Toxicant Initial Conditions and Loadings, Mean Weights, Lipid
Fractions and Mortality Coefficients. Click on each tab name to move from one tab to the next.
For each tab, the user can enter values for each age class or can choose one of several
distributions, characterized by user-supplied statistics.

The values can be graphed as well to better display trends over the life of the fish.

Parameter screens and trophic interaction screens can be accessed foryoung-of-the-year (YOY)
and older fish. When the "older fish" parameter screen and trophic interactions screens are
entered, the parameters are relevant for all age classes one year old  and older.

To add a multi-age clsas fish select to Add a state variable from the main study window and then
select "Multi.Age-Class Fish" at the bottom of the available list of variables to add. A series of
dialogs follows allowing you to designate the number of age-classes modeled (Maximum Fish
Age) and to load parameters into the new fish. Alternatively, a multi-age fish may be added in
Step 7 of the AQUATOX Wizard .

Site Parameters

The Site  Type Screen

This screen allows a user to select which site type is being simulated. Six site types are
available:  pond, lake, stream (creek or river), reservoir, (experimental) enclosure, and estuary.

For standing water (ponds, lakes,  and reservoirs) site type is not currently a sensitive parameter
(these site-types can be used interchangeably).
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Selecting to model a "stream" has the following effects:

    •   the sand-silt-clay model is enabled;
    •   Manning's coefficient may be used to model water volume;
    •   dynamic temperature stratification is disabled;
    •   periphyton scour and velocity limitation for plants is a function of water velocity;

Selecting "Estuary" produces a salt-wedge model and permanent stratification.

For a linked version, a seventh site type is available, that being a "Tributary Input." State
variables are not solved within this type of segment;  rather, loadings of nutrients, organic matter,
and biota to this type of segment are loaded directly  into the directly modeled system.

Additionally, a user may edit a site's underlying data, remineralization parameters, or
stratification options from this screen, or load default site or remineralization data from libraries.

Parameters that may be edited on this screen in a time series or constant manner are

    •   "Fraction of site that is shaded."  See modeling shade.
    •   "Water Velocity." See velocity.
    •   "Mean  Depth." See mean depth.

Site Data Screen

Each site can be characterized by a relatively small number of site constants. These can  be seen
and edited by clicking on Edit Underlying Data in the Site Data window, or they can be loaded
from the Library. There is some redundancy in that Volume, Area, and Mean Depth all have to
be specified. Based on mean and maximum depth, the bathymetry of the site is computed.
Volume is a state variable and can be computed in a variety of ways (accessible through the
volume loading screen ); however, one option is to set it to remain constant using the value
provided in the site screen.

The Max. Length is the distance, usually the long axis, across which wave buildup can occur; it
determines the depth of mixing in stratified systems.

The epilimnetic and hypolimnetic temperature parameters are only used if the user has selected
to "use annual mean and range loadings" on the Temperature Screen. If the user has selected
to use this manner of calculating loadings, then both epilimnetic and hypolimnetic temperature
parameters have to be specified on the site data screen,  even for streams and ponds, where they
can be set equal.

Likewise, if in the Light Screen a user has selected  to "use annual mean and range loadings,"
that user must provide data about the Average Light and the Average  Light Range, from which
seasonal fluctuations are computed. These  are not computed from the  latitude because of local
and regional differences in elevation, cloud cover, and maritime or continental climatic
conditions.

Latitude is used to compute the seasonal variation in day length, though this can be overridden
in the Light Screen. This override can be useful for laboratory simulations.

Altitude is used in the computation of oxygen saturation.

Enclosure wall area is used for experimental enclosures only and affects a site's morphometry.
(Specifically, it increases the fraction of the site area within the euphotic zone.)


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Baseline Percent Embeddedness is the initial condition percent embeddedness for a site. This
value is used to calculate effects when organisms are sensitive to embeddedness, calculated as
a function of TSS. See the "Interstitial Sediments" portion of section 4.3 of the Technical
Documentation.

The Minimum Volume Frac. multiplied by the initial condition volume for a site represents the
minimum volume that a water body can attain within the simulation. If the water volume drops
below this level, numerical modeling of state variables stops and the simulation skips forward to
the next period in time when  the water volume is calculated to be above the minimum level.

Toward the bottom of the site screen are the parameters for the phytoplankton retention
function. Phytoplankton and zooplankton will quickly wash out of a short reach, but may be able
to grow over an extensive reach of a river, including its tributaries.  To solve this problem,
AQUATOX takes into account the "Total Length" of the river being  simulated, as opposed to the
length of the river reach, or "SiteLength" so that phytoplankton and zooplankton production
upstream can be  estimated.  The assumption is that conditions upstream are similar to those in
the reach being modeled.

For estuaries, a series of tidal range parameters is required (amplitudes and epochs).
These are used to calculate the daily average tidal range for the site, which affects the average
depth of the salt wedge on that day.  These parameters are generally available for download
from NOAA databases.

If a stream is being simulated (set  in the Site Type Screen) the Stream Data button in the upper
right is  enabled. Clicking this button displays a series of important stream parameters regarding
site morphometry, habitats represented, and parameters for the optional sand-silt-clay
sediment model.

To maximize comprehension, parameters on this user interface screen are described with several
English words rather than symbolically. Appendix B of the AQUATOX Technical
documentation contains a full  description of each of the  parameters shown here under "Site
Underlying Data" as well as their manner of referral in the equations of the technical
documentation  (often a shorter variable name). In this manner, a user can use this appendix as a
reference to search the technical documentation and find all equations in which each parameter
is utilized. Advanced users can also easily find the parameters within the AQUATOX source
code as the "internal" variable names are also listed within Appendix B.

Remineralization

The parameters on the remineralization screen primarily govern the settling and degradation of
organic matter and its nutrient stoleniometry.

Many of the parameters on the  remineralization screen may be assumed to remain constant from
one simulation to another. A few highlights that may be worth examination:

   •   Maximum degradation  rates:  If site-specific data are available.
   •   CBODu to BODS conversion factor: This becomes relevant if you have organic matter
       loadings into the simulation in the form of BODS.  The conversion factor is variable
       depending on the source of BOD loadings.
   •   Detrital Sedimentation  Rate (KSed): This governs the deposition of organic matter and
       phytoplankton from the water column to sediment (unless the sand-silt-clay model is
       utilized).

Because of these differences, there are three types of Remineralization records:
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AQUATOX (RELEASE 3)  USER'S MANUAL
    •   Eutrophic - should be used in highly nutrient-impacted high systems;
    •   Default - same as Eutrophic record;
    •   Mesotrophic - should be used in moderately nutrient-impacted systems;
    •   Florida - used for Florida lakes; this record is different because of the stoichiometry of
       labile detritus observed in Florida.

Modeling Shade

Shade can be an important limitation to light, especially in riparian systems. A user input "fraction
of site that is shaded" parameter (also known as FracShade) can be entered either as a constant
or as a time series within the "Site" input screen. (\-FracShade) is a multiplier on the amount of
light reaching the water surface.

This parameter can be left as zero to simulate no shading effects on light.

Velocity

Velocity in cm/s may be calculated  by AQUATOX or entered as a time series. Otherwise, velocity
is calculated as a simple function of flow and cross-sectional area.

Velocity has wide reaching effects including:

    •   deposition of phytoplankton and scour of periphyton;
    •   breakage of macrophytes;
    •   entrainment of zooplankton;
    •   deposition or scour of organic matter;
    •   scour and deposition  in the sand-silt-clay model; and
    •   oxygen reaeration.

Mean Depth

Mean depth may be kept constant or entered as a time series.  Mean depth has significant
effects on  the average light climate that plants are exposed to (especially periphyton, which
reside on the  bottom of the water column.)

As noted on the entry screen, if "Use Bathymetry" is not selected, the mean depth calculation is
irrelevant.  In  this case, mean depth is volume divided by surface area.

Stratification Options

In the case of stratification, a site's thermocline depth may be calculated as  a function of
maximum  depth and site-length. Alternatively, the depth of the thermocline  may be entered as a
constant or a  time series. The model will interpolate between entered dates if necessary.
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AQUATOX (RELEASE 3)  USER'S MANUAL
Inflow of water (and the loadings within that waterbody) can be routed to a specific vertical
segment, as can the outflow of water.

AQUATOX calculates stratification based on the temperature difference between segments.
When the temperature difference is less than three degrees, the system is assumed to be
perfectly mixed, otherwise vertical stratification occurs.  Alternatively, on this screen, users may
enter the specific dates of stratification for their modeled systems.  (A time-series may be
entered. On each date where a "1" is entered, the system will stratify, if not already stratified. On
each date where a "0"  is entered, the system will undergo overturn and become completely mixed
again.)

See section 3.4 of the technical documentation for more discussion of stratification and mixing.

Exploring Model Setup

Study Setup

The Setup Screen specifies important parameters about the simulation itself (as opposed to any
of the components of the simulation.)
Simulation Time and Relative Error

At the top of the setup screen you can  modify the first and last days of the simulation.
defines the simulation period.
This
The Relative Error is the acceptable error in the simulation; if it is not achieved in a particular
time step, the variable Runge-Kutta routine decreases the step size and tries again.  If the relative
error is too large, the results may be erroneous; if it is too small, the run time may be too long.
Usually a value between 0.005 and 0.0005 is appropriate, but you may wish to experiment for a
particular application.

Daily and Hourly Simulations: A user may set the native model time step to one day or one
hour.  If a daily simulation is utilized, average light conditions are utilized throughout the day. If
an hourly simulation is selected, solar radiation is calculated as variable during the course of each
day. Rather than calculating daily average oxygen concentrations, if an hourly simulation is
selected, AQUATOX will simulate  hourly average oxygen concentrations within the water
column. These concentrations will be based on the hourly light climate (and optional hourly
oxygen loadings). These hourly predictions will then be used to calculate lethal and non-lethal
effects due to low oxygen.  The user can output hourly values of DO or set a larger Data Storage
Step and examine the minimum and maximum predictions over that time interval.
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AQUATOX (RELEASE 3)  USER'S MANUAL
Biota Modeling Options

If the model is run in "Spin-up Mode" initial conditions for biota will be set based on the model
results on the last time-step of the simulation. This can be useful in setting biotic initial conditions
when there are insufficient available data. As initial conditions will be automatically overwritten it
is best to first save a simulation before running in "spin-up mode."

Toxicant Modeling Options

If you wish to compute steady-state BAFs, you may wish to keep the freely dissolved toxicant
constant; this was done in an application concerning PCBs in Lake Ontario (see Validation
Studies document). Otherwise toxicant mass is balanced on the basis of loading, partitioning,
washout, and other toxicant loss terms.

Toxic effects may be calculated based on internal or external concentrations. See Chapter 9
of the Technical Documentation for more information about the consequences of this choice.

When calculating effects based on external concentrations, internal concentrations may be less
important. For this reason, a user may wish to estimate toxicant uptake based on  BCF rather
than using the default kinetic model of toxicant uptake.  In many cases this will speed up
simulation run time significantly.

The older literature often did not distinguish between freely dissolved contaminants and those
complexed with dissolved organic matter. You may  choose to include the complexed
contaminant in computing BAFs so that the results are directly comparable with the older
literature values.

Output Options

The Data Storage Step represents the time period over which results are averaged.  This
obviously can have a significant effect on the amount of output that is produced by the model.

In plotting output for stratified systems it is usually more pleasing to plot continuous values for the
hypolimnion, even when the system is not stratified.  This is done by duplicating epilimnion values
for the hypolimnion when the system is well mixed (by selecting the Write Hypolimnion Data
When System not Stratified option); however, this  option takes additional storage, so you may
choose not to duplicate those data points, especially in  systems that may not undergo
stratification.

If you  click on Show Integration Info, you will be able to see what time steps are used in solving
the differential equations and which rates and associated relative errors are causing the
integration to slow down while the model is running.

The default method of averaging results is by Trapezoidal Integration, which calculates the
average value that has occurred since the last data storage step. A user may also bypass
integration altogether and output "instantaneous" concentrations, that is to say, predictions
that occur exactly at a given time-step.

For more information about the other buttons on the screen, please see the Rate Output Screen,
Uncertainty Setup Screen, Control Setup Screen, or Output Setup Screen sections of the
help file.
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AQUATOX (RELEASE 3)  USER'S MANUAL
Rate Output Screen

You may save state variable rates internally, or externally for examination with a spreadsheet
program.

In the Setup Screen, choose Save Rates and click on Rate Specifications to designate those
state variables for which you want the additional output.

If the rates are saved internally, each element of a selected state variable's derivative will be
integrated and saved along with the results for the simulation. They can then be graphed or
viewed in tabular format. Units for rates are "percent" which is short for "percent of state variable
concentration per day."

If rates are saved externally, instantaneous rates will be saved into an Excel file within the
designated AQUATOX "OUTPUT" directory. The output file name will be the name of the study
file, followed by "_Rate.xls"  However, rates saved internally can also later be exported to Excel
(or other format) along with all other model output.

Usually you will wish to save rates for each data-storage step, by choosing "When Writing
Results". However, when exporting to  Excel, you can also save rates for each step in the
solution of the differential equations, that is, "Each Attempted Step". This provides data about
sub-steps within the data reporting interval as well as steps that were thrown out because the
relative error of the numerical analysis was too great.
Uncertainty Setup Screen

AQUATOX can be run using point estimates for all available parameters (deterministic mode).

Alternatively, a Latin Hypercube uncertainty analysis can be utilized, or a nominal range
sensitivity analysis.

Deterministic Mode

Deterministic mode is the default mode for the AQUATOX model.  In this case, a single
simulation is used, and all parameters are represented by point-estimates.

Uncertainty Mode

Another powerful feature of AQUATOX is that it can perform uncertainty or sensitivity analysis to
provide probabilistic results. Latin hypercube sampling is performed, ensuring that all parts of the
chosen distribution are sampled. Therefore, the number of iterations can be kept to a minimum,
which is important because each iteration is a complete simulation. Twenty iterations is the
default, meaning that the distribution is divided into 20 segments for purposes of sampling, and
that twenty simulations will be run (besides the initial "deterministic" run). The number of
iterations should be increased as the number of involved  parameters increases.

The Uncertainty Setup screen enables you to view all of the parameters and loadings that can be
chosen, either singly or in combination.

Nearly all AQUATOX parameters can be modified in an uncertainty analysis. Selecting the top
branch of the tree interface (the "plus" next to All Distributions ") allows you to view all possible
distributions that could be utilized (parameters that can be modeled as a distribution rather than a
point estimate). Alternatively, distributions can be sorted  by parameter type or by state variable.


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The bottom branch of the tree interface shows the distributions that are currently included in the
model's uncertainty run. (Click on the  "+" or "-" symbols to expand or contract the various
portions of the tree interface.)

You may choose to utilize a non-random seed if you wish to be able to reproduce your
simulation's results.  A pseudo-random number generator requires a "seed" to start its sequence
of "random" numbers.  A random seed will be based on the exact clock time in which the
simulation was started and this sequence will therefore not be reproduced in later simulations. A
non-random seed will produce a repeatable sequence of random numbers.

To turn off the uncertainty portion of AQUATOX, select the "Deterministic Mode" radio button,
below the list  of distributions.

By double-clicking on a variable one can access the corresponding distribution information.

Correlations  between modeled distributions may also be modeled.

When you have left this screen and choose to run an uncertainty analysis, you will be prompted
for a database file name in which to store the results. Based on all of the iterations, the minimum,
maximum, mean, deterministic, and standard deviation result for each data-storage step will be
saved in these database files. (Sometimes uncertainty results will not fit in one database file and
will need to be added to one or more additional successively numbered database files.)  For
more information on viewing these files, see Viewing Uncertainty Output.

Also see section 2.5 of the technical documentation for more discussion of the AQUATOX
uncertainty analysis.

Distribution Information

This screen shows whether a parametric distribution has been chosen for a given variable or
whether a point estimate is used. The radio buttons at the bottom of the screen are used
to toggle back and forth.

The default is a normal distribution with a mean of the point-estimate parameter value and a
standard deviation of 60% that value.  In most cases this will need to be overwritten with
parameter-specific information.

A triangular, uniform, normal,  or lognormal distribution may be chosen and the nature of the
parameters changes based on the selected distribution.  Graphs of the distribution will then be
produced and can be used to ensure that the parameters chosen  are producing the desired
distribution. Please check these graphs (probability and cumulative  distribution) carefully.

Often  loadings are well represented by lognormal distributions.  If less is known about the
distribution, but  minimum and maximum values and some central tendency can be defined, then
a triangular distribution may be appropriate.  If only minimum and maximum values are known,
then a uniform distribution may sufficient, and the number of iterations can be decreased.

Note for the lognormal distribution that the parameters are "mean" and "standard deviation" rather
than geometric mean (GM) and geometric standard deviation (GSD). To convert a GM or GSD to
the required parameters, take the natural logarithm of the GM or GSD.

Correlations

This screen allows a user to specify correlations between two or more input distributions. All
selected distributions for the uncertainty analysis are  listed at the top of the screen. The user


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may then add a correlation, edit a correlation or remove a correlation from the list of correlated
distributions.

Within AQUATOX, input distributions for uncertainty analyses are correlated using the method of
Iman & Conover (1982). The Iman and Conover method induces a rank correlation between two
randomly derived  (stochastic) distributions such that the correlation of the two randomly derived
distributions matches the correlation of the input data.  Output distributions can be examined to
verify that they closely match the correlation coefficient of the input distributions.

For the Iman Conover method to work, correlation matrices must be "Positive  Definite." In the
authors' experiments with input matrices, we have found this to mean three things in  practice:

    •   No correlations greater than 0.8 or less than -0.8 may be  used.
    •   When there are multiple correlations for the same variable, often correlations must be
       reduced to less than 0.75 (or greater than -0.75) due to interactions between variables.
    •   The method requires a minimum number of iterations to successfully impose correlations
       on  input distributions.  A specific error will be raised if insufficient iterations have been
       specified (asking the user to increase the specified number of uncertainty iterations).

To examine the input distribution correlation matrix and test for positive definite status, a user
may use the "summarize correlation matrix"  button.  Unless an  error is indicated, the output will
summarize the user-specified correlation matrix followed by the rank correlation of draws
produced by the correlation method.

For more information on this method, please see: Iman, R. L, and W. J. Conover. 1982. A
distribution-free approach to inducing rank correlation among input variables. Communications in
Statistics 611:311 -334.

Sensitivity

"Sensitivity" refers to the variation in output of a mathematical model with  respect to changes  in
the values of the model inputs (Saltelli 2001). It provides a ranking of the model input
assumptions with  respect to their relative contribution to model output variability or uncertainty
(U.S.  Environmental Protection Agency 1997).

AQUATOX can automate a nominal range sensitivity analysis (also known as  a "one-at-a-time"
sensitivity analysis). This analysis assumes independence among tested parameters.

As was the case for the uncertainty analysis, nearly all AQUATOX parameters can be tested for
sensitivity.  Selecting the top branch of the tree interface (the "plus" next to All Distributions  ")
allows you to view all possible parameters that can be  tested for sensitivity.
Alternatively, parameters can be sorted by parameter type or by associated state variable. The
bottom branch of the tree interface shows the distributions that are currently included in the
model's sensitivity analysis. (Click on the "+" or"-" symbols to expand or contract the various
portions of the tree interface.)  If you double-click on any of these  parameters, or  press 
while the parameter is highlighted the parameter will be selected (or deselected) for the sensitivity
analysis.

To run the model in sensitivity mode, the user must

    •   Select one or more parameters to test  using the procedure given above;
    •   Select a set of output to track using the Choose Output to Track button; and
    •   Choose the percentage to vary by editing "Nominal Percent to Vary."


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When the model is then executed, the user will be prompted for an Excel file-name in which to
save model results.  The model will run a deterministic run and then will run the model twice for
each parameter selected (testing sensitivity to changes in each parameter in the positive and
negative directions in the magnitude of "percent to vary.")  In this manner, if a user selects n
parameters, the model will run (2n+1) iterations.  It should be noted that the number of output
variables will not appreciably affect the simulation run time. However, the number of tested
parameters obviously determines the number of iterations and therefore the total simulation time.

One important note about sensitivity analyses: Sensitivity analyses will calculate the percent
difference between the results of the deterministic run and the modified simulation in the last
averaged time-step of the simulation. In most  cases the user will want to utilize a much longer
averaging period within the model setup screen, for example, to calculate averaged sensitivities
over the last year of the simulation or the entire simulation. If sensitivity at an intermediate date is
desired, the simulation should  be shortened so that the intermediate period is reflected in the last
averaged time-step of the simulation.

Periphyton and phytoplankton  may be linked within the AQUATOX model. If the "Link
Periphyton/Phytoplankton" check-box is selected, whenever a linked periphyton or
phytoplankton  parameter is varied, the same parameter will be varied within the linked algal state-
variable by the same percentage.

After the run is complete, the output file can be viewed in Excel. Additionally, AQUATOX can
automatically produce tornado diagrams of the results. See Viewing Sensitivity Output for
more information.  Also see section 2.4 in the technical documentation.

References:

Saltelli, A. 2001. Sensitivity Analysis for Importance Assessment. Paper read at Sensitivity
Analysis Methods, June 11-12, 2001, at  North Carolina State University, Raleigh NC

U.S. Environmental Protection Agency. 1997. Guiding Principles for Monte Carlo Analysis. Risk
Assessment Forum. Washington, DC: U.S. Environmental Protection Agency

Control Setup Screen

There are two separate banks of memory for model results, one which  is labeled "control" and
one which is labeled "perturbed." The default is for the control simulation to have all organic
toxicants zeroed out or omitted. However, there is considerable flexibility in setting up the control
run.  For example, toxicants can be kept and point-source nutrients can be omitted in the control
run.  In fact, it is possible with a few judicial choices to set up a factorial analysis to determine the
effects of various combinations of pollution control scenarios.

If none of the check-boxes are selected, then the  results of the control  and perturbed simulations
will be identical. A user could, however, first run a "control" simulation, then change parameters
or loadings in the simulation as a "perturbation," and finally run a "perturbed" simulation.  In this
manner, the control and perturbed sets of results  can be used with considerable flexibility, without
even utilizing the "control-setup" screen.

Due to the multi-threaded nature of AQUATOX, it is often useful to run  the control and perturbed
simulations simultaneously, especially with multi-core computer processors.
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Output Setup Screen (Results Tracking)

The Output Setup Screen, (available through a button on the setup screen) allows the user to
select which output is saved by AQUATOX, and which output is not saved.

Because AQUATOX has the potential for modeling so many state variables, a user has the option
of not saving given state variables, thus decreasing the memory requirements. This can be
especially important when running simulations on a decadal scale or when outputting many data-
points per day. However, given the RAM capacity of modern computers, this screen rarely needs
to be utilized.

Use the "<" and ">" buttons to move results into the "Results to Track" or "Results NOT to Track"
lists.

If "Save PPB Data" is unchecked, the model will not save concentrations of chemicals in biota
with PPB units.

Note, when stratification is likely to occur, assume that RAM memory utilization will roughly
double.

Running the Simulation

To run an AQUATOX simulation, select the "Control" or "Perturbed" button in the main window.

The model will run  for the amount of time specified in the setup screen.  When the model has
completed you may use the output button to view the model results.

You may work with the  model while it is running and, for example, change parameters or open
different models. If you change parameters though, AQUATOX will warn you when it is copying
results back to the  interface as your model results will no longer match the parameters or
loadings associated with that study.  In this case you will be able to optionally save the model
results to a new file in which the model parameters will match the model output.

Control vs. Perturbed Simulations

A powerful feature  of AQUATOX is that it can run paired simulations for  perturbed and control
conditions. You can use the control setup screen to specify the differences between these two
types of runs, or you can change parameters or loadings in between running the two different
types of simulations.

The graphical output screen allows users to compare control and perturbed graphs and also to
display difference graphs in which percentage differences are displayed.

Viewing  Graphical Output

AQUATOX has extensive capabilities for graphical and tabular output. By clicking on Output on
the main screen one can create a new Output Window.  There are four tabs available at the top
of this window, each representing a different form of output. Click on the tabs at the top of the
output screen to move from one type of output to the next.

The default window is the "Graph Library" window.  Graphical output is often the most useful
manner to quickly view  and interpret model output. The AQUATOX graphing capabilities are
quite powerful as described in the following sections.


                                             35

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AQUATOX (RELEASE 3)  USER'S MANUAL
The first two tabs "Perturbed Simulation" and "Control Simulation" provide tables of output for
the control and perturbed results.

The tab on the right leads to a window in which uncertainty and sensitivity results may be
examined.

Within the output window you may also save the current bank of results and the "graph
library" along with the current set of parameters using the "Save These Results"  button.  This is
no different from exiting the output window and saving the existing "APS" file. Results may also
be loaded using the "Load Results from File" button.

Viewing Existing Graphs

To determine which graphs have been  produced for the given simulation, click on the triangle at
the top left of the  graph. If there is more than one graph included, a drop-down list of graphs will
be shown. The user may select a graph within this list or use the up and down arrows to the right
of the list to scroll through the graph library.
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Control, Perturbed, and Difference Graphs

Each graph may be displayed in perturbed mode, control mode, or difference mode.  By selecting

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AQUATOX (RELEASE 3)  USER'S MANUAL
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AQUATOX (RELEASE 3)  USER'S MANUAL


Editing an Existing Graph

Modifying a Graph

By clicking on the "edit" button above a graph,  one can get to this Change Graph Variables
screen.
 ซ|s|,Chan.gR Srapjh Variables

    Graph Name:  ammonia and detritus]

                                                               Graph Date Range:  reset
      Show All Results
    <~" Filter By Substring:
   I   Exclude Substring
      Selected Set of Results
                                    Results on Y1 Axis (mg/L);
     Tot. Sol. P(mg/L)
     CO2 (mg/L!
     Oxygen (mg/L)
     TSS(mgfL)
     R ctetr sed (g/m2 (Jry)
     L detr sed (gfm2 dry)
     BuryRDetr(kg/m2)
     BuryLDetr (kg/m2)
     Peri Low-Nut D (grtn2 dry)
     Peri High-Nut (gfm2 dry)
     Phyt High-Nut (mg/L dry)
     Phyt Low-Nut D (mg/L dry)
     Phyto, Navicul (mg/L dry)
     Peri, Nitzschi (g/m2 dry)
     Cladophora (g/m2 dry)
     Peri, Green (g/m2 dry)
     Phyto, Green (mg/L dry)
     Phyt, Blue-Gre (mg/L dry)
     Peri, Blue-Gre (g/m2 dry)
                               (V use Automatic Scaling
                               r Use Below Values
Results on Y2 Axis  fmolLdry)
Ldetrdiss
R detr part
                               fซ use Automatic Scaling
                               r Use Below Values
                                                                r Logarithmic Y Axis
                                                                  Auto Scale to Control 8, Perturbed
At the top of this window is the capability to edit the graph's name. This affects how the graph
appears in the drop-down list.

Next, choose the output to place on each axis.  There are potentially hundreds of output variables
associated with each simulation. The filter tools help the user pick out relevant variables from this
considerable list (see Selecting from a  List of Output.)

You may graph variables on either one or two Y axes.  Use the button under the results list to
toggle between one or two Y axes.  Use the "<" and ">" buttons to move results into and out of a
particular axes' results. The "ป" button moves all variables into the selected axis.  Note: all
variables on the same axis must have the same units.

The Graph Date Range (x axis range) may be set to a smaller or larger range of dates than the
AQUATOX simulation by typing dates in the edit boxes. Selecting "reset" above the date range
will set the graph date range to the dates of available model results.

Scaling for each axis (Y1 and Y2) may be entered  manually or automatically calculated  by
AQUATOX. Under the axis scale entry is the option to model a Logarithmic Y Axis.
                                                 38

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AQUATOX (RELEASE 3)  USER'S MANUAL
The check-box to Auto Scale to Control & Perturbed will scale the Y axes to the minimum and
maximum of botfithe control and perturbed simulations (if they both have been run). This
enables the user to switch back and forth between control and perturbed graphs while looking at
axis-scales that remain constant between the two graphs, enabling easier comparison.

By using the Graph Type pull-down menu, you can choose between the four types of plots:

    •   The X-Y plot is the standard, this plots observed results against dates.
    •   Percent exceedance graphs plot percent exceedance on the X axis against model
       results on the Y axes.
    •   Duration graphs plot duration of exceedance on the X axis against model results on the
       Y axes.
    •   Scatter plots allow you to examine correlations between model outputs by plotting one
       model output on the X axis against one or more different model outputs on the Y axis.

If the system has stratified results, you can choose the vertical layer that you wish to display
results for using the Epilimnion and Hypolimnion radio buttons.  This may also be toggled in the
graph menu.

One additional note: if a duration or exceedance graphs is displayed, a "Draw Threshold" button
is displayed on the  lower right of the graph. When pressed, a dialog box comes up that both
displays duration of exceedance above a threshold and draws a line on the graph.

Selecting from a List of Output

There are potentially hundreds of output variables associated with each simulation. To help the
user pick out relevant variables to be graphed, the "Filter By  Substring" option is available at the
top of the screen. Typing a string in the box provided limits the list of variables to those with
names that include  the string. For example:

       Type "peri" to find all periphyton variables
       Type "mg/L" to find all variables with units of mg/L.
       Type "ppb" to find all parts per billion output.
       Type "detr" to find all results that pertain to detritus.
       Type "T1" to find all results that pertain to the first organic toxicant.

You may also exclude any results containing the substring by checking the "Exclude Substring"
box.

Use the "<" and ">" buttons to move results into and out of the "Available Results" and "Results to
Display" columns. Using "ซ" and "ป" moves all results into or out of the relevant column.

Graph Menu

At the top right of the graph is a menu that may be accessed as a  pull-down menu. After the
menu has been used, users may repeat their  last action by clicking the "(repeat)" button that
appears. Note: this menu may also be accessed by right-clicking on any AQUATOX graph.
                                              39

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AQUATOX (RELEASE 3)  USER'S MANUAL
Menu:


	
T 1
Copy \>
Print
Graph Setup
Erase Graph
Other Segment
Help
External Data
Write Stein ha us
Trophic State Indices
Export all MSWord
Copy

The copy option allows a user to copy the current graph as a bitmap or a windows meta-file to be
copied into presentations, documents, logs, or reports. Bitmaps are generally smaller files, but
meta-files give better resolution.

Print

The print option allows the user to print the current graph. A "preview" window will first appear
allowing the user to select printer,  orientation, margins, etc.

Graph-Setup

The graph-setup menu allows the  user to change the colors, symbols, default text, and fonts used
on a graph through a dialog box. (See Graph Setup Screen).

Erase Graph

Erase graph will permanently remove the current graph from the graph library (after prompting).

Other Segment

If there are results for both the  epilimnion and hypolimnion segments, this menu option toggles
between the two segments.  Currently, in linked mode, to view another segment, the user must
first go into that segment and then open the output window for that segment.

External Data

Allows the user to  import observed data. (See Importing Observed Data)

Write Steinhaus

This option enables a user to write a set of Steinhaus similarity indices in Microsoft Excel format.
The Steinhaus index (Legendre and  Legendre 1998) measures the concordance in values
between two samples for each species.  A Steinhaus index of 1.0  indicates that all species have
identical biomass in both simulations (i.e., the perturbed and control simulations); an index of 0.0
indicates a complete dissimilarity between the two simulations.

A time series of indices is written for each day of the simulation representing the overall species
similarity on that date. Separate indices are written out for all plants, all animals, invertebrates
only, and fish only. Completed control and perturbed simulations (of the same length) must have
been completed for this option  to work.
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AQUATOX (RELEASE 3)  USER'S MANUAL
Trophic State Indices

AQUATOX will calculate and display trophic state indices (TSIs) for lakes using the methods
identified in the EPA Nutrient Criteria Technical Guidance Manual
(http://www.epa.gov/waterscience/criteria/nutrient/guidance/lakes/lakes.pdf). Separate TSIs are
calculated based on average concentrations of Total N, Total P, and Chlorophyll a, as well as
average Secchi Depths. The date range over which values are averaged may be specified in
the TSI calculation screen as well as the AQUATOX scenario (control or perturbed) and the
vertical segment (epilimnion or hypolimnion, if relevant).

Export All MSWord

All graphs will be exported into a new Microsoft Word document in "Windows Metafile" format.
When this option is selected, Microsoft Word will start up and each of the graphs will be pasted
into a new document. Microsoft Word 2000 or later must be installed on the user's system for this
option to work properly.  Note, Windows Metafile does not always work properly when computer
monitors are in "portrait mode," due to what is apparently a  minor glitch in Microsoft Windows
(fonts become distorted).

Arrange Graphs

When dealing with a large number of graphs, it can be very useful to arrange them in a particular
order. This also affects the order of the graphs when they are exported to Microsoft Word.  When
the "Arrange Graphs" menu item is selected, a dialog box appears showing all available graphs
for the simulation. The user may drag and drop the graph names in the list to reorganize them or
use "up" and "down" arrows next to the list to reorder them.  Graphs may also be deleted using a
"delete this graph" button or by pressing the  key when a graph is selected. All changes
made in this window may be undone using the "cancel" button.

Statistics from Graph

The statistics  dialog allows the user to examine and export statistics based on the current graph.
(See Output Statistics).
The Graph Setup Screen

This screen allows a user to change the appearance of the current graph.  The graph can be
shown in 3D, and various grid-line options can be selected. The 3D option is somewhat flashy
but not particularly flexible and of limited utility.
      Appearance


   17 Chart Is 3D

   F Show Vertical Gridlines

   17 Show Y1 Ms Gridlines

   r Show Y2,Axis Gridlines
                                             41

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AQUATOX (RELEASE 3)  USER'S MANUAL
Additionally, captions, text, and font can be edited to personalize the graph.  Un-checkthe
relevant "Use Defaults" check-box and edit the text under "Custom Captions."  Fonts may be
modified using the font buttons.
            Use      Custom Captions
          Defaults
     Titlel

     Title2

     X/teis

     Y1 .Axis

     Y2Axis
Finally, by selecting a data-series from the "Series Specific Characteristics" list, a user can
select which color, line-thickness, symbol size, and symbol represents the selected series.  The
"SmallDot" symbol renders like a thin line, and is the AQUATOX default. Sometimes it is useful
to change the symbol or line-thickness when it is desirable to emphasize one of several series on
the graph.

All of the Graph Setup changes will be saved along with the graph when the study (APS) file is
saved.
Importing Observed Data

External "observed" data may be imported into AQUATOX for graphing against model results.
After a data series has been imported, it will be displayed at the bottom of the list of data
available for graphing in the edit graph dialog. You may also use the text filter within that dialog
to find your observed data.

To access the "Attached Observed Data" window select "External Data" from the Graph Menu.
                                             42

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AQUATOX (RELEASE 3)  USER'S MANUAL
     Observed Data Series Currently Attached
       ake C dn\ a (ug/L)
       ake C obs Secchi (m)
       ake C Era DO fm
      C, Tot Diatom Density (*/mL)
             Series Characteristics
                 Series Name
      |c, Tot Algae Density
                  Series Units
           Series includes Error Bars  p
         Series includes hJon-Detects  p
              Series in Hypolimnion  T
              Reference, Comment
       Obs Algae.xls, {Oklahoma State
       University 1996).
                   Import Data
6/4J1992
7/2/1992
8/19/1992
9/12/1992
10/24/1992
3/8/1993
4/18/1993
5/26/1993
6/25/1993
7/23/1993
8/4/1993
8/19/1993
9/2/1993
9/16/1993
9/30/1993
10/22/1993
Data
1865.55
2025.02
8100.1
1616.86
382.07
171.48
2987.96
1676,04
2846.92
4475.08
2894.71
5956.23
4563.4
3155.45
1002.37
2585.97
To add a series, use the Add button; the Delete button deletes the selected series.
When a new series has been added, you may edit the series name and the series units.  The
series units are important as you may only put items with precisely the same units on a single Y
axis. So copy the text format of any units that you wish to plot against your imported data
precisely. A reference or comment may be added below in which the source of the external
data is identified.
Imported series may contain error bars, in which  case vertical bars will be plotted against every
point in which error bars are included.  Imported series may also contain non-detects in which
case red arrows will be plotted at the location of the value (which is assumed to represent the
detection limit). You may also flag whether the  series belongs within the Hypolimnion segment.
After setting these flags, click the Import Data button.
The interface to import external data is almost identical to the one  used to import loadings.  The
significant differences come because non-detects and Error bars may be imported. To change
                                               43

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AQUATOX (RELEASE 3)  USER'S MANUAL
the file type to import, click on the pulldown menu under "List Files of Type." Directions about
the precise file-format required will be found in the rectangle at the bottom of the screen.
             File Name:
              *.xls
                                 Directories:

                                 C:\WortAAQUATOX\Rel3.1\PROGiRAM
                                                 AQUATOX
                                                 • Re 13.1

                                                CD Backups
                                                CD DLL
                                                CD Model Support_aquato;
              List Files of lype:
                                Drives:
              Excel f .xlsj
                             * for        i
greater. Cols. D & E hold optional min. and max. bars,   j
Or the location  of data within Excel can be entered above.!
                                                                   Cancel
Generally, the date appears in the first column of the file, with the values to be imported in the
second column; however when importing from Excel you can change the column designations.  If
you are importing Non Detects check the yellow "Import NDs" box.  Non-detects appear in the
third column.  If a value is not detected in the greater-than direction, use the ">" identifier,
otherwise any non-blank value will be interpreted as a "lesser-than" non-detect which is much
more common.

If you wish to import error bars, check the "Import Err Bars" box.  Minimum bar values will be
imported from the fourth column with maximum bars imported from the fifth column.

After data have been imported, they will be displayed along with their dates in a table.  This table
may be edited by double-clicking on a cell and editing it.  Additional rows may be added or rows
may be deleted from the table using the directions that appear below it. ( and
)
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AQUATOX (RELEASE 3)  USER'S MANUAL
When you have finished importing and editing your observed data series, select the OK button
and it will appear in the list of output available to graph.  The external data, and any plots created
with it, will be saved and loaded along with your AQUATOX simulation.

Output Statistics Screen

This dialog box enables the user to examine the following statistics that pertain to AQUATOX
Output:

    •   n: number of data points evaluated;
    •   Mean: (Sum/ n);
    •   Median: The 50th percentile. If there is an even number of data points, the average of
       the two middle values;
    •   Minimum: Minimum value given the time-period chosen;
    •   Maximum: Maximum value given the time-period chosen;
    •   Variance: A measure of statistical dispersion.  Calculated using the robust two-pass
       algorithm;
    •   Standard Deviation: Square root of the variance;
    •   5th Percentile: 95% of values are greater than this value. The 5th percentile rank is
       calculated of the ordered data and, if the rank is non-integer, linear interpolation is used
       between the two nearest ranks;
    •   95th Percentile: 5% of values are greater than this value. Calculated using linear
       interpolation as above.

The date ranges, scenarios to export statistics for (i.e. "control" vs. "perturbed,") and the vertical
segment (if relevant) may all be edited and the statistics will automatically  update to reflect the
changes from the various inputs and pull-down menus.

AQUATOX displays these statistics in a fixed-width basis and they may be copied and pasted to
any spreadsheet or database software.  The width of numbers and the width of the output names
may also be varied using the entry boxes at the bottom of the screen. Statistics may also be
exported to Excel using a button at the bottom of the screen.

The derivation  of statistics from external "observed" datasets is not enabled at this time.

Viewing Table Output

Tables can be  obtained for both perturbed and control runs.  Click on the tabs at the top of the
output screen to move from one type of tabular output to the next.

Using the Change Variables button, the user can specify what variables are to be tabulated.

Exporting Tabular Output

Using the Save Table to Excel button, the results can be saved to an Excel file. This button
requires Microsoft EXCEL to be installed on the user's machine to work properly. After the export
is completed the user is prompted  as to whether they wish to view the Excel file immediately or
just leave it saved to disk in the specified location.

Viewing Uncertainty Output

An additional form of graphical output is the Uncertainty Graph.  If an uncertainty analysis has
been performed, the results can be potted as a series of lines representing the mean, minimum,

                                             45

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AQUATOX (RELEASE 3)  USER'S MANUAL
maximum, mean -1 standard deviation, mean + 1 standard deviation, and deterministic results as
tracked through the simulation time.  Only one output variable can be viewed at a time, so click
on View a Different Variable to view another.  The multi-purpose uncertainty window may also
used to view the results of a sensitivity analysis .
 Perturbed Simulation  Control Simulation  Graph Library  Uncert. Sensitivity
                                          Copy
Graph Setup
Print Graph
  View a Different Database

Viewing Data in File: C:\work\AQUATOXWocumentsWorkingDocsWncertJ5enslMueEarthtie_Uneert_BMP_FixE
     View a Different Variable
                                            View Biomass Risk Graph
                    v Show Min. & Max
                         NH3 & NH4+ (mg/L)
                         3/27/2008 9:25:25 PM
                                                                   - Mean
                                                                   — Minimum
                                                                   — Maximum
                                                                   - Mean - StDev
                                                                   — Mean + StDev
                                                                   — Deterministic
          3/21/1999  7/19/1999  11/16/1999  3/15/2000   7/13/2000   11/10/2000
Uncertainty output is not saved as part of an APS file but is instead saved in Paradox databases
(*.db).  Sometimes uncertainty results will not fit in one database file (which are limited to 255
columns); the results are then added to one or more successively-numbered database
files. Immediately after running an uncertainty analysis, the uncertainty window will bring up the
primary database file that is associated with the uncertainty run.  However, if the study is closed
and opened again, the pertinent database file will need to be re-opened using the View a
Different Database button. This will also be required if the output variable you wish to graph is in
one of the successively numbered database files.

When many iterations have been run, the minimum and maximum output become less relevant
(existing  far out on the tail of the output distribution, with very little probability of occurrence.)  For
this reason, the minimum and maximum scenarios can be toggled on or off again using the Show
Min & Max checkbox.

Of particular interest to risk assessors is the Biomass Risk Graph, which plots the probabilistic
results as "percent probabilities" against "percent declines" by the end of the simulation.  Any
number of organisms can be plotted simultaneously on the Risk Graph, so that the responses of
both tolerant and intolerant organisms can be analyzed.  If an organism increases in biomass, for
example  because of release of herbivory (for plants) or predation  (for animals), then the percent
declines  are shown as negative values.  Data to support the biomass risk graph are stored in
CSV files with the suffix "_decline" added to the end of the file-name. A different CSV file may be
loaded using the View a different CSV file button, though the CSV file that is relevant to the
simulation will be initially displayed.
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AQUATOX (RELEASE 3)   USER'S MANUAL
100.0
95.0
900
85.0 -
80.0
| 70.0
& 65.0
I 60.0
| 55.0
| 50.0
i%
40.0
35,0-
30.0
25.0
20.0
Biornass Risk Graph
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* Largemouth Ba
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40 SO 60 70 80 90
Percent Decline at Simulation End
Uncertainty graphs may be copied to the clipboard using the "Copy" button, or printed using the
"Print Graph" button.  The "Graph Setup"  button brings up the graph setup window allowing
the appearance of the graph to be edited.

Viewing Sensitivity Output

The same screen that is used to view uncertainty output can automatically produce tornado
diagrams which is a visual manner of interpreting the results of a sensitivity analysis  .
 Peri. Chlorophyll (mg'sq.m)
Show12   Most Sensitive Variables  [jBe
            Sensitivity of Peri. Chlorophyll (mg/sq.m) to 20% change in tested parameters
                                       3/27/20089:46:11 PM
          101% - Peri, Green: Optimal Temperature (deg. C) * Linked *
         91.7% - Phyto, Green: Optimal Temperature (deg. C) * Linked"
        68.5% - Phyto, Green: Max Photosynthetic Rate (1 /d) * Linked *
         61.3% - Peri, Green: Max Photosynthetic Rate (1 /d) * Linked *
        45.1 % - Phyt High-Nut: Optimal Temperature (deg. C) * Linked *
         33% - Peri Low-Nut D: Optimal Temperature (deg. C) * Linked *
       29.2% - Phyt High-Nut: Max Photosynthetic Rate (1 /d) * Linked *
          24.8% - Phyt Low-Nut D: N Half-saturation (mg/L) ' Linked *
             24.8% - Peri, Navicula: N Half-saturation (mg/L) * Linked *
             24.2% - Phyto, Green: N Half-saturation (mg/L)" Linked"
              23.5% - Peri, Green: N Half-saturation (mg/L)" Linked"
   21.2% - Peri, Green: Exponential Mort. Coefficient: (max / d) * Linked *
                                                            80        90        100
                                                               Peri. Chlorophyll (mg/sq.m)
                                                                                          110
                                                    47

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AQUATOX (RELEASE 3) USER'S MANUAL
For a particular model output, a tornado diagram displays the parameters that cause the most
sensitivity, sorted in decreasing order of sensitivity. Red bars plot results in which the parameter
has been reduced by the given percent and blue bars plot results in which the parameter has
been increased. The black vertical line in the middle of the blue and red bars represents
the baseline model result (in which no sensitivity test was performed.)  Clicking on any of the bars
will give the user information about which parameter was tested and the result as compared to
the baseline result.

The statistic shown in front of the parameter names is the "sensitivity" statistic which is a
normalized average sensitivity for that parameter. If a given parameter was varied by 20% in
each direction and the output result also varied by 20% (on average) the "sensitivity" for that
parameter/output pairing would be calculated as 100%.

Sensitivity analysis results are stored in Excel files. Immediately after running a sensitivity
analysis, the proper Excel file will be loaded.  Otherwise, the user will need to select "View a
Different Database" change the file-type to "Sensitivity Results, (*.xls)" and load the file in
which sensitivity results have been saved.   If there are many results in the given Excel file
loading the data from Excel could take several minutes.

All the model outputs that were tested for sensitivity may be scrolled through, using  the drop-
down menu at the top left of the tornado diagram. The user may also scroll through these
tornado diagrams using the up and down arrows immediately to the right of the drop-down menu.
 Peri. Chlorophyll (mg/sq.m)
The number of rungs on the tornado diagram can be changed by changing the "Show	Most
Sensitive Variables" edit box.  For a full accounting of all parameters tested for this particular
endpoint and their ordered sensitivity statistics press the "List Sensitivities" button

Exporting Results

Control and perturbed results may be exported to a variety of formats by clicking the Export
Results or Export Control buttons on the main study window.

Providing there are available results for export, a dialog box will appear allowing you to choose
the output variables you wish to export. After output variables are chosen, the user is prompted
for a file-name in which to save the results. Results may be saved in Excel format (*.xls),
Paradox database format (*.db), DBase IV format (*.dbf), or Comma Separated Value format
(*.csv).  To change the format of the export, choose a different format from the "Save as type"
pull-down menu.

An alternative means of exporting data is to save tabular output (viewed in the output window) to
Excel.

As part of the BASINS linkage, when exporting results,  if GenScn is installed on your machine,
the "Export all Results to GenScn" button  is available.  This automatically starts the GenScn
program with all available AQUATOX output attached.  However, most of the GenScn functions
are now available within the AQUATOX interface.

Clearing Results

AQUATOX study files include results of any control or perturbed simulations that have been run.
When a long simulation has been executed, with a small data storage step, these results can take
                                              48

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AQUATOX (RELEASE 3)  USER'S MANUAL
up a lot of disk space. To clear these results from memory, select Study and choose Clear
Results from the menu bar. The results will be disposed of so that your study file will be smaller
but you will not be able to view output using the output window until the simulation is re-executed.

Export to Release 2.2/3.0 Databases

When a  simulation is open, in the Study menu, there are two options to "Export to 2.2
Databases" and "Export to Rel3 Databases."  This option will export all animal and plant
parameters from the simulation into databases with the chosen file-format. The user will be
prompted as to which databases the data should be written to.  Any available database entries
with the  same plant or animal names will be overwritten as part of this procedure, so it may be
wise to back-up the original databases before exporting.

"Taking Parameters" from Another Study

When a  simulation is open, in the Study menu there is an option to "Take Parameters from
Another Study." This will copy all  "non-site-specific" parameters from another study (which is
required to have exactly the same state variable list).  This procedure allows  a user to test a
single set of parameters at multiple sites.

Parameters that are copied include:
    •   Animal "underlying data" and trophic interactions
    •   Plant "underlying data" and linkages
    •   Chemical and Diagenesis parameters, if relevant
    •   Settings in the Control Setup Window
The user also will be prompted as to whether initial conditions should be copied into the given
simulation.


Other Interface Components

Using the Toolbar

Virtually every function in AQUATOX can be accessed by clicking on the applicable icon on the
toolbar.  For the experienced user this provides a quick way to bring up a particular screen or to
perform a function, such as saving a simulation, without going through several layers of options in
the menu bar or the "big buttons." (The big buttons can be suppressed entirely through the View
menu option.)

To determine the  purpose of a given button, hover the mouse cursor over the button fora
moment and a "hint" which describes the purpose of the button will become visible.

Toolbar icons can be added, deleted, or moved by clicking on Edit Toolbar under the View menu
option. This is also a good way to learn the functions of the icons. There are 32 icons that are
listed and can be  used, compared to the 24 icons shown on the default toolbar. One can also
use dividers to visually group icons representing similar functions.
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Editing the Toolbar

When you have entered this screen, you may edit the toolbar by dragging and dropping buttons
onto the toolbar from the available list. Alternatively, you can drop buttons from the toolbar into
the large trash-can icon and they will no longer appear on the toolbar.

Your custom toolbar will be saved when you exit AQUATOX and  reloaded when you start the
program the next time.

Dividers appear at the bottom of the button list and they can be used to group tool-button
functionality. You cannot drag dividers from the toolbar into the trash, instead, the Remove all
Dividers button (at the bottom of the tool-button list) must be selected and new dividers can then
be dragged onto the toolbar.

The AQUATOX Wizard

Wizard Overview

The AQUATOX Setup Wizard is intended to guide the user through the 19 steps necessary to set
up a new simulation. Even experienced users may find it to be a convenient checklist, providing
a measure of quality assurance not usually found in models.  Most steps consist of several parts,
and one can  move systematically through these by making choices or entering values, then
clicking on Next.

Wizard Components:
    •   Progress Screen
    •   Summary Screen
    •   Step 1: Simulation Type
    •   Step 2: Simulation Period
    •   Step 3: Nutrients
    •   Step 4: Detritus
    •   Step 5: Plants
    •   Step 6: Invertebrates
    •   Step 7: Fish
    •   Step 8: Site Characteristics
    •   Step 9: Water Volume
    •   Step 10: Water Temperature
    •   Step 11: Wind Loading
    •   Step 12: Light Loading
    •   Step 13: Water pH
    •   Step 14: Inorganic Solids
    •   Step 15: Chemicals
    •   Step 16: Inflow Loadings
    •   Step 17: Direct Precipitation
    •   Step 18: Point-Source Loadings
    •   Step 19: Non Point-Source Loads

Wizard: Progress Screen

The Progress Screen lists all the Wizard's steps and shows which step is currently active.

It also provides a means of skipping from one step to another by double-clicking on any step in
order to move there.

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If a user decides to create a new AQUATOX study from scratch, the wizard's progress screen
also displays the status of each step (whether the user has completed entering data for that step
or if data gaps still exist for the step).

The progress screen can be hidden by selecting the Hide Progress button. It can be shown
again at any time by selecting the Show Progress button that is always available on the main
wizard window.

Wizard: Summary Screen

The Summary Screen provides a list of the state and loading variables as they are changed
while going through the Wizard. This is for informational purposes only, to give the user a sense
of how their actions are changing the state variable list they will see when the wizard is complete.
The list is not editable through the summary screen.

The summary screen can  be hidden by selecting the Hide Summary button. It can be shown
again at any time by selecting the Show Summary button always available on the main wizard
window.

Wizard: Step 1- Simulation Type

In order to create a simulation from scratch, you must access the Wizard from the File menu
("New Simulation Wizard"). If you have an existing study open, selecting the Wizard "big button"
means that you wish to edit that particular study with the wizard.

When creating a new simulation with the AQUATOX setup wizard, you have the choice to start a
simulation from scratch, or work with a default study.  When starting a study from scratch, every
parameter within the simulation will be specified by you or loaded  from the data libraries.
Otherwise,  a simulation will be based on an existing simulation and you will have the opportunity
to make modifications to the default parameters.

The name for the simulation will appear on the main screen and will  be used as a  heading in the
output. Six site types are available: pond, lake, stream (creek or river), reservoir, experimental
enclosure, and estuary.

Wizard: Step 2- Simulation Period

This screen allows you to specify the time period for the simulation.

The time period for the simulation may be a few days, corresponding to an experiment, or a year,
or even several decades.  The time period does not have to correspond to the loadings, because
the loadings can be interpolated automatically.  However, it is advisable to consider the
correspondence between the start date and the initial conditions; if the initial conditions are poorly
known then a start date in the middle of winter may allow the simulation to "spin up" before going
into the growing season.  (This is different than  "spin-up mode" from the model-setup window
which sets biotic initial conditions to the value at the end of the simulation)

Wizard: Step 3- Nutrients

Initial conditions for dissolved nutrients must be entered. Phosphate can be considered as
soluble reactive phosphate; by going into the phosphate loading screen phosphate can be
adjusted for availability. Because of interchange with the atmosphere, the model is not very
sensitive to the initial conditions for carbon dioxide and dissolved oxygen.

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Wizard: Step 4- Detritus

This set of screens allows you to specify initial conditions of detritus in the sediment bed and in
the water column.

Labile detritus is readily decomposed and assimilated, refractory is resistant to decomposition.

Initial conditions and loadings of detritus in the water column can be input as Organic Matter (dry
weight), Organic Carbon, or Biochemical Oxygen Demand (BOD) and the model will make the
necessary conversions. Suspended and dissolved detritus initial conditions and loadings are
divided into four compartments: particulate refractory and labile detritus and dissolved refractory
and labile organic matter.  Initial conditions and loadings are parsed by specifying % particulate
and % refractory.

Wizard: Step 5- Plants

The user is provided with a list of plants within each taxonomic group from which to choose.
These taxonomic groups can be scrolled through using the "next ป" and "ซ back" buttons.
Drag the plant name from the list provided into the simulation list to include it in the simulation.

After the plants  have been specified, an initial conditions entry screen is reached. Initial
conditions should  be entered for each  plant group; as with any biotic group, a value of "0" will
keep the group from being simulated.  Note that the units are sensitive to whether the plant is
planktonic or benthic.

The list of available plants corresponds to those species that have entries in the Plant Library.
As the library continues to expand with additional applications, the set of plants available within
the wizard also expands.

Wizard: Step 6- Invertebrates

The user is presented with a list of invertebrates for each ecological guild from which to choose.
Some are general taxonomic groups and some are genera and species.   These groups can  be
scrolled through using the "next ป" and "ซ back" buttons. Drag the invertebrate name from
the list provided into the simulation list to include it in the simulation.

After the invertebrates have been specified, an initial-conditions entry screen is reached. The
initial conditions are either mg/L or g/m2, depending on the mode of life (pelagic or benthic).

The list of available invertebrates corresponds to those species that have  entries in the Animal
Library. As the library continues to expand with additional applications, the set of invertebrates
available within the wizard also expands.

Wizard: Step 7-Fish

Fish are classified as forage fish, bottom fish, and game fish.  At least two species can be
modeled for each  general class. Furthermore, two size classes can be modeled for each
species, and one species  can be modeled as multiple year classes.

For each trophic guild (forage fish, bottom fish,  and gamefish) the user can choose from a list of
appropriate species in the database.
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After fish species are specified, an initial conditions screen can be reached by selecting "next
ป." Ir
basis.
ป."  Initial conditions are given as g/m2 because it is easiest to express biomass on an areal
The list of available fish corresponds to those species that have entries in the Animal Library.
As the library continues to expand with additional applications, the set offish available within the
wizard will also expand.

More information on Fish Type and Fish Class follow.
Wizard: Select Type of Fish Compartment(s)

You can choose to model a fish species either as a single state variable, as two state variables
representing size classes, or as up to fifteen state variables representing age classes.

Wizard: Select Class of Fish

Each fish species that is modeled within AQUATOX must be classified as a forage fish, bottom
fish, or game fish.

If you are modeling this species as a single state variable you must also choose whether it is of
large or small size. When modeling a species as a single state variable this has little effect on the
model's results but food preferences for any predators of the species must be set  accordingly.

Wizard: Step 8- Site Characteristics

The most important morphometric characteristic is mean depth  because that controls light
penetration, volatilization, and attached plant distribution. Mean annual evaporation is used for
computing the water balance. Latitude is used to compute photoperiod for photosynthesis.

The data entered here is used to parameterize the site underlying data.

The wall area is important for an experimental enclosure because it represents additional area for
attachment of periphyton. This is only visible if your site type is an experimental enclosure.

Wizard: Step 9-Water Volume

This screen presents the options for modeling water flow in an AQUATOX simulation.  Depending
on the method chosen, inflow or discharge values may be required.  The Manning's  equation can
be used to compute changing volumes in a stream.  The simplest procedure is to  hold volume
constant at the initial condition. Volume can be computed dynamically, given the inflow and
outflow (and  factoring in evaporation).  Finally, time series of known values can be entered.

For more information about modeling water volume in AQUATOX see Water Volume Data.

Wizard: Step 10-Water Temperature

A constant temperature,  annual mean  and range in temperature or time series can be entered.
Depending on your selection, you will be required to enter additional information about
temperature  patterns in the next screens.

For more information about modeling temperature in AQUATOX see Temperature Data Screen.


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Wizard: Step 11-Wind Loading

Wind loadings can be constant, a time series can be entered, or a default time series can be
used.  The default is a 365-day record taken from the Buffalo NY airport.  This series is
represented by a Fourier series, with a mean value that can be specified by the user (the default
is 3 m/s).

For more information about modeling wind in AQUATOX see Wind Data.
Wizard: Step 12- Light Loading

Constant, time series, and annual mean and range may be given for light in Langleys/day.

When in a simulation, light loadings may be altered by double clicking on "Light" within the state
variables list.  This will bring you to the light loading screen.

Wizard: Step 13-Water pH

The pH may be specified by the user,  either as a constant or as a time series. It has various
effects on simulations (see section 5.7 of the Technical Documentation).

When a time series is utilized the rules for dynamic loadings apply.

Alternatively, through the main pH entry screen, the user may specify that AQUATOX should
calculate pH values using a simple semi-empirical formulation (this requires a Mean Total
Alkalinity input, see section 5.7 of the technical documentation for more information.)

Wizard: Step 14- Inorganic Solids

There are three options for simulating  water-column inorganic solids within AQUATOX.

When the concentration of total suspended  solids (TSS) is known, TSS can be input to provide a
measure of inorganic solids.  The model subtracts phytoplankton and detritus from the TSS to
estimate the inorganic solids; therefore, care should be taken to use contemporaneous TSS and
nutrient time series.

The second option, the sand-silt-clay model, is only relevant for streams. This option requires
considerably more parameters and simulates the scour, deposition and transport of sediments
and calculates the concentration of sediments in the water column and sediment bed within a
river reach.  More information about the sand-silt-clay model can be found in the section titled
Sand-Silt-Clay.

The third option is to utilize the multi-layer sediment model, though this cannot be accessed
through the AQUATOX wizard.

Wizard: Step 15- Chemicals

AQUATOX can simulate as many as 20 different organic chemicals simultaneously. The
assumption is that the toxic effects are additive. Initial concentrations for each toxicant are
required for all associated state variables; as a check, the model calculates the total mass.
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After a chemical has been selected for a simulation, you may specify initial conditions for that
chemical in the water column and in all biotic and detrital compartments.

Wizard: Step 16- Inflow Loadings

The Wizard compiles a list of all variables that may be loaded as concentrations in inflowing
water. The units are sensitive to each given variable.

Click on one  of the loadings from the "Inflow Loadings in Simulation" box and you will be able to
select whether this is a constant or dynamic loading and edit the details about the loading.

Wizard: Step 17- Direct Precipitation

The Wizard compiles a list of all variables that may be loaded as atmospheric deposition ("direct
precipitation" and dry fall). The units are on an areal basis because deposition is on the surface
of the water.

Click on one  of the loadings from the "Direct Precipitation Loadings" box and you will be able to
select whether this is a constant or dynamic loading and edit the details about the loading.

Wizard: Step 18- Point-Source  Loadings

Point-source loadings are entered  as mass per day (g/d) to the water body.

Click on one  of the loadings from the "Point-Source Loadings" box and you will be able to select
whether this is a constant or dynamic loading and edit the details about the loading.

Wizard: Step 19- Nonpoint-Source Loads

Nonpoint-source loadings are also entered as mass per day (g/d) to the water body.

Click on one  of the loadings from the "Nonpoint-Source Loadings" box and you will be able to
select whether this is a constant or dynamic loading and edit the details about the loading.

Wizard Data Incomplete

This screen appears if the user originally selected to create a study from scratch. In order to do
this, the user must populate all of the parameters within the model.

At this point you have selected to leave the wizard before providing all of the required
parameters.  You must go back and enter values for these parameters or accept default
parameters provided by the program. The wizard progress screen can help you to determine in
which areas you have not entered  data.  Arrows appear by the steps in which you have not
entered  information.

Exporting  Parameters as Text

In either linked mode or single-segment mode, a user  may choose to "Export Parameters as
Text" from the Study menu. This procedure will save all parameters and, optionally, all time-
series loadings that make an AQUATOX simulation unique. This procedure can be useful for
several reasons:
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    •   It makes all modeling choices transparent when documenting your model application, as
       an appendix, for example.
    •   Two AQUATOX simulations may be compared to determine how they differ, using a
       word-processor "compare documents" function. Any differences in parameters or
       loadings will be highlighted.

After this function has been selected, the user will be prompted as to whether to save time-series
inputs to the file as well. This will increase the size of the text export, but will completely
document the input variables that are driving simulation results. The file-name in which to save
results will then be solicited. In linked mode, segment-specific parameters and initial conditions
for all segments will be included in the text output.

Aquatic-Dependent Vertebrates

This simple submodel pertains to shorebirds and other terrestrial animals that feed exclusively
from the water column. The model will estimate chemical concentrations in tissues based on
dietary uptake from food-sources in the water column.

In this screen, the user can specify a biomagnification factor (BMP) for each chemical, and the
preferences of the animal for various food sources so that alternate exposures can be computed.
Dietary preferences will be normalized to 100% by the model if they do not sum to 100% within
this screen.

The concentration of each chemical is based on the chemical concentration in prey at  a given
time-step.  Uptake of toxicant is assumed to be instantaneous, but depuration of the chemical is
governed by the user-input clearance rate.

After a run, when the aquatic-dependent vertebrates model is used, the list of model output will
include one or more entries titled: "[Toxicant in] Birds etc. (ug/kg wet)."
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         -- Afluatjc tef^ndent Vertebrates
    Trophic Interactions and BMFs for Aquatic-Dependent Vertebrates

Acteocina (gastropod
Oyster Drill
Penaeus (Shrimp)
Callinectes (Crab)
Anchoa (anchovy)
Brevoortia (menhaden
Micropogonias {croak
Mugil (mullet)
^ . . .
Preference (ratio) References:
0.05
0
0.1
0.05
0.1
0.1
0.05
0.05
n n*\
<" View all data * View Organisms in Current Study Only
A
V
III Write To Excel
          Chemical in Model
     Biomagnification Factor (unitless)
                                                            References
obs for PCBs
                           Load Troph, Int. from a File
Batch Mode

There are applications where it is desirable to run a series of studies automatically. This can be
done by creating a Batch subdirectory under the Study directory and placing studies with
appropriately chosen options in it. The subdirectory should also contain a text file labeled
"batch.txt" that lists the names of the studies to be run, one to a line. On the menu bar you should
click on Run Batch on the pull-down File window. That will open a window that allows you to
Run in Batch Mode. You also can save the BAFs and organic-matter partition coefficients
(KOMs) to a comma-separated text file batchout.csv. (See chapter 8 in the Technical
Documentation for discussion of bioaccumulation of organic toxicants, BAFs and KOMs.)

Batch mode automatically runs each simulation in "perturbed" mode.  If "Run Control Simulations
Too" is selected then control and perturbed simulations are both run (with differences reflecting
the options selected in the Control Setup Screen.)

Important Specifications for Batch Mode:

jTo run a program in batch mode, you must have a subdirectory off of your Studies
Subdirectory named "batch"

In that "batch" subdirectory must be all studies you wish to run along with a file named
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"batch.txt".

The batch.txt file must include each study name that you wish to run on a separate line.
No blank lines may be included.

The program will then execute each of the specified studies one by one and save them
along with their results.

To output the last BAF datapoint for each organism in each of the batch files, select the
Output button.  This will open each study and write all of the BAF data from each study file
into a CSV file named batchout.csv.
Setting Up Simulation, Single Segment

Setting up a single segment simulation generally has the following types of steps

    •   Locate a surrogate simulation that can be modified.
    •   Produce a reasonable model of water volume for your site.
    •   Choose appropriate biotic state variables for your site.
    •   Examine the trophic interactions matrix to ensure that a reasonable food-web has been
       constructed.
    •   Use best available data to set initial conditions for nutrients, biota, sediment, and
       chemicals  (optional).
    •   Use data to determine the external boundary condition loads for nutrients and, optionally,
       chemicals.
    •   Test the model and calibrate against observed chlorophyll a, biomass, and nutrients.
    •   Examine the effects of changing nutrient, sediment, or organic toxicant conditions.

Data Requirements

Data requirements in AQUATOX depend considerably on the site-type that is being modeled and
the goal of the modeling study. A modeling study to assess the effects of changing nutrient levels
would benefit from an excellent accounting of nutrients in the  water column and boundary-
condition loadings  of nutrients.  On the other hand, a study that  is primarily about chemical effects
may not require such detailed nutrient information.  In this case, a user may wish to produce the
required nutrient environment for a stable food-web and then  detailed information about chemical
loadings can be applied to the simulation to assess effects.

Precise accounting of time-series loadings is especially important in rivers and streams. Due to
low retention time,  inflow loadings can be the dominant factor affecting water column conditions.
For standing water, initial  conditions may be more important than a perfect representation of daily
loadings depending on  both simulation time and retention time.

Here is a list of data requirements put together for a single-segment simulation project. This list is
not necessarily exhaustive but contains what are generally the most important time-series inputs.

LIST OF AQUATOX TIME-SERIES INPUTS

    •   Total N or  Nitrate and Ammonia (inflow water,  point source, non-point source)
    •   Total P or  Bioavailable P, (inflow water,  point source, non-point source)
    •   Chemical loadings (inflow water, point source, non-point source).


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    •   Oxygen in inflow water
    •   TSS value in water
    •   Light Loadings (can be calculated based on latitude)
    •   BOD or Suspended Detritus Cones (organic matter inflow, point source, non-point
       source).
    •   Water Volume,  Inflows, or Outflows (some flexibility here)
    •   Water Temperature (mean ranges can be used)
    •   Water Mean Depth (optional)

See Additional Data Requirements for Linked Mode for a more detailed list.  Not all loadings
are required to be precisely accounted for. However, it is useful to have at least some data for
each of the various categories (nutrients, oxygen, organics, inorganic sediments or TSS, etc.)

A user will need to define the  biotic state variables in each segment as well. Default
parameterizations may be used for various organisms "Underlying Data" unless there's a
pressing need to use your own site-specific parameters.

If a chemical is modeled, requirements include initial concentrations in biota, loadings from
various sources, and toxicity data if you wish to model effects.

Site Types

The site type may be set in the site data screen.

The following site-types are included.

    •   Pond, Lake, Reservoir:  These standing-water types may stratify based on temperature
       inputs. These site-types may currently be used interchangeably but different site-types
       are provided for user  clarification.
    •   Stream:  Assumed to be moving water that will not stratify.  Streams have the following
       additional characteristics
           o  the sand-silt-clay model may be enabled;
           o   Manning's coefficient may be used to model water volume;
           o  dynamic temperature stratification is disabled;
           o   periphyton scour and velocity limitation for plants is a function of water velocity;
           o  oxygen reaeration is a function of stream velocity.
    •   Enclosure: An artificial experimental enclosure within a pond or lake.  As noted in the
       technical documentation, the littoral fraction is modified based on the area of
       the encosure's wall.
    •   Estuary: Assumed to be governed by  permanent salt-wedge stratification.  See Estuary
       Version Notes.
    •   Tributary Input Segment:  This is a specialized segment for linked-mode only. See
       Tributary Input Segments.

Estuary Version  Notes

When an estuarine  model is selected, the modeled segment is assumed to be composed of two
well-mixed layers; stratification is governed by  salinity. Water balance between the two layers is
computed using a salt-balance approach. For this reason, the salinity in each layer at the mouth
of the estuary must be specified as a constant  value or as a time series.  Time series of salinity is
especially important for the upper layer as this  will permit a reasonable and variable set of water
flows to be derived.
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Water-column salinity can have effects on animal mortality and gamete loss and algal
photosynthesis, ingestion, respiration, and sinking.  Estuary reaeration is also affected by
salinity, especially through calculation of the level of saturation at which oxygen saturation occurs.
Salinity can also affect the rate of volatilization of organic toxicants.

Various additional Tidal Range Model parameters are required at the bottom of the site
"underlying data" screen to properly model the timing and extent of tidal amplitude (which
especially affects salt-water intrusion). These parameters are generally available for download
from NOAA databases on line.

Water volume is modeled on a daily basis, rather than an hourly basis; hourly tidal fluxes are not
modeled.  For this reason, the overall daily-average water volume of the estuary is assumed to
remain constant. The simplifying assumption is that the water volume of the estuary is not
sensitive to the fresh-water inflow.  The water volume input screen is therefore different for the
estuary model, with inflow water representing fresh-water inflow. Salt-water inflow is derived from
the tidal range model as discussed above. The size and depth of the fresh and salt wedges then
vary as a function of the tidal range and fresh-water flows.

Although  Release 3 of AQUATOX can now run on an hourly basis, the estuary version of the
model has not been set up to model tidal inputs and withdrawals on  an hourly basis

Nutrient Inputs to the Lower Layer

Nutrient concentrations in seawater flowing into the lower (saline) layer are set to temporally
constant levels. The nutrients in seawater may be edited using a button available in the initial
conditions and  loadings screen for each relevant variable (Ammonia, Nitrate, Phosphate, Oxygen,
and Carbon Dioxide).

Chapter 10 of the technical documentation covers the AQUATOX estuary model in detail.

Starting with a Surrogate Simulation

When applying AQUATOX to a new site it is usually most efficient to find a surrogate site that
best matches the characteristics of the site to be modeled. The user can then modify that site's
characteristics so that  it matches the modeled site with respect to Nutrients, Organic Matter,
Turbidity, Biota, and  Organic Chemicals (if relevant).

To assist in this process the file "Study descriptions.pdf" has been added to the STUDIES
directory that describes the characteristics of each of the example model applications included
with the model.

After the surrogate simulation has been loaded, the user should rename the file, and then
examine the following components for compatibility with the site being modeled

    •  Physical characteristics (site-underlying data)
    •  Water volume  setup (water volume loadings and initial condition screen.)
    •  Nutrients, organic loadings, and turbidity in the loadings and initial conditions screens
    •  Simulation time and other model setup
    •  Water temperature settings
    •  Biotic compartments included and food web (trophic interaction matrix)

Stepping through each step of the AQUATOX Wizard can be a useful method of examining the
"key" parameters in a given simulation.


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Starting from Scratch

To start a simulation "from scratch" within AQUATOX, use the "New Simulation Wizard" in the
file menu and then select "Create Simulation from Scratch" in Step 1 of the wizard.

Some users will want to enter each and every parameter and loading "from scratch" (that is to
say, not to use  a surrogate simulation). In some senses this may be the "safest" approach as the
user is required to assess the appropriateness of each and every parameter in the model for their
individual site.  However, there are many parameters that can safely be assumed to be "global"
within the model and the user will lose the benefit  of having those parameters automatically
brought in to their simulation. Because of this, the model creators suggest that users who are
concerned about bringing in external parameters examine each and  every parameter of the
surrogate simulation closely.

Water Volume Modeling Options

As noted  in the Water Volume Data screen, there  are many options as to how to compute or
specify water volume; each  requires a different set of input data. Often, the selected volume-
modeling option is a function of the available data  for the site being modeled.

Time series of stream volumes are quite rare whereas discharge data are more often available.
For this reason, the Mannings Equation Method, which derives river  volumes based on discharge,
slope, and width data, is often used for streams.

Standing water volumes are sometimes assumed  to be  kept constant;  alternatively time series of
known volumes may be used. See the Water Volume Data section  for more information about
modeling water volume.

Adding a State Variable

Chemicals and biotic state variables may be added to the model, either through the AQUATOX
wizard or through the main interface screen  (by clicking the "Add" button  under the list of state
variables). When adding fish, the wizard is especially useful as size-class and age-class linkages
will be automatically produced through a user-friendly interface.

Adding a Chemical

To add a  chemical in AQUATOX select one of the 20 available toxicant compartments (usually
"Dissolved org. tox 1").  You will then be prompted as to which  chemical to load from the chemical
library. Loading this chemical brings in "Underlying data" for the chemical and also animal and
plant toxicity databases.

To properly model toxicity for the chemical, each and every animal and plant in the simulation
must be properly linked to one of the rows in these toxicity databases.  If  any plant or animal
lacks the  link to toxicity data an error message with be returned. In the animal and plant
underlying data records, toxicity data may be entered individually or "Edit AN" may be selected,
allowing you  to link these organisms all at once.

Adding a Plant

To add a  plant  in AQUATOX, select one of the available plant compartments (Diatoms, Greens,
Blue-greens, Other Algae, or Macrophytes) and then choose the plant data you wish to load from


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the plant library.  Initial conditions for the plant and any external loadings must then be specified.
A "seed" loading may be specified (generally as a constant load) to prevent permanent extinction
from occurring. Seed loadings are generally very small loadings (e.g. 1e-5 mg/L) that allow for
reintroduction of an organism after environmental conditions have improved to allow that
organism to be viable in the system  again. The Wizard interface may also be used to add plant
variables.

Adding an Animal

To add an animal in AQUATOX, select one of the available animal compartments (Shredders,
Sediment Feeders, Suspended Feeders, Clams, Grazers, Snails, Predatory Invertebrates, Large
and Small Forage Fish, Large and Small Bottom Fish, and Large and Small Game Fish) and then
choose the animal data you wish to  load from the plant library.  Initial conditions for the animal,
any external loadings, and trophic interactions within the food web must also be specified. Size-
class fish may be specified using the Animal Linkages interface.  A "seed" loading may be
specified as a constant load to prevent permanent extinction from occurring. As mentioned
above, the Wizard interface is especially useful for adding fish state variables.

Adding a Multi-Age Fish

To add a multi-age fish, select "Multi. Age-Class Fish" from the  bottom of the list of state
variables. The user will be prompted for the number of age-classes to add and sources of
parameters for the young of year (YOY) fish and all older fish. You will then be able to access the
Multi-Age Fish interface  by double-clicking  on the "Age-Class  Fish" item at the bottom of the
state variable list.

Using  Sediment Bed Models and Data Requirements

There are four primary options for modeling  the sediment bed within AQUATOX.

The simplest manner is to use the AQUATOX "default" sediment bed model in which organic
matter is modeled in the sediment bed as a food source and toxicant-sorption site but inorganic
matter in the bed is ignored.

In combination with the default sediment bed model, TSS concentrations may be entered as a
time  series to properly represent the light climate in the water column of the modeled system.

The sand-silt-clay model adds inorganic sediments to the default sediment bed model and also to
the water column.  Additional data are required to properly characterize boundary conditions and
deposition and erosion parameters are required. This model is  only relevant for streams and
rivers.

The multi-layer sediment  model was designed to model long-term sequestration or exposure of
chemicals within a sediment bed. It is a complicated model with considerable additional data
requirements.

The sediment diagenesis model implements Di Toro's sediment diagenesis model as presented
in Sediment Flux Modeling (2001). This model tracks the decomposition of organic matter in the
sediment bed with more precision and predicts nutrient flux based on this decomposition (and the
oxygen climate in the water column). This model is best suited  to represent predominantly
anaerobic sediments in eutrophic lakes.
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A non-reactive active layer may also be modeled, which allows the user to specify the
concentrations of toxicants in pore-water and sediments for uptake into biota. This model would
generally only be useful when linking model results from a linked sediment model. The non-
reactive layer may not be run in combination with the sand-silt-clay, multi-layer, or sediment
diagenesis models.

Default Sediment Bed  Model

The default sediment bed model includes the following components

   •    Labile Sed. Detritus (Labile Detritus in the Sediment bed) (g/m2 dry);
   •    Refract. Sed. Detritus (Refractory Detritus in the Sediment bed) (g/m2 dry);
   •    Toxicants sorbed to both of those compartments (ug/kg dry);

These detrital compartments are assumed to be located in the active layer and available for
consumption, and sorption of chemical from (and desorption of chemical to) the water column.
Through long-term deposition, these compartments may become deeply buried and join the
buried detritus state variables.  Nutrients within these detrital compartments are subject to release
to the water column when the organic matter decomposes. For more information about this
model, please see section 5.1 of the AQUATOX technical documentation.

Data requirements for this model are rather minimal, primarily being the initial condition of detritus
in the sediment bed. Setting the initial condition for these state variables may be facilitated by
using an Excel spreadsheet (AQUATOX_Sed_Bed_lnputs.xls) that is included in the Studies
directory. This spreadsheet will convert data about the fraction organic carbon, depth, sediment
density, and fraction labile into initial conditions for these two state variable compartments.

TSS

AQUATOX allows a user to input time-varying concentrations of TSS so that the light climate of
the system being modeled may be accurately represented. TSS may represent either total
suspended solids or total suspended sediments.  A user can choose which of these options is
being specified using the radio buttons on the upper right portion of this screen (accessed by
double-clicking TSS on the state variable list):
 TSS Values Represent

 (•  Solids flncl. organics}
 •T  Inorganic Segment
If total suspended solids are chosen, AQUATOX will subtract phytoplankton and detritus loadings
from the TSS loadings to estimate the inorganic solids loadings, therefore, care should be taken
to use synoptic TSS and nutrient time series.

When TSS is included in the model, it is a driving variable that is not in any way linked to the
sediment bed unlike the inorganic sediments in the Sand-Silt-Clay model.
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Sand-Silt-Clay Model

The sediment transport component of AQUATOX simulates scour, deposition and transport of
sediments and calculates the concentration of sediments in the water column and sediment bed
within a river reach.  For running waters, the inorganic sediment model within AQUATOX is based
primarily on Hydrologic Simulation Program in Fortran (HSPF, US EPA 1991). Within river or
stream simulations, sediment is divided into sand, silt, and clay. Wash load (primarily clay and
silt) is deposited or eroded within the channel reach depending on the daily flow regime.  Sand
transport is also computed within the channel reach.  Inorganic sediments in standing water are
computed  based on total suspended solids loadings, and not by means of the sand-silt-clay
model.

Within AQUATOX, inorganic sediment concentrations affect the extinction coefficient for water,
and therefore change the light climate for algae.  Chemicals are assumed not to sorb to inorganic
sediments within the AQUATOX sand-silt-clay model  but chemicals sorbed to organic matter are
assumed to scour and deposit with the same characteristics as the inorganic silt within the
model. The fraction of detritus that is being scoured or deposited within a river reach is assumed
to equal the fraction of silt that is being scoured or deposited.

There are  additional data requirements for this model. AQUATOX requires loadings information
for each of the inorganic sediment categories as well  as an initial condition. The  initial fraction in
bed sediments must also be specified. Additional parameters for the inorganic sediment model
are found in the site underlying data (stream data) or  can be found in Step 14 of the setup
Wizard. These parameters include

    •      Critical shear stress for scour for silt & clay
    •      Critical shear stress deposition for silt & clay
    •      Fall velocity for silt & clay

To add the Sand-Silt-Clay model to a simulation,  select to "Add" a state variable  from under the
state variable list and select Sand, Silt, or Clay from the list of variables (directly under the
chemicals) or use Step 14 of the AQUATOX Wizard.

Multi-Layer Sediment Model

AQUATOX also includes a complex multiple-layer sediment model.  This model can simulate up
to ten bottom layers of sediment. Within each sediment layer, the state variables consist of
inorganic solids, pore waters, labile and refractory dissolved organic matter in pore waters,  and
sedimented detritus.  Each of these state variables can also have up to twenty organic toxicant
concentrations associated with it.

Data requirements for this model are significant

    •   Thickness of each layer;
    •   Densities of organic matter and inorganic matter in the sediment bed;
    •   Pore water quantities and initial toxicant concentrations;
    •   Organic matter initial conditions and initial toxicant concentrations;
    •   Inorganic matter initial conditions;
    •   Deposition and Scour may be inputs or the Sand-Silt-Clay model may be used (with its
       own data requirements);
    •   Boundary condition loadings of inorganic sediments in the water column.
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This model was designed primarily to capture the long-term sequestration or release of chemicals
from the sediment bed. As such, linkage to a complex model of sediment scour and deposition
may be warranted.  For more information on this model, please see section 6.2 of the AQUATOX
technical documentation.

To add the Multi-Layer Sediment model, select "Add Multi-Layer Sediment" from the AQUATOX
Sediment menu. It cannot be added with the AQUATOX Wizard.

Sediment Diagenesis Model

Di Toro's model assumes a small aerobic layer (L1) above a larger anaerobic layer (L2). For this
reason, it is best to apply this optional submodel in sites where anoxic sediments are prevalent,
otherwise the assumption of a very thin aerobic layer may not be appropriate.

The additional 22 state variables added when the sediment diagenesis model is enabled are as
follows:

    •   POC (Particulate Organic Carbon) in sediment: three state variables to represent three
       reaction classes in the anaerobic layer.
    •   PON (Particulate Organic Nitrate) in sediment: three state variables to represent three
       reaction classes in the anaerobic layer.
    •   POP (Particulate Organic Phosphate) in sediment: as with POC, three state variables to
       represent three reaction classes in the anaerobic layer. The third modeled component of
       POM.
    •   Ammonia: two state variables to represent two layers (Layers 1  and 2).
    •   Nitrate: two state variables (in Layers 1 and 2).
    •   Orthophosphate: two state variables (in Layers 1 and 2).
    •   Methane: modeled in Layer 2
    •   Sulfide:  two state variables (in Layers 1  and 2).
    •   Bioavailable Silica:  modeled in Layer 2.
    •   Non  Biogenic Silica: two state variables (in Layers 1 and 2).
    •   COD: Driving variable for chemical oxygen demand in the water column that affects the
       flux of sulfide to the water column.

The sediment diagenesis model is only relevant for depositional regimes. No direct scour of
sediment from the sediment bed is modeled.

Data requirements for this model are moderate. Initial conditions  for all variables specified above
are required but most rapidly become a function of organic matter deposition and may be derived
by "spinning up" the model (running the model with arbitrary initial conditions and then setting the
initial conditions based on model results which  are a function of organic matter deposition).

A large set of "Diagenesis Parameters" govern the behavior of the model as well. Most of these
may be kept constant from site-to-site but Di Toro suggests a few parameters are site-specific.
Partitioning to phosphorus, in particular, is subject to considerable site specificity (kdPO42
and dKDPO41f).  For reporting and  archival purposes, parameters may be saved to Excel using  a
button on this screen.
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    jm1          10.5     kg/L

    m2          0.5     kg/L

    H1          0.001   m

    Dd          0.001   m2/d

    w2          3E-5    m/d

    H2          0.1     in

    KappaNHSf   0.131   m/d

    KappaNHSs   0.131   m/d

    KappaN03_1f  0.1     m/d

    KappaN03_1s 0.1     m/d

    KappaN03_2  0.25    m/d
Solids concentration in layer 1

Solids concentration in layer 2

Thickness of sediment aerobic layer 1    1 mm default, may be increasec

pore water diffusion coefficient

Deep burial velocity                 (Q2K uses 0.000005}

Thickness of sediment anaerobic layer 2
Freshwater nitrification velocity

Saltwater nitrification velocity

Freshwater denitrification velocity

Saltwater denitrification velocity

Denitrification in the anaerobic layer 2
(Cerco and Cole suggest value
(Cerco and Cole suggest value
                   Save Table to Excel
Setting the initial condition for the organic matter sediment diagenesis state variables may be
facilitated by using an Excel spreadsheet (AQUATOX_Sed_Bed_lnputs.xls) that is included in
the Studies directory.  This spreadsheet will convert data about the fraction organic carbon,
depth, sediment density, fraction labile and nonreactive sediment, as well as stoichiometry
assumptions, into initial conditions for these three state variable compartments (POC, PON, and
POP).

For more information about the equations and concepts within the Sediment Diagenesis model,
please see chapter 7 of the Technical Documentation.

To add the Sediment Diagenesis model, select "Add Sediment Diagenesis" from the AQUATOX
Sediment menu.  It cannot be added with the AQUATOX Wizard.

Non-Reactive Active Layer

In some cases, it may be worthwhile to see the effects of different sediment chemical
concentrations on the modeled set of biota.  This would usually only be relevant when working
with linked models (to provide a time-series  of chemical concentrations in the sediment bed).  By
selecting to model a "Non-Reactive Active  layer" under the "Sediment" menu, toxicants within
pore water and organic matter in the active layer of sediment may be input as a time series and
biotic  uptake and toxic effects will be calculated as a result. Uptake equations from the multi-layer
sediment model are used,  but chemical concentrations in the sediment bed are externally forced
rather than calculated.

Model Calibration

Model calibration is the process of modifying model parameters within their range of uncertainty
(reasonable ranges as defined by literature if possible) such that model results match observed
data or observed conditions.
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The Import Observed Data function is quite useful for model calibration as all external data that
is relevant to model output may be imported into the model and compared to model results. The
graph gallery can be filled with plots of model results compared to observed data, and then
parameters can be modified to best  match all available data sets.

Parameters will have a varying degree of uncertainty depending on how measurable a parameter
is, variability in field measurements,  and  uncertainty in lab equipment. Parameters will also have
a varying degree of sensitivity depending on how much model outputs change as a function of the
parameter changing. Within AQUATOX, parameter sensitivity may be ascertained through an
automated sensitivity analysis. "Calibration Parameters" might be defined as those parameters
that simultaneously have some range of  uncertainty and to which important output endpoints are
sensitive.

Calibration can be like trying to solve a puzzle.  It requires creative thought and a real
understanding of why the model is behaving like it is.

The first step in AQUATOX model calibration is often to ensure that the food-web is stable and
matches current conditions.  This may be quantitative if biomass data exists for a site or it may be
qualitative.  For example, if the user knows that largemouth bass exist at the modeled site but this
category does not persist in model results then some type of respecification of the trophic
interactions (or the lower portions of the food-web) will be required.  Once the food-web is stable
and reasonable then calibration to organic toxicant data may be undertaken.

It is recommended that the user start by trying to specify all of the important components of the
food-web simultaneously.  Calibrating the primary producers alone might be misleading as this
model would not include grazing pressures from higher in the food web.  Next, produce a plot of
all the plants and animals and ensure that there are no inappropriate crashes or expansions of
biomass beyond what would be supported by observed data or, lacking that, common sense.

Some notes about calibrating plants:

    •   Algae are differentiated on basis of:
           o   nutrient half-saturation values (P half-saturation and  N half-saturation)
           o   light saturation values (Light Saturation or Adaptive Light option), and
           o   maximum photosynthetic rate (PMax).
    •   Phytoplankton sedimentation rates differ between running and standing water.
    •   Some parameters are fairly site-specific, probably due to adaptation by the algae: these
       include critical force for periphyton scour (FCrit) and optimum temperature (TOpt).
    •   AQUATOX biotic state variables  are quite sensitive to temperature parameters.  These
       parameters include "optimal temperature," "maximum temperature," and "temperature
       response slope."
    •   "Percent lost in slough event" is a sensitive parameter for periphyton biomass.

Some notes about calibrating animals:

    •   The trophic interaction matrix is usually subject to uncertainty; proper specification of
       feeding preferences is critical.
    •   Simpler food-web models are more sensitive to effects from food-web interactions (less
       dietary substitution is possible resulting in more intense indirect effects).
    •   Biomass predictions are sensitive to "Maximum Consumption" rate and "Endogenous
       Respiration" rates when not calculated based on weight (allometrics); when allometric
       formulations are used, the model is sensitive to those parameters.
    •   Mortality is often a site-specific response and is therefore subject to calibration;
    •   The optimum temperature can have a significant effect on biomass  computations;

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    •   The minimum prey for feeding affects the efficiency of foraging behavior;

Additional information about model calibration may be found in the following document:

EPA, 2009. AQUATOX Technical Note 1, A Calibrated Parameter Set for Simulation of Algae in
Shallow Rivers, EPA-823-R-09-003 February 2009

Also see Section 2.6 on "Calibration and Validation" in the AQUATOX Technical Documentation
and Section 2.4 on Sensitivity Analysis.

Model Validation

Model validation is a numerical cross-examination of the model against new data sets. By
definition, a calibrated model is effective given conditions it was calibrated against, but there is
uncertainty as to whether it will correctly model changing conditions.  If the model can be tested
against a different data set than was used for calibration, this increases confidence that the model
has the capability to effectively capture alternative scenarios.

Model validation can consist of extrapolating the model

    •   to new time-periods,
    •   to new sites and exposure conditions, or
    •   to different chemicals.

Model validation exercises are often defined by, or limited by, the data available for validation.

If a model validation is not successful then the user can go back to the model calibration and
seek an alternate calibration that will adapt to changing conditions more successfully. The model
calibration and validation data sets may be combined for a more robust calibration, but at the
expense of the loss of the validation data sets.

Also see Section 2.6 on "Calibration and Validation" in the AQUATOX Technical Documentation.


Working with a Linked Simulation

A linked AQUATOX simulation  is essentially a series of single segment simulations that are linked
together with flow-fields. All linked segments must have an identical set of state variables. (State
variables that do not occur in an upstream segment may be set to zero there.)  State variables
can pass from segment to segment through active migration, passive drift, diffusion, or bedload
transport. Mass balance of all state variables is maintained throughout a multi-segment
simulation.

Linked AQUATOX simulations have a file extension of *.ALS.  When a linked simulation is
loaded, and show segment data is checked, all segments are displayed  in a list. Double clicking
any of the segments will bring the user into the Main Window for viewing that particular segment.
Segment-specific parameters and output may be viewed through that window.

Clicking on Show Link Data will provide the user with a list of water-flow  linkages between
modeled segments.  Double clicking on any of the linkages will bring a user to the Edit Linkage
Between Segments window.
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Linked  Output brings a user to a screen that will provide graphical or tabular output for all
segments simultaneously.

For reference, an optional bitmap map may be attached to the simulation, though this map will
have no practical interface functionality.

Another option, within Linked mode, is to run several single-segment simulations together without
any linkages to one another. This is an efficient way of calibrating several single-segment
simulations with the same set of parameters; it can be used to calibrate across sites or
experiments (such as artificial streams).  In a linked simulation, changing a biotic parameter in
one segment changes that parameter for all segments. Therefore, if it is desirable to test
parameter changes over several simulations,  loading these simulations into a "linked simulation"
but specifying no water linkage between the segments may be possible. However, linked mode
also requires that so the state variable list is identical in every segment.

For more information on linked simulations, see section 3.8 in the Technical Documentation.

Viewing and Editing Linkages

There are two types of linkages that may be specified between individual segments: "cascade
links" and "feedback links." A cascade link is unidirectional; there is no potential for water or state
variable flow back upstream. Segments that are linked together by cascade linkages are solved
separately from one another moving from upstream to downstream. This is particularly useful
when modeling faster flowing rivers and streams.

"Passive" state variables such as nutrients and organic chemicals dissolved in water and
plankton, move between segments as a function of water flow and diffusion. Benthic state
variables and fish are assumed to stay in the reach in which they were initiated with the exception
of specified fish migration and specified bed-load between segments (for benthic state variables).

A feedback link allows for water or state variables to flow in both directions. For bookkeeping
purposes, water flows are required to be unidirectional (i.e. entered water flows over a feedback
link must not be negative).  However, two feedback links may be specified simultaneously (in
opposite directions) to allow for bidirectional water flows. Segments that are linked together by
feedback links are solved simultaneously. There may only be one contiguous set of segments
linked together by feedback linkages within a simulation but there is no limit on the number of
segments that may be linked together in this manner.

Each segment's water volume inflow is a function of any boundary condition loadings that are
specified in the water volume screen, and water from one or more linked segments. Each
segment's water volume outflow is a function of any boundary condition outflows,  as specified in
the water volume screen, and outflow of water to one or more linked segments.

Feedback linkages also allow for passive diffusive exchange of state variables between
segments.  To model this type of state variable movement, the dispersion coefficient for each
interface must be specified along with the cross section area and the characteristic length.

Finally bed loads for inorganic sediments may be specified over a given linkage if the multi-layer
sediment model is included in time series of g/d.

Linkages between segments should be assumed to be horizontal unless two segments have
been specified  as a stratified pair in which case vertical linkage is possible.
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AQUATOX (RELEASE 3) USER'S MANUAL
Edit Linkage Between Segments
Type of Link:
Link Name: F

i T 	 i 	
(• Cascade Link •'" Feedback Link i 1 i

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Link From Segment:
|[R]: Riverine





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j-J | [TE]: Trans. Epi. ^j
Help Edit Bed Loads
Water flow data: cti. m/d
























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Date
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12/21/1993
12/22/1993
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Examining Linked Output

The linked output window displays results from all linked segments simultaneously.

As in the single-segment output window, a library of graphs may be specified and observed data
may be plotted against them. The only difference between the single-segment and linked output
screens is that every segment is plotted simultaneously, differentiated by segment ID.

To observe results from a single segment only, go into a specific segment by double-clicking on
the list and then go into the output window from there.

Currently there is no way to specify a sub set of linked segments, and the graph-setup window is
disabled in linked-output mode.

The tabular data in this screen will also display all segments' results simultaneously.

Examining a Single Segment

When a linked AQUATOX simulation (*.ALS) is loaded, and show segment data is checked, all
segments are displayed in a list.  Double clicking any of the segments will bring the user into the
Main Window for viewing that particular segment. Segment-specific parameters and output may
be viewed through that window.
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"Local" and "Global" Parameters

Double-clicking on any of the state variables in the state variable list will display segment-specific
initial conditions and segment-specific inflow loadings.  Inflow loadings are associated with inflow
boundary conditions as specified in the water volume screen.  Inflow loadings from another
modeled segment are a function of the internal processes in that segment and the flow of water
over the linkage between segments.

Moving into underlying data for any of the state variables shows parameters that are global to
all the animals, plants, or chemicals in a simulation.  Changing any of these parameters will
change the parameters for all segments in the system.  Similarly, if a state variable is added  or
removed when the model is run in Linked-Mode, that change to the state variable list will occur
within every modeled segment.

On the other hand, going into the site type screen within a linked segment will shows
parameters that are unique to the given segment. Site underlying parameters, remineralization
parameters, water velocity, shade, and water depth information are all unique to the  segment
being modeled.

Exporting All Graphs to  Word (Linked)

When the main linked interface is shown (map and list of segments) a user may export all of the
graphs associated with the simulation into Microsoft Word at the same time, by going to the Study
menu and selecting "Export All Graphs to MSWord" AQUATOX will step through each and
every graph in the graph library of every linked segment and paste those graphs into MSWord in
Windows Metafile format.

This can be a powerful tool if summary graphs and graphs of observed data vs. model results are
available in each segment. The selection of a single  menu item will enable a graphical summary
of the entire system.

Tributary Input Segments

For the linked version of AQUATOX "Tributary Inputs" are available as a site-type.  State
variables are not solved within this type of segment, rather, loadings of nutrients, organic matter,
and biota to this type of segment are loaded directly into the directly modeled system.

In the single-segment formulation, point sources and  non-point source loadings have no water
associated with them, and  loadings are given in grams per day. The tributary input segment
allows a user to  enter a set of nutrient, organics,  toxicants, and/or biotic loadings as
concentrations in water that are associated with a water flow. The loadings are entered as
loadings to the tributary input segment itself.  The water flows that those loadings are
associated with are entered as linkages from the tributary input segments into the modeled
segment.

Tributary inputs  may be used to model tributaries, point sources, non-point sources,  and even
groundwater inputs into the modeled system.

Because tributary input segments are "dummy" segments, output from these segments are not
included in the linked output screen, or when linked results are exported.

Tributary input segments provide the user with considerable flexibility for playing "what-if games
regarding a given source of loadings.  For example, if a management practice  could

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AQUATOX (RELEASE 3)  USER'S MANUAL
conceivably reduce both the amount of water released and the concentrations of pollutants within
that water from a point-source, the effects of this change can be directly modeled.


Setting Up a Linked Simulation

Additional Data Requirements for Linked Mode

The primary data requirements difference between linked mode and single-segment mode is that
a water volume and water movement model for the entire linked system must be specified and
input as a combination of boundary condition loadings, boundary condition withdrawals, and
movement of water from segment to segment.  This model may be derived from an external
model or through simple assumptions and Manning's equation for linked streams. Additionally,
initial conditions and boundary conditions are required for the entire modeled system rather than
a single modeled segment.

The overall data requirements for a linked project are  represented by the following list. For each
segment modeled, the following time  series may be required:

    •   Water Flow (m3/day)
           o   Boundary Condition Inflow (Tributary / main stem flow into first segment, m3/day)
           o   Boundary Condition Outflow (Withdrawals m3/day)
           o   Flow from One Modeled Segment to Another (m3/day)
    •   Nitrogen (TotN, or Ammonia & Nitrate)
           o   Nitrogen Boundary Condition Inflow (Tributary / main stem 1st, mg/L)
           o   Nitrogen Point Source Loadings (g/d)
           o   Nitrogen Nonpoint-Source Loadings (g/d)
    •   Phosphate (TotP, or TSP)
           o   Phosphate Boundary Condition Inflow (Tributary / main stem 1st, mg/L)
           o   Phosphate Point Source Loadings (g/d)
           o   Phosphate Nonpoint-Source Loadings (g/d)
    •   Oxygen Loadings
           o   Oxygen Boundary Condition Inflow (Tributary / main stem 1st, mg/L)
           o   Oxygen Point Source Loadings (g/d)
           o   Oxygen Nonpoint-Source Loadings (g/d)
    •   Organic Matter Loadings (BOD, OC, or Organic Matter)
           o   Organic Matter Boundary Condition Inflow (Tributary / main stem 1st, mg/L)
           o   Organic Matter Point Source Loadings (organic matter, g/d)
           o   Organic Matter Nonpoint-Source Loadings (organic matter, g/d)
    •   pH or Total Alkalinity in Each Reach (pH or ueq CaCO3/L)
    •   TSS Concentration (optional) in Each Reach (mg/L)
    •   Water Temperature in Each Reach (deg C)
    •   Boundary Chlorophyll a loadings (mg/m3 = ug/L)
    •   Mean Depth Time Series  (m)
    •   Fraction Shaded (Const, or Time Series)
    •   Water Velocity (Optional,  in cm/s)
    •   Sand / Silt / Clay Loadings, Optional (separated for each sediment category, TSS not
       required if SSC used)
           o   Sand / Silt / Clay (optional) Boundary Condition Inflow  (Tributary / main stem 1st,
              mg/L)
           o   Sand / Silt / Clay (optional) Point-Source Loadings (g/d)
           o   Sand / Silt / Clay (optional) Nonpoint-Source Loadings (g/d)
    •   Toxicant boundary condition loadings and initial conditions (ug/L, optional)
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Note, this list is fairly exhaustive and includes many optional items so the user should not be put
off by it- there are many ways to fill these parameters using observed water quality data. In
some cases, point-source and nonpoint-source loadings may not be required.

Using  a Template Segment

To create a new linked simulation, a user must first select an existing single-segment AQUATOX
simulation to serve as a template for the rest of the simulation. This will be the first segment in
the linked system. The state variable list within this segment must be matched by any additional
segments that are added. However, state variables may be added  and subtracted in linked-
mode; they will simply be added or removed from all linked  segments simultaneously.

Excel Template Import Capability

Using the AQUATOX interface to set-up or modify a large linked-mode AQUATOX simulation can
be very time-consuming. This process is also  not "auditable," that is to say, it is very difficult to
ascertain whether a mistake was made in the time-consuming click-intensive process. The  wrong
loadings could be imported to the wrong segment and the only way of knowing this would be by
clicking into the loadings for that segment or carefully examining the "text output"  of AQUATOX
parameters.

To facilitate the process of setting up large  simulations and  understanding the parameters that go
into them, a function to import loadings and parameters from an "Excel template"  was added to
the linked mode of AQUATOX.  This enables a user to perform the following tasks:

   •  Create a multi-segment simulation  from a single template segment;
   •   Modify site-specific parameters that are likely to change between segments;
   •   Input or modify state-variable loadings within  each segment of the simulation, including
        non-detects handling;
   •   Input or modify water-flow linkages between segments;
   •   Define tributary or groundwater inputs  into the linked-mode system;
   •   Import observed data that can be used to compare  against model results;
   •  A log  of all simulation modifications is written  including warnings and errors.

The Excel template function can be utilized when first setting up a linked simulation (the user is
prompted when selecting "File,  New Linked Simulation"). In this case, the user will need to
specify an existing single-segment simulation to reproduce  as multiple segments  of the new
simulation. Alternatively, a template can be used to modify an existing linked simulation ("Add
Time series from Excel" in the "Study" menu).  The user will need to provide the file name and
location of the Excel template in either case.

AQUATOX reads from the Excel template on a column by column basis. Depending on the type
of input being read, and whether non-detects are included,  each model input can  be from one to
three columns in width.   The first column (column "A") of each worksheet is not read from; it
usually contains a "cheat-sheet" of allowed template input strings (for reference only). This
"cheat-sheet"  is included in the AQUATOX  "STUDIES" directory as "import_template.xls." In cell
"B1"  of the spreadsheet is the first "command"  to AQUATOX and it explains what type of model
input is located in its columns (and potentially in the next one or two columns to the right). After
those columns have been processed, AQUATOX reads the top row (Row "1") of the next column
to obtain its next input or command.
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AQUATOX (RELEASE 3)  USER'S MANUAL
A
1 Addition Type
2 Addition Name |ND Flag?)
3 Seg/Link ID |seg ID from)
4 Length km fseg /D to) (commentl)
5 Vol 1C m3 f/m/f type) (comment2)
6 SARA m2 (time series header)
^TjMean Depth m (time series)
3 Max Depth m
9 Slope m/m 
11
12 {set inflow, disch to zero)
13 {calc dynamic}
14
15
B
NewSeg
Seg 1
31
4.50
113,231
289.620
2.558
3.837
0.0023
0.043





C
NewSeg
Seg 2
32
13.03
243,304
456.152
1.875
2.812
0.0024
0.043




D
Inflow
FALSE
31
E ~ F " G

Upstream Boundary
Boundary Condition
Date:
1/1/1398
1/2/1 958
1/3/1398
1/4/19-98
1/5/1998
1/6/1998
1/7/1 9-98
1/8/1 S98

Flow (m3/d'
604,304
ฃ94,518
596,964
592,071
594,518
599,411
604,304
596,964

31 to 32
31
32
Cascade
Date: Flow (m3/d)
1/1/1398 605,408
1/2/1998 594,241
1/3/1995 597,518
1/4/1998 591,734
1/5/19-98 593,965
1/6/1398 598,860
1/7/1998 605,131
1/8/1998 596,413

The type of model inputs, their parameters, and their effects on a linked simulation are
documented below.

    •   "NewSeg" (One column in width) : This command either creates a new segment or, if the
       Segment ID already exists, overwrites relevant parameters for the segment with those in
       the spreadsheet. If there is no segment with the given Segment ID, the template
       segment is copied to a new segment and the parameters listed below are used to
       populate the segment. The rows below the header contain the following parameters:
          o   Row 2: Segment Name (string)
          o   Row 3: Segment ID (string)
          o   Row 4: Length of Segment in km (number)
          o   Row 5: Initial Condition Volume in cubic meters (number)
          o   Row 6: Surface Area in square meters (number)
          o   Row 7: Mean Depth of segment in meters (number)
          o   Row 8: Maximum Depth of segment in meters (number)
          o   Row 9: Optional, Slope of segment in m/m (number)
          o   Row 10: Optional, Manning's N in s/m1/3  (number)

    •   "NewLink" (Two columns in width): This command creates or modifies a water-flow
       linkage between two existing segments.
          o   Row 2: Linkage Name (string)
          o   Row 3: Segment ID, water volume from (string)
          o   Row 4: Segment ID, water volume to (string)
          o   Row 5: Link Type, "cascade" or "feedback"
          o   Row 6: Time-series header.  Strings to help provide context within the template.
              Usually "Date" in the first column and "Flow in m3/d" in the second column.
          o   Rows 7-End:  Dates in the first column and flows in m3/day in the second
              column. AQUATOX will read downward until it finds the first empty cell.

    •   "NewTrib" (Two columns in width): This command creates or modifies a tributary input
       into the AQUATOX simulation. The tributary segment will not be a modeled segment but
       any loadings into the tributary are loaded directly into the relevant modeled segment.
       Any water volume linked from the tributary input into the modeled segment will also be
       added to the modeled segment.  This command specifies the water loading and the
       linkage to the modeled segment but other loadings (nutrients, chemicals, etc.) must be
       specified later using the nutrient input format documented below.
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           o  Row 2: Tributary Name (string)
           o  Row 3: Tributary Segment ID (string)
           o  Row 4: Segment ID to link tributary to (string)
           o  Row 5: Link Type,  irrelevant, must be "cascade."
           o  Row 6: Time-series header. Strings to help provide context within the template.
              Usually "Date" in the first column and "Flow in m3/d" in the second column.
           o  Rows 7-End:  Dates in the first column and flows in m3/day in the second
              column. AQUATOX will read downward until it finds the first empty cell.

    •   "Inflow" or "Disch" (Two columns in width): This command creates or modifies a
       boundary condition inflow or boundary condition discharge from an existing segment.
           o  Row 2: Non-Detect Flag, generally "false" for water flows (Boolean)
           o  Row 3: Segment ID, water volume to or water volume from (string)
           o  Rows 4 and 5: Optional comment fields that will be loaded into the AQUATOX
              "Notes" fields.
           o  Row 6: Time-series headers.  Strings to help provide context within the template.
              Usually "Date" in the first column and "Flow (m3/d)" in the second column.
           o  Rows 7-End:  Dates in the first column and flows in m3/day in the second
              column. AQUATOX will read downward until it finds the first empty cell.

    •   Nutrient Import / Parameter Valuation Format (two or three columns in width): This is
       relevant to
           o  "NO3:"" Nitrate loadings in mg/L;
           o  "NH4:" Ammonia loadings in mg/L;
           o  "TSP:" Total Soluble P loadings in mg/L;
           o  "OM:"  Organic Matter loadings in mg/L
           o  "BOD:" Biochemical Oxygen Demand loadings in mg/L;
           o  "O2:" Oxygen loadings in  mg/L;
           o  "TP:" Total P in mg/L;
           o  "TN:" Total N in mg/L;
           o  "pH:" pH values in water;
           o  "TSS:" Total Suspended Solids within segment water, mg/L;
           o  "Temp:" Temperature of segment water, degrees C;
           o  "ZMean:" Mean Depth of segment, meters;
           o  "Shade:"  Percent shaded  for segment, (0-100);
           o  "WaterVel:" Water velocity of segment, cm/s;

       Loadings or valuations are added to the linked simulation or modified if they already
       exist. Loadings in water are relevant to boundary conditions only, as nutrients moving
       from modeled segment to modeled segment are internally calculated. As noted above,
       loadings applied to unmodeled "tributary input" segments will be passed directly through
       to the modeled segment that the tributary is linked to.

           o  Row 2: Non-Detect Flag.  If "false," two columns will be input with the first column
              holding date  and the second column  holding loadings values. If "true," three
              columns will  be input with the first column holding the date, the second column
              holding the non-detect flag, and the third column holding the loading in the proper
              units. If the non-detect column is not empty, the sample is assumed to be a non-
              detect and the loading is set to one half of the loading in the third column
              (detection limit).
           o  Row 3: Segment ID or Tributary Segment ID to  load nutrients into (string)
           o  Rows 4 and 5: Optional comment fields that will be loaded into the AQUATOX
              "Notes" fields.
           o  Row 6: Time-series headers.  Strings to help provide context within the template.
              Usually "Date" in the first column and "Loading / Value" in the second column.


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AQUATOX (RELEASE 3)  USER'S MANUAL
              Alternatively "Date" in the first column, "Non-Detect Flag" in the second column
              and "Loading / Value" in the third column.
           o  Rows 7-End: Dates in the first column and loadings or valuations in the
              appropriate unit in the second column.  AQUATOX will read downward until it
              finds the first empty cell.

    •   "Chla" (two or three columns) Chlorophyll a in ug/L: Chlorophyll a is processed the same
       as are nutrient loadings above with two exceptions. To convert chlorophyll a data to
       biomass units, the "multiply loading by" field within AQUATOX is set to 0.0532 (mg/L
       biomass to ug/L chla). Secondly, the chlorophyll a data must be assigned to a particular
       phytoplankton species within the simulation. The import template first looks for an
       assignment in the second column of Row 2 (to the right of the non-detect flag).  The
       following strings are valid:
           o  "diatoms'!" to "diatoms6"
           o  "greens'!" to "greens6"
           o  "blgreensl" to "blgreens6"
           o  "otheralgl" to "otheralg2"

       If no valid assignment is found in this cell, the software finds the phytoplankton
       compartment highest on the list of state variables.  If there are no phytoplankton
       compartments in the simulation, the linkage raises an  error.

    •   "Observed" (Two or three columns in width): This command creates or modifies external
       data imported into  AQUATOX for use in graphing model results against observed data..
           o  Row 2: Non-Detect Flag.  If "false," two columns will be input with the first column
              holding date and the second column holding loadings values.  If "true," three
              columns will be input with the first column holding the date, the second column
              holding the non-detect flag, and the third column holding the loading  in the proper
              units.  Non-detects are plotted as red arrows.
           o  Row 3: Segment ID, observed data is relevant to this segment (string)
           o  Row 4: Description of data and source (string)
           o  Row 5: Units for data (string) in first column, optional reference in second
              column.
           o  Row 6: Time-series header.  Strings to help provide context within the template.
              Usually "Date" in the first column, "Non-Detect flag" if relevant in the second
              column, and "Value" in the column that follows.
           o  Rows 7-End: Dates in the first column, non-detects in the second column, if
              relevant, and values in the second or third column. Non-detects are assumed
              "lesser than" unless a ">" is used as the non-detect flag in which case it is
              assumed "greater than" (e.g. the denominator of a normalized data field is non-
              detected).

    •   "NextPage" Command to move to column B in the next Excel worksheet (next tab in the
       spreadsheet). Used when the user runs out of rows in the given sheet.

    •   "End" Command to stop processing the Excel spreadsheet.

Adding a New  Segment

To add a new segment to an AQUATOX linked simulation, the "Add" button may be used on the
main linked study screen when "show segment data" is selected.
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    New Segment ID:
    Segment Name:    New Segment
        r Add new "blank" segment
        f" Load segment from *.aps file
                Help
Cancel
OK
A dialog box will appear that allows the user to specify a segment ID (which is used
organizationally, and to label output), a segment name (which describes each segment), and
three options as to how to add a segment. The Segment ID must be four characters or less.

    •   An existing segment may be copied, in which case initial conditions, loadings, and site-
       specific data will then need to be modified to match the new segment to be modeled.
    •   A "blank" segment may be added, in which case initial conditions and inflow loadings are
       set to zero.
    •   A segment may be loaded from another single-segment mode simulation *.APS (single-
       segment mode) file. In this case, all state variables must match the existing segments in
       the linked simulation or the study won't be loaded.

Linking Segments Together

To link two segments together, the two segments must first be included in a simulation. Then
"Show Link Data" must be selected in the main  linked interface window. When "Add" is
then selected, the Edit Linkage window will be visible which will enable the user to specify a
"cascade" or "feedback" linkage and also to enter water flows and other relevant parameters and
time series. For more information on editing linkages see "Viewing Linkages."

Adding "Tributary" Inputs

To add a tributary input (which can also be used to model point and non-point sources and
groundwater inputs) a new segment must first be added. Adding a "blank" segment will work
fine. Then, the site type must be designated as a "tributary input" within the site-type input
screen.

Within this tributary-input segment,  initial conditions, site underlying data, and boundary-condition
water volume inflows and outflows should all be ignored as these are  not relevant. Instead, the
user should focus exclusively on the nutrient, oxygen, biota, and chemical loadings into the new
tributary segment.  These loadings will be passed right through this tributary segment and into the
modeled segment that it is linked to. For this reason, a linkage of the  tributary input into the
modeled segment must also be added.  Add a new linkage from the tributary input segment to the
modeled segment and specify an associated water flow. This linkage must be a "cascade"
linkage, as no feedback to the tributary input is possible.
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On a given date, the water flow of the linkage multiplied by the concentration of the loadings
specified into the tributary-input segment will provide an accounting of the mass of the loading to
the modeled segment.  The volume of water specified, of course, will also be added into the
modeled segment.

Tributary inputs can also be added through the Excel Template Import, which can simplify the
process. For more information about tributary inputs see "Tributary Input Segments"

Setting up Linkages, Complex Systems

Setting up linkages in truly complex systems can be a time-consuming process. For this reason,
the Excel Template  Import Capability is especially powerful for these systems. This allows
complex linkages to be set up in an auditable fashion and also allows these linkages to be
modified and re-imported.  Alternatively, boundary-condition loadings may be modified in the
Excel template and then the template may again be re-imported to test alternative scenarios.

It is important to remember that inflow water for each segment is governed by two sources of
model input:

   •    Boundary-condition inflows as specified in the water volume screen.
   •    Inflows from other linked segments as specified in the list  of linkages.

Similarly, the outflow water for each segment has two sets of loadings:

   •    Boundary-condition outflows or withdrawals as specified in the water volume screen.
   •    Outflows to  other linked segments as specified in the list of linkages.

Water Balance Considerations

New  users of the AQUATOX linked-mode often have difficulties with water volumes going to zero,
or water volumes going to unreasonably high levels.

Other than evaporation (which is often set to zero in linked-systems for simplicity sake),
AQUATOX  linked segments are modeled as "closed systems" where no water enters or leaves
the system unless it is specifically accounted for. The outflow for each segment is defined as
"Inflow - d Volume + inputs - withdrawals."  The inflow of the next segment may be defined as
the ouffitowfrom the previous segment.

For systems that take a long time to run, any changes to the water balance model may be tested
by first saving the simulation, then saving the simulation to a different name, then removing all
state variables and ensuring that the water volumes are behaving  as required before returning to
the version with all of the biotic state variables.

An external "closed  system" accounting of water flow is often quite useful for the use of the
linked-mode model, whether it be a complex hydrological model or a simple spreadsheet model.

Stratification in a Linked System

"Dynamic stratification" does not apply to the multi-segment model. Instead, a user must specify
two unique linked segments as a vertically stratified pair. In this case, the segments must be
linked together with  a feedback linkage. A stratification screen within each segment's main
interface allows a user to specify whether a segment is part of a vertically stratified pair and, if so,


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whether it is the epilimnion or the hypolimnion segment.  (The model needs to know if one
segment is located "on top" of another segment so that sinking organic matter and plants are
routed to the lower segment rather than moving directly to the sediment bed. This also affects
light climate in the lower segment.)

Unlike the single-segment model, water flows must be specified between the two segments.
Overturn may be specified by a high degree of mixing between the two segments whereas
periods of stratification will have considerably lower flow.

When two segments are set up as stratified together, the thermocline area is defined by the user-
entered cross section between them. As noted above, annual cycles of stratification and overturn
may be specified using the time-varying water flows and dispersion coefficients.  As was the
case in the dynamic stratification model, fish automatically migrate to the epilimnion in the case of
hypoxia in the lower segment. Sinking phytoplankton and suspended detritus in the epilimnion
segment pass into the designated hypolimnion segment. The light climate of the bottom segment
is limited to that light which penetrates the segment defined as the epilimnion.
            Hypothetical linked model setup with four model segments
          includingtwo reservoir segments as a vertically stratified pair


       Upstream River            Reservoir             Downstream River
                          i
Epilimnion
                                Hypolimnion
           Notes: Blue arrows indicate required time-series of waterflows;
   This reservoir has epilimnetic withdrawal though the model is flexible in this regard
Tutorials

Simple Tutorial

This basic tutorial introduces basic concepts and gets started using the AQUATOX interface.

Tutorial- Step 1:  Deleting and Adding a Plant

Open the file FarmPond MO Esfenvalerate.aps .  (see Loading a Study)

First remove the macrophyte compartment by highlighting it in the list, clicking on the Delete
button, and confirming the deletion. The variable will disappear from the state variable list.
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Likewise, state variables can be added by clicking on the Add button and choosing from the list.
Let's add macrophytes back to the list of state variables.

Click on the Add Button and scroll down the list until you see the Macrophytel to MacrophyteB
compartments. AQUATOX has the capacity to model up to six unique macrophyte species.
Select Macrophytel and click on OK.

You will then see another dialog box open asking you to "Select Plant Entry to Load Into
Macrophytel." Note that the names of the taxonomic groups and ecologic guilds on the main
study screen are followed by the names of the specific groups in brackets. We therefore have to
specify the type of macrophyte we are adding.  Select Myriophyllum on the list at the left and click
on OK.  If you do not select a species when you run the simulation you will receive an error
message indicating that there is no data associated with the state variable Macrophyte.

next, Setting an Initial Condition

Tutorial- Step 2:  Setting an Initial Condition

To continue with our macrophyte example, we should enter a value for the biomass of
macrophytes present at the beginning of the simulation; if the value is left as  0 and there is no
loading, then macrophytes would not be simulated. The initial condition will depend on when the
simulation starts (which is specified in Setup).  In this example we will enter a value of 0.1 g/m2,
which is appropriate for Myriophyllum in a temperate  pond at the beginning of the growing
season.

Double click on the state variable list on Macrophytel: Myriophyllum. Enter 0.1 as shown below.
                          Macrophytel: [Myriophyllum]

                        Initial Condition:
                       |M           grtn2 dry

                        Loadings from Inflow:
                       (f Use Constant Loadng of
                        J1E-4MW5      gifm2dry

                       c Use Dynamic Loadings
As you can see from the picture above, AQUATOX also automatically entered a loading of 1e-5
(0.00001) for macrophytes in this simulation. Ordinarily we think of loadings as pertaining only to
chemicals and freely moving organisms such as plankton and fish. However, it
is usually desirable to enter a small constant loading to serve as a "seed" if the population or
group is killed off entirely by a toxicant or adverse environmental conditions. The "seed" will allow
the biota to recover,or recolonize if and when conditions become favorable. This is especially
important for macrophytes that suffer winter die-back

For all biota, AQUATOX uses a value of 1E-5 g/m2 as a constant loading.  This is small enough
that it will not affect the results during the growing season, but it is large enough to prevent
extinction.

Click the OK button to return to the Main screen.
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See Also: Initial Conditions and Loadings

next, Viewing Parameters

Tutorial- Step 3:  Viewing Parameters

In the following examples we will examine a record from each of the other libraries. A record can
be down-loaded into a study from a library  by choosing Load data on the Edit State Variable
Data screen.

We will examine first the parameter screen for plants. From the main screen, choose Plants from
the Library menu at the top of the screen (or click on the big button "P" on the toolbar), view the
default library file, then move to Cyclotella nana by clicking on the arrows at the top left of the
screen.  We are  now moving  through a database of  parameters that is provided with AQUATOX.
These parameters are not necessarily associated with the study file that has been loaded, but
can be loaded into the simulation.  (To see the parameters associated with the given study,
double-click on any of the animals or plants in the state variable list and then click the
"Underlying Data" button.)
     N
         Plant
Cyclotella nana              Find
New
H
             Plant Type:   (Phytoplankton   J       Toxicity Record:

                                                  Taxonomic Type:

                                      Plant Data:
                                                  Diatoms
                                                  Diatoms
Two fields near the top of the screen require explanation. If you click on the triangle to the right of
Plant type, you will be given a choice.  The choice of Plant type is important because different
types have different physical or biological processes that apply to them. For instance,
phytoplankton are subject to sinking, but not periphyton, which are attached to a surface.
Conversely periphyton are limited somewhat by very slow current velocity; but not phytoplankton,
which are adapted to still water.

Less obvious is the Toxicity Record; again, clicking on the arrow to  the right of the field will give
you several choices. The intent is to associate the organism record with one of the limited
number of organisms for which there might be toxicity data or procedures for estimating toxicity.
In this instance, if you choose Diatoms the model will utilize the toxicity data (e.g., EC50) for
Esfenvalerate to Diatoms, as listed on the Toxicity Record portion of the Chemical Properties
screen.

The given  parameter values are provided to get you started; if you have more appropriate values,
you should use them.  Those parameters that do  not pertain to phytoplankton are dimmed. If you
try to enter a value for Reduction in still water, for example,  you will find that the field cannot be
edited.
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Next we will locate the record for Chironomid from the Animal Library. Hit Cancel to leave the
plant screen and again go through the Library menu to get to the animal library. (Alternatively
you can select the "A" for animal on the toolbar.)  Again, scroll using the arrow buttons to find the
Chironomid record or you can click the find button and enter "Chironomid." (This is case
sensitive so use a capital letter C)

Click on Animal Type to see the pull-down menu. Chironomids have aquatic larvae, so Benthic
insects is chosen; this is important because emergence is simulated by AQUATOX for insects as
a loss term, but does not apply to other animals.
    Animal  Chironomid
                   Find
New
Help
          Animal Type:   ] Benthic Insect

 Taxonomic Type or Guild:
                         Toxicity Record:  jChironomid
Fish
Pelagic Invert.
Benthic Invert.
                                                             Trophic Interactions
                               Animal  Data:
                                                             References:
Click the button to see the associated trophic interaction table.  The trophic interaction table is
important because it defines the food-web relationships and assimilation efficiencies. Here we
see that Chironomids  primarily feed on labile detritus, and that they assimilate 70% of it. See also
Trophic Interactions.

After exiting the trophic interactions screen, click on the scroll bar to the right to see the rest of the
Animal screen. The Bioaccumulation Data section contains parameters relevant to
bioaccumulation of organic toxicants, only one of which (Fraction that is Lipid) is sensitive in
insects. The model is  not sensitive to the longevity of the insects because emergence is a
function of growth rate, which  depends on local, seasonally varying conditions. Likewise, mean
weight can only be approximate across all instars because it will vary greatly during the growing
season.

Finally, we will examine the remineralization screen for this study.  To do this, return to the main
screen, select the Site Button and then select the Remineralization button from within that
dialog. Many of the parameters on this screen are global and there is little need to change them
for a site, unless the organic material is quite different or there is some reason that the microflora
might have adapted to abnormal conditions, such as a thermal spring or acid mine drainage. For
more information  about the three different types of Remineralization records included with
AQUATOX see the section titled Remineralization.
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      Default Remin Record
                 Find
              New
                                          Help
                    Remineralization Data:
                                                          References:
 Max. Degrdn. Rate, Labile

Max. Degrdn. Rate, Refrac.
           (ColonizeMax)

    Optimum Temperature

   Maximum Temperature
0.25  g/g-d


     g / g • d

     ฐC
               Wetzel 2001, p. 514 (max for bl-grj; diatc
0.007
Collins and Wlosinski 1933
               temperature at which measured
  65
Alexander'61
Next, Viewing Toxicant Loadings

Tutorial- Step 4:  Viewing Toxicant Loadings

Following the tutorial to this point, we should still have FarmPond MO Esfenvalerate.aps loaded
from back in step 1.  Return to the main screen.

Double-click on Dissolved org. toxicant or click on the Chemical button on the main screen to
bring up the Edit Chemical Data screen, and to examine the various options for loadings to the
system.  Pollutant loadings can be entered as constant or dynamic loadings in several different
forms. The pollutant can be entered as a concentration in the dissolved phase (or loosely bound
to suspended sediment); the water inflow and the site volume are then used by the model to
compute the loading  per unit volume. The gas-phase concentration is used to compute
atmospheric exchange; ordinarily concentration in the atmosphere can be considered to be 0,
although some pollutants such as PCBs may have significant atmospheric concentrations.
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Dissolved org. tox 1 : [Esfenvalerate] H
Initial Condition: Gas-phase cone.:
0
Loac
ff USซ
0
r us<
ug/L JO g/mJ
mgs from Inflow:
Biotran sformation

ug/L
3 Dynamic Loadings

I Date JLoading
Toxicity Data
15

I
I
| Change
Multiply loading by 1
ug/L
Help

Notes: jloadings are output from PRZM for adjacent
cornfield; 2/10 worst case scenario
                                                      Loadings from Point Sources
                                                   C  U se Const. Loafing of ]         g/d  Convert j
                                                   (•  Use Dynamic Loadings
Date
12/26/1994
12/27/1994
12/28/1994
>

Loading
4.21006-02
1.32896-01
8.5170e.02
O.OOOOeOO

A,




V
                                                                           gfd
                                                       •f    —
                                                                 A  Change
                                                   Multiply loading by [02


                                                    Loadings from Direct Precipitation

                                                   ff Use Const. Loading of jo        g/m2-d
                                                   r use Dynamic Loadings

                                                                Loading   f j
                                                                            g/m2-d
                                                                     Change J
                                                    Multiply loading by
I  Load Data  ||| Edit Underlying Data fa
                                                                  ,ซ/ Q.K.  I X Cancel

Point-source loadings are mass per day (g/d) for the entire site; they are divided by the site
volume to obtain the loading per unit volume. In this example, dynamic loadings from a point
discharge as calculated by the PRZM model are entered. Note that the dynamic loadings are
interpolated, so if the intent is  to represent a spike such as from storm runoff on a  particular day,
then the loading should be bracketed by "0" loadings.  The model assumes that the loadings
"wrap around" with an annual  cycle and that the last loading can be interpolated to the first
loading as if it were in the succeeding year.  Exercise caution when modeling multiple years, but
only have loadings data form one or a few years. Sporadic loadings, which would only be
expected in that one particular year may inappropriately be repeated in successive years. If you
do not wish loadings to be repeated, enter values ("0" or otherwise) for the first and last days of
the simulation.  The dynamic loadings in this example were entered by hand; an excellent
alternative is to download or prepare a file external to the model and import it into the study using
the Import button.  See also: Dynamic loadings.

Another potential pollution loading source is from direct precipitation. These are given as g/m2
day because AQUATOX does not explicitly consider precipitation.  Click on N.P.S. to toggle to the
non-point source screen, which is  in  g/d.

Next, Running the Simulation

Tutorial- Step 5: Running the Simulation

To run the simulation, select the Perturbed or Control buttons from the main screen.
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Depending on the control setup selected, a control run will omit toxicants or nutrients from the
run.

Tutorial- Step 6: Viewing Output

When a study has been executed, results for various state variables are stored in memory. To
view this output, you must either export the output to an appropriate file-type using the Export
buttons on the main screen or select the Output Button from the main screen.  The Output
Button will spawn an additional output window which will allow you to view output in graphical or
tabular format.

Also See: Viewing Graphical Output
Stream Tutorial

This tutorial uses the AQUATOX Wizard to make a number of changes to a "template" study.
This is the usual approach taken when applying the model.

Stream Tutorial- Step 1: Selecting an Existing Study as a Template

An existing study that is as close as possible to the site you are trying to model should be chosen.
In this case, open the file Rum R MN.aps (see Loading a Study), save as LBR Middleton
ID.aps.

It is best to immediately save the template study with a new name so that you do not overwrite
the original with your changes.
Stream Tutorial- Step 2: Modifying the Study with the AQUATOX Wizard

Click on large Use Wizard button on main screen

    Step 1: Change the study name to "L. Boise Middleton" This will be the heading for output.

    Step 2: Change the simulation period to 1/1/2000 to 12/31/2001.

    Step 3: Keep the nutrient initial conditions; stream simulations are not sensitive to nutrient
       initial conditions but rather to loadings

    Step 4: Also keep the sediment detrital initial conditions for lack of better data and  keep the
       water-column BOD (you can change this after importing loadings)

    Step 5: Keep the following algal state variables and initial conditions; generally these will be
       applicable to any stream; however, remove the macrophyte Fontinalis, which is a moss.
       To do this click "Next ป" until you reach the "Macrophytes" screen.

    Step 6: Keep the invertebrates without change until "Pred Inverts" when you should drag
       Stonefly to the simulate column on the right.  Keep the initial conditions, but use 0.01  for
       Stonefly

(Note, any changes that are made in the wizard may be undone using the "cancel" button.
However, any changes made in any single wizard session must be undone at the same time.
Therefore you may wish to periodically click on "finish" to save your partial changes and then
reenter the wizard.  You may jump to whatever screen is appropriate by double clicking on that
step on the Wizard Progress window.)
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    Step 7: Although plant and invertebrate groups do not vary appreciably over large geographic
       areas, fish are often restricted to specific drainage basins; therefore, you will need to
       make numerous changes. First, select and remove: Bluegill, Smallmouth Bass (YOY
       and Lg) and Walleye.

    Next add:

       Single compartment, small forage fish Chiselmouth and Dace;
       Single compartment, large forage fish Pikeminnow; and
       Size-class gamefish Largemouth Bass, Mtn Whitefish, and Rainbow Trout (Use YOY
       data record for small size class).

       When adding a size-class fish, you are first prompted to load parameters for the small or
       young-of-year fish.  Then you are asked if you wish to use the same parameter record for
       the large fish or not.  In this tutorial, load a different parameter set for the large size-class
       fish.

       Change all fish initial conditions to 0.1; we will let the model adjust these during iterative
       simulations.
    Step 8: Change the site name; enter
       5 km for length (arbitrary);
       surface area = 1.8e5 m b
                         based on 36 m width determined from Google Earth;
    mean depth = 0.42 m;
    max depth = 0.63 (1.5 X mean depth);
    evaporation = 0 (is not important for a short stream reach);
    latitude = 43.57.

    Keep channel slope = 0.002
    Keep Manning's coefficient estimated based on natural stream;
    Riffle = 47% and pool = 0% based on site observations and pebble count.

Step 9: Continue to Use Manning's Equation for simulating water volume and select "Next."

    For initial volume use 7.56E4 m3 based on mean depth of 0.42 and surface area given
    earlier; discharge data should be entered by clicking on Change and Import and
    choosing Midd discharge.xls, which is based on USGS data from the Middleton gage
    converted from cfs to cu m/d.

    Note:  All Excel files to be imported in this tutorial may be found in the "Tutorial" directory
    off of the AQUATOX "Studies" directory.  Whenever one time-series is imported over
    another, the original data is deleted prior to the import.

Step 10: Select to use time-varying temperature and then use time series imported from
    Midd temp.xls; these observed data are almost a year in  length and are from a later
    period, but they represent the variable nature of temperature and are interpolated by the
    model. The initial temperature for January 1 may be set to 5 degrees based on the
    imported data.

Step 11: Wind is not important for a stream, so keep it constant at 1 m/s

Step 12: Select "Annual Mean and Range" then use observed mean = 380 Ly/d and range =
    516 Ly/d; Internet sources for these data change frequently. One  research approach
    would  be to  Google the nearest airport and "solar" to try to find approximate values
    (which may require units conversion).

    Select to compute photoperiod from latitude.


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    Step 13: Import observed pH from Midd pH.xls; keep initial pH at 8, which is approximate for
       January 1 based on the imported data.

    Step 14: Sporadic observed TSS values are available from the USGS NWIS for the Middleton
       gage; a regression was obtained for TSS as a function of discharge (see Midd
       discharge TSS.xls), but there was no significant relationship, so the raw observed
       values in Midd TSS.xls should be imported.

    Step 15: No toxic chemicals will be simulated.

    Step 16: Ignore loadings for ammonia; import nitrate loadings from Midd TN.xls and P from
       Midd TP.xls (you will have an opportunity to verify that these are specified as TN and TP
       later); keep CO2 loadings constant at 0.7; For oxygen, import Midd DO.xls; import
       suspended and dissolved detrital loadings from Midd BOD.xls (here too you will check
       later to make sure it is specified as BOD); all the biotic loadings are seeds to prevent a
       group from becoming extinct and should be 1 -E5.

    Step 17: Direct precipitation is unimportant to a  stream such as this, so all values should be
       either 0 or ignored.

    Step 18: Point-source loadings are unimportant for this reach and should be 0; these would
       be set for effluent such as from a wastewater treatment plant.

    Step 19: Nonpoint-source Loads are also unimportant and should be 0 for this reach.
Stream Tutorial- Step 3: Additional Study Modifications and Running

You have now finished changing state variables, site constants, and loadings with the Wizard.
Now check to make sure that N is as TN, P as TP, and Suspended and Dissolved Detritus as
BOD by double-clicking on each in the state variable list in the main screen ("Nitrate," "Total
Soluble P," and "Susp. and dissolved detritus.")  In the last of these three screens, the %
refractory and % particulate detrital allocations are 20% and  10%, which do not need to  be
adjusted; however, they could be quite different if a factor such as leaf fall were important.

Click on Site and then Edit Underlying Site Data to review and change other site constants.
Altitude should = 760  m; Total Length for Phytoplankton Retention = 63 km (distance to  Diversion
Dam).  This last parameter is located toward the very bottom of the screen.

Click on Setup and choose Run model in spin-up mode so that the biotic end conditions will be
used as the initial conditions  in the next simulation.  (Turn this off when you have a fully calibrated
model.) Also, unless storage space is a problem, choose all the state variables for saving rates
(click on Save Rates and Rate specifications and then on ป to move all variables into the  save
column).

Be sure to save the study! Then click on Control to run the model. You will get a reminder that it
is in spin-up mode, which you should ignore; you will also get a warning that there is zero habitat
for adult largemouth bass—cancel, double-click on  bass in the state variable list,  and set % Pool
= 50%. The once again click on Control.

Stream Tutorial- Step 4: Examining Model Output and Importing External Data

When run is complete, click on Output.  The observed data from Rum River will still display,  so
choose External Data from the pull-down menu in the upper right corner of the screen.  Delete
"Obs. chlorophyll a" and select "Obs. periphyton chl a" (we will keep the title and units).  Click on
Import Data and choose "Mid peri chl norm.xls." You may wish to open the file first in Excel  to
make sure where the data are located; in this case the default sheet 1, row 1, columns a and b
are satisfied, so choose Import and overwrite data. When the data are plotted the simulation
results will span only a short  period compared to the observed data. Click on Edit and Graph


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date range reset (in the upper right corner). By choosing Statistics from Graph in the pull-
down menu, you can get the statistics for the plotted variables. Also, from the pull-down menu (or
right-clicking on the graph itself) you can Copy, Print, and alter the graph (using Graph Setup).

Click on New, choose Custom Graph and then label as "Depth" and select "Zmean (Dynamic)"
to plot depth. You will see that a constant, initial depth is used. This is not realistic for a stream
that is hydraulically regulated for purposes of seasonal irrigation.  Therefore we will change the
way that mean depth is modeled.

Exit Output and click on the large Site button and Show Mean Depth Panel; choose Import and
Change, then Import "Midd Depth.xls" and re-run simulation (perhaps saving results as
"perturbed" to help see the effects of this change). "Midd Depth.xls" was created  using  Manning's
equation with observed discharge  in an Excel spreadsheet originally calibrated for the Glenwood
Bridge site on the Lower Boise River (Glenwood Depth Discharge calibration.xls). The  new model
result fits the sparse periphyton data equally well, but is more defensible.

You  may wish to run the simulation over again  to obtain better initial conditions for the animals
due to spin-up mode. Remember to turn off spin-up mode when you have obtained a calibrated
model.

Also, click on the "save-as" icon often to save the study. Saving iterative versions of the model
under different names along with a log file that  describes changes between versions is
recommended.
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