United States       ' Office of
           Environmental Protection    Water Enforcement EN-338
           Agency          Washington, DC 20460
&EPA     NPDES Compliance
           Monitoring Inspector
           Training

           Biomonitoring

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      NPDES COMPLIANCE MONITORING




       INSPECTOR TRAINING MODULE




             BIOMONITORING









  U.S. ENVIRONMENTAL PROTECTION AGENCY
          ENFORCEMENT DIVISION




OFFICE OF WATER ENFORCEMENT AND PERMITS




           COMPLIANCE BRANCH




               JUNE 1980

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     U.S. ENVIRONMENTAL PROTECTION AGENCY
       COMPLIANCE MONITORING INSPECTOR TRAINING
                          DISCLAIMER






     This module has been reviewed by the Office of Water



Enforcement and Permits, U.S. Environmental Protection Agency, and



approved for publication.  Mention of trade names or commercial



products does not constitute  endorsement or recommendation for use.

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     U.S. ENVIRONMENTAL PROTECTION AGENCY
       COMPLIANCE MONITORING INSPECTOR TRAINING

                        ACKNOWLEDGEMENT

     These modules  were developed by Barbara A.  Schick,
Claire M. Gesalraan,  Duane Geuder, Edward Bender, and with  contri-
butions by Dave Shedroff, all of whom are staff  members  of the
Enforcement Division, Office of Water Enforcement and Permits.  The
Compliance Branch,  Enforcement Division, Office  of Water Enforcement
and Permits, wish to express their appreciation  to the secretarial
staff for the assistance provided in the preparation of  this module,
especially Mrs. Mary F. Rogers and Mrs. Wilma Haney.
                              II

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U.So ENVIRONMENTAL PROTECTION AGENCY
  COMPLIANCE MONITORING INSPECTOR TRAINING




                          LIST OF HANDOUTS


      Handout 1:  Glossary

      Handout 2:  21 Primary  Industrial Categories

      Handout 3:  Compounds Considered to be Toxic

      Handout 4:  Priority  Pollutants

      Handout 5:  Clean Water  Act of 1977

      Handout 6:  Recommended  Test Organisms

      Handout 7:  Equipment Checklist

      Handout 8:  Preinspection Questions

      Handout 9:  Effluent  Sampling Procedures

      Handout 10: Definitive  Test Requirements

      Handout 11: Data Sheet  for Toxicity Test

      Handout 12: Methods for  Calculating LC50,  EC50

      Handout 13: Reporting Test Results

      Handout 14: Day-to-Day  Activity Guide

      Handout 15: Acute Toxicity Laboratory Evaluation Form

      Handout 16: NPDES Compliance Inspection Report form

      Handout 17: Instruction  for Completing Compliance
                 Inspection  Report form
                         TTT

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     U.S. ENVIRONMENTAL PROTECTION AGENCY
        COMPLIANCE MONITORING INSPECTOR TRAINING
                             FOREWORD
      The  National Pollutant Discharge Elimination System  (NPDES)
 Compliance  Monitoring Inspector Training Modules were  developed
 by the  Environmental Protection Agency (EPA),  Office of Water
 Enforcement and Permits (OWEP)f to instruct NPDES inspectors  in
 various aspects of conducting NPDES Compliance Monitoring
 Inspections.

      The  EPA Regions have identified a need for training  materials
 to instruct new employees in conducting NPDES  inspections.  Train-
 ing seminars that are currently offered either do not  address  the
 training  needs of an NPDES inspector or are not available due  to
 limited resources or conflicting course schedules.  These training
 modules were developed to fill the Regions' need for  in-house
 inspector training.

      The  objectives of the training modules are:

 1.   To acquaint new inspectors with the NPDES Compliance
      Inspection program;

 2.   To serve as a refresher course for experienced NPDES
      Inspectors;

 3.   To assure consistency in the NPDES Compliance Inspection
      program; and

 4.   To inform and instruct inspectors concerning new  inspection
      procedures.

      The  modules were designed to be used as a self-taught  course
 or as the basis for a lecture course to supplement on-the-job
 training.   The modules should be presented by  experienced and
 knowledgeable Regional staff who can answer any questions,  discuss
 Regional  policies regarding the topic being presented, and  conduct
 on-the-job  training.

      The  module format was chosen for this training program because
 of its  flexibility.  Each module covers a specific aspect of a com-
 pliance inspection.  Instructors for a particular module  may be
 selected  according to their expertise, and training sessions could
 be scheduled based on the needs, the resources, and the time avail-
 able to the Region.  The modules can be presented individually or
 as a complete package.

      An outline of information contained in the individual  training
 modules is  listed below.  There are currently  five NPDES  Compliance
 Monitoring  inspector Training modules:

      1.   The Overview module gives the inspector an overview
          of the compliance program and a brief summary of the
	different types of compliance inspections.     	
                               IV

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     U.S. ENVIRONMENTAL PROTECTSON AGENCY
       COMPLIANCE MONITORING INSPECTOR TRAINING
                       FOREWORD  (Continued)

     2.  The Legal Issues  module outlines  the legal issues which
         must be addressed during  an  inspection and legal
         information that  will assist  inspectors in performing
         their duties.

     3.  The Biomonitoring module  outlines  the principles of bio-
         monitoring and the role of biological testing in the
         inspection program.

     4.  The Sampling module details  the sampling procedures that
         an inspector uses when  conducting  a sampling inspection.

     5.  The Laboratory Procedures module  outlines the procedures
         and information necessary for an  inspector to perform an
         effective evaluation of a permittee laboratory.

The layout of the text  of  each module  is on a half page so that
students may include their notes with  the  text.

     These training modules were developed  for the Regions and are
designed to be used by the Regions for in-house training.  If these
modules are to be a success,  the Regions must participate in their
ongoing development.  This can be  accomplished by providing EPA
Headquarters with changes  or information which Regional instructors
or managers.believe would  improve  the modules.  The format of the
modules can be updated  and revised at OWEP  as the need arises as
they were developed and produced at EPA Headquarters.  Cooperation
and commitment to training by the  Regions will promote the
development of a useful training document.

     These training modules were developed  primarily for Regional
NPDES inspectors; but they are also available to other interested
parties such as State offices, attorneys, other program offices,
facility owners and operators, and members  of the general public.

     Comments, information, and  suggestions to improve the modules
should be addressed to  the:

     Technical Evaluation  and Support Section (EN-338)
     Office of Water Enforcement and  Permits
     U.S. Environmental Protection Agency
     401 M Street, SW
     Washington, D.C.  20460

Modules covering new topics may  be added to the existing ones as
the need arises.  Subject  suggestions for  future modules should
be sent to the above address.

     Requests for training modules will be  handled at the above
address depending on available supplies.

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U.S. ENVIRONMENTAL PROTECTION AGENCY
  COMPLIANCE MONITORING INSPECTOR TRAINING
                 TABLE OF CONTENTS


DISCLAIMER                                        p. I

ACKNOWLEDGEMENT                                   p. II

LIST OF HANDOUTS                                  p.

FOREWORD                                         p. IV

TABLE OF CONTENTS                                 P. VI

I.     INTRODUCTION                               p. 1

       A.   Scope                                  p. 1
       B.   Definitions                            p. 2
       C.   Background                             p. 2
       D.   Acute Toxicity Testing                  p. 5
       E.   Advantages of Biomonitoring             p. 7
       F.   Disadvantages of Biomonitoring           P. 7
       G.   Objectives of Biomonitoring             p. 8
       H.   Inspectors Responsibilities             p. 9
       I.   Phases of CBI                          p. lo

II.     PHASE I PREINSPECTION PLANNING              P. 10

       A.   Selecting Permittees for a CBI           p. 10
       B.   Equipment Requirements                  P. 11
       C.   Preinspection Coordination              p. 20

III.    PHASE II    INSPECTION PROCEDURES           P. 21

       A.   Evaluation Inspections                  p. 21
       B.   Sampling Inspections                    P. 24

IV.     POST-INSPECTION PROCEDURES                  P. 30

       A.   Data Evaluation                         p. 31
       B.   Data Interpretation                     P. 32
       C.   Inspection Report                      p. 34
                         VI

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          U.S. ENVIRONMENTAL PROTECTION AGENCY
COMPLIANCE MONITORING INSPECTOR TRAINING  MODULE:
           NOTES
                  LESSON   BIOMONITORING
    Effluent Biomonitoring
I.  INTRODUCTION
    Purpose of This Module
Reference: The Interim NPDES
Compliance Biomonitoring
Inspection Manual
Effluent Biomonitoring
    As a result of  this training, the
trainee will  be able  to:

(1)  Define  the  terms  effluent
    biomonitoring,  LC50 and EC50, and
    explain how they  relate to the National
    Pollutant Discharge Elimination System
    (NPDES) permit  program

(2)  Describe  the methods of determining the
    toxicity  of pollutants through acute
    bioassays

(3)  Describe  the NPDES compliance
    biomonitoring inspection process

(4)  Distinguish between a Compliance
    Biomonitoring Inspection, a Performance
    Audit Inspection, and a Compliance
    Evaluation  Inspection (CBI, PAI, and
    CEI) for  biomonitoring

(5)  Determine how a CBI can best be used

I.   Introduction

     A.   Scope

          The purpose of this training
     module is  to provide all inspectors
     with an  understanding of the tech-
     niques and potential applications
     for Compliance Biomonitoring
     Inspections (CBIs).  As a result of
     this training, the inspector should
     learn  the  purposes of the CBI and
     recognize-some of the resources and
     technical  requirements necessary to
     use a  CBI.  Our  discussion will cover
     the background of effluent biomoni-
     toring,  definitions, and general
     procedures for conducting biomoni-
     toring inspections.  In addition,
     we will  discuss  choosing facilities
     for biomonitoring inspections,

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          U.S. ENVIRONMENTAL PROTECTION AGENCY
COMPLIANCE MONITORING INSPECTOR TRAINING  MODULE:
           NOTES
              LESSON   BIOMONITORING
    Definitions

    - biomonitoring
    - acute  toxicity testing
    Handout  1 - Glossary
    EPA  Experience
interpreting  the  results of bioassays,
and preparing  the  inspection report.
The interim NPDES  Compliance
Biomonitoring  Inspection Manual MCD-62
(October 1979)  should be used in
conjunction with  this training module
as a reference for sample collection
and toxicity  testing procedures.  For
brevity this  reference will be called
the "Manual"  in this module.

B.   Definitions

     The term effluent biomonitoring
or effluent biological monitoring,
means measurements of the biological
effects (e.g.,  toxicity, bioaccumu-
lation, and biostimulation) of
effluents on  populations of indigenous
organisms.

     Acute toxicity tests  (acute
bioassays) are biological monitoring
techniques which measure the lethal
effects of a  compound, mixtures of
compounds, or effluents on  an organism.
We will discuss the use of  acute bio-
assays to evaluate the  toxicity of
effluents.  A glossary of  these terms
is provided in Handout 1.

C.   Background

     EPA has  used many different
biological monitoring techniques to
determine the effects of compounds
and wastewater discharges  on receiving
streams.  These techniques  range from
ambient {instream) monitoring of
aquatic communities (e.g.,  species
composition,  indicator  species, and
community structure)  to effluent moni-
toring (e.g., bioassays).   Section
101(a)(3) of the Clean Water Act  (CWA)
states that it is a national goal to
prohibit the discharge of  toxic
pollutants to our waterways in  toxic
amounts.  Bioassays have been used to

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          U.S. ENVIRONMENTAL PROTECTION AGENCY
COMPLIANCE MONITORING  INSPECTOR TRAINING  MODULE:
           NOTES
LESSON   BIOMONITORING
                                 develop effluent limitations  and  water
                                 quality criteria for some  toxic
                                 chemicals, particularly under Section
                                 307(a) of the CWA.   However,  toxicity
                                 data are available for only a limited
                                 number of compounds and in most cases,
                                 these data do not consider the
                                 interactions of compounds  in  the
                                 effluent which affect toxicity.   No
                                 chemical test can detect toxicity;
                                 only living organisms can  be  used to
                                 determine toxicity.

                                      Biomonitoring is an important
                                 part of EPA's strategy to  control the
                                 discharge of toxic materials. Bio-
                                 monitoring is used because no other
                                 technique can predict the  biological
                                 effect of chemicals and changes in
                                 water quality.  This strategy was
                                 developed from EPA's experience with
                                 toxicity testing and prompted by
                                 Sections 101, 301,  and 307 of the
                                 CWA and recent judicial history.

                                      The consent decree of June 7,
                                 1976, between the Natural  Resources
                                 Defense Council, Inc., and EPA
                                 (Natural Resources Defense Council et
                                 al. v. Train 8 E.R.C. 2120 (D.D.C.
                                 1976)) required that the Administrator
                                 develop and promulgate regulations
                                 establishing effluent limitations and
                                 guidelines, new source performance
                                 standards and pretreatment standards
                                 for new and existing sources.
                                 Effluent limitations and new  source
                                 performance standards require that
                                 Best Available Technology  (BAT)
                                 economically achievable be considered
                                 in  their development.

                                      All three of the regulations must
                                 take into account the list of toxic
                                 pollutants categories given in
                                 Appendix A of the consent  decree  and
                                 commonly referred to as the list  of
                                 65  categories of toxic pollutants.

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          U.S. ENVIRONMENTAL PROTECTION AGENCY
COMPLIANCE MONITORING INSPECTOR TRAINING  MODULE:
           NOTES
                                LESSON  BIOMONITORING
    Handout 2:  21 Primary
    Industrial Categories
    Handout 3:  Compounds
    Considered to be Toxic
    Handout 4:
    Pollutants
Priority
     The consent decree also required
that these  limitations be applied to
21 primary  industrial categories (see
Handout 2)  named in  the consent
decree.  In the course of developing
the technical  data base required to
formulate limitations for the 65 cate-
gories of pollutants, EPA developed a
list of specific compounds  in ten
categories  which are considered to
be toxic (Handout 3).  This list
(May 1977)  included  129 compounds
(Handout 4) which are known as
priority pollutants.

     The consent decree prompted the
1977 amendments to the CWA which out-
lined EPA's approach to control toxic
pollutants.  This approach  included
BAT quidelines and biological monitor-
ing.  BAT guidelines will be the most
stringent level of treatment for any
industrial  category. However,
effluents which meet BAT may still be
toxic because  the limitations have
been developed based on available
treatment technology without toxicity
data for the effluents.  Effluent
guidelines  for some  industries may
include limits for  individual priority
pollutants, however, the effluents
from specific discharges may still
contain other priority pollutants and
other toxic compounds which are not
limited by the permit.  For some
industrial  categories, BAT may be
equivalent  to Best Practicable
Technology (BPT) which  is now used
to issue permits.   For  these cate-
gories, wastewater  treatment will not
change.  In addition, some  effluent
guidelines will  not  be  promulgated
for several years.   Effluent biomoni-
toring can be used   now to  evaluate
the toxicity of effluents and to
assure that BAT mitigates the adverse
biological  effects of toxic effluents.
The CBI uses acute  toxicity tests to

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          U.S. ENVIRONMENTAL PROTECTION AGENCY
COMPLIANCE MONITORING INSPECTOR TRAINING  MODULE:
           NOTES
              LESSON   BIOMONITORING
    Acute Toxicity
    Testing
    Definitions:
     -Effluent Biomonitoring

     -LC50
    -Range-Find ing Test

    -QA Bioassay

    -Definitive Test
identify effluents  which are toxic.
The alternative  is  to  identify all
compounds or mixtures  of compounds
in the effluent  which  are toxic,
and to establish limits on each com-
pound.  This is  impractical and much
more costly than a  direct screen of
the effluent for toxicity.

D.   Acute Toxicity Testing

     Now it is necessary to define
some other terms commonly associated
with effluent biomonitoring
(Handout 1).  Acute toxicity tests
are used to measure the effluent con-
centrations, expressed as a percent
volume, that are lethal to or have
some other adverse  effect on 50% of
the population of organisms within a
prescribed period of time.  If mor-
tality is the effect being measured,
the toxicant concentration is
expressed as a median  lethal concen-
tration, or LC50.   In  other instances,
mostly with invertebrates, death is
not easily detectable, and indices
such as immobilization or reduction of
shell growth must be used to measure
an adverse effect.   The concentration
of an effluent,  expressed as a percent
volume, that causes a  defined adverse
effect other than death in 50% of the
test organisms within  a prescribed
exposure period  is  termed the median
effective concentration, or EC50.

     Acute bioassays described in
the Manual, consist  of  static or
flow-through tests  performed as a
1) range-finding test, 2) a Quality
Assurance (QA) bioassay, or 3) a
definitive test.  Range-finding
tests are short  term bioassays
(8-24 hours) used to approximate

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          U.S. ENVIRONMENTAL PROTECTION AGENCY
COMPLIANCE MONITORING INSPECTOR TRAINING  MODULE:
           NOTES
              LESSON   BIOMONITORING
    Handout 5: Clean Water
    Act of 1977
the range of the  LC50 for a defini-
tive test.   Usually  five organisms
are tested in each concentration of
the effluent with a  range such as
50%, 10%, 1%, and 0.1%  volume/volume
dilution.  The effluent concentrations
used in the definitive  test should
include the LC50  concentration
estimated from the preliminary range
finding test.  A  QA  bioassay with a
standard toxicant (supplied by
Environmental Monitoring and Support
Laboratory {EMSL) -Cincinnati) is
conducted to determine  the sensitivity
of the test organisms.  The LC50
determined from the  QA  bioassay is
compared to a standard  range LC50 of
the reference toxicant  for the test
organism.  If the LC50  from the QA
bioassay falls within the range, that
batch of organisms can  be used for
testing.  The definitive test consists
of five or more effluent concentra-
tions and a control  with 20 organisms
in each concentration.  The definitive
test is run for 24,  48, or 96 hours
under static or flow-through
conditions.  This test  provides an
estimate of the effluent dilution that
kills half the test  population, the
LC50.  These procedures are described
in more detail later in the module.

     The acute toxicity test  is the
basis for the NPDES  biomonitoring
inspection.  The biomonitoring  inspec-
tion is authorized under Section
308(a) of the Clean  Water Act of 1977
(Handout 5).  This Section authorizes
the Administrator of EPA to require
the owner or operator of any  source
discharge to:

     1.  Establish and  maintain
         records

     2.  Make reports

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          U.S. ENVIRONMENTAL PROTECTION AGENCY
COMPLIANCE MONITORING INSPECTOR TRAINING  MODULE:
           NOTES
              LESSON   BIOMONITORING
    Advantages of
    Biomonitoring
    Limitations of
    Biomonitoring
     3.   Install,  use, and maintain
         monitoring equipment and
         methods  (including, where
         appropriate, biological
         monitoring methods)

     4.   Sample the effluent

E.   Advantages of Biomonitoring

     Biomonitoring has several
advantages over chemical by chemical
monitoring.   Because  the acute
toxicity test uses living organisms
which are indigenous  to the receiving
water,  it provides an estimate of the
potential 'effects  of  the effluent on
the survival  of that  species in the
receiving stream.  Living organisms
respond  to the collective effects of
all chemicals in  the  effluent.
Therefore, acute  toxicity tests
measure  the short  term effects of
compound interactions (i.e synergism
and antagonism).   The test measures
the lethal effects of the effluent.
Static bioassays  of effluents can be
run for  as little  as  $400.00, while
detailed chemical  analysis can cost
several  thousand  dollars.  On-site
96-hour  flow-through  bioassays can be
performed for about $5,000.00.

F.   Disadvantages of Biomonitoring

     Effluent biomonitoring is not a
panacea  to solve  all  problems with
toxic effluents.   The test has several
limitations which may require support-
ing chemical  analysis or repeated
testing  to overcome.  One limitation
is that  the composition of some
effluents is  highly variable.  This
problem  also  applies  to sampling
effluents for chemical analysis.  The
variability of the effluent can affect
the results of a  static bioassay, so

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          U.S. ENVIRONMENTAL PROTECTION AGENCY
COMPLIANCE MONITORING INSPECTOR TRAINING  MODULE:
           NOTES
              LESSON   BIOMONITORING
    Objectives of CBI
the sample should be collected as a
24-hour composite (described in the
Manual) to minimize the variability.
Flow-through  bioassay techniques may
be used to continuously monitor
toxicity of an  effluent.  Another
problem is that the acute toxicity
test does not measure the persistence
of the toxic  effect; however, special
procedures are  included in the Manual
to evaluate persistence.  The primary
complaint about effluent biomonitoring
is that it does not identify the cause
of toxicity,  only the effect; but, as
mentioned earlier, chemical analysis
does not identify biological effects,
it only identifies the chemicals
present.  Thus, if we wish to know the
cause-effect  relationship associated
with an effluent, we must use chemical
analysis to support biomonitoring.  If
the wastewater  varies in composition
over a period of time, it is best to
perform chemical analysis (i.e., a
Compliance Sampling Inspection (CSI))
concurrently  with a CBI.  If this is
not possible, past CSI results should
be reviewed and discussed in the
inspection report.  The persistence
and acute toxicity of the effluent
are used to extrapolate out results
to establish  safe concentrations of
the effluent in the receiving stream
(i.e., concentrations which are not
chronically toxic).  This procedure
will be discussed later under
Post-Inspection Activities.

G.   Objectives of CBI

     As stated in the Manual, the
objectives of the CBI are:

     1.  To serve as a screening
         mechanism, isolating toxic
         conditions in effluents
         which may not have been

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          U.S. ENVIRONMENTAL PROTECTION AGENCY
COMPLIANCE MONITORING INSPECTOR TRAINING  MODULE:
           NOTES                   V          LESSON  BIOMONITORING
   Inspectors
   Responsibilities
         detected through routine
         chemical analyses

     2.   To  evaluate compliance with
         State water quality standards

     3.   To  monitor toxics which may
         or  may not be controlled
         through BCT (Best
         Conventional Technology)
         /BAT (Best Available
         Technology)

     4.   Evaluate permit limits

     5.   Develop enforcement cases

     6.   Investigate probable cause
         violations

     7.   Develop data for establishing
         permit limits

H.   Inspectors Responsibilities

      The inspector has certain
obligations  and responsibilities in
conducting a biomonitoring inspection.
These include:

     1.   Knowledge of biomonitoring
         permit conditions,  effluent
         toxicity limitations, and
         related interim and final
         requirements set forth in
         the latest NPDES permit

     2.   Knowledge of EPA policies and
         procedures for conducting,
         interpreting, and reporting
         biomonitoring of wastewater
         effluents

     3.   Developing the inspection
         plan and scheduling the
         inspection

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          U.S. ENVIRONMENTAL PROTECTION AGENCY
COMPLIANCE MONITORING INSPECTOR TRAINING  MODULE:
           NOTES
                   LESSON   BIOMONITORING
    Phases of a CBI
II.  PHASE I-  PREINSPECTION
    PLANNING
    Criteria  used to Select
      Permittees  for
      Biomonitoring
         4.  Conducting the on-site
             biomonitoring  evaluation
             inspection

         5.  Preparing or assembling
             complete and accurate records
             of self-monitoring  practices
             and other issues  addressed
             during an evaluation
             inspection

          6. Follow-up with the  permittee
             and interested parties after
             the audit with regard to
             biomonitoring  performance,
             quality control,  and  related
             compliance activities evalu-
             ated during the inspection

     I.   Phases of a CBI

         The CBI will be discussed in
     three phases:  Phase I  - Preinspec-
     tion Planning, Phase II -   Inspection
     Procedures, and Phase III  -   Post-
     Inspection Activities.   You  should
     refer to the Manual for detailed
     procedures for each phase.  The
     description of these phases  will
     help a  new inspector understand
     biomonitoring.

     Phase I  Preinspection Planning
[I.
     This phase includes both adminis-
trative  procedures to select sites,
notify permittees, and review files;
and  technical procedures to evaluate
the  sensitivity of test organisms and
verify that  equipment materials are
ready for the inspection.

     A.   Selecting Permittees  for  a  CBI

         All inspectors should be aware  of
     the criteria used to select  permittees
     for biomonitoring inspections.   CBI's
                                    10

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          U.S. ENVIRONMENTAL PROTECTION AGENCY
COMPLIANCE MONITORING  INSPECTOR TRAINING MODULE:
           NOTES
              LESSON   BIOMONITORING
    Equipment Requirements
    Equipment
    a)  types of labs
    b)  equipment
    c)  water
        types
        specifications
are frequently performed  at  facilities
with major discharges of  process water
that are suspected  to be  toxic.  This
evidence may include inadequate self-
monitoring reports,  fish  kills in the
receiving stream, and citizen
complaints which suggest  that the
effluent causes some health  hazards.
These are examples  of probable cause.

     CBI's may also be  performed on
a class of facilities defined by
certain criteria in  a neutral
inspection scheme.   For example, EPA
Headquarters,  a Region, or a State
agency may suspect  that a particular
industrial category has a toxic
effluent because the Effluent
Guidelines Division, EPA, has found
priority pollutants in  the waste
stream of some plants within this
category.  Therefore, CBI's  could be
performed on all major  dischargers
with a certain Standard Industrial
Category (SIC) code.  Ambient data
(e.g., toxic hot spots) could be the
basis to perform CBI's  on all major
discharges within a section  of a
stream.  When  classes of  dischargers
are inspected, individual permittees
have been selected  by a neutral
scheme.

B.   Equipment Requirements

     Effluent  toxicity  tests may be
performed in either  a mobile
laboratory or  a permanent facility.
Permanent laboratory facilities are
used primarily for  static bioassays
of effluents;  mobile labs are usually
equipped with  dilutors  to run flow-
through tests.  Permanent facilities
are required to maintain  cultures of
test organisms.  Depending upon the
scope of the bioassay program and the
equipment needed for testing, typical
facilities include  aquaria equipment,
dilution systems, chemical-physical
                                    11

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          U.S. ENVIRONMENTAL PROTECTION AGENCY
COMPLIANCE MONITORING INSPECTOR TRAINING MODULE:
           NOTES
              LESSON   BIOMONITORING
    Construction Materials
    Dilutees

    Flow-Through Dilutions

    Design Considerations:
    1) location
    2) space
    3) application
    4) reliability
    5) performance
    6) cost
monitors,  and  delivery systems.  The
source of  the  dilution water used in
the tests  may  be ground-water, sur-
face water,  reconstituted water, or
dechlorinated  tap water.  Holding,
acclimation, and dilution water should
all be temperature-controlled and
aerated.   Air  used  for aeration must
be free of oil and  fumes, and test
facilities must be  well ventilated.
During the test, organisms should be
isolated  from  any disturbances.

     Materials used to construct test
equipment  that will come into contact
with the  effluent or test organisms
should be  carefully chosen so that
leaching,  dissolution, or sorption are
prevented.  Glass,  no. 316 stainless
steel, and perfluorocarbon plastic
should be  used whenever possible.
Plastics  made  without a plasticizer
(for example,  polyethylene, polypro-
pylene, and fiberglass), can be used
for holding and acclimating, in
dilution  water storage tanks, and in
the water  delivery  system.  Copper,
galvanized material,  rubber, brass,
and lead  should not be used.  If
materials  such as vinyl garden hoses
are used  to pump effluents into the
trailer,  they  should be rinsed thor-
oughly before  testing and discarded
after the  test.

     1.  Dilutors

         The flow-through,
         proportional-dilutor delivery
         system is  the best system for
         routine effluent toxicity
         testing.   The following
         design considerations should
         be used in planning your
         system:
                                   12

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          U.S. ENVIRONMENTAL PROTECTION AGENCY
COMPLIANCE MONITORING INSPECTOR TRAINING MODULE:
           NOTES
     LESSON   BIOMONITORING
    Operating Requirements
     for the Dilutor
a. Whether  the  apparatus will
   be installed and used in a
   permanent  or mobile
   laboratory

b. Space and/or structural
   requirements for the
   delivery system, test
   chambers,  effluent, and
   dilution-water storage

c. The applicability of
   the delivery system
   to specific  effluent
   characteristics  (high
   suspended  solids,
   volatiles, etc.)

d. The dependability,
   durability,  flexibility,
   and ease of maintenance
   and replacement  of the
   system

e. The ability  of the system
   to perform within accept-
   able flow  rate and
   concentration limitations

f. The cost of  the  system

The dilutor must provide for
at least five complete water
volume changes  in 24 hours
in each test  chamber, plus
sufficient  flow to  maintain
an adequate concentration of
dissolved oxygen (^ 4mg/l).
The flow rates  through the
test chambers should not vary
by more than  10% between
chambers.  The dilutor should
be capable  of maintaining the
test concentrations in each
test chamber  within 5% of
                                   13

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          U.S. ENVIRONMENTAL PROTECTION AGENCY
COMPLIANCE MONITORING INSPECTOR TRAINING MODULE:
           NOTES
        LESSON   BIOMONITORING
    Test Chambers
    Cleaning Equipment
    the starting concentration
    throughout the test.   Two of
    the most popular dilutor
    systems are:  the solenoid
    valve and the vacuum  siphon
    systems.  Note:  a solenoid
    valve system is available
    from Ace Glass, Inc.,
    Vineland, New jersey  08360
    (609-692-3333).

2.   Test> Chambers

    Test chambers vary according
    to the  type of test being
    conducted.  For flow-through
    tests,  test chambers  should
    be constructed of 1/4-inch
    plate glass held together
    with small quantities of
    silicon adhesive.  The size
    of the  chambers may vary
    according to size of  the
    test organisms and the
    facility, but the test
    solution should have  a
    minimum depth of 5 cen-
    timeters in the chamber.
    All chambers should have
    either  a glass or screen
    cover to prevent organisms
    from jumping out.  Wide-
    mouthed, 3.9-liter soft-glass
    bottles are often used for
    test chambers  in static
    tests.

3.   Maintenance

    All test chambers and any
    other equipment that  comes
    in contact with the dilutor
    system  must be washed to
    remove  surface contaminants
    using the following
    procedures:
                                   14

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          U.S. ENVIRONMENTAL PROTECTION AGENCY
COMPLIANCE MONITORING INSPECTOR TRAINING MODULE:
           NOTES
    Test Organisms
     LESSON   BIOMONITORING
a. Soak and  wash all
   equipment in a suitable
   detergent and water,
   preferably heated to a
   temperature of 50°C or
   greater.   The detergent
   (powder or liquid) should
   be entirely synthetic
   (SPARKLEEN or ALCONOX).

b. Rinse with tap water
   (preferably heated to
   50°C or greater).

c. Rinse with a fresh, dilute
   (5 percent) hydrochloric
   acid to remove metals and
   bases.

d. Rinse with tap water
   (preferably heated to
   50°C or greater).

e. Rinse with acetone to
   remove organic compounds.
   When contaminated with a
   pesticide, test  chambers
   must be rinsed with
   acetone before they are
   placed  in the hot
   detergent to soak.

f. Rinse twice with dilution
   water.

It is important to  clean all
equipment thoroughly to avoid
transferring toxicants from
one experiment to another.

Test Organisms

The test organisms  to be used
for toxicity testing depend
on the salinity of  the
effluent.  If you are testing
a freshwater effluent, the
                                   15

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          U.S. ENVIRONMENTAL PROTECTION AGENCY
COMPLIANCE MONITORING INSPECTOR TRAINING  MODULE:
           NOTES
      LESSON   BIOMONITORING
Handout 6:  Recommended
Test Organisms
fathead minnow (Pimephales
promelas)  and  Daphnia
(Daphnia magna)  should be
used.    The  species preferred
for saltwater  testing are the
mysid  shrimp (Mysidopsis or
Neomysis)  and  the  sheesphead
minnow (Cyprinodon
variegatus).  Other species
can be tested  in conjunction
with these species.  The
recommended  fish and inverte-
brate  species  must be used
for consistency  and compari-
sons between sites
(Handout 6).

a. Age
It is  recommended  that
minnows be more  than ten
weeks, but less  than ten
months old and that inverte-
brates be  in the juvenile
stage  of development.  These
are the most sensitive post-
hatch  life stages. If other
fish are being tested, it is
important  that they be taken
from the same  year class; and
the total  length and weight
of the fish  should be
approximately  the  same.

b. Acclimation
Disinfect  holding  chambers
and other  equipment with 0.5
percent commercial bleach for
1 hour. Brush thoroughly
with disinfectant  and rinse.
Holding tanks  should receive
uneontarninated water of
consistent quality at a flow
rate of 2-5  tank volumes per
d'ay.

c. Feeding
Acclimation  procedures are
used to prevent  stress on new
                                    16

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          U.S. ENVIRONMENTAL PROTECTION AGENCY
COMPLIANCE MONITORING INSPECTOR TRAINING MODULE:
           NOTES
LESSON  BIOMONITORING
                                           batches of test organisms
                                           caused by drastic environ-
                                           mental changes.  During
                                           acclimation, environmental
                                           changes should be limited  to
                                           a  change of 3°C in water
                                           temperature or 3 ppt salinity
                                           in any 12-hour period,  or  a
                                           total change of 6°C or  6 ppt
                                           salinity.  Over-crowding
                                           should also be avoided. The
                                           water should be aerated, if
                                           necessary, to maintain  an
                                           adequate dissolved oxygen
                                           supply.

                                           d. Disease Control
                                           Test organisms should be fed
                                           at least once a day during
                                           acclimation and holding, and
                                           excess food and fecal
                                           material should be removed
                                           at least twice a week by
                                           siphoning.  The organisms
                                           should be observed
                                           constantly, and a daily log
                                           of feeding schedules,
                                           behavior, and mortality
                                           should be maintained.

                                           During holding, fish should
                                           be chemically treated to cure
                                           or prevent disease.  However,
                                           if the fish are severely
                                           diseased, all should be
                                           discarded.  Diseased
                                           invertebrates should also  be
                                           discarded.  Tanks that  have
                                           held diseased organisms
                                           should be thoroughly dis-
                                           infected with 0.5 percent
                                           commercial bleach before the
                                           tanks are used again.

                                           Organisms from a permanent
                                           facility are transported to
                                           the test site in the water  in
                                           which they were reared  and
                                    17

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          U.S. ENVIRONMENTAL PROTECTION AGENCY
COMPLIANCE MONITORING INSPECTOR TRAINING  MODULE:
           NOTES
LESSON   BIOMONITORING
                                           held.   If the laboratory is
                                           mobile, the acclimation tank
                                           can  be  used for transporting
                                           organisms from the rearing
                                           and  holding facilities to the
                                           test site.  At the test site,
                                           dilution water (receiving
                                           water)  is pumped to the
                                           laboratory for acclimating
                                           the  organisms.  If dilution
                                           water is not readily acces-
                                           sible to the laboratory, it
                                           can  be  transported to the
                                           laboratory and stored in a
                                           tank for use in the accli-
                                           mation  procedure and the
                                           toxicity tests.  During
                                           transport, the organisms
                                           should  not be subjected to
                                           any  salinity or temperature
                                           changes.

                                           At  the  test site, the organ-
                                           isms are acclimated to the
                                           test dilution water and
                                           temperature by gradually
                                           changing from 100% holding
                                           water to 100% dilution water
                                           over 24 hours.  All organisms
                                           must be exposed to 100% dilu-
                                           tion water and be held at the
                                           test temperature (j^2°C) for
                                           at  least 24 hours before
                                           tests are begun.

                                           A group of organisms must not
                                           be  used for a test if they
                                           appear  to be diseased or
                                           otherwise stressed, or if
                                           more than 5% die during the
                                           48  hours immediately pre-
                                           ceding  the test.  If the
                                           organisms fail to meet these
                                           criteria, the entire group
                                           must be discarded and a new
                                           group obtained.
                                    Ifa

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          U.S. ENVIRONMENTAL PROTECTION AGENCY
COMPLIANCE MONITORING INSPECTOR TRAINING  MODULE:
           NOTES
LESSON   BIOMONITORING
   Dilution Water
                                       5.   Dilution  Water

                                           Dilution  water  is water to
                                           be used  in preparing the
                                           different concentrations of
                                           the effluent.   Dilution water
                                           is acceptable if healthy test
                                           organisms survive in it
                                           through  the  acclimation
                                           period and the  toxicity test
                                           without  showing signs of
                                           stress such  as  discoloration
                                           or abnormal  behavior.  The
                                           dilution  water  should be a
                                           sample of the receiving water
                                           and should be obtained from
                                           a point  close to the outfall,
                                           but upstream and outside of
                                           the zone  influenced by the
                                           effluent.  It is preferable
                                           to pump  the  dilution water
                                           continuously to the accli-
                                           mation tank  and dilutor.
                                           However,  it  may be more
                                           practical to transport
                                           batches of water in tanks to
                                           the testing  site as needed,
                                           and then  continuously pump
                                           water to  these  systems from
                                           holding  tanks.

                                           Pretreatment of the dilution
                                           water should be limited to
                                           filtration through a nylon
                                           sieve that has  2-4 millimeter
                                           mesh to  remove  debris and/or
                                           break up  large  floating or
                                           suspended solids.  The water
                                           should be obtained from the
                                           receiving water as close as
                                           possible  to  the time the test
                                           begins.   It  should not be
                                           obtained  more than 96 hours
                                           prior to  testing.  If
                                           acceptable dilution water
                                           cannot be obtained from the
                                           receiving water, some other
                                           uncontaminated, well-aerated
                                   19

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          U.S. ENVIRONMENTAL PROTECTION AGENCY
COMPLIANCE MONITORING  INSPECTOR TRAINING MODULE:
           NOTES
              LESSON  BIOMONITORING
    Handout 7: Equipment
    Checklist
    Preinspection
    Coordination With
    the Permittee
    Handout 8: Preinspection
    Questions
         surface or ground water, or
         commercial source may be used
         or prepared.  This water must
         have  a total hardness, total
         alkalinity, and specific
         conductance within 25% of the
         receiving water at the time
         of testing.  The pH must be
         within 0.2 units of the
         receiving water at the time
         of testing.

         All biomonitoring teams must
         have  at least one experienced
         biologist who directs the
         preparation of equipment for
         the inspection.  Minor
         equipment items and supplies
         should be duplicated to avoid
         unnecessary delays in cases
         of failure or breakage.  Many
         Regions have developed
         checklists of equipment and
         supplies, such as Handout 7,
         to reduce problems with
         packing.

C. Preinspection Coordination

     Preinspection procedures also
include a review of the permittee's
compliance file, permit application,
and previous inspection data to
develop an understanding of the
permittee's production process,
wastewater composition and possible
problems.  If  an on-site flow-through
bioassay ,is.planned, it is best to
contact the permittee early.  One way
to reduce logistics problems is to
supply the. permittee with a list of
questions before-the inspection
(Handout 8).  In cases where.the
inspection is  announced well in
advance, it is important to compare
the discharge  characteristics during
the inspection period with routine
measurements (particularly for flow)
                                    20

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          U.S. ENVIRONMENTAL PROTECTION AGENCY
COMPLIANCE MONITORING INSPECTOR TRAINING MODULE:
           NOTES
                   LESSON  BIOMONITORING
III.   PHASE II  INSPECTION
      PROCEDURES

    Evaluation  Inspections
    PA I
    Review Procedures
     to be sure the plant was operating
     normally.  Variations of more  than
     ten percent should be identified  and
     explained in the inspection report.

        The latest QA bioassay results
     should be reviewed before the
     inspection.  The purpose of the QA
     bioassay is to determine the sensi-
     tivity of the test organisms to a
     reference toxicant, usually sodium
     lauryl sulfate.  The QA bioassays may
     be performed during the inspection or
     as part of the laboratory routine.   If
     the results are more than three weeks
     old, another QA bioassay should be
     conducted, either concurrently with
     the inspection or before the
     inspection.

[II.  Phase II Inspection Procedures

     A.   Evaluation Inspections

         The inspector will be conducting
     both sampling and nonsampling  inspec-
     tions related to biomonitoring. The
     nonsampling or evaluation inspection
     includes the PAI and the CEI.   The
     procedures for these inspections  are
     described in other training modules
     and specific manuals.
          1.   performance Audit Inspections

              The PAI requires at least one
              inspector with experience in
              bioassays.  The inspector
              must review the performance
              of the permittee's staff and
              evaluate  their test and
              sampling procedures.   The
              inspector should review all
              test and culturing proce-
              dures.  Particular attention
              should be given to the
              following areas:
                                    21

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          U.S. ENVIRONMENTAL PROTECTION AGENCY
COMPLIANCE MONITORING INSPECTOR TRAINING  MODULE:
          NOTES
     LESSON   BIOMONITORING
    Condition  of Test
    Organisms
    Test Procedures
    Recording  Results
a.
b.
  What  is  the source, age,
  weight,  and species of
  test  organism?

  Do they  appear to be
  healthy?

  At what  temperature are
  they  maintained?

  What  and how are they
  fed?

  Are they isolated from
  test  chambers and vola-
  tile  components of the
  effluent?

  How are  they treated for
  disease?

How are test concentra-
tions established?

Are flow,  temperature,
D.O., etc. controlled or
monitored  during tests?

What is the source of
dilution water?

Is there any chemical
analysis to verify that the
dilution water is a good
control?
c. How are  the test observa-
   tions  made and recorded?

   Is the data in a bound
   log?

   Are test conditions/
   failures described?

   Do the log results
   correspond to DMR data?
                                 22

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          U.S. ENVIRONMENTAL PROTECTION AGENCY
COMPLIANCE MONITORING INSPECTOR TRAINING  MODULE:
          NOTES
     LESSON   BIOMONITORING
    Data Analysis
    Sampling Procedures
d.  What  method is used to
   calculate LCSO's?

   Do the  survival levels
   correspond to the
   requirements for a
   definite test?

e.  When  conducting the
   performance audit, the
   inspector must evaluate
   the permittee's effluent
   sampling practices and
   verify  that:

   Samples are taken at
   locations prescribed in
   permit

   Sampling locations
   specified in the permit
   are adequate to provide  a
   well-mixed and represen-
   tative  sample

   The frequency of sam-
   pling is done in
   accordance with the
   permit

   Grab  sample devices
   are clean and properly
   operated

   Sample  containers are
   clean and appropriate

   Automatic sample col-
   lectors are operating
   properly

   Samples are properly
   preserved and shipped

   Effluent samples for bio-
   assays  are used without
   preservatives or
   adulteration
                                  23

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          U.S. ENVIRONMENTAL PROTECTION AGENCY
COMPLIANCE MONITORING INSPECTOR TRAINING  MODULE:
           NOTES
             LESSON   BIOMONITORING
    CEI
     Sampling CBI's
           There is no more  than  a
           24-hour maximum holding
           time before sample
           is used for biomonitoring

           Cross-contamination  of
           samples is prevented

           Testing equipment is
           routinely calibrated

        Furthermore, a performance
        audit must be conducted for
        the permittee's laboratory.
        •The inspector is responsible
        for evaluating facilities,
        test organisms, dilution
        water, test procedures, and
        test results.

     2.  Compliance Evaluation
        Inspections

        The CEI is the simplest and
        least resource-intensive
        type of 'evaluation inspec-
        tion.  At facilities that
        conduct biomonitoring,  a  CEI
        consists of an examination
        of the permittee's self-
        biomonitoring files and
        records, analytical and
        bioassay laboratory, and
        production*facilities  along
        with the routine review of
        sampling records and reports.

B.  .  Sampling Inspections

     Sampling inspections are an
evaluation of an effluent based  on
sampling and testing."^ Sampling
inspections can be conducted on-site
or off-site.  On-site* biomonitoring
sampling inspections include:
                                    24

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          U.S. ENVIRONMENTAL PROTECTION AGENCY
COMPLIANCE MONITORING INSPECTOR TRAINING  MODULE:
           NOTES
        LESSON   BIOMONITORING
    Sampling Procedures
    An  8-  to 24-hour range-
    finding bioassay

    A 96-hour  flow-through
    bioassay

    A 24-hour  QA bioassay with a
    reference  toxicant

    Careful examination of the
    permittee's self-monitoring
    program

1.   Effluent Sampling

    When  static bioassays are
    performed, samples of the
    effluent must be collected.
    Grab  samples may be used if
    the composition of the
    effluent is very consistent
    (see  Manual).  However, com-
    posite samples should be used
    if  the effluent is variable
    or  retention time of the
    effluent in the treatment
    system is  less than seven
    days.  Composites should be
    collected  for one full opera-
    ting  day.  If the discharge
    is  continuous, a 24-hour
    composite  is required.

    Effluent samples should be
    taken  at the point specified
    in  the NPDES discharge per-
    mit.   It is important that
    the sample represent the
    "normal or typical" discharge
    and operating conditions of
    the facility.  Sampling
    should be  based on an under-
    standing of the short-term
    operations and schedules of
    the discharger.  Samples
    should not be altered prior
    to  testing, although they
    may be filtered through a
                                   25

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          U.S. ENVIRONMENTAL PROTECTION AGENCY
COMPLIANCE MONITORING INSPECTOR TRAINING  MODULE:
           NOTES
        LESSON   BIOMONITORING
    Handout 9:  Effluent
    Sampling procedures

    Sampling Handling
     Range-Finding Tests
   2-4 millimeter screen  made
   of stainless steel  or  Teflon.
   Handout 9 lists effluent
   sampling procedures.

   Once the effluent grab sample
   has been collected, it must
   be stored in a covered but
   loosely sealed container.
   Violent agitation must be
   avoided, but gentle agitation
   may be necessary to disperse
   suspended solids before
   dilutions are made.  Samples
   should be stored at 4°C in
   a refrigerator, a constant-
   temperature water bath, or  an
   environmentally controlled
   room at the test temperature.
   The test should be conducted
   within 48 hours after
   collection.

   Acute toxicity testing
   procedures  include both range
   finding and definitive tests
   which were  defined earlier.
   Although toxicity testing may
   be done under static or flow-
   through conditions, static
   tests are used primarily
   because they are less  expen-
   sive and easier to perform.

2.  Definitive  Tests

   The flow-through testing
   procedure is preferred for
   the definitive test because
   it eliminates concerns about
   holding time and wastewater
   variations.  However,  the
   flow-through  is more expen-
   sive because  it is done at
   the permittee's site.
   Therefore,  it requires more
   planning and  resources than
                                    26

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          U.S. ENVIRONMENTAL PROTECTION AGENCY
COMPLIANCE MONITORING INSPECTOR TRAINING MODULE:
           NOTES
     LESSON   BIOMONITORING
    Criteria for Valid LC50
    Handout 10:  Definitive
    Test Requirements
    Conditions
a static  test performed in a
permanent laboratory.  In
flow-through tests, the
delivery  system should be in
operation for 24 hours prior
to adding the test organisms;
for static test, the effluent
should  be mixed well by
stirring  with a glass rod.
The test  conditions needed
for determining an LC50 must
include at least five
concentrations and a control.

a. Three  criteria must be met
   in order to calculate a
   reasonably accurate LC50:

   Concentrations must be
   at least 50% of the
   preceding effluent
   concentration.

   One  concentration must
   have affected 65% of the
   organisms exposed to it,
   and  one concentration, other
   than the control, must have
   affected less than 35%
   of the organisms. If 100%
   effluent does not affect
   more than 65% of the exposed
   organisms, it is then
   necessary to report the
   numbers that are affected.
   Toxicity must increase with
   higher effluent concentra-
   tions.

b. Handout 10 lists the
   definitive test require-
   ments.  These are:

   Twenty test organisms of a
   given  species must be used
   for  each concentration or
                                   27

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          U.S. ENVIRONMENTAL PROTECTION AGENCY
COMPLIANCE MONITORING INSPECTOR TRAINING  MODULE:
           NOTES
LESSON  BIOMONITORING
                                             ten for each  chamber
                                             (replicate  test chambers).

                                             If two or more species are to
                                             be used in  the test chambers,
                                             they should be separated with
                                             Nitex screens.

                                             Randomization of  treatments
                                             is desirable  (i.e., organisms
                                             should be assigned to concen-
                                             trations based on statistical
                                             randomization techniques).

                                             Loading must  not  exceed 5
                                             grams of test organism per
                                             liter at temperatures of
                                             20°C or less, or  2.5 grams/
                                             liter at temperatures above
                                             20°C for flow-through tests.

                                             Loading for static tests must
                                             not exceed  0.8 grams per
                                             liter-at temperatures of 20°C
                                             or less, and  0.4  grams/liter
                                             at temperatures above 20°C.

                                             Temperatures  must be held to
                                             within +2.0°C of  the
                                             acclimation temperature for
                                             both static and flow-through
                                             tests.

                                             Dissolved oxygen  concentra-
                                             tion should not be permitted
                                             to fall below 40%
                                             saturation  for warm water
                                             species and 60% for cold
                                             water species.

                                          The  test period  begins when
                                          the  organisms  are  first
                                          exposed to the effluent.

                                          Organisms are  not  fed during
                                          testing, unless  they are
                                          mature or newly  hatched.
                                          The  duration of  the  test
                                   28

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          U.S. ENVIRONMENTAL PROTECTION AGENCY
COMPLIANCE MONITORING INSPECTOR TRAINING MODULE:
           NOTES
         LESSON   BIOMONITORING
    Test Results
    Data Reporting

    Handout 11:  Data
    Sheet for Effluent
    Toxicity Test
    is 24 hours for static tests
    and 96 hours for flow-through
    bioassays.

3.   Test Results

    Three types of results can
    be obtained from an  acute
    toxicity test done on
    effluents:

    Biological data

    Chemical and physical data

    LC50 calculations

    a. These data are recorded
      on the data sheet shown  •
      in Handout 11.  Biological
      data include:

      Length and weights of
      representative test
      organisms, and age,
      where applicable

      The number of affected
      (dead)  organisms  in each
      test container after each
      day of testing

    b. Physical and chemical data
      include:

      Dissolved oxygen, tempera-
      ture, and pH measurements
      taken at the beginning of
      the test and each day
      thereafter in the control
      and in each effluent
      concentration

      Specific conductivity,
      total alkalinity,
      hardness, and salinity
      measurements taken at the
      beginning of the  test in
                                   29

-------
          U.S. ENVIRONMENTAL PROTECTION AGENCY
COMPLIANCE MONITORING INSPECTOR TRAINING MODULE:
           NOTES
                  LESSON   BIOMONITORING
    Handout 12:  Methods  for
    Calculating LC50,  EC-50
    Handout 13: Reporting
    Test  Results

    Handout 14: Day-to-Day
    Activity Guide

IV.  POST-INSPECTION
    PROCEDURES
    Handout 15: Acute
    Toxicity Laboratory
    Evaluation Form

    Handout 16: NPDES
    Compliance Inspection
    Report Form
                the  control and in each
                effluent concentration.
                Total  ammonia nitrogen
                measured at the beginning
                and  end of each static
                test in the control and
                in all effluent concen-
                trations

                LC50 and EC50 calculations
                and  their 95 percent con-
                fidence limits must be
                obtained for each set of
                data based on the initial
                volume percent of the
                effluent.  Several
                statistical methods are
                available.  Handout 12
                discusses two of the more
                common methods:
                Litchfield and Wilcoxon,
                and  log concentration
                versus percent survival.

                Finally, information that
                should be included in the
                report of the results is
                shown in Handout 13.
                Handout 14 provides a
                guide for day-to-day
                inspection activities.
IV.   POST-INSPECTION PROCEDURE

     Post-inspection activities described
     below include:

     Data evaluation

     Completing the Acute Toxicity
     Laboratory Evaluation Form
     (Handout 15)

     Completing the NPDES Compliance
     Inspection Report (Handout 16).
                                    30

-------
          U.S. ENVIRONMENTAL PROTECTION AGENCY
COMPLIANCE MONITORING INSPECTOR TRAINING  MODULE:
           NOTES
              LESSON   BIOMONITORING
    Data Evaluation
A.   Data Evaluation

     The primary  purpose of a
compliance biomonitoring bioassay is
to establish  compliance status with
bioassay requirements  in a permit.
Bioassays can also  be  used to deter-
mine if a particular effluent is
acutely toxic.   Evaluation of biomoni-
toring data is  based on the following
parameters:

     LC50 of  the  waste

     In-stream  waste concentration

     Permit limits

     Chemical parameters of the
     effluents  associated with the
     bioassay,  such as:

     -Dissolved oxygen
     -Temperature
     -pH
     -Conductivity
     -Metals
     -Organics

     The LC50 can be determined by
probit analysis or  graphical technique
as shown in Handout 12.  The graphical
technique does  not  provide confidence
intervals for the LC50;  therefore,
statistical methods such as probit,
moving average  angle, or log it are
preferred.   The Manual provides a
discussion of how to determine
compliance using  the LC50, the
instream waste  concentration, and
an application  factor.  Since this
is critical,  it is  reviewed here.
Computer programs are available
for the statistical methods from
Mr. Charles Stephen, ERL-Duluth,  MN
(8-783-9510).
                                   31

-------
          U.S. ENVIRONMENTAL PROTECTION AGENCY
COMPLIANCE MONITORING  INSPECTOR TRAINING  MODULE:
           NOTES
             LESSON   BIOMONITORING
    Data interpretation
B.   Data Interpretation

     Bioassay  results are evaluated
by verifying that  the test conditions
(e.g.,  water quality, survival of
controls, and  loading rates) were
acceptable and by  estimating the
impact  of the  effluent on the receiv-
ing stream.   Continuous measurements
of water quality in  flow-through tests
monitor changes in wastewater composi-
tion.  Changes in  the flow-rate during
sampling may indicate that changes
occurred in the manufacturing
operation or stormwater runoff.  Acute
toxicity may be caused by water
quality variation  such as low dis-
solved  oxygen  concentrations (4.0
mg/1) or changes in  pH.  Water tem-
perature and conductivity measurements
can detect changes caused by different
manufacturing  operations.  The
biologist should examine the pattern
of survival during the test.  If the
LC50 is the same at  24 hours as one
estimated for  96 hours, the effluent
probably contains  toxic compounds
which are degraded or volatile.  In
some cases,  this toxicity could be
reduced by increasing the retention
time of the treated  effluent.  If the
effluent causes deaths throughout the
test period, toxic compounds in the
effluent are probably bioaccumulated
by the test organisms.  The toxicity
of the effluent is based on the LC50
and the instream waste concentration.
The effluent may be  highly  toxic (LC50
1.0%),  moderately toxic (1-25%),
somewhat toxic (25-50%) or marginally
toxic (50%).  Highly toxic effluents
require immediate action- to reduce the
toxicity.  In most cases the permittee
is probably violating permit limits
already.  Permittees with moderately
toxic effluents should be considered
for biomonxtoring requirements in
their permits.  Permittees  with
somewhat toxic effluents and
marginally toxic effluents  should
                                    32

-------
          U.S. ENVIRONMENTAL PROTECTION AGENCY
COMPLIANCE MONITORING INSPECTOR TRAINING  MODULE:
           NOTES
              LESSON  BIOMONITORING
    Example
be retested  if  they  fail the IWC
criterion.

    The instream  waste concentration
(IWC)  is the  ratio of the effluent
flow to the  7 day, 10 year low flow of
the receiving stream.  IWC = Q effluent
                               7Q10

(Note:   In some cases, where 7Q10 is
not available,  the annual low flow
value  may be  acceptable.)

     The IWC  is compared to the product
of the LC50,  expressed as the percent
concentration of  the effluent, and an
Application  Factor (AF).  The
Application  Factor is intended to
protect aquatic organisms from chronic
toxicity outside  the mixing zone.  If
the toxicity of the  effluent is
considered persistent (i.e., the
effluent contains persistent compounds
or the toxicity persists), AF = 0.01;
if non-persistant, AF = 0.05.  Thus,

                 pass

non-persistent  iWC^LCSOxO.05
persistent      IWCM.C50xO.01

                 fail

non-persistent  !WC
-------
          U.S. ENVIRONMENTAL PROTECTION AGENCY
COMPLIANCE MONITORING INSPECTOR TRAINING  MODULE:
          NOTES
             LESSON   BIOMONITORING
    Reporting  Results
    Handout 15  & 17:
    Instruction for
    Completing  Forms
than 1000, additional treatment may be
needed.   For lakes or estuaries, a
mixing zone volume is used  instead of
the 7 day, 10 year low flow.

C.   Inspection Report

     The  Acute Toxicity Laboratory
Evaluation form and the NPDES
Compliance Inspection Report should
be filled out according to  the
instructions provided in Handout 15
and 17.   Each trainee should review
the instructions prior to filling
them out.

     Although biomonitoring  is not
currently required under the NPDES
Program,  it undoubtedly will be in
the future.  Thus, it is important
for NPDES inspectors to be  thoroughly
familiar with the procedures for
conducting a biomonitoring  inspection
and performing subsequent toxicity
tests.
                                   34

-------
                                                  HANDOUT 1

                             GLOSSARY
Acclimation — The process of adjusting to a new environment.  The
               test organisms should be acclimated to the dilution
               water before the test begins.

Acute Toxicity — Short-term effect of a toxicant on test
               organisms.  Death is the end point in acute
               toxicity tests.

Acute Toxicity Testing — Short term bioassays to evaluate
               lethal effects.

Announced Inspection — An inspection in which the permittee is
               informed of the exact dates the inspection will be
               conducted.

Audit (or Performance Audit Inspection - PAI) — A nonsampling
               type of inspection to assess all the elements of
               a permittee's self-monitoring program, such as
               testing procedures and methodology while being
               implemented, quality assurance, data gathering and
               interpretation, files, and laboratory data
               facilities.

Bioassay — A test to detect or measure the effect of one or more
               substance, waste, or environmental factors on living
               organisms.

Effluent Biomonitoring — Measurements of the biological effect of
               effluents (such as toxicity, biostimulation, and
               bioaccummulation) and their effect on the abundance,
               composition, and functions of indigenous aquatic
               organisms in receiving waters (biological integrity),
               For purposes of this manual, effluent biomonitoring
               refers to acute toxicity bioassays.

Definitive test — A full-scale bioassay consisting of at least
               five different concentrations of effluent and a
               control each containing 20 or more organisms of a
               given species.

Dilution Water — Mixing water to be used for preparing dilutions
               of the effluent.  This water is usually collected
               from a point as close as possible but outside the
               zone of influence of the effluent.  This water is
               also used for the control test chambers.

-------
                                                HANDOUT 1
                                                (Continued)
EC50 — Median effective concentration; the concentration producing
               a specific response, other than death, in 50% of the
               test organisms.  Responses can be behavioral, a
               developmental abnormality, or a deformity.

Effluent — For the purposes of this module, an outflow from a
               point source which is regulated by an NPDES permit.

Evaluation Inspections — Inspections that do not include any type
               of sampling of the effluent or" the receiving waters.
               These inspections assess some aspects of a permit-
               tee's self-monitoring program.

Flowthrough Bioassay — Continous flow bioassay; the type of test
               where different concentrations of the effluent are
               prepared by mixing with adequate quality dilution
               water and are then tested by allowing the effluent
               mixtures to flow at predetermined rates into
               chambers containing the text organism.

LC50 — Median lethal concentration.  The concentration which
               kills 50% of the test organisms.

Rangefinding Test — A short-term (8-24 hours) flowthrough or
               static bioassy (usually static) used for determining
               the approximate concentrations, above and below the
               LC50, to be used in the definitive test.  In this
               test, groups of five organisms are exposed to from
               three to five widely spaced effluent dilutions.

Sampling Point — Particular site whose location may be specified
               in a permit and from which effluent samples are to
               be collected for testing and evaluation.

Standard Toxicant — A reference toxicant used for quality
               assurance purposes in the biomonitoring program.
               It is used in a bioassay to determine the repro-
               ducibility of test results and differences in
               senstivity among batches of test organisms.

-------
                                                             HAMDOUT 2
                                                             (1 of 8)
                       Point Source Categories
1.   TIMBER PRODUCTS PROCESSING

     SIC'  2411 - Logging Canps and Logging Contractors (Camps Cnly)
     SIC  2421 -'Sav Mills and Planing  Mills, General
     SI-C  2426 - Hardwood Dincncior. and Flooring Mills
     SIC  2429 - Special Purpose Sawmills, Not Elsewhere Classified
     SIC  2431 - Millwork
     SIC  2434 - Wood Kitchen Cabinets
     SIC  2435 - Hardwood Veneer and Plywood
     SIC  2436 - Softwood Veneer and Plywood
     SIC  2439 - Structural Wood Members,  Not Elsevhere Classified
     SIC  2491 - Wood Preserving
     SIC  2499 - Wood Products, Not Elsewhere Classified ^Furniture
                 Kills)
     SIC  2661 - Building Paper and Building Boz.ro Mills (Kardboard
                 Only)

2.   STEAM ELECTRIC POWER PLANTS

     SIC  4911 - Electric Services (Limited to Stezrc-Eloctric Pover
                 Plan-ts)

3.   LEATHER TANNING AMD FINISHING

     SIC  31   - Leather and Leather Products

4.   IRON AND STEEL XA-NUFACTURIKG

     SIC  3312 - Blast Furnaces (Including Coke Ovans),
                 Steel Works and Rolling Mills.
     SIC  3313 - Elcctrometallursical Products.
     SIC  3315 - Steel Wire .Drawing and Stesl Nails and Spikes.
     SIC  3316 - Cold Rolled Steel Sheet,  Strip and Bars.
     SIC  3317 - Steel Pipe and Tubes.

5.   PETROLEUM RF.FK.'INC

     SIC  2911 - Petroleum Refining (Including 1) Topping Plant; 2)
                 Topping and Cracking Plants; 3) Topping, Crsckinr; and
                 Pctro-chcmical Plants; 4) Integrated Plants; and, 5)
                 Integrated and Pctro-chc:»ical Plants)

-------
                                                              HANDOUT  2
                                                              (Concinued 2 of 8)

 .           C cnr.MiCAi.5 MA::UKACU:KT:.T.

     SIC  2812 - Alkclics and Chlorine
     SIC  2813 - Industrial Casscs .
     SIC  2816 - Inorganic Pigir.cnts
     SIC  2819 - Industrial Inorganic Chemicals, Not Llscwhcre
                 Classified

7.   TEXTILE HILLS

     SIC    22 - Textile Mill Produces
     SIC    23 - Apparel and Other Finished Products Made fron Fabrics and
                 Similar Materials

3.   ORGANIC CHEMICALS MANUFACTURING

     SIC  2865 - Cylic (Coal Tar) Crudes, and Cylic
                 Intermediates, Dyes, and Organic Pigments (Lakes :md Toners)
     SIC  2869 - Industrial Organic Chemicals, Not Elsewhere Classified.
9.   NONFERROUS METALS MANUFACTURING

     SIC  2819 - Industrial Inorganic Chemicals, Not Elsewhere
                 Classified (Hauxite Refining Only)
     SIC  3331 - Primary Smelting and Refining of Copper
     SIC- 3332 - Primary Smelting and Refinir.g of Lcsc
     SIC  3333 - Prirary Sr.elting and Refining of Zinc
     SIC  3334 - Primary Production of .'.luninua .
     SIC  3339 - Primary Smelting and Refinir.g of Xonferrous Metals,
                 Not Elsewhere Classified
     SIC  3341 - Secondary Smelting and Refining of Kcnferrous l-Jerals

10.  PAVING ANT) ROOFING XATERIALS (TAT.S ANT) ASPHALT)

     SIC  2951 - Paving Mixtures and Blocks
     SIC  2952 - Asphalt Felts ar.d Coatings
     SIC  3996 - Linoleum, As?haitec-relt-Ease, and Other Hard
                 Surface Floor Coverings, Not Elsewhere Classified

11.  PAINT AND INK FORMULATION A>D TRiNTING

     SIC  2711 - Newspapers: Publishing, Publishing and Printing
     SIC  2721 - Periodicals: Publishing, Publishing snd Printing
     SIC  2731 - Books: Publishing, Publishing and Printing
     SIC  2732 - Book Printing
     SIC  2741 - Miscellaneous Publishing
     SIC  2751 - Cocncrciol Pointing, Lett.crprcss and Screen
     SIC  2752 - Commercial Printing, Letterpress and Lithographic
     SIC  2753 - Engraving and '1'lice Printing
     SIC  2754 - Commercial Printing, Gravure
     SIC  2761 - Mainfold Business Forms
     SIC  2771 - Greeting Card Publishing
     SIC  2793 - Photoengraving
     SIC  2794 - Elcctrotyping and Stereotyping

-------
                                                                  HANDOUT  2
                                                                  (Continued 3 of 8)
     SIC  2795 - Lithographic 1'J.itcmnk.irc and 'icOoCccl D
     SIC  2851 - Taints, Varnishes, Lacquers, Lnairsls, and Allied  Products
     SIC  2893 - Printing Ink
     SIC  3951 - Pens, Mcchajiicnl pencils, and Parts and  Stamp
                 Pads (Inked Materials Only)
     SIC  3952 - Lead Pencils, Crayons, and Artists' Materials
     SIC  3955' - Carbon Paper and Inked Ribbons

12.  SOAP AND DETFRCEKT MA::LTACTuT>ll!G

     SIC  2841 - Soap and Other Detergents, except Specialty  Cleaners

13.  AUTO AND OTHER LAUNDRIES

     SIC  7211 - Power Laundries, Family and Commercial
     SIC  7213 - Linen Supply
     SIC  7214 - Diaper Service
     SIC  7215 - Coin-operated Laundries and Dry Cleaning
     SIC  7216 - Dry Cleaning Plants, Zxcept Rug Cleaning
     SIC  7217 - Carpet and Upholstery Cleaning
     SIC  7218 '- Industrial Laundries
     SIC  7219 - Laundry and Carrrent Services, Not Elsewhere  Classified
     None      - Auto Wash Establishments

14.  PLASTIC AXD SYNTKLTIC MATERIALS riA-XU
     SIC   282 - Plastic Materials and Synthetic F.esir.s,  Synthetic and
                 Other Kanraade: Fibers, except Glass

15.  PULP AND PAPERBOASD >:tLLS ; AMD CONVERTED PA?E?. PRODUCTS

     SIC  2611 - Pulp Mills
     SIC  2621 - Paper Mills, except Buildir.g Paper Hills
     SIC.  2631 - Paperboard. Mills
     SIC  2641 - Paper Coatirvg and Clawing
     SIC  2642 - Envelopes
     -SIC  2643 - Bags, Except 'Textile Bags
     SIC  2645 - Die-Cut 'Paper and Paperboarc and Cardboard
     SIC  2646 - Pressed ar.d Molded Pulp Goods
     SIC  2647 - Sar.itcry Paper Iroducts
     SIC  2648 - Stationery,  Tablets, ana Related Products
     SIC  2649 - Converted Paper and. Paperboard Products, Not  Elsewhere
                 Classified
     SIC  2651 - Folding Papexboard Boxes
     SIC  2652 - Set-up Paperboarc Boxes
     SIC' 2653 -.Corrugated end, Solid Fiber Boxes
     SIC  2654 - Sanitary Food Containers
     SIC  2655 - Fiber Cans, '.Tubes, Drums, and Similar Products
     SIC  2661 - Building Paper and Building Board Mills
     SIC  '2782 - Blankbooks,  Looselcaf Binders and Devices

16-  RUBBKK PP.OCr.SSIMC

     SIC  2822 - Synthetic Rubber  (Vulcanizablc Eljstovcir.)
     SIC  269] - Rubber Cement

-------
                                                              HANDOUT  2
                                                              (Continued 4 of 8)


     SIC  3011 - Tires and Inner Tubes
     SIC  3021 - Kubbcr unci I'lji/tics Footwear (Kubbcr UnJy)
     SIC  3031 - r.oclair.cd Kubbcr
     SIC  3041 - Rubber and Plactics Hose and Belting (Rubber Only)
     SIC  3069 - Fabricated Rubber Products, Hot Elsewhere Classified
     SIC  3293 - Caskets, Packing, and Sealing Devices
                 (Rubber Packinc Only)

17.  MISCELLANEOUS CHEMICALS

     SIC  2831 - Biological Products
     SIC  2833 - Medicinal Chemicals and Botanical Products
     SIC  2834 - Pharmaceutical Preparations
     SIC  2861 - Cun and Wood Chemicals
     SIC  2879 - Pesticides and Agricultural Chemicals, Not Elsewhere Classified
     SIC  2891 - Adhesive and Sealants
     SIC  2892 - Explosives
     SIC  2895 - Carbon Black
     SIC  2899 -. Chemicals and Chemical Preparation, Not Elsewhere
                 Classified
   •  SIC  3861 - Photographic Equipment and Supplies

18.  MACHINERY AM3 MECHANICAL PRODUCTS :i^CUF/.C7LT.I?:C

     SIC  3021 - Rubber and Plastics Footwear (Balance)
     SIC  3041 - Rubber and Plastics Hose and Belting (Balance)
     SIC  3079 - Miscellaneous Plastics Produces
     SIC  3293 - Caskets, Packing, and Sealing Devices (3alar.ce)
     SIC  3321 - Gray Iron Foundries
     SIC  3322 - Malleable Iron Foundries
     SIC  3324 - Steel Investment Foundries
     SIC  3325 - Steel Foundries, Not Elsewhere Classified
     SIC  3351 - Rolling, Drawing, and Extruding of Copper
     SIC  3353 - Aluminum Sheet, Plate, and Foil
     SIC  3354 - Aluminum Extruded Products •
     SIC  3355 - Aluminum Rolling and Drav.-ing, Not Elsewhere Classified
     SIC  3356 - Rolling, Drawing, ar.d Extruding.of Konferrous
                 Metals, except copper and 'aiur.inum
     SIC  3357 - Drawing and Insulating of Nonferrous Wire
     SIC  3361 - Aluminum Foundries (Castings)
     SIC  3362 - Brass, Bronze, Copper, Copper Base Alloy Foundries (Castings)
     SIC  3369 - Nonferrous Foundries (Castings),'Mot Elsewhere Classified
     SIC  3398 - Metal Heat Treating
     SIC  3399 - Primary Metal Products, Not Elsewhere Classified
     SIC  3411 - Metal Cans
     SIC  3412 - Metal Shipping Barrels, Druns, Kegs, and Pails
     SJC  3421 - Cutlery
     SIC  3423 - Hand and Edge Tools, Except Machine Tools and Hand Saws
     SIC  3425 - Hand Saws and Saw Blades
     SIC  3429 - Hardware, Not Elsewhere Classified
     L1C  3431 - Enameled Iron and Metal Sanitary tore
     SIC  3432 - riurr.bir.g Fixture Fittings and Trin (Brass Goods)
     SIC  3433 - Heating Equipment, Except Klectric and Karc Air Furnaces
     .SIC  3441 - Fabricated Structural Metal
     SIC  3442 - Metal Doors, Sash, Frames, Moldiiv}, and Trim

-------
                                                               HANDOUT  2
                                                               (Continued 5 of 8)
SIC  3443 - 1'abriciitud Pljicwork  (lioilcr  Shops)
SIC  3444 - Sheet .Metal l.'ork
SIC  3446 - Architectural end Orn.-imcntal  Metal Work
SIC  3448 - Prefabricated Metal I'.uildin^s  and Components
SIC  3449 - Miscellaneous Metal V.'ork
SIC  3451 - Screw Machine Products
SIC  3452 - Bolts, Huts, Screws,  Rivets,  and Washers
SIC  3462 - Iron and Steel Forcings
SIC  3463 - Konfcrrous Forcings
SIC  3465 - Automotive Stampings
SIC  3466 - Crowns and Closures
SIC  3469 - Metal Stampings, "ot  Elsewhere Classified
SIC  3482 - Small Ar.T.s AnT.unition
SIC  3483 - Annumtion, Except for Sr.all  Anrs, tfot  Elsewhere Classified
SIC  3484 - Sir.all Arrs
SIC  3489 - Ordnance and Accessories, Not  Elsewhere Classified
SIC  3493 - Steel Springs, Except Wire
SIC  3494 - Valves and Pipe Fittings, Except Plur.bers''Brass Goods
SIC  3495 - Wire Springs
SIC  3496 - Miscellaneous Fabricated Wire  Products
SIC  3497 - Metal Foil and Leaf
SIC  3498 - Fabricated Pipe ar.d Fabricated Pipe  Fittings
SIC  3499 - Fabricated Metal Products, I'.ot Elscvherc Classified
SIC  3511 - Scean, Gas, and Hydraulic Turbines ar.d  Turbine
            Generator Set Units
SIC  3519 - Internal Combustion Engines,  Sot Elsev.-here Classified
SIC  3523 - Farn Machinery and Equipncr.t
SIC  3524 - Garden Tractors ar.d Lawn and  Garden  Equipment
SIC  3531 - Construction Machinery and Equipment
SIC  3532 - Mining Machinery and  Equipment, Except  Oil Field
            Machinery snd Equipnent
SIC  3533 - Oil Field Machinery ar.d Equiprent
SiC  3534 - Elevators and Moving  Stairways
SIC  3535 - Conveyors and Conveying Equiprier.t
SIC  3536 - Hoists, Industrial Cranes, ar.d Xcrorail Systcrs
SIC  3537 - Industrial Trucks, Tractors,  Trailers,  and Stackers
SIC  3541 - Machine Tools, Metal  Cutting  Types
SIC  3542 - Machine Tools,, Metal  Fcriring  Types
SIC  3544 - Special Dies ar.d Tools, Die Sets, Jigs  and
            Fixtures and Industrial Molds
SIC  3545 - Machine Tool Accessories and  Mea.-jrir.g  Devices
SIC  3546 - Power Driven Hand Tools
SIC  3547 - Rolling Mill Machinery and Equiprent
SIC  3549 - Metalworking Mcchinery, Not Elsewhere Classified
SIC  3551 - Food Produces Machinery
SIC  3552 - Textile Machinery
SIC  3553 - Woodworking Macninory
SIC  3554 - Paper Industries Machinery
SIC  3555 - Printing Trar.rs Machinery and  Equipment
SIC  3559 - Special Indcsrry Kaclur.cry, Not F'.sewhcre  Classified
SIC  3561 - Punps .ind Pur-ping Equipment
SIC  3562 - Ball and Roller Hearings
SIC  3563 - Air and Gas Coripressors
SIC  3564 - Elov.crs and Exhaust nnd Vc.ntilat.icii  Funs
SIC  3565 - Industrial PaLic-ins

-------
                                                              HANDOUT 2
                                                              (Continued  6 Df 8)
C1C   3566 - Speed  Changers,  Industrial  High S;>C-L'(| Drives,  uiid Gears
SIC   3567 - Industrial  l'i OCL-.S  1'urn.jcc:-. ,nid 0-,-. "s
SIC   3568 - Mcch^niLil  POY.UI  Tron^uic&ion hquipi.icnt ,  Not Elsewhere
SIC   3569 - Ccncrnl  Industrial  ti.ichincry ;md llquipnicnt ,  Hot Elsuuhcrc
            Classified
SIC   3572 - Typewriters
SIC   3573 - Electronic  Computing  Equipment
SIC   3574 - Calculating and  Accounting  Machines,  Lxccpt  Electronic
            Computing Equipment
SIC   3576 - Scales and  Balances,  Except Laboiatory
SIC   3579 - Office Machines,  Not  Elsewhere Classified
SIC-  3581 - Automatic Merchandising  Machines
SIC   3582 - Coriroercial  Laundry, Dry  Cleaning, and Pressing Machines
SIC   3585 - Air Conditioning and  l.'arir Air ilc..';ing Equipment
            and Commercial «jnd  Industrial Refrigeration  Equipncnt
SIC"  3586 - Censuring and Dispensing Pur.ps
SIC   3589 - Service  Industry  .''achines ,  Mot Elsewhere  Classified
SIC   3592 - Carburetors,  Piston,  Piston Rings,  and Valves
SIC   3599 •* Machinery,  E;-cept Electrical, Not Elsewhere  Clcs.siiie.ci
SIC   3612 - Pou-cr, Distribution,  and Specialty  Transformers
SIC   3613 - Switchgear  and Switchboard  Apparatus
SIC   3621 - Motors and  Generators
SIC   3622 - Industrial  Controls
SIC   3623 - Welding  Apparatus,  Elecrric
SIC   3624 - Carbon and  Grapnite Products
SIC   3629 - Electrical  Industrial Apparatus, Not  Elsewhere Classified
SIC   3631 - Householc Cooking Equipment
SIC   3632 - Household Refrigerators  and Home and  Farrn Fieczers
SIC   3633 - Household Laundry Equip-.cnt
SIC  .3634 - Electric Housewares and  Fans
SIC   3635 - Household Vacuum Cleaners
-SIC   3639 - Household' Appliances, Not Elsevhers Classified
SIC   3641 - Electric Lar.ps
SIC   3643 - Current-Carrying Wiring  Devices
SIC   3644 - Noncurrent-Carrying '.firing  Devices  .
SIC   3645 - Residential Electric  Lighting Fixtures
SIC   3646 - Corjnercial, Industrial,  and Institutional Electric
            Lighting Fixtures
SIC   3647 - Vehicular Lighting  Equipment
SIC   3648 - Lighting Equip-ier.L, Noc  Elsew.iere Classified
SIC   3651 - Radio  and Television  Receiving Sets,  Except  Corrrunicatior.
            Types
SIC   3652 - Phonograph  Records  and Pre-recorded Magnetic Tape
SIC   3661 - Telephone and Telegraph  Apparatus
SIC   3662 - Radio  and Television  Transmit^ir.^,  Signaling,  and Detection
            Equipment and Apparatus
SIC   3671 - Radio  and Television  Receiving Type Electron Tubes, Except
            Cathode  Ray
SIC   3672 - Cathode  Kay Television Picture Tubes
SIC   3673 - Transir.it ting, Industrial, ar.d Special Purpose  Electron Tubes
SIC   3674 - Semiconductors and  Related  Devices
SIC   3675 - Electronic  Capjcitors
SIC   3676 - Resistors,  for Electronic Applications
SIC   3677 - Electronic  Coils, Trnnsf orr.icrs ar.3  Other  Ii.d-jctors
SIC   3678 - Connectors, for  Electronic  Appl icc:io:is

-------
                                                             HANDOUT 2
                                                             (Continued 7 of 8)
SIC  3C79 - Electronic Conponcnts, Not Elsewhere Clabbificd1
SJC  3691 - Stor.ij;c K.it icr j os
SIC  3692 - Primary JJ.TCLcricc, Dr> and Vet
SIC  3693 - Rndiographic X-rny, i'luoroscopic !'.-ray,  Therapeutic  X-ray,
            and Other X-ray Apparatus and Tubes.; Electroi.iudical  and
            Elcctrothorapcutic Apparatus
SIC  3694 - electrical Lquipn.ont for Internal Conbustion  Engines
SIC  3699 - Electrical Machinery, Equipment, and Supplies,  Not Elsewhere
            Classified
SIC  3711 - Motor Vehicles and Passenger Car Bodies
SIC  3713 - Truck and Bus  bodies
SIC  3714 - Motor Vehicle  Parts and Accessories
SIC  3715 - Iruck Trailers
SIC  3721 - Aircraft
SIC  3724 - Aircraft Lnr;incs and Engine Parts
SIC  3728 - Aircraft Pares and Auxiliary Eq-Jipnenc ,  Not Elsewhere'Classified
SIC  3731 - Ship Building  ana Repairing
SIC  3732 - Boat Building  and Repairing
SIC  3743 - Railroad Lquioineiit
SIC  3751 - Motorcycles, Bicycles, and Parts
SIC  3761 - Guided Missiles ana Space Vehicles
SIC  3764 - Guided Missile and Space Vehicle Propulsion Units and
            Propulsion Unit Parts
SIC  3769 - Guided Missile and Space Vehicle Parts and Auxiliary Equipment,
            Not Elsewhere  Classified
SIC  3792 - Travel Trailers and Campers
SIC  3795 - Tanks and Tank Components
SIC  3799 - Transportation Equipment, ;;ct Elsc'.j'nera  Classified
SIC  3811 - Engineering, Laboratory, Scientific, and Rcsearcli.Instrur.tnts
            and Associated Equipment
SIC  3822 - Automatic Controls for Regulating Residential and Corj-ercial
            Environner.ns and Appliances
SIC  3823 - Ir.dustnal Irstrurents for >:a53ur;r,er.t ,  Display and  Control
            of Process Variables; and Related Procucts
SIC  3824 - Totalizing Fluid Meters and Counting Devices.
SIC  3825 - Instrunents, for measuring ar-.c Testing of Electricity and
            Electrical Signals
SIC  3829 - Measuring and  Controlling Devices, Not Elsewhere Classified
SIC  3S32 - Optical Ir.strur.cnts ar.c Lenses
SIC  3841 - Surgical and Medical Instrur.-er.ts sr.d Apparatus
SIC  3842 - Orthopedic, Prosthetic, and 5-jrgiral Appliances and  Supplies
SIC  3843 - Dental Equipment and Supplies
SIC  3351 - Ophthalmic Goods
SIC  3873 - Watches, Clocks, Clockwork Operated Devices and Parts
SIC  3911 - Jewelry, Precious Metal
SIC  3914 - Silverware, Plated l.are, ar.c Stiir.less Steel '.'arc
SIC  3915 - Jewelers' Fin-dings and Materials, snd Lapid.nry VorL
SIC  3931 - Musical Instruments
SIC  3942 - Dolls
SIC  3944 - Gar.es, Toys, and Children's Vehicles; Except Dolls and
            Bicycles
SIC  3949 - Sporting and Athletic Goods, Not Ilscwnere Classified
SIC  3951 - Pens, Mechanical Pencils., and P.i-ts (Balance)
SIC  3961 - Costunc Jewelry and Costu.no Kovolties, E::ccpt
            Precious Metal

-------
                                                              HANDOUT 2
                                                              (Continued 8 of 8)
     SIC  399]  - Broons and. Brushes
     SIC  3993  - Sifcns and Advertising Displays
     SIC  3995  - Burial Caskets

19.   ELECT P.OrLATII.'O

     SIC   347  - Coating,  Engraving, and Allied Services


20.   ORE MINING AMD DRESSING

     SIC  1011  - Iron Ores'
     SIC  1021  - Copper Ores
     SIC  1031  - Lead and  Zinc Ores
     SIC  1041  - Cold Ores-
     SIC  1044  - Silver Ores
     SIC  1051  - Bauxite and Other Aluminum Ores
     SIC  1061  - Ferroalloy  Ores,  Except Vanadium
     SIC  1092  - Mercury Ores
     SIC'  1094  - Ursniun-Radiur.-Vanadiu:n Ores
  *   SIC  1099  - Metal Ores, Not Elsewhere Classified

21.   COAL MINING

     SIC  1111  - Anthracite
     SIC  1112  - Anthracite  Mining Services
     SIC  1211  - Eituninous  Coal and Lignite
     SIC  1213  - Bituminous  Ccal and Lignite Kininy Services

-------
                                                                                                                    HANDOUT   3
                                        THE  1*9  PRIORITY  TOXIC  POLLUTANTS
           Pollutant
                                           CharactanaM*
                                                                                                                 Remarks
 Generally
Readily  attimilaled  by
  MMMlft.  fM  Mluble.  Cancan
  WBIed  tniouin the food  eluln
  (ManufmflM). peranient In HU
                                                                     Direct tppllcaiKM  IB  farm.  »id
                                                                       •nd fardana,  urban runoff.  41*
                                                                       chorfe In  Industrial wntawaiar
                                          cMonnated  hydreearaon
                                    poueMea already natrtcted  BIT
                                    EPA: eldrm. dlaMnn. COT. 000.
                                    efldrin. htpucnlor,  Mono, md
POfycMonnoted fcprlenjrla (POM
Uoad In  electrical copaclton  and
  trorafomwra. palm*, platllca. to-
  lectteidet. ettiar indualrial prod-
                                   Readily  oialmiUale*  By  eovettc
                                     anifnol*« fot MluMd SUBJ^CI to
                                     btenufnRcitia*.
                                     cftmtieiUy timllir n MM
                                     kutatf hrdrocareont
                                  Mimclptl  *nd  Indintrlil
                                    4hchif(n dltpncd of In
                                    •nd landfill*
                                  TSCA ban  on piMucMn  attar
                                    6/1/79 but will wnitt In taal-
                                    mam: fatlnoiaiw  on  many
                                    fmhwatar dihaHaa aa a raauit
                                    ef K8  oalkitlon  («*.  taw
                                    HfjtfMn, uppvr NovMtOfii& pvftv
                                    ef Uka
  nckal. Mlcnium. arivar. thallium.
  and line
 nn»r inerfantea
                                   Mt  bMdMraaa*1*. paniatant  hi
                                     MdUnwn. nuc m teiuben. MB-
                                     lact n bwmawilneaMn
                                  Indumlrtal  dlachanjaa. mbilnf ac-
                                    bvity, urDtfi nuttff,  •vocton of
                                    maMMch Mil,  eartaln  utttul-
                                    ami  na*  (ax. manairy  aa  a
                                   May cam* cwicar whan Inhalad,
                                     Muatic Bateity not wall imdtr-
                                   Cyamda
                                   Variably panlatant. uinibiM eiygan
                                  AtbMHU
                                  Manufacnm and waa aa a rataid-
                                    ant roofinf matadvl. Biaka lln-
                                    bid ate*: mnoff fmn mlnlnaj
                                  CyanM*
                                  Wlda nnaiy of  InduMitel inn
 Natofanafad aMpnafka
 Utad M flf* aiMncuiihan. rafrifar-
  anft. BiopallaMjL aatMcMat. an-
  v«nta for oilv. and ffvaaaa and
  hi dry claanta(
Ur|a« ungla elata  af  "Brtsrtty
  tone*.*  can causa damaia  to
  cantral namoua ayuam and Hoar.
  not vary pmfelant
Producad by ehleiinaben of wator.
  \rioortiabon durinf uaa
                                                                                                       Lanja valuma induatrlal ckamksali.
                                                                                                        wUaly ilipanad. but laaa thraat
                                                                                                        IB Ilia aiNlionniaM tfun paniii-
 Clnan
 Uud mainly n tahmis far pair
  mar plntica
                                   PatBAC caNinodan* aouatic wncity
                                                                     beapa durtni pnMuctien and uaa
                                                                     Thousti torna aia  «ola«la.
                                                                      hava baan  Identified  In
                                                                      natural watin.
mmafala Mtan
Uaad ehiafiy m praductwn af pair-
  vnyl cnianda and tnarmoplaanca
  ai ptaaiiclzan
  rmjiacenw prooertin in tow can.
                                     braiaa ara pamcuiarly  Hnvtln
                                     to toate aflactK parvlalant: and
                                     can bo bwmacniflad
                                  Waate diaeotal mponzenen during
                                    vita (In nonpla*M»
 Uonocrd'C  erometica  (ucfuduif
 Uaad « ma mamfacnini af ottur

  p«cm«nta, and m
         and
                                   Cantral  nanoua  lyxam
                                     tant eon damaca Ihrar and Uo>
                                  CMar  anvironmant  during praduc.
                                    twn  and  byDroduct pioducnoii
                                    (tain  by  dlraet  vaUaiaMon.
Ursa  volunia  induatrlal
  pound* uaad driafiy aa
                                   Tulclty incrtaaat with dafraa of
                                     cluaruiaiion ef tha phanollc met.
                                     eblactlanabla  Oder  and taat*
                                     to drinaint watar dlfneult to n>
                                     ma** horn Mler by eanvanHaoal
                                  Occur  naturally In  fowl
                                    mm
                                    ou
                                      m
                                        pintle  manufactunnc
                                    all
Palrcyaiic aramafie n>dro«»tbona
Uaad a* dyaaniff*. chamiail Intaf-
                                                                       duettont. Inaemplal* CBmbuauen
                                     oowotM  MaMHy of Bieae  a
                                     pound*: not pentatent and
U»d in the pnMuctNjn af ortanle
  ehamlcala  and rvbbw; patant*
  •ml en pracntn ywif ttina
                                     anowH IH* nimaaminn IB
                                                                       apontanaauily In feed
Reference:    Council  of  Environmental  Quality,  Environmental  Quality  1978,
                   USGPO  #041-11-0040-8,  December,  1978

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                                              Recommended List of Priority
                                                        Pollutants
Compound Name
 1.  *acenaphthene
 2.  *acrolein
 3.  *acrylonitrile
 4.  *benzene
 5.  *benzidene
 6.  *carbon tetrachloride (tetrachloromethane)
     •Chlorinated benzenes (other than
          dichlorobenzenes)
 7.       chlorobenezene
 8.       1,2,4-trichlorobenzene
 9.       hexachlorobenzene
     *Chlorinated ethanes(including 1,2-
          dichloroethane, 1,1,1-trichloro-
          ethane and hexachloroethane)
10.       1,2-dichloroethane
11.       1,1,1-trichloroethane
12.       hexachloroethane
13.       1,1-dichloroethane
14.       1,1,2-trichloroethane
15.       1,1,2,2-tetrachloroethane
16.       chloroethane
     *Chloroalkyl ethers  (chloromethyi,
          chloroethyl and mixed ethers)
17.       bis  (chloromethyi) ether
18.       bis  (2-chloroethly) ether
19.       2-chloroethyl vinyl ether (mixed)
     •Chlorinated naphtalene
20.            2-chloronaphthalene
     •Chlorinated phenols (other than those
          listed elsewhere;  includes trichloro-
          phenols and chlorinated cresols)
21.       2,4,6- tr: chloroohenol
22.       oarachlorometa cresol
23.  *chloroform (trichloromethane)
24.  *2-chlorophenol
     *Dichlorobenzenes
25.       1,2-dichlorobenzene
26.       1,3-dichlorobenzene
27.       1,4-dichlorobenzene
     *Dichlorobenzidine
28.       3,3'-dichlorobenzidine
     *Dichloroethylenes (1,1-dichloroethylene
          and 1,2-dichloroethylene)
29.       1,1-dichloroethylene
30.       1,2-trans-dichloroethylene
31.  *2,4-dichlorophenol
     *Dichloropropane and dichloropropene
32.       1,2-dichloropropane
33.       1,2-dichloropropylene  (1,3-dichloropropene)
34.  *2,4-dimethylphenol
       *Dinitrotoluene
35.       2,4-dinitrotoluene
36.       2,6,-dinitrotoluene
37.  *l,2-diphenylhydrazine
38.  *ethylbenzene
39   *fluoranthene
     *Haloethers (other than those listed
          elsewhere)
40.       4-chlorophenyl phenyl ether
41.       4-bromophenyl phenyl ether
42.       bis(2-chloroisopropyl) ether
43.       bis(2-chloroethoxy) methane
    *Specific compounds and chemical classes as listed
    in the consent degree.

-------
     *Halomethanes (other than those listed elsewhere
44.       methylene chloride (dichloromethane)
45.       methyl chloride (chloromethane)
46.       methyl bromide (bromomethane)
47.       bromoforni (tribromomethane)
48.       dichlorobromomethane
49.       trichlorofluoromethane
50.       dichlorodifluoromethane
51.       chlorodibromomethane
52.  *hexachlorobutadiene
53.  *hexachlorocyclopentadiene
54.  *isophorone
55.  *naphthalene
56.  'nitrobenzene
     *Nitrophenols (including 2,4-dinitrophenol
          and dinitrocresol)
57.       2-nitrophenol
58.       4-nitrophenol
59.      *2,4-dinitrophenol
60.       4,6-dinitro-o-cresol
     *Nitrosamines
61.       N-nitrosodimetlv/lamine
62        N-nitrosodiph6i\ylamine
63.       N-nitrosodi-n-propylamine
fi4.  *pentachlorophenol
65.  *phenol
     *Phthalate esters
66.       bis(2-ethylhexyl)  phthalate
67.       butyl benzyl phthalate
68.       di-n-butyl phtalate
69.       di-n-octyl phtalate
70.       diethyl phtalate
71.       dimethyl  phthalate
     *Polynuclear aromatic hydracarbons
72.       benzo  (a)anthracene (1,2-benzanthracene)
73.       benzo  (a) pyrene  (3,4-benzopyrene)
74.       3,4-benzofluoranthene .
75.       benzo(k)fluoranthane (11,12-benzofluoranthene)
76.       chrysene
77.       acenaphthylene
78.       anthracene
79.       benzo(ghi)perylene (1,12-benzoperylene )
80.       fluorene
81.       phenanthrene
82.       dibenzo  (a,h)anthracene  (1,2,5,6-dibenzanthracene)
83.       indeno  (1, 2, 3-cd) pyrene  (2,3,-o-phenylenepyrene)
84.       pyrene
85.  *tetrachloroethylene
86.  *toluene
87.  *trichloroethylene
88.  *vinyl chloride  (chloroethylene)
Pesticides and Metabolites
89.       *aldrin
90.       *dieldrin
91.       *chlordane (technical mixture s metabolites)
*DDT and metabolites
92.       4,4'-DDT
93.       4,4'-DDE(p,p'DDX)
94.       4,4'-DDD(p,p'TDE)
*endosulfan and metabolites
95.       a-endosulfan-Alpha
96.       b-endosulfan-Beta
97.       endosulfan sulfate
*endrin and metabolites
98.       endrin
99.       endrin  aldehyde
*heotachlor and' metabolites

100.      heptachlor
101.      heptachlor epoxide
*hexachlorocyclohexane  (all isomers)
102.      a-BHC-Alpha
103.      b-BHC-Beta
104.      r-BHC  (lindane)-Gamma
105.      g-BHC-Delta
*polychlorinated  biphenyls  (PCB's)
106.
107.
108.
109.
110.
111.
112.
113.
114.
115.
PCB-1242 (Arochlor
PCB-1254 (Arochlor
PCB-1221 {Arochlor
PCB-1232 (Arochlor
PCB-1248 (Arochlor
PCB-1260 (Arochlor
PCB-1016 (Arochlor
*Toxaphene
*Antimony (Total)
*Arsenic (Total)
1242)
1254)
1221)
1232)
1248)
1260)
1016)
(-> 3>
O -3Z
P0>
o

-------
116.      'Asbestos (Fibrous)
117.      *Beryllium (Total)
118.      *Cadmium (Total)
119.      *Chromium (Total)
120.      'Copper (Total)
121.      'Cyanide (Total)
122.      *Lead (Total)
123.      *Mercury (Total)
124.      'Nickel.(Total)
125.      'Selenium (Total)
126.      'Silver (Total)
127.      'Thallium (Total)
128.      'Zinc (Total)
129.      "2,3,7,8-tetrachlorodibenzo-p-dioxin  (TCDD)

" This compound was specifically listed in  the  consent
degree.  Because of the extreme toxicity  (TCDD).   He  are recommending
that laboratories not acquire analytical standard  for the compound.
                                                                                                              o -z.
                                                                                                              = o
                                                                                                              i-f O
                                                                                                              n
                                                                                                              CL
                                                                                                              OJ
                                                                                                              o

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                                                         HANDOUT 5
             SECTION 308(aJ  OF  THE  CLEAN  WATER  ACT
Section 308(a)  of the Act  states  in  part:

     (A)  the Administrator  shall  require  the  owner  or operator
          of any point source  to  (i)  establish and maintain
          such  records,  (ii) make  such  reports,  (iii) install,
          use and maintain such monitoring equipment or methods
          (including  where appropriate,  biological monitoring
          methods),  (iv) sample such  effluents (in accordance
          with  such methods, at such  locations,  at such intervals,
          and in such manner as the Administrator shall prescribe),
          and (v) provide  such other  information as  he may
          reasonably  require;  and

     IB)  the Administrator or his authorized  representative
          upon  presentation of his credentials--
          (i) shall have a right of entry to,  upon or through any
          premises in which an effluent  source is located or in
          which any records required  to  be maintained under
          clause (A)  of this subsection  are  located, and
          (ii)  may at reasonable times have  access to and copy
          any records, inspect any monitoring  equipment or
          method required  under clause  (A),  and  sample any
          effluents which  the  owner or operator of such source
          is  required to sample under such clause.

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                                                                       HANDOUT  6
            TABLE 1.   RECOMMENDED  SPECIES  AND  TEST  TEMPERATURES
Species                                          Test Temperature (°C)
Freshwater
   Vertebrates
      Coho salmon, Oncorhynchus kisutch                       12
      Rainbow-trout,  Salmo gairdneri                          12
      Brook trout, Salvelinus fontinalis                      12

      Goldfish, Carassius auratus                             22
     9 Fathead minnow, Pimephales promelas                     22
      Channel catfish, Ictalurus punctatus                    22
     ' Bluegill, Lepomis macrochirus                           22
   Invertebrates
    i Daphnids, Daphnia magna or D_.  pulex                     17
      Amphipods, Carcmarus lacustris, £.  fasciatus,  or         17
                 (». pseudolimnaeus                            17
      Crayfish, Orconectes sp., Cambarus sp. ,  Procambarus     22
                 sp.,  or Pacifastacus leniusculus             22
      Stoneflies, Pteronarcys sp.                             12
      Mayflies, Baetis sp. or Epheiierella sp.                  17
                Hexagenia limbata or H.  bilineata             22
      Midges, Chironomus sp.                                  -22
Marine and estuarine
   Vertebrates
     « Sheepshead minnow, Cyprinodon variegatus                22
      Mummichog, Fundulus heteroclitus                        22
      Longnose killifish, Fundulus similis                    22
      Silverside, Henidia sp.                                  22
      Threespine stickleback,  Casterosteus aculeatus          22
      Pinfish, Lagodon rhonboides                             22
      Spot, Leiostomus xanthurus                              42
      Shiner perch, Cymatogaster aggregata                    12
      Pacific staghorn sculpin, Leptocottus armatus           12
      Sanddab, Citharichthys stigmaeus                        12
      Flounder, Paralichthys dentatus, P_. lethostigma         22
      English sole, Parophrys vetulus                         12

-------
                                                              HANDOUT 6
                                                              (2  of 2}
TABLE 1.  (Cont'd)
Species                                         Test Temperature (DC)a
Marine and estuarine
   Invertebrates
      Shrimp, Penaeus setiferus,  J?.  duorarum,  or             22
         IP. aztecus
      Grass shrimp, Palaemonetes  sp.                         22
      Shrimp, Crangon sp.                                     22
      Oceanic shrimp, Pandalus jordani                       1<2
      Blue crab, Callinectes sapidus                         22
      Dungeness crab, Cancer magister                        12
     /Mysid shrimp, Mysidopsis sp.,  Neomysis sp.             22
      Atlantic oyster, Crassostrea virginica                 22
      Pacific oyster, Crassostrea gigas                      20


 Freshwater amphipods, daphnids,  and midge larvae and shrimp should be
cultured and tested at the recommended test temperature.   Other in-
vertebrates should be held and tested within 5°C of the temperature of
the water from, which they were obtained.  If the recommended test tempera-
ture is not within this range, they should be tested at the temperature
from the series 7, 12, 17, 22, and 27°C that is closest to the recommended
test temperature and is within the allowed rang:.

-------
                                   Mobile Bioassay .Equipment
                                           Checklist
                                                                       HANDOUT  7
     Permittee^
     Permit No.
General Equipment

	 company file
	 data file
	maps
_____ field data sheets
	 chemical analysis request
	pens
	 pencils
_____ marker pens
	 paper
_____ clipboard
	 Kim-wipes
_____ plastic garbage bags
	 electrical tape
	tape
	 flashlights
	 camera
	 film
                                 Chemical Sampling Equipment
                                                                    Hardness
                                      cube containers - 1 qt.
                                      cube containers - 1 gal.
                                      cube containers - 2 1/2  gal.
                                      TOC test tubes
                                      volatile organic jars
                                      nitric acid (metals)
                                      sulfuric acid (nutrients,TOC)
                                      Kemmerer sampler
                                      buckets
                                 Chemical Analyses Equipment
                                                                     buffer solution
                                                                     Enochrome black T
                                                                    ' EDTA 0.01M
                                                                Residual Chlorine
                                                                     phenylarsene oxide
                                                                     (titrant)
                                                                     pH 4 buffer
                                                                    \ PH 7 buffer
                                                                     potassium iodide
                                                                     electrolyte tablets
 rope
 rain gear
 waders
_ half boats
 250 ft. extension cords (3)
 short extension cords
 tools
 solder and soldering gun
 silicone glue
 battery charger
 jumping cables
 oil and funnel
' gas cans (1 N.J.) 2. N.Y.
                                      1000 ml beakers (7)
                                      41. graduate (1)
                                      11. graduate (2)
                                      500 ml graduate (2)
                                     ] 250 ml graduate (1)
                                      100 ml graduate (2)
                                     \ 50 ml graduate (2)
                                      funnels
                                      plastic cups (4.5  oz)
                                      stirring bars
                                      distilled  ater
                                      composite jampler
                                      3/8" vinyl nalgene  tubing
                                      ice chest
Bioassay Equipment
     static bioassay jars
     funnels with screening
     3/16" silicone tubing
    _ 5/8" silicone tubing
     silent giants
    _ oxygen tank
     air stones
    ' 1/8" Latex tubing
     specimen jars
     40% isopropyl
    ' submersible pumps (3)
     dilution water pump
     black hose line
     output lines
    \ fish food
    \ artemia eggs
     glycerin
     stoppers
                                 Dissolved Oxygen
                                 _ manganous sulfate
                                  alka-azide
                                  cone,  sulfuric acid
                                  sodium thiosulfate
                                 _ starch
                                  buodate
                                 ' 500 ml wide mouth
                                  erlenmeyer flask
                                  automatic pipetts (3)
                             Alkalinity and pH
                                  0.02 N sulfuric acid
                                  pH meters
                                 . pH recorder
                                 ' pH 4 buffer
                                  pH 7 buffer
                                 ' pH 10 buffer

-------
                                                  Handout 8

                     Preinspection Questions
     The following list of questions can be used to establish
points of contact, request utilities, and to determine special
equipment requirements for on-site flow-through bioassays.
Because of the logistics involved, on-site bioassays must be
announced by 308 letter.  These questions can be posed to the
permittee in the 308 letter.  The permittee would be requested
to answer the following questions:

     1.  Who is the point of contact in the facility (name and
         telephone number)?

     2.  Is electrical power available near the NPDES outfall
         to operate the trailer (State operating requirements
         for amperage, voltage, and number of circuits)?

     3.  Is there space adjacent to the outfall to park a
         trailer (give dimensions)?

     4.  What safety equipment are required to enter the
         facility?  Is this supplied?  (Indicate the number
         of people on the inspection team.)

     5.  What is the source of process water ?

-------
                                                                HANDOUT  9
                                                                (1  of 2)
                       EFFLUENT SAMPLING PROCEDURES
Flowthrough Test

     If the industrial or municipal  facility discharges continuously,
     the effluent should be pumped directly and continuously from ^he
     discharge line to the dilutor system for the duration of the test.
     The use of effluent grab samples should be avoided.   However, if
     the effluent cannot be pumped directly and continuously to the
     dilutor system, the following alternative methods may be employed
     for collecting the effluent:

          (1)  When the measured minimum retention time of the effluent
               is less than 96 hours, a 6-hour composite  sample consis-
               ting of equal volumes taken every 30 minutes must be
               collected and transported to the diTutor every 6 hours
               for the duration of the test.

          (2)  When the measured minimum retention time of the effluent
               is between 4 days (96 hours) and 14 days,  a 24-hour
               composite sample consisting of equal  volumes taken every
               hour may be collected daily for the duration of the test.

          (3)  When the measured minimum retention time of the effluent
               is greater than 14 days, a single grab sample may be
               collected daily for the duration of the test.

     If the industrial or municipal  facility discharges intermittently,
     i.e., where the waste is discharged over a single 8-hour workshift
     or is accumulated and discharged at the end of the shift or the
     week, a composite sample consisting of equal  volumes collected
     every 30 minutes, may be taken  for an 8-hour operating shift or
     for the duration of the plant operating schedule, or a single grab
     sample may be taken in the case of a batch discharge.
Static Test

     If a flowthrough test cannot be used,  a  static  test may be conducted
     with effluent collected by one of the  following methods:

          (1)  When the measured minimum retention time of the  effluent
               is less than 96 hours,  four  consecutive 6-hour composite
               samples, each consisting of  equal  volumes taken  every
               30 minutes, are collected and  used in setting up four
               separate static tests.

-------
                                                     HANDOUT  9
                                                     (Continued  2  of  2)
(2)  When the measured minimum  retention  time  of  the  effluent
     is between 4 days (96 hours)  and  14  days,  a  24-hour
     composite sample consisting of  equal  volumes taken every
     hour is  collected daily  and used  in  the test.

(3)  When the measured minimum  retention  time  of  the  effluent
     is greater than  14 days, a single grab sample may be
     collected and used in the  test.

-------
                                                HANDOUT 10
            -DEFINITIVE TEST REQUIREMENTS
•  Ten organisms of a given species must be present
   in each test chamber,  for a total of twenty per
   concentration.

•  If more than one species are to be used in the test
   chambers, segregation  is necessary.

•  There should be randomization of organisms in test
   chambers.

•  Loading must not exceed 5 grams per liter at
   temperatures of 20°C or less, or 2.5 grams/liter
   at temperatures above  20°C for flowthrough tests.

•  Loading for static tests must not exceed 0.8 grams
   per liter at temperatures of 20°C or less, and 0.4
   grams/liter at temperatures above 20°C.

•  Temperatures must be held to within t2.0°C of the
   acclimation temperature for both static and flow-
   through tests.

•  Dissolved oxygen concentration should not be
   permitted to fall below 40 percent saturation for
   warm water species or  60 percent for cold water species.

t  The test begins when the organisms are first exposed
   to the effluent.

•  Organisms are not fed  during testing, unless newly
   hatched or immature.

•  The duration of the definitive test is 48 or 96
   hours.

-------
                                         DATA  SHEET FOR EFFLUENT  TOXICITY TEST
Industry/Toxicant.
Address _____
Contact	
Effluent Serial Number.
NPDES Permit Number
Beginning:  Date.
Ending:    Date.
Test Organism	
.Time.
.Time.
Test Temperature Range.
Cone.
or
K
















Test
Container
Number
















Number ol
Live Organisms
0
















24
















48
















72
















96
















Dissolved Oxygen
|mg/l)
0
















24
















48
















72
















96
















pH
0
















24
















48
















72
















96
















Total Alkalinity
(mg/l asCaC03)
0
















24
















48
















72
















96
















Total Hardness
(mg/l as CuCOs)
0
















24
















48
















72
















96

















0
















24
















48
















72
















96

















-------
                                                              HANDOUT 12
                                                              (1 Of 7)
                      LC5Q DETERMINATION METHODS



A.  LITCHFIELD AND WILCOXON ABBREVIATED METHOD OF DETERMINING THE LC50

General Procedure

Step 1;  Tabulate the data (see sa3ipl=j dzt: shaat, Fig.  1,  p. 33) showing
t'.ie percent-effluent volumes used,  tha total number of organisms exposed to
each percent-effluent volume, the number of affected organisms, and the
observed percent-affected organisms (see Example 1 below).   Do not list
more than 2 consecutive 100 percent affects at the higher  percent-effluent
volumes or more than two consecutive 0 percent affects at  the lower percent-
effluent volumes.

Step 2:  Plot the parcen;-affected organise against the percent-effluent
volume on 2 cycle, logarithmic probability paper (Fig. 2),  except for
0 percent or 100 percent affect values.  With a straight edge, fit a
temporary line through the points,  particularly those in tha region of
40 percent to 60 percent affects.

Step 3:  Using the line drawn through the points, read and  list an
"expected" percent affect for each percent-effluent volume  tested.  Disre-
gard the "expected" percent value for any of the percent volumes less tr^.n
0.01 or greater than 99.99.  Using the expacted-percent-affact, calculate
froT Table 7 a "corrected" value for each 0 percent or 100  percent affect
obtained in the test.  (Since the expected values in the table are whole
numbers, it will be necessary to obtain intermediate values by interpolation.)
Plot these values on the logarithnic probability paper (F-g. 2) used in
Step 2 and inspect the fit of the line to the co^.pletsly plotted data.  If
after plotting the corrected expected values for 0 percent  and 100 percent
affected, the fit is obviously unsatisfactory, redraw the  line and obtain
a new set of expected values.

Step 4:  List the difference between each observed (or corrected) value and
the corresponding expected value.  Using each difference and the corres-
ponding expected value, read and list the contributions to  Chi-square (Chi2)
from Fig. 3 ( a straight edge connecting a value on tha Exgected-Percent
Affected scale with a valua on the Observed-Minus-Expscced  scale, will
indicate at the point of intersection of tha Chi^ scale,the contribution Co
Chi-. SUTI the contributions to Chi^ and multiply the total  by tha average
number of organisms per effluent volume, i.e., the number  of organisms used
in K concentrations divided by K, where K is the nu-aber of  percent-affected
organism values plotted. The product is the "calculated" Chi2 of the line.
The degrees of freedom (N) are 2 less than the number of points plotted,
i.e., N =K-2.  If the calculated Chi2 is less than the Chi2 given in Table 8
for N degrees of freedom, the data are non-heterogeneous and the line is
a good fit.  However, if the calculated Chi^ is greater than the Chi2 given
in Table 8 for N degrees of freedom, the data are heterogeneous and the line
is not a good fit... In the event a line cannot be fitted (the calculated
Chi2 is greater than the tabular Chi2), the data can not be used to calcu-
late a LC50 or EC50.  Litchfield and Wilcoxon provided an  alternate
irec'r.od for calculating the 95 percent confidence limits under these cir-
cupstances.  However, the toxicity test should be repeated.

-------
                                                                  HANDOUT  12
                                                                  (Continued 2 of 7)
Step 5:  Determine the confidence limits of the LC50.

a.  Read from the fitted line (Fig. 2), the percent effluent volumes for
    the corresponding 16, 50, 84 percent affects (LC16,  LC50 and LC84).
b.  Calculate the slope function, S, as:

                  S = LC84/LC50 + LC50/LC16
    From the tabulation of the data determine Nf, which is defined as tha
    total n-idiber of test organisms used within the percsnt-af fected-
    organism interval of 16 percent and 84 percent. Calculate the exponent
    (2.77//N1) for the slope function and the factor, fLC50» used to
    establish tha confidence limits for the LC50 (or EC50) .
                               .(2.77//N1)
                       LC50
        ^LC50 can be obtained directly from t'r.a r.csogram in Fig. 4 by laying
    a straight-edge across the appropriate base and exponent values and
    reading the resultant "f" value.

e.  Calculate the confidence limits of the LC50 as follows:

    (1) Upper Unit for 95% probability = LC50 X
    (2) Lower linit for 95Z probability = LC50/f
                                                LC50
Example
   Steps 1-4:  The data were tabulated and plotted (Fig.  2) and the
expected values were read froa
STZ? ONE
7,
Effluent
Volume
3.2
5.6
10.0
13.0
32.0
56.0
100.0

Number of
Organisms
20
20
20
20
20
20
20
Number of
Affected
Organisms
0
1
11
7
12
18
20
Observed %
Affected
Organisms
0(.2)b
5
55
35
60
90
100 (99.0)
ch* graph.
STZ? THREE

Expected 7,
(Fig. 2)
.6
3.5
(14.5)*
38.0
67.0
87.5
97.0


STIP FOOH
Observed
Klaus
Expected
0.4
1.5
Aberrant
3.0
7.0.
2.5
2.0
Total

Chi2
0.005
0.006
Value
0.004
0.024
0.006
0.014
0.059
a. Percent-affected organisms at the 10 percent effluent volume is obviously
   an aberrant value and should be omitted when fitting the line in Step 2.

b. Step 3 "Corrected" affected values from Table 7.

-------
                                                                HANDUUT 12
                                                                (Continued 3 of 7)
Step 4 (Cont.);

Calculation of Chi

a.  Total number of organisms used in 'K1 concentrations - 120  • 20
                                                            6

b.  Calculated Chi2 - 20 x 0.059 = 1.18

c.  Degrees of Freedom (N)-K-2-6-2-4
                         2
d.  From Table 8, the Chi  for 4 degrees of freedom = 9.49
                      2                             2
e.  The calculated Chi  is less than the tabular Chi .   Therefore, it is
    assuaed the line is a good fit, and the data are non-haterogeneous.

Step 5;

a.  From the fitted line in Fig. 2, determine the (percent) effluent
    concentrations corresponding to the 16%, 50% and 84% affected
    organism values:

b.  LC84 effluent concentrations  • 50.0%
    LC50     "          "         =• 23. OZ
    LC16     "          "           10.OZ

c.  Calculate the slope function, 'S1, as:

     S = LC84/LC50 + LC50/LC16    50.0/23.0 + 23.0/10.5
                   2           =            2

       = 2.17 + 2.19     4.36     2.18
         	2	  '  T  =

d.  N1 = 40  (From Figure 2)

e.  Calculate the exponent (ft1) and factor, firrn
             2.77//N'      2.77//40       2.77/6.32     ,R0.433>   ,  ,,  .
    I  __ = 5        «= z.lo         = Z.lo          « Z.j.8     • = 1.41  •
f.  Calculate the confidence liaits of the LC50
    (1)  Upper limit for 95Z probability - LC50 X fLC5Q - 23.0 X 1.4 = 32.2%

    (2)  Lower linit for 95Z probability  = LCSO/f     - 23.0/1.4 - 16.4%

-------
                                                   HANDOUT 12
                                                   (Continued 4 of 7)
 EXPECTED
    %
 AFFECTED
  SOf-30
  70 —W
                                                       Chi
  80
    — 20
    — 10
  S3-—2
  99
    — 1.0
 99.34—5
 99.6
 39.7
 39.8
 99.9
99.35

99.9*-

9S9T-


99.98-
     -3
    -.2
• Oi

.04

.03


.02
                      OBSERVED MINUS
                      EXPECTED

                          - 50

                          -40

                          - 30


                          -20
                               -10
                               - 2
  .5
  .4

- -.3


- -.2
 -.03
                            — 2.0




                            — 1.0



                            — .30
                            — .40

                            — .30


                            — .20



                            — .10
                                                       .03
                                                       .04

                                                      — 03


                                                      - .02
                                                          — .01
                                                        .003

                                                        .004

                                                      — .003


                                                      — .002
                            — .001
       FIG. 3.  NOMOGRAPH FOR OBTAINING Chi2  FROM
              EXPECTED  % AFFECTgD AND
              OBSERVED-MINUS-EXPBCTED (STEP 4).

-------
                                                 HANDOUT 12
                                                 (Continued 5 of 7)
 BASES
EXPONENT
lo-




ss-

40 —


3,0-
                           LC50
2.0—
1.50 —



1.40 —
_
_
-
—
-

-

-
-






- 100
- 50
— 10.0
— 5.0
— 3.0
— 2.0
— 13
- 1.4
- 1.3
— 1.2
-1 10
- 1 05
— 1.04
- 1.03
- 102
— 4
_ i


— 2
— 1

— 1
-
-__
—
-
;
.
"•

                                                     — 2.0
                                                     -1.5
         1.0

         .90

         .BO


         .70


        — .60



        — .50




        — .40





        — .30
                                                       .20
     FIG. 4.  NOMOGRAPH FOR RAISING BASE S TO
             A FRACTIONAL EXPONENT

-------
                                    HANDOUT 12
                                    (Continued 6 of 7)





5





SJ i

_J
o
>
}-
2:
LJ
-J I
_J
4

li i ,
^^
0



3


IUU-


CJL
- JO -



- 32 -



13

10



C i




.2-





i '
1 ' 1 : '
-T-- -i ;--j- -T
1
i
ft j '
|
1 x. !
i • ^Sji

LC50-25.4 : /^
: ; i
' 1 1
1
1
1
1 !_ i
.. . _.j _. .-l_. (
1 ; ;
i ! . 1
1 i 1 i -fy

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1 j ! •

i
i i !

	
i i
T- - : T;
i
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: i
i '
r





\
\

1





.















1
1

1
1


!



• -

*x

























•














f



1
1
I
1
1 . • .
• 1
I
I
1
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i



i '
1
:

i -
-«
' •

i


!•-.:; ! ' L L r
-'::; -.'v
" - --; -"- "•
.^ . ,,
    0  10  20   30  40   50  60  70  80  90 100

             % SURVIVAL

FIG. 5.  PLOTTED DATA AND FITTED LINE FOR

       LOG-CONCENTRATION VERSUS

       % SURVIVAL METHOD

-------
                                                                 HANDOUT 12
                                                                 (Continued 7 of 7)
B.  LOG-CONCENTRATION VERSUS PERCENT-SURVIVAL METHOD OF DETERMINING
    THE LC50

General Procedure

Step 1:  Plot che percent effluent volumes and the corresponding
percent survival on semi-logarithmic paper (Fig.  5).

Step 2:  Locate the 2 highest points on the graph which are separated
by the 50 percent survival line and connect then with a diagonal straight
line.  However, if one of the points is an aberrant value,  the next
lowest or highest percent-effluent volune is used.

Step 3:  Read on the scale for percent-effluent volu=e, the value of
the point where the diagonal line and the 50 percent survival line
intersect.  This value is the LC50 percer.t-effluent volume  for the
test.  If by chance one of the effluent concentrations happens to have
50 percent survival, no graphins is necessary.

Example

Step 1:  The percent-effluent volumes and the corresponding percent
survival data front the Litchfield and Wilcoxon example are  plotted in
rig. 5.

Step 2;  The two highest points which are separated by the  50 percent
survival line (65 percent and 40 percent) are located ard connected with a diagonal
diagonal straight line.  The percent survival in the 10 percent-effluent
volume was considered an aberrant value and, therefore, was omitted from the
evaluation.

Step 3:  An LC50 of 25.4 percent-effluent volume for the test was
derived fron the point where the diagonal line and the 50 percent
survival line intersected in Fig.5.
SOURCE:  Methods for Measuring the Acute Toxicity of Effluents to Aquatic
         Organisms.  EPA  600/4 78-012, revised July, 1978.

-------
                                                                   HANDOUT 1 3
                             REPORTING TEST RESULTS


A report of the results of a test must include the following:

  o  The name of the test method, investigator and laboratory,  and the date
     the test was conducted.

  o  A detailed description of the effluent, including  its  source, date and
     time of collection, composition,  known physical  and  chemical  properties,
     and variability.

  •  The source of the dilution water, the date and time  of its collection,
     its chemical characteristics, and a  description  of any pretreatment.

  •  Detailed information about the test  organisms, including  scientific name,
     length and weight, age, life stage,  source,  history, observed diseases,
     treatments, and acclimation procedure used.

  0  A description of the test procedure and the test  chambers,  including the
     depth and volume of solution; the way the test was begun;  the number of
     organisms per treatment; and the  loading.   For the flowthrough system,
     the water volume changes per 24 hours in each test chamber must be
     calculated and  reported.

  0  The definition  of the adverse effect (death,  immobility,  etc.) used in
     the test, and a summary of general observations  on other  effects or
     symptoms.

  0  The number and  percentage of organisms in each test  chamber (including
     the control chambers) that died or showed the "effect" used to measure
     the toxicity of the effluent.

  0  A 24-, 48-, 72-,  and 96-hour LC50 or EC5n value  for  the test  organisms,
     depending on the duration of exposure.   If 100 percent effluent did not
     kill  or affect  more than 65 percent  of the test  organisms, report the
     percentage of the test organisms  killed or affected  by various
     concentrations  of the effluent.

  0  The 95-percent  confidence limits  for the IC™ and  EC™ values and the
     method used to  calculate them.

  0  The methods used  for the results  of  all  chemical analyses.

  0  The average and range of the acclimation temperature and  the  test temperature,

  0  Any deviation from this method.

  0  Any other relevant information.

-------
                                                                HANDOUT  14
                                                                (1 Of 2)
                   GENERAL BIOMONITORING COMPLIANCE SAMPLING
                          INSPECTION DAILY ACTIVITIES
A.  Day 1
    1.  Get power connected to mobile laboratory.
    2.  Level and stabilize laboratory.
    3.  Collect dilution water.
    4.  Begin acclimation.
    5.  Set up static rangefinding test
    6.  Hake necessary entries in logbooks  and fill  in  necessary  forms.

B.  Day 2
    1.  Check results of rangefinding test  and make  necessary  logbook
        entries.
    2.  Assemble  dilution board and delivery system.
    3.  Calibrate dilution board.
    4.  Activate  diluter and begin filling  test tanks.
    5.  Cease flow to acclimation tank.
    5.  If a composite sample is to be used for the  static  test,  the
        compositer should be set up this  day.
    7.  Collect dilution water.
    8.  Make all  necessary logbook entries.

C.  Day 3
    1.  Check all  systems to ascertain that all  have worked  overnight.
    2.  Check temperatures to see if the  acclimation temperature  and
        test temperature are approximately  the same.
    3.  Start the pump in the circulating water bath and  turn  the thermal
        equilizing unit on.

-------
                                                              HANDOUT 14
                                                              (Continued  2 of 2)
    4.   Collect the sample for the static  test,  whether  it  is  grab
        or composite.

    5.   Set up static  test tanks.

    6.   Perform temperature,  dissolved oxygen,  pH,  and conductivity
        readings in all  test  containers.

    7.   Introduce the  test organisms  to both  the static  and flowthrough
        test containers.

    8.   Collect additional dilution water.
D.  Days 4,  5,  and 6

    1.  Check all  systems to ascertain  that  all  have  worked  overnight.

    2.  Record test organism mortality  in  all  test  containers  and
        remove dead organisms where appropriate.

    3.  Perform length and weight measurements on dead  fish  (make
        necessary  logbook entries).

    4.  Calibrate  the appropriate meters and take meter readings.

    5.  Collect dilution water.

    6.  When scheduled, conduct  a compliance biomonitoring  evaluation
        inspection.

    7.  Make all  necessary logbook entries.


E.  Day 7

    1.  Check all  systems to ascertain  that  all  have  worked  overnight.

    2.  Record test organism mortality  in  all  test  containers  and
        remove dead organisms where appropriate.

    3.  Calibrate  the appropriate meters and take meter readings.

    4.  Recalibrate diluter board.

    5.  Make all  necessary entries in logbooks.

    6.  Dismantle  laboratory and secure equipment.

    7.  Inform permittee of your departure and sign out with gate
        security guard.

-------
                                                            HANDOUT 15
                                                            (1 of 9)
                 INSTRUCTIONS  FOR  COMPLETING  THE  ACUTE
                  TOXICITY LABORATORY  EVALUATION  FORM

1.        Laboratory or Industry  - Enter  the  complete name  of  the
laboratory or industry conducting  the  acute  toxicity test.
l.a.      Industry SIC Code -   Enter this number  and briefly describe
the type of industry,  raw materials used  and  estimated effluent
composition if available.
2.        Location - Enter the address of the laboratory or industry
conducting the acute toxicity  test.
2. a.      NPDES Permit No. - Enter the corresponding number and other
necessary permit identification such as date  of issuance and
expiration.
3.        Date • Enter date of evaluation.
4.        Investigator - Enter name and title of  person, conducting
evaluation.
5.        Company Representative  - Enter  name of person(s)  interviewed
and telephone'number (if available).
6.        Test Method - Enter  brief narrative of the test being
conducted and the reference where  written instructions on the
methodology apptars (i.e. 96-hour static  bioassay; or reference: EPA
660/3-75-009 April, 1975).
7.a.      Dilution Water - Source  - Enter the source of the dilution
water; the date and time of its collection.
7.b.      Diltuion Water: Chemical Analyses  Performed - Enter specific

-------
                                                              HANDOUT 15
                                                              (Continued 2 of 9)
chemical tests performed on dilution water if any.   Also enter
chemical characteristics recorded by the analyst (average and/or range
values).
7.c.      Dilution Water: Pretreataient - Enter a description of my
pretreatment of dilution water.
8.a.      Effluent Water: Source - Enter the source of the effluent to
be tested,  the date and time of its collection.
8.b.      Effluent Water; Variability - Enter a description of the
physical or chemical variability of the effluent (i.e. constant flow of
effluent from a lagoon with 14-days detention time  ^r batch process
releasing effluent having variable flow and chemistry directly into the
receiving water).
8.c.      Effluent Water; Sampling Technique - Enter a brief
description of the method used to collect the sample(s)  of effluent.
8.d.      Effluent Water: Holding time and Conditions -  Enter the
amount of time and conditions under which the test  effluent is held
before being used in the toxicity study.
8.e.      Effluent Water; Pretreatment - Enter a description of any
pretreatment of the effluent.
8.f.      Effluent Water: Chemical Analyses Performed -  Enter specific
chemical tests performed on effluent.  Also enter chemical
characteristics recorded by the analyst (average and/or range values).
9.a.      Test Organism: Species - Enter the conmon and  scientific
name of the test organism.
9.b.      Test Organism: Life State - Enter the age,  life stage, as
well as length and weight (if apporpriate) of the test organism.

-------
                                                              HANDOUT 15
                                                              (Continued 3 of 9)
 9.c.      Test Organism: Source - Enter the specific source of the test
 organism; the date and time of the collection (i.e. Brown Fish
 Hatchery, Central City, Iowa; collected 0800 hours on January 10,
 1978).
 9.d.      Test Organism: Holding Facilities - Enter a brief description
 of the  facility used to hold test organisms prior to the biomonitoring
 study (i.e. 500-gallon Minnow-Kool tank with flow-through dechlorinated
 tap water).
 9.e.      Test Organism: Acclimation Procedure - Enter a brief
 description of the procedure used to acclimate the test-organism  to
 laboratory conditions prior to biomoni toring tests.
 9.f.      Test Organism: Treatment - Enter any observed diseases  and
 specific treatment rendered if any.  State the number of treatments and
 dates.
 10.a.     Experimental Design: Equipment Cleaning Procedure - Enter a
 brief description of step-by-step pre-cleaning procedure' for equipment
 (tanks,  etc.) used in biomonitoring tests.  Li^t trade name and
 scientific name of cleaning compounds (if  available).
                                                 i
 lO.b.(l)  Experimental Design: Test Chambers:  Construction1 Material
 Enter the type of material  used in constructing  the test chanoers.
 10. b. (2)  Experimental 'Design: Test Chambers:  Dimensions -.Inter  the
 specific size of the test  chambers (length,  width,  height).
 10.b.(3)  Experimental Design: Test Chambers:  Volume -  .Enter1 the
 designated volume of the test chambers  as  well  as the specific depth
and volume of solution used during the  biomoni toring test.
 10. .     Experimental Design;  Test Chambers:  Volumetric Exchange.Rate
- Enter  the rate of exchange of test solution  in flow
through/continuous-flow test chambers.

-------
                                                              HANDOUT 15
                                                              (Continued 4 of 9)
10.c.     Experimental  Design;  Test  concentrations - Enter  a  list  of
solution concentrations in which  test  organisms were exposed.
10.d.     Experimental  Design:  Number  of organisms per concentration  -
Enter the number of test organisms exposed  to each concentration of
test solution.
10.e.     Experimental  Design:  Loading Rate - Enter  the weight of  test
organisms per liter of  test  solution (i.e.  5 grams/liter).
19.f.     Experimental  Design:  Test  Temperature - Average and Range -
Enter the temperature  (average  and range) of the solution in  which test
organisms are exposed  during the  biomonitoring study.
   .g      Experimental  Design:  Chemical Parameters Monitored  and
Frequency - Enter the  type of chemical tests performed and  the
frequency which each chemical  test is  performed during the
b iomoni to rin g stu dy.
lO.h.     Experimental  Design;  Duration and Frequency of Test - Enter
the time period of the  biomonitoring test and the number of times  tJhe
biomonitoring test is  performed each year.  (Record  both as
"performed" and "as required in NPDES  permit11).
10.i.     Experimental  Design:  Definition of adverse effect - Define
the endpoint of the biomonitoring test (i.e. death).
lO.j.     Experimental  Design:  Frequency of Observations -  Enter  the
time intervals  when test organisms were observed during the
biomonitoring study (i.e. observed each 12-hour period).
   .k.     Experimental  Design:  Method  of calculating EC50 or LC50  -
         •
Enter the name  of the  calculation procedure used and the reference
citation.

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                                                             HANDOUT 15
                                                             (Continued  5 of 9)
10.1.     Experimental Design; Special Conditions - Briefly describe
test conditions not addressed elsewhere in this questionnaire (i.e.
dead organisms not removed during the test; or test chambers aerated
continuously with pure 02 during test).  Attach a supplement sheet if
needed.
11.       Methods Used for All Chemical Analyses - Enrar the reference
cited for the chemical analyses performed during the biomonitoring
study.
12.       Other Relevant Information - Enter explanations of infor-
mation provided elsewhere in the Acute Toxicity Laboratory Evaluation
questionnaire or other pertinent information not presented in the audit
questionnaire (i.e.,  quality assurance program,  training and experience
of analyst,  adequacy of laboratory equipment and facilities,  etc.).

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                                                               HANDOUT  15
                                                               (Continued 6 of 9)
               ACUTE TOX1CITY LABORATORY EVALUATION FORM
     Laboratory or industry
     a.   Industry or SIC Code_
     Location
     a.   NPDES Permit No.
3.   Date
4.   Invest! gator_
5,   Company Representative^
6. .  Test Method
7.   Dilution Water
     a.   Source
     Date:              Time
     b.   Chemical Analyses Performed

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     c.   Pretreatment
8.   Effluent Water

     a.   Source
     b.   Variability
     c.   Sampling Technique_
     d.   Holding time and conditons_
     e.   Pretreatment
     f.   Chemical analyses performed_
9.   Test Organism

     a.   Species_
     t>.   Life stage
     c.   Source
     d.   Holding facilities.
                                                              HANDOUT  15
                                                              (Continued  ? of 9)

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                                                             HANDOUT 15
                                                             (Continued 8 of 9)
     e.   Acclimation Procedure
     f.   Treatment (schematic or-flow chart. If available)
10.  Experimental Design

     a.   Equipment Cleaning Procedure_
     b.   Test Chambers

          (1)   Construction material^

          (2)   Dimensions	

          (3)   Volume
          (4)  Volumetric exchange rate_

     c.   Test concentrations
     d.   Number of organisms per concentration
     e.    Loading rate_
     f.   Test temperature - average and range
     g.    Chemical  parameters monitored and frequency
     h.    Duration and frequency  of test

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     i.   Definition of adverse effect
     j.   Frequency of observations_
     k.   Method of calculating EC50 or LCSO_
     1.   Special  conditions_
11.  Methods used for all  chemical  analyses
12.  Record Keeping_
13.  Other relevant information
                                                              HANDUUI 15
                                                              (Continued 9 of 9)

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                                                             HANDOUT 16
Form Approved
OMB No  158-R0073
NPDES COMPLIANCE INSPECTION REPORT (Coding Instructions on back of last page)
TRANSACTION
CODE
U liJ
1 2
1 1 1 1

21
1 1 1 1

55 70
1 1 1
NPDES YR MO DA T
1 1 1 1 1 1 1 1 1 1 1
3 11 12 17
1 1 1 1
REMARKS
II 1 1 1 1 1 1 1 1 1 1 1 1
INSPEC- FAC
YPE TOR TYPE
U U U
IB 19 2O
1 1 1 1 1 1 1
TIME
am I p m.
1 1 1 1 1 1 1
64
ADDITIONAL
SECTION A • Permit Summary
NAME AND ADDRESS'OF FACI LITY (Include County. State and ZIP code)


RESPONSIBLE OFFICIAL TITLE
FACILITY REPRESENTATIVE TITLE
EXPIRATION DATE
SSUANCE DATE
PHONE
PHONE
SECTION B . Effluent Chwietariities (Additional sheen attached )
PARAMETER/
OUTFALL






SAMPLE
MEASUREMENT
PERMIT
REQUIREMENT
SAMPLE
MEASUREMENT
PERMIT
REQUIREMENT
SAMPLE
MEASUREMENT
PERMIT
REQUIREMENT
SAMPLE
MEASUREMENT
PERMIT
REQUIREMENT
SAMPLE
MEASUREMENT
PERMIT
REQUIREMENT
MINIMUM










AVERAGE










MAXIMUM










ADDITIONAL










SECTION C • Facility Evaluation (S = Satisfactory, U = Unsatisfactory. N/A = Not applicable)
EFFLUENT WITHIN PERMIT REQUIREMENTS OPERATION AND MAINTENANCE SAMPLING PROCEDURES
RECORDS AND REPORTS COMPLIANCE SCHEDULE LABORATORY PRACTICES
PERMIT VERIFICATION FLOW MEASUREMENTS OTHER:
SECTION D- Comments
SECTION E • lns|Metion/R«vi«w
SIGNATURES AGENCY
INSPECTED BY
INSPECTED BY
REVIEWED BY
DATE



ENFORCEMENT
DIVISION
USE ONLY
COMPLIANCE STATUS
D COMPLIANCE
DNOMCOMfUANCE
EPA FORM 3560-3 (9-77)
                          REPLACES EPA FORM T-S1 (9-76) WHICH IS OBSOLETE.
                                                                                             PAGE 1 OF 4

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                                                              HANDOUT  16
                                                                                    Form Approved
                                                                                    OMB No 158-R0073
Sections F thru L: Complete on all inspections, as appropriate. N/A = Not Applicable
PERMIT NO.

SECTION F • Facility and Permit Background
ADDRESS OF PERMITTEE IF DIFFERENT FROM FACILITY DATE OF LAST PREVIOUS INVESTIGATION BY EPA/STATE
(Including On. County and ZIP code)
FINDINGS
SECTION G • Records and Reports
RECORDS AND REPORTS MAINTAINED AS REQUIRED BY PERMIT. DYES D NO D N/A (Further explanation attached 	 )
DETAILS
(a) ADEQUATE RECORDS MAINTAINED OF.
(i) SAMPLING DATE, TIME, EXACT LOCATION
III) ANALYSES DATES. TIMES
(,!,) INDIVIDUAL PERFORMING ANALYSIS
liv) ANALYTICAL METHODS/TECHNIQUES USED
(v) ANALYTICAL RESULTS (e g., consistent with self-monitonng report data)
(b) MONITORING RECORDS (e.g.,flow, pH, D.O , etc.) MAINTAINED FOR A MINIMUM OF THREE YEARS
INCLUDING ALL ORIGINAL STRIP CHART RECORDINGS (eg. continuous monitoring instrumentation.
calibration and maintenance records).
(c) LAB EQUIPMENT CALIBRATION AND MAINTENANCE RECORDS KEPT.
(d) FACILITY OPERATING RECORDS KEPT INCLUDING OPERATING LOGS FOR EACH TREATMENT UNIT.
(e) QUALITY ASSURANCE RECORDS KEPT
If) RECORDS MAINTAINED OF MAJOR CONTRIBUTING INDUSTRIES (and their compliance Statist USING
PUBLICLY OWNED TREATMENT WORKS.
D YES D NO
D YES D NO
D YES D NO
D YES D NO
D YES D NO
D YES D NO
D YES D NO
D YES D NO
D YES D NO
D YES D NO
DN/A
ON/A
DN/A
DN/A
DN/A
DN/A
DN/A
DN/A
DN/A
DN/A
SECTION H - Permit Verification
INSPECTION OBSERVATIONS VERIFY THE PERMIT. DYES DNO D N/A (Furliier explanation
DETAILS
(a) CORRECT NAME AND MAILING ADDRESS OF PERMITTEE.
(b) FACILITY IS AS DESCRIBED IN PERMIT.
(c) PRINCIPAL PRODUCT(S) AND PRODUCTION RATES CONFORM WITH THOSE SET FORTH IN PERMIT
APPLICATION.
(d) TREATMENT PROCESSES ARE AS DESCRIBED IN PERMIT APPLICATION.
(el NOTIFICATION GIVEN TO EPA/STATE OF NEW, DIFFERENT OR INCREASED DISCHARGES
If) ACCURATE RECORDS OF RAW WATER VOLUME MAINTAINED
(g) NUMBER AND LOCATION OF DISCHARGE POINTS ARE AS DESCRIBED IN PERMIT.
(h) CORRECT NAME AND LOCATION OF RECEIVING WATERS.
d) ALL DISCHARGES ARE PERMITTED.
ittaehrd )

D YES D NO
D YES D NO
D YES D NO
D YES D NO
D YES D NO
D YES D NO
D YES D NO
D YES D NO
D YES D NO

D N/A
DN/A
DN/A
DN/A
DN/A
DN/A
DN/A
DN/A
DN/A
SECTION 1 • Operation and Maintenance
TREATMENT FACILITY PROPERLY OPERATED AND MAINTAINED. ID YES DNO D N/A (Further explanation attached 	 ;
DETAILS
(a) STANDBY POWER OR OTHER EQUIVALENT PROVISIONS PROVIDED.
(b) ADEQUATE ALARM SYSTEM FOR POWER OR EQUIPMENT FAILURES AVAILABLE.
(e) REPORTS ON ALTERNATE SOURCE OF POWER SENT TO EPA^TATE AS REQUIRED BY PERMIT.
(d) SLUDGES AND SOLIDS ADEQUATELY DISPOSED.
(e) ALL TREATMENT UNITS IN SERVICE.
(f) CONSULTING ENGINEER RETAINED OR AVAILABLE FOR CONSULTATION ON OPERATION AND
MAINTENANCE PROBLEMS.
(e) QUALIFIED OPERATING STAFF PROVIDED.
(h) ESTABLISHED PROCEDURES AVAI LABLE FOR TRAINING NEW OPERATORS.
Ill FILES MAINTAINED ON SPARE PARTS INVENTORY, MAJOR EQUIPMENT SPECIFICATIONS. AND
PARTS AND EQUIPMENT SUPPLIERS.
!|) INSTRUCTIONS FILES KEPT FOR OPERATION AND MAINTENANCE OF EACH ITEM OF MAJOR
EQUIPMENT.
Ik) OPERATION AND MAINTENANCE MANUAL MAINTAINED.
II) SPCC PLAN AVAILABLE.
m) REGULATORY AGENCY NOTIFIED OF BY PASSING. (Dates )
In) ANY BY-PASSING SINCE LAST INSPECTION.
(a) ANY HYDRAULIC AND/OR ORGANIC OVERLOADS EXPERIENCED.
D YES D NO
D YES D NO
D YES D NO
D YES D NO
D YES D NO
D YES D NO
D YES *D NO
D YES D NO
D YES D NO
D YES D NO
D YES D NO
D YES D NO
D YES D NO
D YES D NO
D YES D NO
DN/A
DN/A
D N/A
DN/A
DN/A
DN/A
DN/A
DN/A
DN/A
DN/A
DN/A
DN/A
DN/A
DN/A
ONMJ
EPA FORM 3560-3 (9-77)
                                                                                          PAGE 2 OF 4

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                                                                            HANDOUT  16
                                                                                                     Form Approved
                                                                                                     OMB No 158-R0073
                                                                                             PERMIT NO
SECTION J • Compliance Schedules
PERMITTEE IS MEETING COMPLIANCE SCHEDULE            DYES  D NO    DN,A   {^urltn'r <
   CHECK APPROPRIATE PHASE(S)
   D (a) THE PERMITTEE HAS OBTAINED THE NECESSARY APPROVALS FROM THE APPROPRIATE
        AUTHORITIES TO BEGIN CONSTRUCTION
   D (b) PROPER ARRANGEMENT HAS BEEN MADE FOR FINANCING (miirteajlC Cinnnlllllll'llti iini'lf i'ft
   EZ (cl CONTRACTS FOR ENGINEERING SERVICES HAVE BEEN EXECUTED
   D (d) DESIGN PLANS AND SPECIFICATIONS HAVE BEEN COMPLETED
   D (el CONSTRUCTION HAS COMMENCED.
   D 'fl CONSTRUCTION AND/OR EQUIPMENT ACQUISITION IS ON SCHEDULE
   D Igl CONSTRUCTION HAS BEEN COMPLETED
   D (h) START UP HAS COMMENCED
   D d) THE PERMITTEE HAS REQUESTED AN EXTENSION OF TIME
                                                                                                      lrJ
SECTION K • Self-Monitoring Program
Pjrt  1 - Flow niejsiircnicnl 11 urthcr explanation attathrd	)
PERMITTEE FLOW MEASUREMENT MEETS THE REQUIREMENTS AND INTENT OF THE PERMIT
   DETAILS
                                                                                           D YES    D NO
ON/A
(al PRIMARY MEASURING DEVICE PROPERLY INSTALLED
                                                                                            D YES    D NO
     TYPE OF DEVICE    DwEIR   D PARSH ALL F LUME  DMAGMETER  D VENTURI METER  D OTHER (Specify.
 I)) CALIBRATION FREQUENCY ADEQUATE (Dalf (Jl last callhralittn .
                                                                                           O YES    D NO
                                                                                                                 DN/A
DN/A
 cl PRIMARY FLOW MEASURING DEVICE PROPERLY OPERATED AND MAINTAINED
                                                                                           D YES    D NO
                                                                                                                 DN/A
 alSECONDARY INSTRUMENTS /totalizers, recuiiltrs. del PROPERLY OPERATED AND MAINTAINED     DYES    DNO     DN/A
 el FLOW MEASUREMENT EQUIPMENT ADEQUATE TO HANDLE EXPECTED RANGES OF F LOW FJATES  D YES    D NO     DN/A
?jrl 2 - Sampling (Further e\planati j>
                                                                                           D YES    D NO
DN/A

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                                                                            HANDOUT  16
                                       Form Approved
                                       OMB No. 158-R0073
                                                                                               PERMIT NO.
SECTION L • Effluent/Receiving Water Observations (Further explanation attached.
 OUTFALL NO.
                  OILSHEEN
                                 GREASE
                                               TURBIDITY
VISIBLE
 FOAM
  VISIBLE
FLO AT SOL
                                                                                             COLOR
                                                                                                            OTHER
                                (Sections M and N  Complete as appropriate for sampling inspections)
SECTION M - Sampling Inspection Procedures and Observations (Further explanation attached	)
   D GRAB SAMPLES OBTAINED
   D COMPOSITE OBTAINED
   D FLOW PROPORTIONED SAMPLE
   D AUTOMATIC SAMPLER USED
   D SAMPLE SPLIT WITH PERMITTEE
   D CHAIN OF CUSTODY EMPLOYED
   D SAMPLE OBTAINED FROM FACILITY SAMPLING DEVICE
COMPOSITING FREQUENCY 	
                                                                       PRESERVATION
SAMPLE REFRIGERATED DURING COMPOSITING     DYES      D NO
SAMPLE REPRESENTATIVE OF VOLUME AND NATURE OF DISCHARGE —
 SECTION N • Analytical Results (Attach report ij necessary)
EPA Form 3560-3 (9-77)
                                                                                                          PAG* « PF 4

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                                                    HANDOUT 17
                                                    (1 of 5)
                  INSTRUCTIONS FOR COMPLETING THE
                NPDES COMPLIANCE INSPECTION REPORT
                        (EPA Form 3560-3)
 Overview

      The intent of the NPDES Compliance"Inspection Report>
-form is to provide standard, reviewable information about an
 inspection to Enforcement.   All inspections will be conducted
 and  all reports will be completed as if they may lead to
 enforcement action.  The form defines the minimum amount of
 information that Enforcement should receive.  Regional and
 State inspectors may elect  to include additional information,
• as the circumstances warrant.

      Both Compliance Evaluation Inspections (CEIs) and
 Compliance Sampling Inspections (CSIs)  of municipal and non-
 municipal facilities will be conducted using the same type
 of Compliance Inspection Report form (EPA Form 3560-3).
 Using the same form and format will minimize the reporting
 burden on inspectors and permittees because identical elements
 of compliance (e.g., permittee records and self-monitoring
 program,  etc.)  are examined in both CEIs and CSIs.  Although
 the  form may be used for either inspection, a completed form
 will be credited in only one category of the Formal Program
 Reporting System (FPRS). A completed form contains all the
 information appropriate, to  the accomplished inspection.  A
 completed form is, by definition,  also what will be accepted
 by the Enforcement Director of the agency responsible for
 enforcing the permit.  Procedures for the distribution of
 the  completed form should be planned with the Enforcement
 Director.   Users should note that the top of page 1 serves
 as a coding sheet for entries into the Water Enforcement
 National Data Base (WENDB).

      The Compliance Inspection Report consists of two major..
 parts.  The first part, sections A-L, is completed for all
 inspections, as appropriate.  The second part, sections
 H-N, is completed only for  CSIs.  For the checklists,
 sections G through K, each  lead statement will 'summarize
 deficiencies covered in the section.   Each item in the
 checklist (except Section I items (n) & (o)) is written so
 that a "yes" answer is positive, indicating some degree of
 permit compliance.  If there are no problems in a section,
 all  the answers will be yes (with the exceptions noted
 above).

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                                                    HANDOUT 17
                                                    (Continued (2 of 6)
     Throughout the form, numerous opportunities exist to
attach additional explanations.  These explanations should
be attached only when necessary.  Although a narrative is
not appropriate when a simple yes, no, or N/A will do,
inspectors must adequately document their observations.
However, lengthy narratives will defeat the purpose of the
checklists.  Nonetheless, if further explanation is deemed
necessary, it should be attached and noted on the form.

     A brief review of each section follows.  For further
explanation of Compliance Inspections, consult the appro-
priate manuals.  (NPDES Compliance Evaluation Inspection
Manual - U.S. EPA, Office of Water Enforcement and NPDES
Compliance Sampling Manual - U.S. EPA, Office of Water
Enforcement).

Keypunch Summary

     This lead information is used to identify the facility
and the inspection date, type and agency.  The data can be
keypunched on one card and entered directly into the Water
Enforcement National Data Base (WENDB) .  Entries in WENDB
will assist tracking of inspection results and will be used
for reporting in FPRS.  To be part of the WENDB, the data
should be reported as follows:

     Column 1       Transaction Code - Use N, C, or D for
                    New, Change or Delete.  All inspections
                    will be new unless there is an error in
                    the data keypunched into WZNDB.

     Column 2       Card Code - always 5 for this card.

     Columns 3-11   NPDES - The NPDES permit number. (The
                    State permit number may be accommodated
                    in the remarks or additional spaces).

     Columns 12-17  Inspection Date - entered in the year/
                    month/day format  (e.g. 77/06/30 = June 30,
                    1977).

     Column 18      Inspection Type - An inspection will
                    fall into one of two possible categories:
                    'C1 for Compliance Evaluation or 'S1 for
                    Compliance Sampling.

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                                                  HANDOUT 17
                                                  (Continued 3 of 6)
Column 19
Column 20
Columns 21-70
Inspector Code - An inspection may be
performed by the Region, State or NEIC
(U.S. EPA National Enforcement Investi-
gations Center).  It may also be the
result of a joint effort. (Credit in
FPRS for a joint inspection is given to
the lead agency.)   Acceptable codes for
WENDB are:

  R - EPA Regional inspections
  S - State inspections
  J - Joint EPA and State inspections -
      EPA lead
  T - Joint EPA and State inspections -
      State lead
  N - NEIC inspections

Facility Type - This code describes the
type of. facility that: was inspected.
Acceptable codes are:

  1 - Municipal - Publicly-Owned Treat-
      ment Works (POTWs) with 1972 Standard
      Industrial Classification (SIC) 4952

  2 - Industrial - Other than Municipal,
      Agricultural, and Federal facilities.

  3 - Agricultural - Those facilities
      classified with 1972 SIC 0111-0971.

  4 - Federal - Those facilities identi-
      fied as Federal by EPA Regional
      office.

Remarks - This remarks field provides
the inspector with a vehicle to store
descriptive information about the in-
spection.  There is no set format within
this 50-position field.  Individual
Regions or States may choose to set
aside portions of this field for their
own specific needs.

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                                                    HANDOUT 17
                                                    (Continued 4 of 6)
     The "Time" and "Additional" boxes can also describe the
inspection, but will not be keypunched.  Supplementary in-
formation that the performing agency or Region needs may be
entered in the Additional box, e.g., STORET numbers, basin
codes, etc.

Section A - Permit Summary

     This'section provides the summary information required
to further identify the inspected facility.  Most of the
elements are self explanatory; however, the last two lines
may require explanation.  "Responsible Official" is the
individual required to sign the Discharge Monitoring Report
or is responsible for wastewater management at the facility.
"Facility Representative" is the individual who acted as a
contact during the inspection.

Section B - Effluent Characteristics

     Effluent Characteristics contains a summary of those
parameters (e.g. BOD, pH, flow) that are regulated by the
permit and any other parameters that are measured but not
regulated by the permit.  If more than one outfall is
inspected, the parameter and outfall should be indicated and
additional sheets attached as required.  If the inspection
will not include samples, it may be advisable, but is not
required, to substitute the data from the latest Discharge
Monitoring Report in the "Sample Measurement" row before
performing the inspection.  However, if self-monitoring data
are entered in the spots for sampling data, they should be
clearly identified as such to avoid confusing the reviewer.
The column marked "Additional" is for the performing agency's
or Region's own requirements, e.g., design data, comments or
explanations of the measurements.

Section C - Facility Evaluation

     The Facility Evaluation provides a summary evaluation
of the inspection results.  The evaluations made in this
section should be documented and supported by notation in
the appropriate checklist portions of the form and by any
additional comments as required.

Section D - Comments

     Little space is allowed for comments here.  Rather than
fragmenting the narrative detailing comments and possible
recommendations, the form allows detailed comments in an

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                                                  HANDOUT i?
                                                  (Continued  5 of 6)
attachment, on the back of the form, or in Section N.  The
Section D comments should be used to flag lengthy comments
(e.g. "Recommendations on p.4")  or used for those inspections
which only merit abbreviated comments.  Procedures for
making recommendations and comments should be worked out
with the Enforcement Director of the organization responsible
for the permit.  All comments or recommendations that are
made should be documented and supported by the checklist
portions of the form.

Section E - Inspection Review

     This section provides the inspector's and reviewer's
names and agencies.  Compliance status should be determined
only by the Enforcement personnel.

Section F - Facility and Permit Background

     If the permittee's address is different from that of
the facility, it should be so indicated.  If the facility
was inspected previously, the date and findings summary
should be noted before performing the current inspection.

Section G - Records and Reports

     This portion of the form documents that the records and
reports maintained by the permittee are in compliance with
permit requirements.  As mentioned earlier, if the checklist
does not adequately represent the situation, further expla-
ation should be attached and so indicated.

Section H - Permit Verification

     Each inspection should identify discrepancies, if any,
between the issued permit and actual conditions.  Again, if
further explanation is necessary, it should be provided and
so indicated.

Section I - Operation and Maintenance

     Each inspection of an operating facility should evaluate
its operation and maintenance.  Operating facilities include
those on final limits and those in the process of being
upgraded.

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                                                  HANDOUT 17
                                                  (Continued 6 of 6)
Section J - Compliance Schedule

     The compliance schedule progress should be evaluated
when the permittee is on a compliance schedule.  Any grant-
related inspections of facilities should be coordinated with
Regional Construction Grants personnel.  The current phase
of compliance schedule status should be marked on the form.

Section K - Self-Monitoring Program

     The permittee's flow measurement, sampling, and labora-
tory procedures should be checked, as appropriate, on all
inspections.  If deficiencies are noted, additional pertinent
information should be provided, if necessary.  For example,
if the laboratory is not calibrating or maintaining the
equipment satisfactorily, the calibration or maintenance
intervals should be noted.  If parameters other than those
required by the permit are analyzed, the parameters and
analytical methods should be noted.  If the permittee
laboratory, flow-measurement, or sampling procedures are not
inspected, an explanation should be provided (e.g., contract
lab off the premises).

Section L - Effluent/Receiving Water Observations

     Visual observations made during the inspection should
be noted, as applicable, for each outfall.  The inspector's
observations are subjective and qualitative, but serve to
focus attention on potential treatment problems.  Discharge
of floating solids or visible foam in other than trace
amounts is prohibited by the permit.  Thus, observations of
greater than trace amounts represent permit violations and
indicate poor treatment.

Section M - Sampling Inspection Procedures and Observations

     The performing agency's or Region's sampling procedures
should be noted for each sampling inspection.  Details docu-
menting the procedures should be provided (e.g., the composite
time interval).

Section N - Analytical Results

     If the analytical results or laboratory report from a
sampling inspection provides more information than can be-
inserted in Section C, the additional information should be
noted in this part or attached to the report form.  This
section also offers more space for comments or additional
materials (e.g. flow diagrams)  as the situation merits.

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