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
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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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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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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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COMPLIANCE MONITORING INSPECTOR TRAINING MODULE:
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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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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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COMPLIANCE MONITORING INSPECTOR TRAINING MODULE:
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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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COMPLIANCE MONITORING INSPECTOR TRAINING MODULE:
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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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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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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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COMPLIANCE MONITORING INSPECTOR TRAINING MODULE:
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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
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COMPLIANCE MONITORING INSPECTOR TRAINING MODULE:
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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
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COMPLIANCE MONITORING INSPECTOR TRAINING MODULE:
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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:
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U.S. ENVIRONMENTAL PROTECTION AGENCY
COMPLIANCE MONITORING INSPECTOR TRAINING MODULE:
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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
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COMPLIANCE MONITORING INSPECTOR TRAINING MODULE:
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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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COMPLIANCE MONITORING INSPECTOR TRAINING MODULE:
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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
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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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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
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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:
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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
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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
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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
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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
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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
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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
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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
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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
-------
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
-------
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.
-------
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
99.345
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
! : i
i ;
1 j !
i
i i !
i i
T- - : T;
i
1
: i
i '
r
\
\
1
.
1
1
1
1
!
-
*x
f
1
1
I
1
1 . .
1
I
I
1
1
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.
-------
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.).
-------
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
-------
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)
-------
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
-------
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)
-------
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
-------
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
-------
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
-------
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
-------
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).
-------
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.
-------
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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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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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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