United State*
 Environmental Protection
 Agency
Permits Division
EN-336
Washington, DC 20460
August 1987
 Water
Program Survey--
Biological Toxicity Testing
in the NPDES
Permits Program

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             PROGRAM SURVEY
BIOLOGICAL TOXICITY TESTING IN THE NPDES
             PERMITS  PROGRAM
              August  1987


            Permits Division
Office of Water Enforcement and Permits
  U.S. Environmental Protection Agency
        Washington, D.C.  20460

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                        TABLE OF CONTENTS


                                                             Page


INTRODUCTION ......................       1

RESULTS OF SURVEY  ...................       2

EPA REGIONAL PROGRAMS AND ENVIRONMENT CANADA'S PROGRAM .      11

STATE PROGRAMS .....................      22
TABLE 1.   Biological Toxicity Testing in NPDES
           Industrial and Municipal Permits  ......       5

TABLE 2.   State Toxicity Reduction Evaluations  ....       6

TABLE 3.   State Biological Testing Annual Programs  . .       7
FIGURE 1.  Major Industrial Permits with Toxicity Testing
           Requirements  ................       8

FIGURE 2.  Major Municipal Permits with Toxicity Testing
           Requirements  ................       9

FIGURE 3.  Permits with Toxicity Limits  ........      10

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                           INTRODUCTION


In 1986, EPA undertook an informal telephone survey of State and
EPA Regional personnel to determine the nature and extent of
biological toxicity testing in the NPDES permit program.  A
program survey summary report containing the information col-
lected in that survey, and bearing the date of July 1986, was
reproduced and distributed.

In the 12 months since that study was made, EPA Regions and many
States have formalized policies relating to NPDES toxicity
testing.  There have been concerted efforts in many States to
introduce biological testing into a greater number or all of
major and many minor permits being considered for reissuance.
The use of chronic toxicity effluent testing has increased, and
this has led to an increase in toxicity determinations and a
requirement on many permittees to conduct toxicity reduction
evaluations.  Because of these developments in the permit pro-
gram, it was appropriate to again contact State and EPA Regional
personnel to ascertain the present program status.  This report
summarizes the results of those contacts.

An informal telephone survey of NPDES State and EPA Regional
personnel was conducted in mid-1987 to determine the current use
of effluent biological testing.  To conduct this survey, EPA
contracted with WESTEC Services, Inc., Reston, Virginia.  Regions
and States were asked the number of NPDES permits they adminis-
tered, the number of permits with a biological toxicity testing
requirement, the number of permits with expressed toxicity limits
and with requirements for toxicity reduction evaluations. Region-
al and State programs were discussed, including the use of bio-
logical testing such as acute and chronic toxicity testing
techniques, instream biotic assessments, water quality trend
monitoring at fixed locations, and aquatic organism flesh
analyses for bioaccumulative materials, as well as any other use
of biological investigative techniques.

EPA has not verified the results of this survey independently.
Nor did EPA or the contractor look at individual permits, State
regulations, or procedures referenced in the report.  The intent
of this summary is to give general information on the use of
biological testing in Regional and NPDES State toxic control
programs.

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                      RESULTS OF THE SURVEY


Biological testing methods used to characterize environmental
effects of toxic pollutants discharges take various forms.
Generally, biological methods are segregated into two groups:
Effluent testing and receiving water testing.  State permit
programs use both methods, but permit-required effluent testing
is by far the more prevalent.

This survey focused on toxicity tests where organisms are exposed
in the static or flow-through environment of undiluted or diluted
wastewater for 96 hours or less to simulate acute exposure, or up
to 7 or more days to simulate chronic exposure.  Various organ-
isms are used in effluent toxicity tests; the EPA published a
list of appropriate organisms in Table 1 of "Methods for
Measuring the Acute Toxicity of Effluents to Freshwater and
Marine Organisms* (EPA/600/4-85/013).  The EPA publication,
"Short-Term Methods for Estimating the Chronic Toxicity of
Effluents and Receiving Waters to Freshwater Organisms"
(EPA/600/4-85/014), provides details for conducting chronic
toxicity tests.  Various biological methods are appropriate for
ambient or receiving water testing, and these provide the
capability to identify environmental damage from toxic pollutants
to some extent.  Methods employed by States include studies and
assessments related to macroinvertebrates, fishes, algae,
periphyton, protozoa, primary productivity, sediment toxicity
tests, fish flesh tainting, fish and mussel flesh analyses for
bioaccumulated substances, caged organism toxicity, fish and
invertebrate chronic toxicity tests, sediment analyses, and fish
avoidance reactions.

The survey showed that:

     o    38 States require industries to conduct toxicity
          testing (Table 1).

     o    27 States require municipalities to conduct toxicity
          testing (Table 1).

     o    1,343 industrial permits require toxicity testing
          (Table 1).

     o    597 municipal permits require toxicity testing  (Table
          1).

     o    The midwestern and western States have the fewest, if
          any, requirements for biological toxicity testing
          (Figures 1 and 2).

     o    Effluent toxicity limits are in industrial permits in
          14 States and are found in 39 percent of the industrial
          permits that require biological testing  (Table  1,
          Figure 3).
                                2

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     o    Effluent toxicity limits are in municipal permits in
          10 States and are found in 78 percent of the municipal
          permits that require biological testing
          (Table 1, Figure 3).

     o    16 States have a toxicity testing requirement in
          industrial permits only (Table 1).

     o    8 States do not require effluent biological testing
          (Table 1).

     o    19 States project that as new permits are issued, or as
          existing permits are reissued, all major and signifi-
          cant minor permits will contain biological testing
          requirements (Table 1).

     o    Using major permits as a means of comparison, 15
          States require biological toxicity testing in 50 per-
          cent or more of their industrial permits.  Califor-
          nia, Virginia, and West Virginia require biological
          testing in all of their major industrial permits and
          some minor permits (Table 1; Figure 1).

     o    Using major permits as a means of comparison, 6
          States require biological toxicity testing in 50 per-
          cent or more of their municipal permits.  New Jersey
          requires biological testing in all of their municipal
          major permits and some minor permits (Table 1; Figure
          2).

     o    10 States have toxicity reduction evaluations programs
          underway, and 13 additional States are beginning such
          programs (Table 2).

     o    5 States have (or will have within one year)
          biological laboratory certification programs (Table 3).

     o    16 States operate mobile biological laboratories for
          flow-through or chronic biological testing (Table 3).

     o    29 States have capability for conducting definitive
          acute or chronic toxicity tests (Table 3).

     o    33 States conduct receiving water macroinvertebrate or
          fish assessments related either to special study
          pollution investigations or to provide trend monitoring
          determinations (Table 3).

It is believed that the information herein is more exact than
that contained in the 1986 report.  Each contact listed in the
address of a State or Federal program page was given the
opportunity to view and comment on the appropriate draft
narrative.  Host took the opportunity to refine numbers or
narrative connotations.  Although the differences between this

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and the 1986 report may be subtle in respect to numerical values
contained in the report, there is a noticeable difference in
future projections by States regarding use of biological toxicity
testing in the NPDES programs; nineteen States responded that as
new permits are issued, or as existing permits are reissued, all
major permits will require biological testing.  Six of the
responding States now have five or fewer permits with such
testing requirements.

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                 TABLE 1.  BIOLOGICAL EFFLUENT TESTING IN HPDE8 PERMITS
                             mUSTHIAL
                                                   •UUCIPAL
IHDUSTH2AL
    *

MUNICIPAL

ALABAMA
ALASKA
ARIZONA
ARKANSAS
CALIFORNIA
COLORADO
CONNECTICUT
DELAWARE
DISTRICT OF co.
FLORIDA
GEOR6IA
HAWAII

INDIANA
IOWA
KANSAS
KENTUCKY
LOUISIANA
MAINE
MARYLAND
MASSACHUSETTS
MINNESOTA
MISSISSIPPI
MISSOURI
MONTANA
NEBRASKA
NEVADA
NEW HAMPSHIRE
NEW JERSEY
NEW MEXICO
NEW YORK
NORTH CAROLINA
NORTH DAKOTA
OHIO
OKLAHOHA
OREBON
PENNSYLVANIA
80UTH CAROLINA
SOUTH DAKOTA
TEXAS
UTAH
VERHONT
VIRGINIA
WASHIN6TON
WISCONSIN
WYOMING
TOTALS
/ *
80
0
40
184
1
88*
4
40
88
8

8
0
13
100
11*
88
88
1
80
0
a
185
B
88*
0
7
88
IB
0
84
0
88
133
B
0
180
87
70
BB*
0
1343
/
81
0
71
100
8
73
18
88
48
88

7
0
10
BB
87
71
83
4
80
D
11
83
81
B
88
B
B
81
78
0
3D
0
•B
88
88
0
1BO
B8
100
BB
0

/
BO
0
.0
143
0
8
0
40
0
0

0
B
IB
0
1
0
17
.0
10
0
D
100
0
0
88
0
0
0
8
0
0
0
0
0
0
0
BO
0
0
0
478
/*
•
«
*
'•
:

0
0
«
'
•
«

*
,
f
f
*
f
*
0
f
*
*
1
f
:

/
0
0
0
178
B
0
1
40
1
1

0
0
4
0
8
80
18
0
0
0
8
178
0
14
70*
1
B
0
0
0
14
B*
1
0
7
0
80
B
B
8
887
/
B
0
0
87
11
0
7
100
88
1
B

0
0
7
0
3
84
18
8
0
0
8
00
0
B
B4
7
B
B
0
0
78
4
8
B
IB
0
48
B
B
18

/ "


18

4










17
7





4B4
"
»
«
*
0
*
•
0
0
0
t
t
1
t
0
f

t
t
1
0
f
f
B
*
•
*
1
f
*

/ *
80
0
40
8BB
B
88
B
1
80
88
B
4
B
B
0
17
IBB
18
48
84
IB
1
80


BOO
B
88
138
1
18
88
IB
B
80
•8
1
88
188
13
0
1BO
87
70
BB
8
1B40
/ m
88
0
74
100
B
41
14
100
83
14
13




48
14
BB
IB
B
1
84
0
0
B
B
B3
14
B
87
B
4
•3
81
44
IB
a
18
•7
88
0
100
41
74 .
84
4

/ "
BB
B
.8°.
a
0
1
80
0
0
0
1
0
0
B
17
0
8
B
83
0
0
IB
0
- B
B
B
878
B
B
188
0
0
B
8
B
B
0
B
0
0
0
0
80
B
0
0
848
f **
.
„
;
0

•
f
B
B
0
f
f
'
1

f
•
f
#
*
t
0
*
*




           any riqulrcainti vl* •da1n1(tr»t1*«  Icttir.
1 NuBbcr clth ticttng ••» fnelud* ••Jot and  Miner  p«r«1t>.
8 Percentage reletee  to ••Jor permits.
• Peralti Bonuin toilelf. ilaiu.
4 When ••• or cnUtlng peratte ere Ucuetf or relouedi
  * • Blologleil toitclty taxing In ell ••Jor  perBlti
  f • Tilting rcqulrenent* mill be pieced In eeleeted pereilte.
  0 • He projected greeth In tooting requirement progren.

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TABLE B.  STATE PR06RANS TNAT NEOUIRE TOXICITT REDUCION fVALUATION

ALABAMA
ALASKA
ARIZONA
ARKANSAS
CALIFORNIA
COLORADO
CONNECTICUT
DELAWARE
FLORIDA
SEOR6IA
MAVAII
IDAHO
ILLINOIS
INDIANA
IOWA
KANSAS
KENTUCKY
LOUISIANA
NAINE
NARYLANO
NASSACHU6ETTS
NICHI6AN
NINNEBOTA
MISSISSIPPI
NX8SOURI
NONTANA
NEBRASKA
NEVADA .
NEV NANPBHIRE
HEN JERSEY
NEW NEXICO
NEW YORK
NORTH CAROLINA
NORTH DAKOTA
OHIO
OKLAHOMA
ORE60N
PENNSYLVANIA
RHODE ISLAND
SOUTH CAROLINA
SOUTH DAKOTA
TENNESSEE
TEXAS
UTAH
VERMONT
VIRGINIA
VABHIN6TON
•E6T VIRGINIA
WISCONSIN
•YONIN8
TOTALS
f
•
•
*
•

•
•
•


•

•
•

•
IB


•
•
*
•


•
•

•
•
•

•

•
•
•
18

•
•
•

•
•
•
•
•
•
•
•
•
•

•
•
•
•
•
•
•
•

•
87

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                  TABLE
                            BIOLOGICAL TESTING PROGRAMS CONDUCTED BY
                               BTATE PEMOMMEL ANNUALLT

ALABANA
ALASKA
ARIZONA
ARKANSAS
CALIFORNIA
COLORADO
CONNECTICUT
DELAWARE
DISTRICT OF CO.
FLORIDA
GEOR6IA
HAWAII
IDAHO
ILLINOIS
INDIANA
IOWA
KANSAS
KENTUCKY
LOUISIANA
MAINE
MARYLAND
MASSACHUSETTS
NICHI6AN
MINNESOTA
MISSOURI
MONTANA
NEBRASKA
NEVADA
NEW NANPSHIRE
MEN JERSEY
NEW MEXICO
MEW YORK
NORTH DAKOTA
OHIO
ORE60N
PENNSYLVANIA
60UTN CAROLINA
SOUTH DAKOTA
TENNESSEE
TEXAS
UTAH
VIRGINIA
WASHINGTON
WISCONSIN
WYOMING
TOTALS

80
0
0
12
e
it
0
>80
0
0
•0
17
0
44
186
ISO
BO
80
100
BO
es
B
0
B
0
0
180
0
0
100
0
78
0
180
0
0
0
BO
eo
0
0
B8
1C
76
0
0
1428

B
0
0
0
e
10
0
0
0
0
0
0
0
0
B4
112
ao
a
is
IB
IS
IB
0
0
0
0
0
80
100
0
10
0
IBS
0
0
0
10
0
o
0
10
IE
0
0
0
BBS

•0
0
0
86
a
10
0
0
•0
0
0
ao
18
0
40
as
a
BB
60
0
0
400
80
0
0
0
18
40
BO
BO
0
BO
0
B40
840
0
IB
100
BO
0
BO
100
480
0
80B6

a
0
0
0
B
•0
e
0
BB
0
0
e
88
0
eo
BO
0
80
0
ao
0
8B
100
a
0
ae*
100
0
0
0
84
100
10
80
80
0
80
ao
178
4S
80
0
1084
r
•
0
0
0
0
•
•
0
0
•
•
0
0
•
•
0
0
0
0
0
•
•
•
•
0
0
0
0
0
•
0
•
•
0
0
0
0
0
0
•
0
•
0
0
•
0
0
0
0
18
r
a
0
o
0
•
0
0
0
o
0
0
0
0
0
0
0
o
a
0
0
0
0
0
0
0
0
0
0
0
•
0
0
•
0
0
0
0
0
0
•
0
0
0
•
0
0
0
0
e
0
B
- Information not ovolloblo
1 Nuobor of loctttoni Briar* oiocrolnvortobroto or floh  ••••••••nt« roloto to
  point court* dlochargoi
8 Nacrolnvortobrot* or floh locotlon ••oooomonto rolotod to  othor thon point
  •oureo dlochirgo*.
a Btota oporotoa o nobll* laboratory to oorvloa point  ooureo dlochorgo*.
4 Hoo or >1ll nova 1n IB nonttia o biological  laboratory  certification progroau
                                      7

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                   FIGURE 1. PERCENTAGE OF MAJOR INDUSTRIAL PERMITS WITH
                            TOXICITY TESTING REQUIREMENTS
CO

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FIGURE 2. PERCENTAGE OF MAJOR MUNICIPAL PERMITS
         WITH TOXICITY TESTING REQUIREMENTS

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FIGURE 3. NUMBER OF PERMITS WITH TOXICITY LIMITS

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                                                          EBkJREGION I
U.S. Environmental Protection Agency, Region I
John F. Kennedy Federal Building
Room 2203
Boston, MA  02203

Mr. Jerry Potamis, Chief             Mr. Clyde Shufelt, Chief
Industrial Permits Section           Municipal Permits Section
(617) 565-3512                       (617) 565-3516
Biological testing is required for industrial or municipal dischargers scheduled
for permit issuance where technology based pollutant limitations diluted by the
receiving water  (7Q10 for acute toxicity and 30Q2 for chronic toxicity) cannot
meet acute or chronic water quality criteria specified in the water quality
criteria documents (45 FR 79318, November 29, 1980) or more recently updated
drafts of final criteria documents.  All municipalities with pretreatment pro-
grams are required to perform effluent toxicity testing as permits are reissued.
Where permit reissuance is not imminent, Section 308 letters are used to
institute toxicity testing.  Discharges containing chemicals for which criteria
have not been developed are evaluated on a case-by-case basis to determine the
likelihood of water use impairment.

Industrial dischargers to fresh water must conduct four acute toxicity tests
using a daphnid and the fathead minnow within a 60-day period.  Permits
containing chemical specific limits based upon biological toxicity testing will
include a requirement for periodic chronic or acute toxicity testing.  The
permittee may accept water quality criteria based, chemical specific permit
limits in lieu of biological toxicity testing if such can be met with available
treatment technology.  Toxicity Reduction Evaluations may be required to bring an
effluent into compliance with Water Quality Standards provisions prohibiting
discharges of toxic substances in amounts toxic to human health or aquatic life.
The Regional policy currently contains no provisions for testing related to human
health.
                                     11

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                                                          EPA REGION II
U.S. Environmental Protection Agency, Region II
26 Federal Plaza
New York, NY  10278

Ms. Ruth Adelman, Chief            Mr. Keith Tingberg
Permits Management Section         Permits Management Section
(212) 264-2911                     (212) 264-2936
The Region's toxicity testing policy considers new permit applications and
reissuances of prior permits.  In addition, toxicity testing is required of
permittees, where necessary, via Section 308 letters.  The Region determines a
"toxicity potential assessment" for a permittee which is an evaluation using:

   o    Dilution of effluent by receiving water
   o    Instream toxic impacts potential
   o    Use classification of receiving water
   o    Industrial category and specific processes/products (industrial
        permits)
   o    Percentage of industrial contribution in municipal permits
   o    Existence of toxic chemicals

Testing is conducted on a two-tier basis.  Tier 1 testing requires 48-hour acute
static renewals with an least two species, a vertebrate and an invertebrate,
using the fathead minnow and Daphnia magna for freshwater discharges and the
silverside  (Mgnidia_s£.) and nqysiAipsjg hahia for discharges to saline waters.
At least four tests are to be conducted over a time period representative of all
facility processes.

Tier 2 testing "should be directed at obtaining data necessary to refine the
impact assessment."  This may include chronic toxicity testing or instream
toxicity testing.  Compliance monitoring of the discharge may be required to
ensure that the toxicity of the effluent does not change.  Failure to pass
toxicity testing  (Tier 1) would lead to Tier 2 testing.  The issuance of a
toxicity permit limit with a monitoring requirement and/or a toxicity reduction
evaluation may be required of the permittee pending the results of the Tier 2
testing.

Region II's program objective of requiring toxicity testing or limitations in
NPDES permits is  "to prevent toxics from being discharged in amounts which are
acutely and chronically toxic to aquatic organisms."  The policy does not address
human health effects, which should be addressed through chemical-specific
approaches.
                                    12

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                                                          EGA REGION III
U.S. Environmental Protection Agency, Region III
841 Chestnut Street
Philadelphia, PA  19107

Mr. Robert Koroncai
Environmental Engineer
Water Quality Control Section, Water Management Div.
(215) 597-0133
Region III has developed guidance relating to the biological testing of effluents
for toxic pollutants.  Such guidance is used in further negotiations with the
states related to procedures for toxic wastes and in the review of those NPDES
permits subject to regional review.  All states within the region have delegated
NPDES authority.

The guidance provides for monthly toxicity testing unless it can be demonstrated
that an effluent has particular chemical constituents that can be adequately
assessed through chemical testing.  For effluents with little variability, less
frequent testing may be acceptable.  Where the wastewater dilution in receiving
water is less than 10:1, the 7-day chronic fathead minnow and Ceriodaphnia
toxicity tests are required.  Where such dilution is greater than above, acute,
static or static renewal 96-hour two species toxicity tests may be used.
Appropriate saltwater species may be substituted as test organisms in marine
waters.  The biomonitoring results are compared with instream waste dilution and
when potential toxicity is suspected as a result of this comparison, a toxicity
reduction evaluation is implemented.

For chemicals specifically identified in permits, a mass balance calculation is
made based upon a design flow of 7Q10 for chronic effects and a 1Q10 for acute
effects.  The results are compared with EPA water quality criteria.

For human health effects, the mean annual flow is used.  Health effects risks are
based upon most recent EPA water quality criteria and upon the EPA Integrated
Risk Information system, which provides computerized health effects risk data for
non-carcinogens and, in the future, for carcinogens.  For carcinogens, a 10-6
risk level is recommended.
                                     13

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                                                          EPA REGION IV
U.S. Environmental Protection Agency, Region IV
345 Courtland Street, NE
Atlanta, GA  30365

Mr. Marshall Hyatt, Environmental Scientist
Facilities Performance Branch
Water Management Division
 (404) 347-3012
With the exception of the State of Florida, all of the states within Region IV
have delegated NFDES authority.  Region IV administers the NPDES program for the
State of Florida.  For those states with NPDES authority, Region IV has issued
guidance in the form of proposed permit toxicity limits and monitoring
requirements.  This guidance follows that set forth in EPA's 1985 "Technical
Support Document for Water Quality-based Toxics Control," in "Methods for
Measuring the Acute Toxicity of Effluents to Freshwater and Marine Organisms"
(EPA/600/4-85-013), and in "Receiving Waters to Freshwater Organisms" (EPA/600/4-
85/014) .  The biomonitoring guidance is summarized as follows:

When instream waste concentration is greater than or equal to 1 percent at
critical low-flow conditions, the permittee shall conduct a 7-day Ceriodaphnia
survival and reproduction test and a fathead minnow larval survival and growth
test on a 24-hour composite sample on a series of effluent concentrations,
including one equivalent to the instream waste concentration, with test solutions
renewed daily.  Toxicity tests shall be conducted every 2 months for a period of
1 year following initiation of tests and once every 6 months thereafter for the
duration of the permit.  A permit violation occurs when toxicity is found in the
initial test.  Generally, a toxicity reduction evaluation will be required only
when toxicity is found in a confirmatory test.  Test procedures are those
recommended in EPA/600/4-85/014.

When the instream waste concentration is less than 1 percent at critical con-
ditions, the permittee shall conduct 48-hour static toxicity tests on three
appropriate species including a fish, an invertebrate, and one species selected
from EPA 600/4-85/013, Table 1.  Tests shall be conducted once every 2 months on
100 percent effluent for a period of 1 year following the initiation of the test
and once every 6 months thereafter for the duration of the permit.  Four separate
grab samples of final effluent shall be collected at evenly spaced intervals over
a 24-hour period and used in four separate tests in order to catch any peaks of
toxicity and to account for daily variations in effluent quality.  Test pro-
cedures are those recommended in EPA/600/4-85/013.
                                      14

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                                                          EPA REGION V
U.S. Environmental Protection Agency, Region V
230 South Dearborn Avenue
Chicago, EL  60604

Ms. Linda Anderson-Carnahan
Permits Section
(312) 886-0136
All of the states in Region V are delegated with the NPDES program.  Under the
compliance monitoring program, the region performs a battery of toxicity tests on
an average of 80 discharges a year,  tine battery of tests includes Daphnia pulex
and Pimephaieg promelaa acute toxicity tests, a 96-hour algal growth inhibition/
stimulation test, and the Ames Assay (conducted with five strains of
with and without enzyme activation) .  In some cases the Daphnia and Pimephalea
acute toxicity tests are replaced by the Ceriodaphnia survival and reproduction
test and the Pimpphaiea promelaa embryo-larval survival and teratogenicity test.
The region also is currently investigating a variety of other toxicity tests
including the rat hepatocyte sister chromatid exchange assay, the mammalian
mitochondria assay, the Pimephalea promelaa larval survival and growth test, and
the effluent toxicity tests performed by Environment Canada.

Upon receiving regional test results and/or the results of toxicity tests
performed by state agencies, the region may send a letter to the state requesting
that the permit for the facility tested be reopened and modified or the draft
permit sent to the region for review contain certain provisions.  Depending on
whether significant toxicity was observed or state water quality standards appear
to have been violated, the region may request that the permit contain biomoni-
toring with a trigger to include provisions for a toxicity reduction evaluation
and final whole effluent toxicity limits with post-control biomonitoring.  When
data are lacking, the region may require biomonitoring alone.  The region also
may use this opportunity to request that chemical specific limits and/or moni-
toring be contained in the modified/reissued permit, depending on the effluent
chemical analyses data available.  In cases where a state has failed to protect
water quality adequately through the use of toxicity/toxicant limits and
monitoring, the region is objecting to the permits.
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                                                          EPA REGION VI
U.S. Environmental Protection Agency, Region VI
1445 Ross Avenue
12th Floor, Suite 1200
Dallas, TX  75202-2733

Mr. Craig Weeks
Environmental Engineer
(214) 655-7180
Except for Arkansas, none of the states within Region VI currently has NPDES
permitting authority.  The Region VI toxics policy is designed to support and
implement the national policy.  The Regional policy is that no source will be
allowed to discharge any wastewater which results in the endangerment of a
drinking water supply; results in aquatic bioaccumulation which threatens human
health; results in instream acute or chronic aquatic toxicity,; or causes a
violation of an applicable general or numerical state water quality standard.

In order to accomplish these objectives, the Region will as part of the third
round permit issuance procedures ensure that no sources will cause or signifi-
cantly contribute to an exceedence of EPA's Maximum Contaminant Level require-
ments for any public drinking water supply; identify and address, with either
state or EPA action, sources which may cause or significantly contribute to any
exceedence of EPA's water quality criteria for human health protection; address
aquatic toxicity by either applying appropriate limitations or toxicity reduction
requirements when toxic conditions are known or requiring biological screening
for all other facilities having any potential for causing ambient toxicity; and
incorporate into permits any applicable specific numerical state water quality
standard.

Permits issued to dischargers with a potential for causing ambient toxicity will
require that the permittee perform periodic toxicity screening using whole
effluent biomonitoring techniques.  Permittees typically will be required to
monitor on a monthly frequency.  As a general rule, discharges which are
substantially diluted by the receiving stream will be evaluated using acute
methods and low dilution discharges will be evaluated using chronic tests.
Discharge samples used by biomonitoring analysis will consist of flow weighted
composite samples of all dry weather flows discharged into a receiving stream.

Where ambient toxicity is identified as a problem the Region will proceed with
validation, limits, or toxicity reduction evaluation (TRE) requirements as soon
as possible.
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                                                          EB& REGION VII
U.S. Environmental Protection Agency, Region VII
Water Management Division
726 Minnesota Avenue
Kansas City, KS  66101

Mr. Don Toensing, Chief
Permits Compliance Section
(913) 236-2817
Region VII is in the process of drafting regional policy relating to the
management of toxic pollutants; the policy is expected to be completed within the
next few weeks.  Draft policy will be discussed with the applicable states, and
it is expected that the ultimate effect will be to increase the degree of
toxicity testing in state NPDES programs.  Currently the states within the Region
have not embraced the concept of effluent toxicity testing.

The Region has tested about 35 facility effluents with daphnid and fathead minnow
acute static toxicity tests and expects to continue this program at 30 facilities
within the next year.  Testing is done via a contract with the University
Hygienic Laboratory in Iowa City, Iowa.  The Region does not have in-house
biological testing capabilities.

The Region continues its Regional Ambient Fish Tissue analyses program at a
significant number of locations in each of the states.  Principal analyses are
for pesticides and some metals.  Chlordane and PCBs have occurred at levels
higher than those recommended for edible flesh in some areas; the source of such
concentrations has not been identified.
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                                                              REGION VTIT
U.S. Environmental Protection Agency, Region VIII
One Denver Place
999 18th Street, Suite 1300
Denver, GO  80202-2413

Mr. James Lazorchak
Water Management Division
(303) 293-1581
A regional policy has been distributed that addresses an NPDES whole effluent
toxics control program.  The policy provides that all major permits must require
two-species testing, completion of a toxicity reduction evaluation if toxicity is
determined, and an appropriate limitation of whole effluent toxicity after
approximately 3 years.  The definition of when chronic or acute toxicity is
"demonstrated" is left to the permit issuing authority.

The testing protocol requires two species acute tests when receiving water low
flow dilution is greater than 100:1, and two species chronic tests when receiving
water low flow dilution is less than 100:1.  Quarterly testing or monthly testing
is governed by the volume of facility discharge; at greater than 20 HOD for a
PO1W or 10 HOD for an industry, monthly testing is required.
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                                                          EPA REGION IX
U.S. Environmental Protection Agency, Region IX
Water Management Division
215 Fremont Street
San Francisco, CA  94105

Mr. Phil Woods
Water Quality Standards Coordinator
(415) 974-8307
A regional policy on biological toxicity testing is in the final stages of
development.  Region IX is pushing toward implementation of the Technical Support
Document for Water Quality-Based Toxics Control but, as yet, has not universally
translated such concepts into permit language for all permit reissuances.  Water
quality standards of states within the region generally express the mandate that
the survival of aquatic life in surface waters subjected to a waste discharge or
other controllable water quality factors shall not be less than that for the same
water body in areas unaffected by the waste discharge as determined, at a
minimum, by use of a 96-hour toxicity test.

Many discharges are to the region's marine and tropical waters.  Guidance
provided in current toxicity testing procedural documents is believed to be less
than adequate in recommending methodology and test organism species for such
waters.

In early February, 1986, Region IX issued a letter to each of the states in the
region that directed them to begin to fully develop the effluent toxicity data
base directed for water bodies where aquatic toxicity problems have been
documented or are suspected and to readjust permit requirements accordingly.
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                                                          EPA REGION X
U.S. Environmental Protection Agency, Region X
1200 Sixth Avenue
Seattle, WA  98101

Mr. Roger Mochnick, Chief             Ms. Amber Wong
Permits Section - Water Div.          Permits Section - Water Div.
(206) 442-4817                        (206) 442-1647
Region X has been requiring biological toxicity testing in all major and new
industrial and municipal permits, in municipal permits where pretreatment