EPA-600/2-76-241
October 1976 Environmental Protection Technology Series
AOUTE TOXIC EFFECTS OF
PETROLEUM REFINERY WASEWAIERS
ON
Robert S. Kerr Environmental Research Laboratory
ifftee «l Researcfc
-------
RESEARCH REPORTING SERIES
Research reports of the Office of Research and Development, U.S. Environmental
Protection Agency, have been grouped into five series. These five broad
categories were established to facilitate further development and application of
environmental technology. Elimination of traditional grouping was consciously
planned to foster technology transfer and1 a maximum interface in related fields.
The five series are:
1. Environmental Health Effects Research
2. Environmental Protection Technology
3. Ecological Research
4. Environmental Monitoring
5. Socioeconomic Environmental Studies
This report has been assigned to the ENVIRONMENTAL PROTECTION
TECHNOLOGY series. This series describes research performed to develop and
demonstrate instrumentation, equipment, and methodology to repair or prevent
environmental degradation from point and non-point sources of pollution. This
work provides the new or improved technology required for the control and
treatment of pollution sources to meet environmental quality standards.
This document is available to the public through the National Technical Informa-
tion Service, Springfield, Virginia 22161.
-------
EPA-600/2-76-241
October 1976
ACUTE TOXIC EFFECTS OF PETROLEUM REFINERY
WASTEWATERS ON REDEAR SUNFISH
by
John E. Matthews
Leon H. Myers
Robert S. Kerr Environmental Research Laboratory
Ada, Oklahoma 74820
ROBERT S. KERR ENVIRONMENTAL RESEARCH LABORATORY
OFFICE OF RESEARCH AND DEVELOPMENT
U.S. ENVIRONMENTAL PROTECTION AGENCY
ADA, OKLAHOMA 74820
, • ->;
, T /8 F J /'i
..,^'.,,,, iioy/ii iwyw
-------
DISCLAIMER
This report has been reviewed by the Robert S. Kerr Environmental
Research Laboratory, U.S. Environmental Protection Agency, and approved
for publication. Mention of trade names or commercial products does
not constitute endorsement or recommendation for use.
11
-------
ABSTRACT
Static bioassays of 24 hours' duration were performed on samples of
wastewaters provided by 22 domestic petroleum refiners. These waste-
waters! represent three types of water discharges prevalent to this
industry: process wastewaters prior to dilution with other streams;
API separator effluents which are a conjugate of various streams within
a refinery; and wastewaters following treatment by activated sludge
systems.
Bioassays were performed using redear sunfish (Lepomis microlophus)
as test organisms. Selection of this species was due to its being a
member of the important freshwater family, Centrarchidae, its wide-
spread distribution, and the availability of a plentiful supply with
a controlled background.
Twenty-four hour 50 percent tolerance limits (TL ) of the various
wastewaters are compared with results of chemical analyses performed
during the same study. Toxicity varied considerably both between
refineries and for waste streams from within a single refinery. Results
of these analyses and observed behavioral symptoms of distressed fish
revealed that ammonia, sulfides, and phenolics, alone or in combination,
were major contributors to toxicity exerted in most samples. Three
refineries had samples which were more toxic than anticipated based
on results of chemical analyses, indicating the presence of other toxic
compounds in unknown quantities; e.g., various hydrocarbons.
Results of bioassays of samples from various wastewater treatment stages
of five refineries with activated sludge treatment systems revealed
a decrease in toxicity as the degree of treatment increased indicating
a reduction in concentration of contributing biodegradeable toxicants.
This report was submitted in fulfillment of ROAP 21 AZP, Task 050 by
the Robert S. Kerr Environmental Research Laboratory of the U. S.
Environmental Protection Agency. Work was completed as of January 1974.
111
-------
CONTENTS
Sections Page
I Conclusions 1
II Recommendations 3
III Introduction 4
IV Methods 7
V Results 11
VI Discussion 20
VII References 29
VIII Appendices 31
-------
ACKNOWLEDGMENTS
The U. S. Environmental Protection Agency, Robert S. Kerr Environmental
Research Laboratory (RSKERL), gratefully acknowledges the cooperation
of the Bureau of Sports Fisheries and Wildlife, National Fish Hatchery,
Tishomingo, Oklahoma, in providing experimental organisms for use in
this project.
The following people at the RSKERL are recognized for their assistance
in conducting these experiments: Tommy Redman, James McNabb, Bob Newport,
Kenneth Jackson, and Roger Cosby.
Particular appreciation is extended to Dr. William A. Brungs, National
Water Quality Laboratory, Duluth, Minnesota; Mr. David N. Peters,
Region VI, Dallas, Texas; and Mr. Marvin Wood, RSKERL, for their
consultation and suggestions during the study and review of the
experimental results.
VI
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SECTION I
CONCLUSIONS
Short-term static bioassays of 24-hours1 duration can be an effective
tool for screening industrial process wastewaters to locate sources of
toxic agents; these tests can also be used to evaluate effectiveness
of industrial waste treatment processes.
Static bioassays cannot be used to obtain reliable 50 percent tolerance
limit (TL™) values for low toxicity wastes that exert a high oxygen
O \J
demand although a range can often be established by an experienced
observer. Dissolved oxygen (DO) usually becomes critical at about 12
hours; low DO plays a major role in a mortallity after this time.
Activated sludge treatment processes tend to reduce the oxygen demand
beyond the critical stage.
Raw wastewaters from different oil refineries vary greatly in their
toxic characteristics; wastewaters from different processes within a
single refinery also vary greatly in their toxic characteristics.
Toxicity of oil refinery wastewaters varies considerably at different
treatment stages in the activated sludge process; toxicity appears to
decrease following each stage of treatment.
Toxicity of oil refinery wastewaters cannot always be predicted from
results of chemical analyses; the toxic effect of the waste is dependent
on the synergistic or antagonistic activity of toxicants present.
The most common toxic constituents of untreated oil refinery waste-
waters are: ammonia, sulfides, phenolic compounds, and cyanides. Raw
wastewaters also may contain other toxic compounds including various
hydrocarbons.
-------
Due to the volatile and unstable nature of some toxic components of
oil refinery wastewaters which may have led to a reduction in concen-
trations during sample transportation, storage, and handling, TL,_0
values obtained during these tests may be higher than the actual value.
Acute toxic effects of raw wastewaters from oil refineries are generally
exerted within the first 12 hours of the static test; therefore, a
24-hour test will provide good positive results under static conditions.
If samples containing toxic compounds, other than those mentioned above,
toxic effects may be exerted over an extended period. Tests should
then be continued for at least 48 hours to obtain more positive results.
Toxicity of the final clarifier effluent from oil refineries with
activated sludge treatment systems is dependent on toxic constituents
present in the influent and their concentrations. Results of chemical
analyses conducted during these tests indicate that concentrations of
most toxicants other than ammonia are reduced by activated sludge treat-
ment systems and at least a four-fold decrease in toxicity can be ex-
pected after treatment.
Although activated sludge treatment systems have very little effect
on ammonia concentrations, the toxic effect of ammonia is lessened
due to a decrease in pH of the treated effluent.
-------
SECTION II
RECOMMENDATIONS
With the many and varied processes employed by oil refineries in the
refining of crude oil, the results of this study and others indicate
the need to further document the effect of treated refinery wastewaters
and individual wastewater constituents on fish and other aquatic life
in receiving waters under a wide range of environmental conditions.
This project was conducted to determine short-term acute toxic effects
of refinery wastewaters on a single fish species. A research study
to investigate long-term cumulative effects of treated refinery waste-
waters on fish and other aquatic life should be conducted.
The oil refining industry should be encouraged to use short-term static
bioassays to assist in their pollution control effort. Some of the
ways in which this type of bioassays can be used are:
1. Acute toxicity of final effluents can be determined.
2. Toxicity of individual process wastewaters can be determined.
These effluents, if highly toxic, may be modified, eliminated,
or treated.
3. Location of processes which contribute high toxicity waste-
waters make possible alternative methods of controlling the
potential problem. The smaller quantities of wastewater
may be pretreated to reduce toxicity or stored with regulated
release into the refinery wastewater treatment system.
4. Leaks, spills, or other losses of toxic compounds into waste
streams can be detected, located, and remedied.
5. The effectiveness of treatment systems on removing toxic
compounds can be established.
-------
SECTION III
INTRODUCTION
Industrial wastes may be detrimental to aquatic life by direct toxic
action or by altering natural qualities of the receiving water. Petro-
leum refinery wastewaters can be characterized as having a high bio-
chemical oxygen demand (BOD) and significant toxicity.1 Adverse effects
from acute concentrations of refinery effluents during several days of
2
exposure have been reported. Experiments with five species of fish
have shown that the emulsified effluent from American Petroleum Insti-
tute's (API) gravity oil separators is toxic to fish unless diluted
or otherwise treated. Treatment of refinery effluents to reduce tox-
icity also results in improvement in other objectionable characteristics
including oil content, oxygen demand, chemical composition, turbidity,
4
and odor.
products and wastes from oil refineries are extremely complex in their
composition and their effects on water quality. While it is recognized
that each particular oil refinery poses its individual problems, certain
pollutants are common to all refineries. Process wastewaters are high
in phenolics, ammonia, and hydrogen sulfide and pose the primary water
pollution problem. All of these compounds are relatively high in
toxicity to aquatic life. In addition to these common constituents,
process wastewaters may contain various hydrocarbons toxic to aquatic
life. ' Findings during a cooperative American Petroleum Institute/
Environmental Protection Agency (API/EPA) waste characterization study
of petroleum refinery wastewaters conducted at the Robert S. Kerr
Environmental Research Laboratory (RSKERL) concurrently with this
-------
project indicate that these wastewaters may contain a large number
of substances which if discharged without treatment into receiving
g
waters could pose a serious threat to fish and other aquatic life.
It was also found, however, that refinery treatment processes sub-
stantially reduce the acute toxic effect of these substances.
Refinery wastewaters often contain a number of these different toxi-
cants which, when mixed or under the influence of other characteristics
of the effluent or receiving water, may produce an entirely different
toxicity level from that of the pure components. Results of chemical
analyses for possible toxic components compared with toxicity levels
reported in the literature for each particular component can not always
be expected to produce satisfactory answers.
The concentration of these pollutants in a particular waste stream is
dependent on the process from whichT the stream flows as well as in-
process procedures for reduction of a potential troublesome pollutant.
The concentration in the final effluent is dependent on the type and
efficiency of treatment to which the wastewater is subjected.
The use of some type of bioassay to determine the toxicity of a complex
wastewater can be the most effective and accurate method of assessing
potential danger. The bioassay provides a direct satisfactory method
for evaluating acute toxicity of chemically complex wastes. Toxicity
determinations rather than complete reliance on BOD and chemical
determinations are necessary for the safe disposal of complex wastes.
Two phases of a study of oil refinery wastewaters were conducted at
the RSKERL during 1972. During" Phase I of the study, samples were
collected by EPA and API personnel from in-plant wastewaters at 17
selected refineries located throughout the United States and shipped
to the RSKERL for analysis of 28 parameters. A single set of samples
was received for each of the refineries. Samples were preserved in
tightly capped cubitainers and refrigerated at 4° C until tested. The
-------
number of sample points tested for each refinery ranged from 1 to 13.
During Phase II of the study, samples were collected from various stages
of the activated sludge treatment process at five selected oil refineries.
Samples were collected for 14 consecutive days at each refinery.
On March 28, 1972, a series of 24-hour static bioassays were started
at the RSKERL to screen samples collected during Phase I of the study.
The purpose of these tests were (1) to evaluate and compare toxicity
of in-plant wastewaters of selected oil refineries throughout the
United States; (2) to compare actual toxicity results with anticipated
toxic action derived from results of chemical analyses of the samples
for potential toxicants normally contained in process wastewaters;
(3) to check for the presence of unknown toxic compounds; and (4) to
observe and catalog behavioral activities of fish exposed to these
wastewaters.
On May 10, 1972, a series of 24-hour static bioassays were started on
samples collected during Phase II of the study. Two or three bioassays
were conducted on composites of the 14 samples collected at each loca-
tion. The purpose of these tests were (1) to evaluate and compare
toxicity at different stages of treatment of selected refineries with
activated sludge treatment systems; and (2) to evaluate the efficiency
of activated sludge treatment for removal of toxic constituents from
refinery wastewaters.
-------
SECTION IV
METHODS
Bioassay procedures used were modified from those described in Standard
9
Methods for the Examination of Water and Wastewater, 13th Edition.
Modifications were necessary because of the small volume of sample
available for use, the number of samples to be tested, and the resources
available. Test procedures outlined in this paper can be expected to
measure short-term effects only. The long-term effect is not measured.
Thus, there may be a cumulative effect of the waste over a longer
period of time.
A series of three to five dilutions and a control were set up for each
sample or composite tested. Test concentrations were prepared on a
percent-by-volume basis in the diluent water. Two, one-gallon glass
jars containing three and one-half liters in each or one, five-gallon
glass jar containing ten liters were used per dilution. Five test
organisms were placed in each jar after the necessary volume of sample
was mixed with the diluent water. This gave either ten fish in seven
liters per dilution or five fish in ten liters per dilution. The
latter was used to try to alleviate problems of low oxygen in some
of the tests and to ensure that stress from crowded conditions did
not affect the test results.
Test organisms used were redear sunfish (Lepomis microlophus) that
were 20-50 millimeters in length and weighing 0.5-1.5 gram each.
Selection of this species was due to its being a member of the
important family Centrarchidae, its widespread distribution, and
the availability of a plentiful supply with a controlled background.
This species was easily maintained under laboratory conditions and
-------
proved highly satisfactory for conducting short-term static bioassays.
Fish were obtained from the National Fish Hatchery, Tishomingo, Oklahoma,
as needed. Fish were held in stainless steel holding tanks at a
constant temperature and saturated dissolved oxygen 16vel until used.
Only fish that appeared to be in excellent condition were used in the
testing.
The dilution range selected for each sample was based on chemical data
for that sample compared with reported toxicity values for measured
chemical constituents. If fish died in all dilutions for a test, it
was assumed that unknown toxic agents were present in the sample and
lower series of dilutions were tested until a TL^ value was obtained.
Logrithmic or geometric series of dilutions were used in all tests.
Dilution water in all tests was tap water from the Ada, Oklahoma,
water supply system (pH 8.3-8.7, Hardness 300-350 mg/1 as CaCO_).
O
Use of a common dilution water made it possible to compare toxicity
levels of samples from different refineries. Dilution water in each
test container was aerated to saturation prior to adding the sample
and test organisms. Aeration was not used during the test period
because of the volatile nature of many of the constituents of the
wastewaters.
Preliminary tests indicated that most toxicity of oil refinery wastes
in static tests is exerted during the first 12 hours. This may be
due to the volatile nature of many of the toxic constituents. Per-
sonal communication with other scientist who have tested similar
wastes confirmed this. ' Therefore, 24 hours was chosen as the
test period. The 24-hour test period tended to eliminate low DO as
a contributor to death in latter stages of the tests during Phase I.
Low DO during the latter stages was still a problem during Phase II
because of the higher waste volumes used in the tests.
-------
Bioassay record sheets were maintained for each sample tested. Obser-
vations and selected analysis were made initially at six hours, twelve
hours and twenty-four hours. Survival percentage was recorded at
each dilution. The 24-hour survival percentage for each dilution
was plotted on semi-log paper and a straight line connected between
successive dilution percentages at which greater than and less than
50 percent survival occurred (Figure 1). The 24-hour TL^ value
(percentage dilution at which 50 percent of test organisms survived
for 24 hours under conditions of the test) was then interpolated from
the straight line. All TL^ values were calculated in terms of the
initial percentage volume of wastewater; undoubtedly, effective concen-
trations of the various toxic constituents became lower with time.
If more than 10 percent of the control organisms had died during a
test, results for that test would have been invalidated.
Within test precision was checked by plotting survival percentages
and establishing a TL5Q individually for the two series of test
containers used for each test (Table 1 and Figure 1).
Table 1. WITHIN TEST CHECK OF PRECISION
Dilution
Test Containers
A
B
Total
10%
5
5
10
5.6%
5
5
10
3.2%
5
5
10
1.8%
5
5
10
1.0%
5
5
10
Control
5
5
10
-------
1000
800
600
400
200
100(5)
80
60
40
20
10
0
Log
Scale
1000 Sample No. 6992:RW-5
Starting Date: 5-29-72
Hour: 9:00 a.m.
Final
Results:
rL50
Time Intervals
A-24
0.042
B-24
0.045
24 hr
0.043
320
Concentrations expressed as percent
0.1% -> 0.01%
180
0.100
0.056
0.018
OOIO
50
Percent
Survival
Test species: Redear Sunfish
Temperature range: 21.5°-25°C
Dilution water source:
Ada City Water
Notes: WilH
t;wi mmi no
and loss of equilibrium noted
at 5 minutes in 0.1% dilution.
All fish dead in 50 minutes.
Eight fish dead in 2 hours in
in 0.056% dilution. Two live
fish in obvious distress.
Tests continued for 48 hours
with no change in TL value.
John E. Matthews
100
Bioassay
Concentrations
A-Front row of test containers
B-Back row of test containers
Figure 1. Bioassay record sheet
10
-------
SECTION V
RESULTS
PHASE I
Bioassays were conducted on 50 samples of in-plant wastewaters from
17 oil refineries during this phase of the study. The 24-hour TL5Q
results are presented in Table 2. Because only a limited amount of
each sample was available for use, 24-hour TL5Q values were not obtained
for all samples tested. The 24-hour TLg for those samples exhibiting
greater than 50 percent survival in all dilutions tested was reported
as greater than the highest dilution tested.
A 24-hour TL™ ranging from 0.04%-13.5% was established for 20 samples
from eight refineries under the conditions of these tests. Bioassay
data for the toxic samples are presented in Appendix A. No 24-hour
TL™ could be established for the remaining 30 samples. Nine refineries
had no samples which exhibited toxicity under these conditions at the
dilutions tested.
Three refineries had one or more samples toxic at <0.3%. These same
refineries also had at least one other sample toxic at <5%. Three
additional refineries had one or more samples toxic at <5%. The
remaining two refineries for which a 24-hour TL,.., was established
had single in-plant wastewater streams with a TL,-n of 7.5% and 13.5%.
Three refineries had one or more samples which were considerably more
toxic than indicated by results of chemical analyses. Toxicity of
these samples was most likely due to the presence of toxic hydrocarbon
compounds. The TL,_n for one sample was attributed to low DO and one
sample to low pH.
11
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Table 2. BIOASSAY RESULTS - PHASE I
Sample
6992 RW-1
11 RW-2
" RW-3
11 RW-4
11 RW-5
11 RW-6
11 RW-7
11 RW-8
" RW-9
" RW-10
11 RW-11
" RW-12
" RW-13
59991 RW-1
69991 RW-1
693 RW-1
" RW-2
" RW-3
69995 RW-1
11 RW-2
22 RW-1
" RW-2
69994 RW-1
" RW-2
6993 RW-1
24-hour TL5Q
(% by volume) Comment
>10
0.042 Same at 48 hours
0.043 Same at 48 hours
>10
>10
>18
>15
>10
>18
0.05 Same at 48 hours
3.8 3.2% at 48 hours
>10
>10
7.5
13.5
0.23a .08% at 36 hours
1.8
7.5 Low DO
>10
>10
>18
>18
4.5
1.9 Low pH
>10
12
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Table 2 (Continued). BIOASSAY RESULTS - PHASE I
Sample
595
it
6995
ti
it
u
II
II
II
It
II
It
92
u
59
59993
u
106
56
67
6996
u
u
u
ti
RW-1
RW-2
RW-1
RW-2
RW-3
RW-4
RW-5
RW-6
RW-7
RW-8
RW-9
RW-10
RW-1
RW-2
RW-1
RW-1
RW-2
RW-1
RW-1
RW-1
RW-1
RW-2
RW-3
RW-4
RW-5
24-hour TL
(% by volume) Comment
10
18
6.5
18
13 Distress
13,5 Distress
4.2
7.2
14
16
15
18
10
2.4
10
10
10
18
10
10
10
10
4.2 2.4% at 48 hours
4.2
0.05
Toxicity greater than indicated by results of chemical analyses.
13
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Estimated concentrations at the TL^ value were made for common toxic
constituents found in refinery wastewaters for purposes of identifying
the most likely toxic agent(s) in each sample (Table 3). These esti-
mations were made using results of analyses performed on corresponding
water samples. Results upon which these estimations are based are
presented in Appendix C. These estimations cannot be assumed to be
actual concentrations present at the measured TL • however, analyses
of water samples from some of the bioassay test containers after 24
hours indicate that the estimated values are close enough approximations
of the real values to use for the purposes intended.
These projected data along with the actual measurements confirm that
ammonia, sulfides, and phenolic compounds are the major contributing
toxic constituents present in oil refinery wastewaters. Fourteen of
the samples for which a TL,_n was established during this study con-
tained these constituents individually, or in combination, at concen-
trations reported in the literature to be toxic to fish and aquatic
life. ' ' Individual or contributing toxic levels of ammonia
was estimated in 12 samples, sulfide in nine samples, and phenolics
in eight samples. Five samples did not contain sufficient concentra-
tions of these three constituents, individually or in combination, to
account for the toxic level found. It is assumed, therefore, that
toxicity of these five samples was due to the presence of unknown toxic
compounds, e.g., hydrocarbons.
The behavior of distressed and dying fish is often an indication of
the type or category of the agent(s) responsible for death. ' ' '
Continuous observations were made during these tests and the exhibited
symptoms noted. Observed symptom were compared with behavioral symptoms
reported in the literature for constituents suspected to be present.
During this series of tests, distressed and dying fish exhibited four
different symptom patterns:
14
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Table 3. ESTIMATED INITIAL CONCENTRATIONS OF COMMON TOXICANTS
AT 24-HOUR TL PERCENTAGE VOLUME (mg/1)
O \J
Refinery/Sample
No.
6992
it
ii
ii
59991
69991
693
it
69994
69995
ii
ii
it
ii
92
6996
ii
ii
RW-2
RW-3
RW-10
RW-11
RW-1
RW-1
RW-1
RW-2
RW-1
RW-1
RW-3
RW-4
RW-5
RW-6
RW-2
RW-3
RW-4
RW-5
Ammonia
1.50
0.10
1.68
3.61
8.85
5.40
0.22
3.58
5.63
5.27
0.17
0.20
5.04
0.06
2.04
0.22
14.7
1.35
Sulfide
1.67
0.02
1.92
4.90
6.30
1.89
0.12
0.20
0.86
7.15
0.05
0.12
3.28
0.01
0.08
0.06
-
1.50
Phenol Cyanide
0.22 0.003
6.24 0.007
0.25 0.001
0.05 0.013
0.57 0.056
0.39 0.19
0.15 0.001
0.01 0.003
0.20 0.017
0.35
0.07
0.07
2.60 0.004
0.03
3.36
0.13
-
0.21 0.061
pH
8.6
8.6
8.7
8.5
8.7
8.2
8.4
9.1
8.9
9.0
8.6
8.6
8.9
8.4
9.0
7.8
8.3
8.6
TL50
% by Vol
0.042
0.043
0.05
3.8
7.5
13.5
0.23
1.8
4.5
6.5
13
13.5
4.2
7.2
2.4
4.2
4.2
0.05
Measured Concentrations
6992
693
it
69994
69995
92
RW-2
RW-1
RW-2
RW-1
RW-1
RW-2
2.40
3.55
5.40
5.85
3.95
1.00
0.20
2.71
15
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1. Wild erratic swimming; loss of equilibrium with periodic
spurts; death very shortly with gills swollen (ammonia, low
pH,- some phenolic compounds).
2. Wild erratic swimming; loss of equilibrium; lethargic condi-
tion; highly irritable; death after long period (petrochemicals,
phenolics).
3. Some hyperactivity prior to loss of equilibrium; no irritation;
state of anaesthesia for long period prior to death; gills
swollen (cyanides, phenols, sulfides).
4. Slow circular swimming at surface; irritable; quick spurts
from lethargic condition; state of anaesthesia on bottom
for long period prior to death; gills swollen (petrochemicals,
phenols, cyanides, sulfides).
The two most common patterns were those in numbers 1 and 4.
PHASE II
During Phase II, 30 bioassays were conducted on composite samples
collected from various stages of the treatment systems of five oil
refineries (Figure 2). A total of 12 samples were tested. Two or
three bioassays were run on each sample. Results of these toxicity
tests are tabulated in Table 4. Low dissolved oxygen in four of the
tests prevented establishment of reliable 24-hour TL-,, values for
DU
those samples; hence, 12-hour TL5Q values for each sample have also
been included in Table 4. Bioassay data for each sample are presented
in Appendix B. Results of chemical analyses of samples collected
from the same sample points concurrent with bioassay samples are pre-
sented in Appendix C.
Toxicity values remained relatively constant for different composites
from the same sample location. API separator effluent samples from
the five refineries exhibited some toxicity. The 24-hour TL_0 of
these samples from the five refineries varied from 4%-50%. Activated
16
-------
Refinery No. 9973
Process
Water
API _(S)
Separator
Equalization
Aeration
Basin
Final (S)
Clarifier
Refinery No. 2115
Oily ^_ API (S)^ Primary
Water Separator Clarifier
Trickling (S) Aeration
Filter Basin
Final (S)
Clarifier
Refinery No. 0288
Oily
Water'
API
Separator
Air (S) Aeration
Floatation
Basin
Final (S)
Clarifier
Refinery No. 6512
All Waste
Water
API
_ Equalization ^ . Chemical (S) Final (S)
Separator M Coagulation riai-i-p-ioT.
„, ....
Clarifier
Refinery No. 6693
All Waste
Water
API (S) Aeration
Separator Basin
Final (S)
Clarifier
(S) Denotes sample point
Figure 2. Sample points from activated sludge refinery wastewater systems
chosen for toxicity tests
-------
Table 4. BIOASSAY RESULTS - PHASE II
Composite Samples
24-hr. TL5Q
Sample Location
Refinery 9973
API Separator
Final Clarifier
ABC
(% Dilution)
h h
29° 19°
>56 >88
h
14°
100
12-hr. TL5Q
ABC
(% Dilution)
42
34 41
Estimated 24- Estimated NH
hr. TL5Q Range Cone, at TL5Qa
(% Dilution) (mg/1)
34-42 3.9 (5.1-3.1)
100 12.5
No mortality or stress
Refinery 2115
API Separator
Trickling Filter
Final Clarifier
Refinery 6512
Equalization
Chem. Coag.
Final Clarifier
No
Refinery 0288
Air Floatation
Final Clarifier
No
Refinery 6693
API Separator
Final Clarifier
No
4.2 4.2
12.5 16
18 19.5
h h
12.5? 18?
n h
12.5° 28°
100 100
mortality or
h
10.5 50
>48 100
mortality or
24 18
>41 100
mortality or
3.5
11.5
20
-
-
-
stress
-
-
stress
22.5
-
stress
4.2
13.5
34
>56
100
Stress
h
34b
>32
4.2 3.7
16 13
35 >28
V,
42b
60
>56
Stress
_
41 22.5
3.3-4.2 4.7 (6.4-3.8)
11-16 12.8 (20.0-10.0)
18-20 19.8 (33.0-9.5)
32-56
42-60
100
24-56
100
18-24
100
values in parenthesis are measured concentrations (at 24 hrs) in test containers bracketing the
interpolated TL,.n value.
i
Low dissolved oxygen contributed to deaths in lower dilutions.
-------
sludge systems of all refineries samples reduced toxicity. Final
clarifier effluents from four refineries illicited no mortality or
stress symptoms during the test period. The API separator effluent
from refinery 2115 exerted a 24-hour TL of 4.2%-3.5% for three
composite samples; corresponding values for the final clarifier effluent
were 18%, 19.5%, and 20%. The major contributing toxic agent in this
refinery is ammonia. Since the concentration of ammonia was essentially
the same at both locations, the decrease in toxicity is most likely
due to removal of contributing biodegradeable toxicants as well as
a decrease in pH of the wastewater as it passed through the treatment
system.
Observations were made of the behavioral patterns of distressed fish.
These observations revealed the same four sypmtom patterns as those
noted in Phase I of this study.
19
-------
SECTION VI
DISCUSSION
PHASE I
Bioassay tests discussed herein were made using untreated process waste-
waters from 17 selected oil refineries. Toxic characteristics of
samples tested varied greatly between the 17 refineries. There is
also a wide variation in toxicity of samples from different processes
within an individual refinery. Results of the tests conducted and
observations made during the tests for each individual refinery are
discussed below.
Refinery 6992
Thirteen samples from this refinery were tested. Four of the samples
exerted a 24-hour TL5Q at dilutions tested: 0.042%, 0.043%, 0.05%,
and 3.8%. Tests were extended over 48 hours with no change in the
first three values and a reduction to 3.2% in the other test. The
DO did not drop below 4.0 mg/1 during the first 24 hours in any of
the tests; therefore, toxicity was due to wastewater constituents.
Most of the toxicity exerted by these samples is due to high concen-
trations of constituents normally found in oil refinery wastewaters
(ammonia, phenolic compounds, sulfide, cyanide) and was predictable
from results of chemical analyses.
Symptoms exhibited by distressed fish were either those described in
numbers 1 or 3 in Section V. These symptoms include those reported
for the most common toxic constituents of oil refinery wastewaters.
Refinery 59991
The sample tested from this refinery exerted a 24-hour TL^n of 7.5%.
*?u
The DO remained greater than 5.0 mg/1 throughout the test period;
20
-------
therefore, toxicity was attributed to constituents in the wastewater.
Symptoms of distressed fish were intermediate between numbers 1 and 4
in Section V, suggesting that the toxic agents were most likely ammonia
and sulfide in combination. The TL<-n value was predictable from results
of chemical analysis.
Refinery 69991
This sample exerted a 24-hour TLt_n of 13.5%. The DO remained greater
than 3.5 mg/1 throughout the test period; therefore, toxicity was
attributed to wastewater constituents. Symptoms of dying fish were
those listed in number 4 of Section V, suggesting that the toxic agents
were most likely ammonia, sulfide, and cyanide in combination. The
TL value was predictable from results of chemical analyses.
Refinery 693
Each of the three samples analyzed from this refinery exerted a 24-hour
TL_ of <10% although toxicity in sample RW-3 was most likely due to
low DO.
Sample RW-1 was much more toxic than predictable from results of chemical
analyses. Dilutions were first set up from 10%-1%. Fish in all five
dilutions became distressed immediately and were dead in less than two
hours. Symptoms were those stated in number 2 of Section V. The
sample had a definite creosote smell; therefore, the most likely toxic
agents was one of the highly toxic phenolic compounds. Additional
dilution series of 1%-0.1% and 0.1%-0.01% were set up to establish a
24-hour TL . In the 1%-0.1% series all test fish were in distress
within four hours with the same symptoms. A 24-hour TL of 0.24%
O \J
was established. Live fish in the remaining dilutions were in obvious
distress. Fish in 0.1% dilution were removed at the end of test period
and placed in clean aerated water. Symptoms appeared to be gone after
six hours. Aeration was stopped and fish observed for 24 hours. All
fish were alive and appeared healthy at the end of 24 hours.
21
-------
Observation time was extended in 0.1%-0.01% dilution series to 36
hours. All fish were alive but in obvious distress at the end of
24 hours. Dissolved oxygen was greater than 4 mg/1 at 24 hours but
had dropped to 2.5 mg/1 at 36 hours and tests were discontinued. The
36-hour TL of 0.05% may have been influenced by the low DO concen-
tration. All live fish were in distress at the end of 36 hours, but
fish at .018% and .010% did not become irritated as they did at higher
dilutions.
Sample RW-2 exerted a 24-hour TL5Q between 1.35%-2.40%. Insufficient
sample was available to set up a 1.8% dilution. Symptoms were those
listed in number 1 of Section V. The TL5 was predictable from the
high ammonia values in the raw sample.
Refinery 69995
A 24-hour TL for these two sample could not be established from the
dilutions tested and was reported as >10%. Some mortality and distress
we-re noted at 10% dilution for sample RW-1; however, low DO definitely
contributed to the mortality. Distress was observed prior to the DO
drop. All fish in RW-2 dilutions were alive and healthy at end of
test period.
Refinery 22
A 24-hour TL for these two samples could not be established from the
dilutions tested and was reported as >18%. Fish at 18% for sample
RW-2 were in distress at 12 hours, suggesting a 24-hour TL of 18%-
O \J
32%. Dissolved oxygen was >4 mg/1 at end of test.
Refinery 69994
Toxicity in sample RW-1 was predictable from results of chemical
analyses. All fish in 10% dilution were dead within 30 minutes with
symptoms described in number 1 of Section V, suggesting ammonia as
22
-------
the major toxic agent. Eight fish were dead in 5.6% dilution within
six hours; the two live fish remained in distress throughout the test.
Dissolved oxygen was >4.0 mg/1 at end of test.
Toxicity for sample RW-2 was mostly due to low pH (raw sample pH 0.9).
All fish in 10% dilution were dead in five minutes with visable bleeding
from gills (pH 2.1). All fish in 5.6% dilution were dead in 25 minutes
and seven fish in 3.2% dilution were dead in 30 minutes (pH 2.6 and 3.25)
One fish was alive in 3.2% dilution at end of test (pH 6.2 at 24 hours).
All fish were alive and in no distress in 1% dilution (pH 7.1).
Since lack of sufficient sample prohibited a set-up at 1.8% dilution,
the 24-hour TL_n was reported to be between 1.35%- 2. 50%.
^u
Refinery 6993
No toxicity was exerted or distress observed at dilutions tested (TL5
Refinery 595
No toxicity exerted or distress symptoms observed during test period
at dilutions tested for sample RW-1 (TL5Q >10%) . Sample RW-2 exerted
no toxicity during 24 hours (TL >18%) . Fish in 18% dilution were
in obvious distress at surface throughout the test and in 10% dilution
after 12 hours, suggesting a 24-hour TL for 18%-32%. Dissolved
oxygen was low at 24 hours .
Refinery 6995
A 24-hour TL was established for five of 10 samples tested. Three
of the toxic samples, RW-3, RW-4, and RW-6, exerted toxicity greater
than that predictable from results of chemical analyses. All live
fish for these three samples were in distress at the end of 24 hours.
Symptoms for all three samples were those listed in number 4 of Section
V. Toxicity of these samples was most likely due to toxic hydrocarbon
compounds. A plasticizer smell was noted.
23
-------
Mortality for samples RW-1 and RW-5 was predictable from results of
chemical analyses. Symptoms exhibited were those listed in numbers 1
and 4 of Section V. The most likely toxic agents were combinations
of ammonia and sulfide in RW-1 and ammonia, sulfide, and phenolics in
RW-5. Low DO may have contributed to mortality in 5.6% dilution for
sample RW-1.
Fish in samples RW-2, RW-7, RW-9, and RW-10 exhibited no stress during
the test period at the dilutions tested. Two fish died in samples
RW-8 in 18% dilution and remaining fish were in obvious distress after
12 hours. Since DO remained >4.5 mg/1 throughout the test, a TL n of
J\J
between 18%-32% could be predicted for this sample.
Refinery 92
Samples RW-1 exerted no toxicity during the test period at the dilutions
tested. Fish in 10% and 5.6% dilutions were in distress at surface at
end of test due primarily to low DO.
Sample RW-2 exerted a TL5Q of 2.4% for the test period with the DO
remaining greater than 5.0 mg/1. Distress was noted in 1.8% dilution
after 12 hours. Symptoms of distressed fish were those listed in
number 1 in Section V. Toxicity in this sample was predictable from
results of chemical analyses (phenol 144 mg/1; NH_ 85 mg/1).
J
Refinery 59
Although a TL™ could not be established at the dilutions tested
(TL^f, >10%), all fish exhibited an abnormal behavior after 12 hours.
Dissolved oxygen remained adequate throughout the test.
Refinery 59995
No toxicity was exerted or distress symptoms exhibited at dilutions
tested during the test period for these two samples (TL >10%).
24
-------
Refinery 106
No toxicity was exerted during test period at dilutions tested. Fish
in 18% and 10% dilutions exhibited distress symptoms at end of test
period primarily due to low DO (TL,.,, >18%) .
Refinery 56
No toxicity was exerted during the test period at the dilutions tested;
however, fish at 10% and 5.6% dilutions exhibited distress symptoms
after six hours (TL5Q >10%).
Refinery 67
No toxicity was exerted or distress symptoms exhibited during test
period at dilutions tested (TL5Q >10%).
Refinery 6996
Three of the five samples tested exerted a 24-hour TL,.,. at dilutions
tested. Sample RW-3 exerted toxicity much greater than indicated by
results of chemical analyses.
No toxicity was exerted or distress symptoms exhibited in sample RW-1
at dilutions tested (TL5_ >10%). Sample RW-2 exerted no toxicity
at dilutions tested (TV >10%); however, fish in the 10% dilution
were in distress at surface at 12 hours with DO >5.0 mg/1 throughout
the test period. A 24-hour TL5Q of 10%-18% is indicated.
Sample RW-3 exhibited symptoms described in number 4 of Section V.
Fish in 3.2% and 1.8% dilutions were in obvious distress at end of
24-hour period; no distress was observed in 1.0% dilution. Test was
extended through 48 hours with some mortality in 3.2% and 1.8%; fish
in 1.0% were not in distress at 48 hours. Dissolved oxygen remained
sufficiently high throughout the 48-hour period to have no effect on
results. The toxic agent is unknown but a hydrocarbon compound is
suspected.
25
-------
Fish in sample RW-4 exhibited symptoms described in number 3 of Section
V, while those in sample RW-5 exhibited symptoms described in number 1.
Dissolved oxygen remained >4.0 mg/1 in both tests throughout the test
period. Toxicity in both samples was predictable from results of
chemical analyses (RW-4: ammonia 355 mg/1; RW-5: ammonia 3,500 mg/1,
phenol 400 mg/1, sulfide 3,000 mg/1, cyanide 128 mg/1). With the
ammonia value of 355 mg/1, RW-4 predicted to be more toxic than the
bioassay indicated.
PHASE II
Bioassay tests discussed herein were conducted using samples from
various treatment stages of activated sludge treatment systems at
five selected oil refineries. Location of sample points in these
refineries are noted in Figure 2. Toxic characteristics of the samples
tested varied considerably between different oil refineries and between
different treatment stages within individual refineries. Toxicity
decreased at all selected refineries as the degree of treatment
increased. Results of the tests conducted and observations made
during the tests for each individual refinery are discussed below.
Refinery 9973
Effluent samples from the API separator and final clarifier were tested
from the treatment system of this refinery. Although low DO after
12 hours prevented establishment of a reliable 24-hour TL^_ for the
separator effluent, some toxicity was exerted. Twelve-hour TL 's
of 42%, 34%, and 41% for the three composites tested were obtained
prior to the DO drop. A rerun of a composite of all samples using
five fish in ten liters gave no mortality at 40%; therefore, the
24-hour TL for this sample is estimated to be approximately the
Ov/
same as the 12-hour value. Symptoms of dying fish were those listed
in number 2 in Section V. Death was most likely due to the combined
toxic action of constituents usually found in oil refinery wastewaters
26
-------
(ammonia, phenolics, sulfides, cyanides). No mortality was exerted
or distress symptoms observed in 24 hours in a 100% composite sample
of the final clarifier effluent.
Refinery 2115
Samples from effluents of all three stages of treatment exerted a
definite 24-hour TL™. Three composite samples for each stage of
treatment were tested with toxicity decreasing as the stage of treat-
ment progressed from approximately 4% for the API separator effluent
to approximately 20% for the final clarifier effluent. The high
ammonia content at all stages of treatment was most likely the major
contributing toxic agent, although symptoms of dying fish indicate
some other toxic agent or agents were present. Stress symptoms were
those listed in numbers 1 or 3 of Section V. Higher concentrations
of ammonia seemed to be necessary for the same level of toxicity as
the level of treatment progressed. A corresponding decrease in pH
as treatment progresses most likely influenced ammonia toxicity in
these tests since toxicity of ammonia increases with an increase in
PH.5' 10
Refinery 6512
Tests were run on two composite samples of effluents from each of three
stages of treatment. Effluent samples from the equalization and chemi-
cal coagulation units exhibited a demand that depleted the oxygen to
a level where test results are considered unreliable. Five fish were
used in ten liters of solution for the test of composite B. This did
not decrease the effect of low DO enough to make the results reliable.
Based on stress symptoms of test fish and early mortality during the
tests, the 24-hour TL,_0 is estimated to be between 32%-56% for equal-
ization unit samples and 42%-60% for chemical coagulation unit samples
discounting effects of low DO. Stress symptoms were those listed in
27
-------
number 4 of Section V. No mortality occurred or stress symptoms were
observed during the test period for the final clarifier effluent sample
in a 100% solution.
Refinery 0289
Tests were run on two composite samples for each of two stages of treat-
ment. Low DO in composite A for the air flotation unit sample prevented
establishment of a reliable TL . By using five fish in ten liters
of solution for composite B, a TL,-n of 50% was obtained. Since the
final DO was 2.5 mg/1 a range of 24%-56% is estimated for this sample.
No mortality occurred or stress symptoms were observed in a 100% sample
solution of the final clarifier effluent.
Refinery 6695
Tests were run on three composite samples of the API separator effluent
and two composites of the final clarifier effluent. The 24-hour TL^_
of approximately 21% (18%, 22.5%, and 24%) for the API separator
effluent was reduced to where no mortality occurred or stress symptoms
were observed in a 100% solution of the final clarifier effluent.
Stress symptoms in the API tests were a combination of those listed
in numbers 1 and 3 of Section V.
28
-------
SECTION VII
REFERENCES
1. Katz, M. The Effects of Pollution Upon Aquatic Life. In: Water
and Water Pollution Handbook, Volume I (L. L. Ciaccio, Editor).
Marcel Dekker, Inc., New York, N.Y. pp. 297-328. 1971.
2. Graham, R. J. and T. C. Dorris. Long-Term Toxicity Bioassay of
Oil Refinery Effluents. Water Research, Pergamon Press (London).
2_:643-663. 1968.
3. Clemens, H. P. and P. Summers. Will Refinery Wastes Kill Fish?
Petroleum Refining. 32_(8): 145-147. 1953.
4. Dorris, T. C., W. Gould, and C. R. Jenkins. Toxicity Bioassay
of Oil Refinery Effluents in Oklahoma. In: Biological Problems
in Water Pollution, 1959 Seminar Transactions. Robert A. Taft
Sanitary Engineering Center, Cincinnati, Ohio. Public Health
Service Technical Report W60-3. pp. 276-285. 1960.
5. McKinney, R. E. Biological Treatment Systems for Refinery Wastes.
Journal Water Pollution Control Federation. _39_(3) :346-359. 1967.
6. McKee, J. E. and H. W. Wolf (Editors). Water Quality Criteria.
California State Water Quality Control Board, Sacramento. Publi-
cation No. 3-A. pp. 229-232, 237-240. 1963.
7. Wilber C. G. Oil Pollution. In: The Biological Aspects of Water
Pollution. Thomas Publishing Co., Springfield, Illinois, pp. 73-92.
1969.
8. Petroleum-Organic Chemicals Wastes Section. National Petroleum
Refining Wastewater Characterization Studies. Robert S. Kerr
Environmental Research Laboratory, U. S. Environmental Protection
Agency, Ada, Oklahoma. (to be published)
29
-------
9. American Public Health Assocation. Standard Methods for the
Examination of Water and Wastewater, 13th Edition. American
Public Health Assocation, New York, N.Y. pp. 562-577. 1971.
10. Brungs, W. A. Personal Communication. National Water Quality
Laboratory, U. S. Environmental Protection Agency, Duluth,
Minnesota. 1972.
11. Burks, S. L. Personal Communication. Zoology Department, Oklahoma
State University, Stillwater. 1972.
12. Jones, J. R. E. Fish and River Pollution. Butterworths Inc.,
London, England, pp. 83-106, 142-152. 1964.
13. Turnbull, H., J. G. DeMann, and R. F. Weaton. Toxicity of Various
Refinery Materials to Fresh Water Fish. Industrial and Engineering
Chemistry. 4i6(2)-.324-333. 1954.
14. Pickering, Q. H., and C. Henderson. Acute Toxicity of Some Impor-
tant Petrochemicals to Fish. Journal Water Pollution Control
Federation. 38(9):1419-1429. 1966.
-------
SECTION VIII
APPENDICES
Page
A. Raw Data for Toxic Samples - Phase I 32
B. Raw Data - Phase II 40
C. Analytical Data - Phase I and Phase II 53
31
-------
APPENDIX A
RAW DATA FOR TOXIC SAMPLES - PHASE I
32
-------
Sample 6992:RW-2
TL5() . 0.042%
Cone . of
Waste
(% by vol.)
0.1
0.056
0.032
0.018
0.010
Control
Sample 6992:
Cone, of
Waste
(% by vol.)
0.1
0.056
0.032
0.018
0.010
Control
Sample 6992:
Cone . of
Waste
(% by vol.)
0.1
0.056
0.032
0.018
0.010
Control
No. of
Test No.
Animals at
10
10
10
10
10
10
RW-3 TL5() - 0
No. of
Test No.
Animals at
10
10
10
10
10
10
RW-10 TL5() -
No. of
Test No.
Animals at
10
10
10
10
10
10
Alive
24 hrs
0
2
8
9
10
10
.043
Alive
24 hrs
0
1
10
9
9
10
0.05%
Alive
24 hrs
0
4
9
8
9
10
D0a
(mg/1)
7.0
6.3
4.3
-
-
3.5
DO
(mg/1)
8.6
-
4.3
-
-
3.7
DO
(mg/1)
5.8
5.2
2.2
-
-
3.8
Temp.
(° C)
23
23
23
23
23
23
Temp.
(° C)
22
-
22
-
23
22
Temp.
(° C)
23
24
22
-
-
23
pH Sulfide
Start (mg/1)
8.6 1.2
8.6 1.1
8.3 0.9
8.4
8.6
8.6
pH
Start
-
-
8.6
-
-
8.65
PH
Start
8.7
-
-
-
-
8.6
aDO, temperature, sulfide, ammonia, and phenol measurements were taken
at 24 hours or at the death of all 10 fish.
33
-------
Sample 6992:RW-11 TL - 3.8%
O \J
Cone, of
Waste
(% by vol.)
5.6
3.2
1.8
1.0
X).56
Control
Sample 59991
Cone . of
Waste
(% by vol.)
10
5.6
3.2
1.8
1.0
Control
Sample 69991
Cone, of
Waste
(% by vol.)
18
10
5.6
3.2
1.8
Control
No. of
Test
Animals
10
10
10
10
10
10
: RW- 1 TL
No. of
Test
Animals
10
10
10
10
10
10
:RW-1 TL
No. of
Test
Animals
10
10
10
10
10
10
No. Alive
at 24 hrs
3
6
10
9
9
10
- 7.5%
No. Alive
at 24 hrs
0
10
10
10
10
10
5Q - 13.5%
No. Alive
at 24 hrs
0
10
10
10
10
10
D0a
(mg/1)
6.0
-
-
_
_
3.7
DO
(mg/1)
7.2
4.0
_
_
_
4.6
DO
(mg/1)
4.0
2.5
2.8
_
_
4.0
Temp.a
(° C)
23
_
_
_
_
23
Temp.
(° C)
23.5
24
_
_
_
24
Temp.
(° C)
24.5
24
24
_
_
24.5
pH
Start
8.5
8.5
_
_
_
8.7
pH
Start
8.7
8.6
_
_
_,
8.6
pH
Start
8.2
8.4
8.4
_
_
8.3
DO, temperature, sulfide, ammonia, and phenol measurements were taken
at 24 hours or at the death of all 10 fish.
34
-------
Sample 693:RW-1
TL5() - 0.23%
Cone, of
Waste
(% by vol.)
1.0
0.56
0.32
0.18
0.10
Control
No. of
Test
Animals
10
10
10
10
10
10
Sample 693:RW-2 TL5Q
Cone, of
Waste
(% by vol.)
10
5.6
3.2
1.8
1..0
Control
Sample 693
Cone, of
Waste
(% by vol.)
10.0
5.6
3.2
1.8
1.0
Control
No. of
Test
Animals
10
10
10
No set up
10
10
:RW-3 TL5()
No. of
Test
Animals
10
10
10
10
10
10
No. Alive
at 24 hrs
0
0
0
9
10
10
-1.8%
No. Alive
at 24 hrs
0
0
0
X
10
10
- 7.5%
No. Alive
at 24 hrs
0
10
10
10
10
10
D0a
(mg/1)
8.0
8.0
7.5
4.1
-
3.7
DO
(mg/1)
8.7
8.8
7.5
X
4.3
4.6
DO
(mg/1)
0.5
4.6
-
-
-
4.3
Temp.a
(° C)
21.5
22
23.5
24
24
24
Temp.
(° C)
22.5
23
24
X
24
24
Temp.
(° C)
25
25
-
-
-
25
pH Phenol
Start (mg/1)
8.5 0.89
8.4
-
_
0.07
- -
pH Ammonia
Start (mg/1)
9.5 16.5
9.2 9.0
9.1 4.8
X X
8.6 2.3
-
pH
Start
8.4
-
-
-
-
-
aDO, temperature, sulfide, ammonia, and phenol measurements were taken
at 24 hours or at the death of all 10 fish.
35
-------
Sample 69994:RW-1
- 4.5%
Cone . of
Waste
(% by vol.)
10
5.6
3.2
1.8
1.0
Control
Sample 69994
Cone, of
Waste
(% by vol.)
10
5.6
3.2
1.8
1.0
Control
Sample 6995
Cone, of
Waste
(% by vol.)
10
5.6
3.2
1.8
1.0
Control
No. of
Test No.
Animals at
10
10
10
10
10
10
:RW-2 TL -
No. of
Test No.
Animals at
10
10
10
No set up
10
10
:RW-1 TL5() -
No. of
Test No.
Animals at
10
10
10
10
10
10
Alive
24 hrs
0
2
10
10
10
10
1.9%
Alive
24 hrs
0
0
1
X
10
10
5.6%
Alive
24 hrs
0
7
9
10
10
10
D0a
(mg/1)
8.5
5.0
4.1
-
-
4.2
DO
(mg/1)
7.8
-
7.2
X
4.5
4.5
DO
(mg/1)
7.5
1.6
4.1
-
-
4.5
Temp . a
(° C)
23
25
23.5
-
-
24
Temp.
(° C)
22
-
24.5
X
24.5
24
Temp.
C C)
23.5
25
24.5
-
-
25
pH Ammoniaa
Start (mg/1)
9.0 12.0
6.5
4.0
-
-
-
PH
Start
2.1
2.6
3.25
X
7.2
8.55
pH Ammonia
Start (mg/1)
9.0 9.0
8.7 5.4
3.3
-
-
-
DO, temperature, sulfide, ammonia, and phenol measurements were taken
at 24 hours or at the death of all 10 fish.
36
-------
Sample 6995:RW-3
- 13%
Cone, of
Waste
(% by vol.)
18
10
5.6
3.2
No. of
Test
Animals
10
10
10
10
1.8 No set up
Control
Sample 6995
Cone . of
Waste
(% by vol.)
18
10
5.6
3.2
1.8
Control
10
:RW-4 TL5Q
No. of
Test
Animals
10
10
10
10
No set up
10
Sample 6995:RW-5 TL5Q
Cone, of
Waste
(% by vol.)
5.6
3.2
1.8
1.0
0.56
Control
No. of
Test
Animals
10
10
10
10
10
10
No. Alive
at 24 hrs
0
9
10
10
X
10
- 13.5%
No. Alive
at 24 hrs
2
8
10
10
X
10
- 4.2%
No. Alive
at 24 hrs
0
10
10
10
10
10
D0a
(mg/1)
7.3
4.5
-
-
X
4.7
DO
(mg/1)
4.5
4.8
-
-
X
4.0
DO
(mg/1)
4.7
3.4
-
-
-
2.6
a
Temp.
(° C)
24
24
-
-
X
24
Temp.
(° C)
24
24
-
-
X
24
Temp.
(° C)
24
24
-
-
-
23.5
pH
Start
8.6
-
-
-
X
~
pH
Start
8.55
-
-
-
X
—
PH
Start
8.85
-
-
-
-
-
aDO, temperature, sulfide, ammonia, and phenol measurements were taken
at 24 hours or at the death of all 10 fish.
37
-------
Sample 6995:RW-6 TL - 7.2%
Cone, of
Waste
(% by vol
18
10
5.6
3.2
1.8
Control
Sample 92
Cone, of
Waste
(% by vol
10
5.6
3.2
1.8
1.0
Control
No. of
Test
. ) Animals
10
10
10
10
No set up
10
:RW-2 TL5Q
No. of
Test
.) Animals
10
10
10
10
10
10
Sample 6996 :RW- 3 TL
Cone, of
Waste
(% by vol
10
5.6
3.2
1.8
1.0
Control
No. of
Test
. ) Animals
10
10
10
10
10
10
No. Alive
at 24 hrs
0
0
9
10
X
10
- 2.4%
No. Alive
at 24 hrs (
0
0
0
10
10
10
50 - 4-2%
No. Alive
at 24 hrs
0
0
10
10
10
10
D0a Temp.a
(mg/1) (° C)
7.0 22
7.0 23.5
5.6 23
-
X X
24
DO Temp . pH
mg/1) (° C) Start
6.5 22.0 9.2
5.7 23.0 9.1
5.3 23.5 9.0
2.5 23.5
_
4.6 23.5
DO Temp .
(mg/1) (° C)
6.9 22.5
7.0 22
4.8 21.5
5.6 21.5
-
6.0 21.5
pH
Start
_
8.4
8.4
-
X
-
Ammonia
(mg/1)
14.2
'8.1
4.6
3.3
2.0
0.7
PH
Start
7.7
-
-
-
-
8.4
Phenol'
(mg/1)
11.7
6.9
4.0
-
-
-
a.
DO, temperature, sulfide, ammonia, and phenol measurements were taken
at 24 hours or at the death of all 10 fish.
38
-------
Sample 6996:RW-4 TL - 4.2%
Cone, of
Waste
(% by vol.)
5.6
3.2
1.8
1.0
0.56
Control
No. of
Test
Animals
10
10
10
10
10
10
No. Alive
at 24 hrs
0
10
9
10
10
10
D0a
(mg/1)
6.5
3.8
-
-
-
3.7
Temp . a
(° C)
23
22
-
-
-
22
pH
Start
8.25
8.50
8.50
-
-
8.40
Sample 6996:RW-5
TL - 0.05%
oU
Cone, of
Waste
(% by vol.)
0.1
0.056
0.032
0.018
0.010
Control
No. of
Test
Animals
10
10
10
10
10
10
No. Alive
at 24 hrs
0
4
10
9
10
10
DO
(mg/1)
7.3
5.6
0
-
-
4.4
Temp.
(° C)
22.5
22
23
-
-
22.5
PH
Start
8.65
8.45
-
-
-
8.45
DO, temperature, sulfide, ammonia, and phenol measurements were taken
at 24 hours or at the death of all 10 fish.
39
-------
APPENDIX B
RAW DATA - PHASE II
40
-------
SAMPLE REFINERY 9973: API SEPARATOR EFFLUENT
Composite A
12-hour TL5 - 42%
Cone . of
Waste
(% by vol)
56
32
18
10
5.6
Control
Composite B
Cone . of
Waste
(% by vol)
48
24
12
6
3
Control
Composite C
Cone . of
Waste
(% by vol)
48
36
25
16.8
12.6
Control
No. of
Test No.
Animals at
10
10
10
10
10
10
12-hour TL
No. of
Test No.
Animals at
10
10
10
10
10
10
12-hour TL
No. of
Test No.
Animals at
10
10
10
10
10
10
alive No.
12 hrs at
3
10
10
10
10
10
50 - 34%
alive No.
12 hrs at
0
10
10
10
10
10
50 - 41%
alive No.
12 hrs at
0
10
10
10
10
10
alive
24 hrs
0
4
10
10
10
10
alive
24 hrs
0
2
10
10
10
10
alive
24 hrs
0
0
2
3
10
10
D0a
(mg/1)
12 hr 24 hr
3.9 3.8
4.0 0.5
4.0 1.2
1.3
-
5.2 3.1
DO
(mg/1)
12 hr 24 hr
3.8
2.0 1.0
1.5
1.5
-
4.8 2.5
DO
(mg/1)
12 hr 24 hr
3.6 0.5
3.2 0.6
3.5 1.0
1.2
1.3
4.9 3.1
Temp.
(° C)
22
21.5
21.5
_
_
21.5
Temp.
(° C)
22
22
22
22
_
22
Temp.
C C)
22
22
22
22
22
22
pH
Start
8.9
8.8
-
_
_
8.7
pH
Start
8.6
8.5
-
_
_
8.4
PH
Start
8.9
8.8
8.7
8.7
8.6
8.6
Ammonia
(mg/D
Start
5.1
3.1
2.0
_
_
-
DO readings at 12 hours, 24 hours, or at death of all 10 fish.
Temperature readings at 24 hours or at death of all 10 fish.
41
-------
SAMPLE REFINERY 9973: FINAL CLARIFIER EFFLUENT
Composite A
24-hour TL5Q - >56%
Cone, of
Waste
(% by vol)
56
32
18
10
5.6
Control
Composite B
Cone . of
Waste
(% by vol)
88
44
22
11
Control
Composite C
Cone, of
Waste
(% by vol)
100
50
25
12.5
6.25
Control
No. of
Test No.
Animals at
10
10
10
10
10
10
24-hour TL
No. of
Test No.
Animals at
10
10
10
10
10
alive No
12 hrs at
10
10
10
10
10
10
50 - >88%
alive No
12 hrs at
10
10
10
10
10
24-hour TL5Q - 100%
No. of
Test No.
Animals at
10
10
10
10
10
10
alive No
12 hrs at
10
10
10
10
10
10
. alive
24 hrs
10
10
10
10
10
10
. alive
24 hrs
10
10
10
10
10
D0a
Og/D
12 hr 24 hr
6.0 3.7
-
_
-
-
5.1 3.0
DO
Og/D
12 hr 24 hr
6.1 3.1
5.5 2.7
-
-
3.8 1.5
Temp.
(° C)
22
-
-
-
-
22
Temp.
(° C)
22.5
22
-
-
22.5
pH
Start
8.3
-
-
-
-
8.2
pH
Start
8.0
8.3
-
-
8.4
Ammonia
(mg/1)
Start
7.0
4.0
_
-
-
-
(no mortality or observed stress)
. alive
24 hrs
10
10
10
10
10
10
DO
Og/D
12 hr 24 hr
7.2 4.0
6.2 3.5
-
_
-
4.0 2.0
Temp.
(° C)
22
22
-
-
-
22
pH
Start
7.6
8.3
-
-
-
8.6
DO readings at 12 hours, 24 hours, or at death of all 10 fish.
'Temperature readings at 24 hours or at death of all 10 fish.
-------
SAMPLE REFINERY 2115: API SEPARATOR EFFLUENT
Composite A 24-hour TL - 4.2%
Cone . of
Waste
(% by vol)
10
5.6
3.2
1.8
1.0
Control
Composite B
Cone, of
Waste
(% by vol)
10
5.6
3.2
1.8
1.0
Control
Composite C
Cone . of
Waste
(% by vol)
7.5
5.6
4.2
3.2
2.4
Control
No. of
Test No.
Animals at
10
10
10
10
10
10
alive
12 hrs
0
0
10
10
10
10
24-hour TL - 4
50
No. of
Test No.
Animals at,
10
10
10
10
10
10
24-hour TL
No. of
Test No.
Animals at
10
10
10
10
10
10
alive
12 hrs
0
0
10
10
10
10
50 " 3
alive
12 hrs
0
0
0
10
10
10
No." alive
at 24 hrs
0
0
10
10
10
10
.2%
No. alive
at 24 hrs
0
0
10
10
10
10
.5%
No. alive
at 24 hrs
0
0
0
8
10
10
D0a
(mg/1)
12 hr 24 hr
7.6
7.7
5.7 2.7
-
_
5.2 3.0
DO
(mg/1)
12 hr 24 hr
7.8
7.6
4.8 2.5
-
_
4.5 2.5
DO
(mg/1)
12 hr 24 hr
8.1
8.1
7.5
5.1 2.2
5.5 1.7
4.5 1.7
Temp.
(° C)
21
21.5
22
-
-
22
Temp.
(° C)
21
21.5
22
-
-
22
Temp.
(° Q
20
20
22
22
22
22
PH
Start
9.4
9.0
8.9
8.6
-
8.4
PH
Start
9.3
9.1
8.8
-
_
8.6
PH
Start
9.3
9.1
9.0
8.8
8.7
8.6
Ammonia
(mg/1)
Start
11.1
6.4
3.8
2.25
1.1
0.2
DO readings at 12 hours, 24 hours, or at death of all 10 fish.
Temperature readings at 24 hours or at death of all 10 fish.
43
-------
SAMPLE REFINERY 2115: TRICKLING FILTER EFFLUENT
Composite A 24-hour TL - 12.5%
Cone . of
Waste
(% by vol)
32
18
10
5.6
3.2
Control
Composite B
Cone, of
Waste
(% by vol)
24
12
6
3
1.5
Control
Composite C
Cone, of
Waste
(% by vol)
24
18
13.5
10.0
7.5
Control
No. of
Test No.
Animals at
10
10
10
10
10
10
2 4 -hour TL
No. of
Test No.
Animals at
10
10
10
10
10
10
alive
12 hrs
0
0
10
10
10
10
50 ' 16!
alive
12 hrs
0
10
10
10
10
10
24-hour TL - 11
oU
No. of
Test No.
Animals at
10
10
10
10
10
10
alive
12 hrs
0
0
4
10
10
10
No. alive
at 24 hrs
0
0
8
10
10
10
\
No. alive
at 24 hrs
0
10
10
10
10
10
.5%
No. alive
at 24 hrs
0
0
0
10
10
10
D0a
(mg/1)
12 hr 24 hr
7.6
-
5.3 2.0
5.3 2.5
-
5.8 2.6
DO
(mg/1) .
12 hr 24 hr
7.0
4.7 1.5
5.3 3.0
-
-
4.7 2.5
DO
(mg/1)
12 hr 24 hr
8.0
6.9
5.9 5.5
5.8 3.3
3.6
5.7 4.0
Temp.
(° C)
18
-
22
22
-
22
Temp.
20
22
22
-
-
22
Temp.
18
21
22
22
22
22
pH
Start
8.8
-
8.6
8.6
-
8.7
PH
Start
8.4
8.3
8.3
-
-
8.5
pH
Start
8.6
8.6
-
-
-
8.6
Ammonia
(mg/1)
Start
33.0
20.0
10.0
6.2
-
-
*DO readings at 12 hours, 24 hours, or at death of all 10 fish.
'Temperature readings at 24 hours or at death of all 10 fish.
44
-------
SAMPLE REFINERY 2115: FINAL CLARIFIER EFFLUENT
Composite A 24-hour TL - 18%
Cone.
of
Waste
(
% by
32
18
10
5.
3.
vol)
6
2
Control
No. of
Test
Animals
10
10
10
10
10
10
D0a
No. alive
at 12 hrs
0
10
10
10
10
10
No. alive
at 24 hrs
0
5
10
10
10
10
(mg/1)
12
6.
5.
5.
-
-
6.
hr
0
6
8
3
24
2
2
3
hr
-
.1
.2
-
-
.1
Temp.
(° C)
22
22
22
-
- -
22
Ammonia
pH (mg/1)
Start Start
8.5 33.0
19.8
9.5
5.8
-
8.7
Composite B 24-hour TL
50
Cone, of
Waste
(% by vol)
48
24
12
6
3
Control
No. of
Test
Animals
10
10 ,
10
10
10
10
DO
No. alive
at 12 hrs
1
10
10
10
10
10
No. alive
at 24 hrs
0
3
10
10
10
10
(mi
12 hr
5.3
5.1
-
-
-
5.0
J/D
24 hr
5.2
2.5
3.2
-
-
3.1
Temp.
(° C)
22
22
22
-
-
22
pH
Start
8.2
8.4
-
-
-
8.6
Composite C
24-hour TL5Q - 20%
Cone.
(
of
Waste
% by vol)
28
21
15
11
8
.5
.5
.7
Control
No. of
Test
Animals
10
10
10
10
10
10
DO
No. alive
at 12 hrs
10
10
10
10
10
10
No. alive
at 24 hrs
1
4
10
10
10
10
(mg/1)
12 hr
5.2
5.3
5.5
-
-
5.5
24
4
3
2
4
hr
.0
.0
.8
-
-
.0
Te
C°
22
22
22
-
22
mp.
.5
.5
.5
-
.5
pH
Start
8.2
8.4
-
-
-
8.5
DO readings at 12 hours, 24 hours, or at death of all 10 fish.
Temperature readings at 24 hours or at death of all 10 fish.
45
-------
SAMPLE REFINERY 6512: EQUALIZATION UNIT EFFLUENT
Composite A Estimated 24-hoUr TL - 32%-56%
Cone, of
c
Waste
% by vol)
56
32
18
10
5.6
Control
No. of
Test
Animals
10
10
10
10
10
10
D0a
No. alive
at 12 hrs
10
10
10
10
10
10
No. alive
at 24 hrs
3
2
1
8
10
10
Ong/l)
12 hr
2.6
2.7
1.6
-
-
2.5
24
0
0
0
0
1
1
hr
.4
.5
.5
.8
.0
.1
Temp. pH
(°
22.
22.
22.
23
23
23
C)
5
5
5
Start
8.
8.
8.
-
-
8.
1
3
5
6
Composite B Estimated 24-hour TL_Q - 32%-56%
Cone, of
Waste
(% by vol)
56
32
18
10
5.6
Control
No. of
Test No.
Animals at
5
5
No set up
5
5
5
DO
, alive
12 hrs
0
5
X
5
5
5
No. alive
at 24 hrs
0
0
X
5
5
5
(mg/D
12 hr
1.0
2.2
X
4.2
5.5
5.5
24 hr
-
1.0
X
2.1
3.0
3.3
Temp.
C° C)
22.5
23.5
X
23.5
23.5
23.5
pH
Start
8.0
8.3
X
8.6
-
8.7
DO readings at 12 hours, 24 hours, or at death of all 10 (5) fish.
Temperature readings at 24 hours or at death of all 10 (5) fish.
46
-------
SAMPLE REFINERY 6512: CHEMICAL COAGULATION UNIT EFFLUENT
Composite A
Estimated 24-hour TL5Q - 42%-60%
Cone, of
Waste
(% by vol)
100
56
32
18
10
Control
No. of
Test
Animals
10
10
10
10
10
10
D0a
No. alive
at 12 hrs
10
10
10
10
10
10
No. alive
at 24 hrs
1
0
2
2
7
10
(mg/D
12 hr
2.2
2.0
2.0
-
-
2.7
24 hr
0.3
0.3
0.3
0.5
0.8
1.1
V,
Temp .
(° C)
22.5
22.5
22.5
22.5
22.5
23
PH
Start
7.6
8.0
8.4
-
-
8.7
Composite B
Estimated 24-hour TL5Q - 42%-60%
Cone, of
Waste
(% by vol)
67
40
20
10
5
Control
No. of
Test
Animals
5
5
5
5
5
5
DO
No. alive
at 12 hrs
2
4
5
5
5
5
No. alive
at 24 hrs
0
0
5
5
5
5
Og/1)
12 hr
2.8
3.2
4.4
-
-
6.4
24 hr
0.5
0.6
1.5
-
-
5.5
Temp.
22.5
22.5
23
_
-
23
pH
Start
7.7
8.0
-
_
-
-
aDO readings at 12 hours, 24 hours, or at death of all 10 (5) fish.
Temperature readings at 24 hours or at death of all 10 (5) fish.
47
-------
SAMPLE REFINERY 6512: FINAL CLARIFIER EFFLUENT
Composite A
24-hour TL,_n - 100% (no mortality or observed stress)
Cone, of
Waste
(% by vo-1)
100
56
32
18
10
Control
No. of
Test
Animals
10
10
10
10
10?
10
No. alive
at 12 hrs
10
10
10
10
10
10
No. alive
at 24 hrs
10
10
10
10
10
10
D0a
(mg/D
12 hr
6.2
5.5
-
_
-
2.2
24 hr
2.4
2.5
2.7
_
_
1.2
Temp.b
C° C)
22
22
22
_
-
22.5
pH
Start
7.4
8.3
-
_
-
8.7
Composite B
24-hour TL - 100% (observed stree and 40% mortality)
Cone . of
Waste
(% by vol)
100
50
25
12.5
Control
No. of
Test
Animals
5
5
5
5
5
DO
No. alive
at 12 hrs
5
5
5
5
5
No. alive
at 24 hrs
5
5
5
5
5
(mg/1)
12 hr
6.5
6.2
6.0
_
6.3
24 hr
3.7
3.5
4.0
_
4.5
Temp.
23
23
23
_
23
pH
Start
7.4
7.9
-
_
-
DO readings at 12 hours, 24 hours, or at death of all 10 (5) fish.
Temperature readings at 24 hours or at death of all 10 (5) fish.
-------
SAMPLE REFINERY 0289: AIR FLOATATION
Composite A
Estimated 24-hour TL5Q - 32%-48%
Cone, of
Waste
(% by vol)
48
24
12
6
3
Control
No. of
Test
Animals
10
10
10
10
10
10
No. alive
at 12 hrs
0
10
10
10
10
10
No. alive
at 24 hrs
0
2
4
10
10
9
D0a
(mg/lD
12 hr
1.2
2.2
1.5
-
-
1.2
24 hr
_
0.9
0.8
1.1
1.1
1.1
Temp . pH
(° C) Start
23
23
23
23
23
23
8.4
8.6
8.7
-
-
8.7
Composite B
Estimated 24-hour TL - 50%
Cone, of
Waste
(% by vol)
56
32
18
10
5.6
Control
No. of
Test
Animals
5
5
5
5
5
5
DO
No. alive
at 12 hrs
5
5
5
5
5
5
No. alive
at 24 hrs
2
5
5
5
5
5
(mg/1)
12 hr
4.2
4.7
5.0
-
-
5.3
24 hr
2.0
2.2
2.9
-
_
3.8
Temp.
23
23
23
-
_
23
PH
Start
8.0
8.4
8.5
-
_
8.7
aDO readings at 12 hours, 24 hours, or at death of all 10 (5) fish.
Temperature readings at 24 hours or at death of all 10 (5) fish.
49
-------
SAMPLE REFINERY 0289: FINAL CLARIFIER EFFLUENT
Composite A
24-hour TL5Q - >48%
Cone, of
Waste
(% by vol)
48
24
12
6
Control
No. of
Test
Animals
10
10
10
10
10
D0a
No. alive
at 12 hrs
10
10
10
10
10
No. alive
at 24 hrs
9
9
10
10
10
(mg/1)
12 hr
3.4
2.5
2.5
-
2.7
24 hr
1.2
1.2
1.4
-
1.9
h
Temp.
(° C)
23
23
23
-
23.5
pH
Start
8.6
8.7
-
-
8.7
Composite B
24-hour TL_n - 100% (no mortality or observed stress)
Cone, of
Waste
(% by vol)
100
50
25
12.5
Control
No. of
Test
Animals
5
5
5
5
5
DO
No. alive
at 12 hrs
5
5
5
5
5
No. alive
at 24 hrs
5
5
5
5
5
(mg
12 hr
7.0
6.6
6.3
-
6.3
A)
24 hf
5.1
5.5
4.5
-
4.6
Temp.
(° C)
23
23
23
-
23
pH
Start
7.7
8.1
-
-
-
DO readings at 12 hours, 24 hours, or at death of all 10 (5) fish.
temperature readings at 24 hours or at death of all 10 (5) fish.
50
-------
SAMPLE REFINERY 6693: API SEPARATOR EFFLUENT
Composite A
24-hour TL5Q - 24%
Cone . of
Waste
(% by vol)
32
18
10
5.6
3.2
Control
Composite B
Cone . of
Waste
(% by vol)
56
32
18
10
5.6
Control
Composite C
Cone, of
Waste
(% by vol)
56
32
18
10
5.6
Control
No. of
Test No.
Animals at
10
10
10
10
10
10
24-hour TL
No. of
Test No.
Animals at
10
10
10
10
10
10
24-hour TL
No. of
Test No.
Animals at
10
10
10
10
10
10
alive
12 hrs
9
10
10
10
10
10
50 ' 18
alive
12 hrs
0
1
10
10
10
10
50 " 22
alive
12 hrs
0
0
8
10
10
10
No. alive
at 24 hrs
0
10
10
10
10
10
%
No. alive
at 24 hrs
0
0
5
9
10
10
.s,
No. alive
at 24 hrs
0
0
8
10
10
10
D0a
(mg/1)
12 hr 24 hr
5.9 4.2
6.4 4.8
-
-
_
6.4 4.1
DO
(mg/1)
12 hr 24 hr
8.5
6.6 6.6
2.0
-
-
5.8 3.7
DO
(mg/1)
12 hr 24 hr
7.7
7.0
6.0 2.8
-
_
6.4 4.8
h
Temp .
(° C)
24
24
-
-
-
24
Temp.
(° C)
22.5
24
-
-
-
24.5
Temp.
C° C)
20
23
23
-
_
23
pH
Start
8.8
8.5
-
-
-
8.4
pH
Start
8.9
8.8
-
-
-
8.8
PH
Start
9.0
8.9
8.8
_
_
-
DO readings at 12 hours, 24 hours, or at death of all 10 fish.
^Temperature readings at 24 hours or at death of all 10 fish.
51
-------
SAMPLE REFINERY 6693: FINAL CLARIFIER EFFLUENT
Composite A 24-hour TL™ - >41%
Cone. of
Waste
No. of
Test
No. alive No. alive
(mg/l)
Temp. pH
by vol) Animals at 12 hrs at 24 hrs 12 hr 24 hr (° C) Start
41
32
18
10
Control
10
10
10
10
10
10
10
10
10
10
10
10
10
10
10
7.6
-
-
-
7.5
5.9
-
-
-
6.6
24
-
-
-
24
8.1
-
-
-
8.7
Composite B 24-hour TL™ - 100% (no mortality or distress observed)
Cone, of No. of DO
Waste Test No. alive No. alive (mg/l) Temp. pH
(% by vol) Animals at 12 hrs at 24 hrs 12 hr 24 hr (° C) Start
100
.50
25
12.5
Control
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
7.0
5.7
4.4
3.8 24.5
3.0 25
7.7
8.4
8.7
DO readings at 12 hours, 24 hours, or at death of all 10 (5) fish.
Temperature readings at 24 hours or at death of all 10 (5) fish.
52
-------
APPENDIX C
ANALYTICAL DATA - PHASE I AND PHASE II
53
-------
RESULTS OF CHEMICAL ANALYSES OF PHASE I SAMPLES
COLLECTED CONCURRENT WITH BIOASSAY SAMPLES
Chemical Parameter
0
Sample No.
6992 RW-2
6992 RW-3
6992 RW-10
6992 RW-11
5991 RW-1
69991 RW-1
693 RW-1
693 RW-2
69994 RW-1
6995 RW-1
6995 RW-3
6995 RW-4
6995 RW-5
6995 RW-6
92 RW-2
6996 RW-3
6996 RW-4
6996 RW-5
Ammonia
(mg/1)
3575
225
3350
95
118
40
95
199
125
81
1.3
1.5
120
0.85
85
5.2
350
2700
Phenolics
(mg/1)
525
14,500
500
1.4
7.6
2.9
64
0.52
4.5
5.4
0.52
0.52
62
0.38
140
3.1
<0.01
410
Sulfide
(mg/1)
3975
37
3846
129
84
14
51
11
19
110
0.4
0.9
78
0.2
3.40
1.4
1.0
3000
Cyanide
(mg/1)
0.6
17.0
1.4
0.34
0.74
1.4
0.22
0.14
0.38
0.03
<0.03
<0.03
0.1
<0.03
0.08
<0.03
<0.03
122
Samples for which definite TL,_n was established.
54
-------
RESULTS OF CHEMICAL ANALYSES OF PHASE II SAMPLES
COLLECTED CONCURRENT WITH BIOASSAY SAMPLES
Chemical Parameter
Sample Location
Refinery 9973
API Separator
Final Clarifier
Refinery 2115
API Separator
Trickling Filter
Final Clarifier
Refinery 6512
Equal . Chamber
Chem. Coag. Unit
Final Clarifier
Refinery 0288
Air Float. Unit
Final Clarifier
Refinery 6693
API Separator
Final Clarifier
Ammonia
(mg/1)
9.25
6.4
127
108.5
106
6.9
6.75
0.375
12.0
11.25
28.4
26.25
Phenolics
(mg/D
11.0
0.01
10.45
0.99
0.035
0.61
0.405
0.01
0.02
0.01
3.4
0.01
Sulfide
(mg/1)
2.88
0.15
24.3
0.75
0.275
1.0
1.5
0.425
0.7
0.5
32.5
0.2
Cyanide
(mg/1)
0.135
0.10
1.50
0.245
0.07
0.22
0.20
0.14
0.06
0.02
0.29
0.185
The median value from results of analyses of samples collected on 14
consecutive days at each location is presented.
55
-------
TECHNICAL REPORT DATA
(Please read Instructions on the reverse before completing)
. REPORT NO.
EPA-600/2-76-241
3. RECIPIENT'S ACCESSION NO.
4. TITLE AND SUBTITLE
ACUTE TOXIC EFFECTS OF PETROLEUM REFINERY WASTEWATERS
ON REDEAR SUNFISH
5. REPORT DATE
October 1976 (IssuingJJate)
6. PERFORMING ORGANIZATION CODE
. AUTHOR(S)
John E. Matthews and Leon H. Myers
8. PERFORMING ORGANIZATION REPORT NO.
. PERFORMING ORGANIZATION NAME AND ADDRESS
Robert S. Kerr Environmental Research Laboratory
Office of Research and Development
U.S. Environmental Protection Agency
Ada, Oklahoma 74820
10. PROGRAM ELEMENT NO.
1BB036
11. CONTRACT/GRANT NO.
12. SPONSORING AGENCY NAME AND ADDRESS
Same as above
13. TYPE OF REPORT AND PERIOD COVERED
Interim
14. SPONSORING AGENCY CODE
EPA-ORD
15. SUPPLEMENTARY NOTES
16. ABSTRACT
Static bioassays of 24 hours' duration were performed on samples of wastewaters
provided by 22 domestic petroleum refiners. These wastewaters represent three types
of water discharges prevalent to this industry: process wastewaters prior to dilu-
tion with other streams; API separator effluents which are a conjugate of various
streams within a refinery; and wastewaters following treatment by activated sludge
systems.
Bioassays were performed using redear sunfish (Lepomis microlophus) as test
organisms. Twenty-four hour 50 percent tolerance limits (TL^) of the various waste-
waters are compared with results of chemical analyses performed during the same study.
Toxicity varied considerably both between refineries and for waste streams from with-
in a single refinery. Results of these analyses and observed behavioral symptoms of
distressed fish revealed that ammonia, sulfides, and phenolics, alone or in combina-
tion, were major contributors to toxicity exerted in most samples. Three refineries
had samples which were more toxic than anticipated based on results of chemical
analyses, indicating the presence of other toxic compounds in unknown quantities;
e.g., various hydrocarbons.
Results of bioassays of samples from various wastewater treatment stages of five
refineries with activated sludge treatment systems revealed a decrease in. toxicity
as the degree of treatment increased indicating a reduction in concentration of
-' - -
ntri hiit i nor hi r.
f nv-i f>gTTf-
17.
KEY WORDS AND DOCUMENT ANALYSIS
DESCRIPTORS
b.lDENTIFIERS/OPEN ENDED TERMS
COS AT I Field/Group
Bioassay
Freshwater fishes
Toxicity
Redear sunfish
Acute toxicity
06A
06T
18. DISTRIBUTION STATEMENT
Release Unlimited
19. SECURITY CLASS (ThisReport)'
21. NO. OF PAGES
62
20. SECURITY CLASS (Thispage)
22. PRICE
EPA Form 2220-1 (9-73)
56
U S. GOVERNMENT PRINTING OFFICE-. 1976-757-056/5^2 Region No. 5-11
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