&EPA
United States
Environmental Protection
Agency
t Environmental Research
Laboratory August
Cincinnati OH 45268
79
Research and Development
Maximum
Utilization of Water
Resources in a
Planned Community
Chlorine and Ozone
Toxicity Evaluation
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RESEARCH REPORTING SERIES
Research reports of the Office of Research and Development, U.S. Environmental
Protection Agency, have been grouped into nine series. These nine broad cate-
gories were established to facilitate further development and application of en-
vironmental technology. Elimination of traditional grouping was consciously
planned to foster technology transfer and a maximum interface in related fields.
The nine series are:
1. Environmental Health Effects Research
2. Environmental Protection Technology
3. Ecological Research
4. Environmental Monitoring
5. Socioeconomic Environmental Studies
6. Scientific and Technical Assessment Reports (STAR)
7. Interagency Energy-Environment Research and Development
8. "Special" Reports
9. Miscellaneous Reports
This report has been assigned to the ENVIRONMENTAL PROTECTION TECH-
NOLOGY series. This series describes research performed to develop and dem-
onstrate instrumentation, equipment, and methodology to repair or prevent en-
vironmental 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.
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EPA-600/2-79-050e
August 1979
MAXIMUM UTILIZATION OF WATER RESOURCES
IN A PLANNED COMMUNITY
Chlorine and Ozone Toxicity Evaluation
by
Brian Hammond and James Bishop, Jr.
Department of Biology
Rice University
Houston, Texas 77001
Grant No. 802433
Project Officers
Richard Field
Anthony N. Tafuri
Storm and Combined Sewer Section
Wastewater Research Division
Municipal Environmental Research Laboratory (Cincinnati)
Edison, New Jersey 08817
MUNICIPAL ENVIRONMENTAL RESEARCH LABORATORY
OFFICE OF RESEARCH AND DEVELOPMENT
U.S. ENVIRONMENTAL PROTECTION AGENCY
CINCINNATI, OHIO 45268
-------
DISCLAIMER
This report has been reviewed by the Municipal Environmental
Research Laboratory, U.S. Environmental Protection Agency, and
approved for publication. Approval does not signify that the
contents necessarily eflect the views and policies of the U.S.
Environmental Protection Agency, nor does mention of trade names
or commercial products constitute endorsement or recommendation
for use.
n
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FOREWORD
The Environmental Protection Agency was created because of
increasing public and government concern about the dangers of
pollution to the health and welfare of the American people.
Noxious air, foul water, and spoiled land are tragic testimony to
the deterioration of our natural environment. The complexity of
that environment and the interplay between its components requires
a concentrated and integrated attack on the problem.
Research and development is that necessary first step in
problem solution and it involves defining the problem, measuring
its impact, and searching for solutions. The Municipal Environ-
mental Research Laboratory develops new and improved technology
and systems for the prevention, treatment, and management of
wastewater and solid and hazardous waste pollutant discharges
from municipal and community sources, for the preservation and
treatment of public drinking water supplies, and to minimize the
adverse economic, social, health, and aesthetic effects of pollu-
tion. This publication is one of the products of that research;
a most vital communications link between the researcher and the
user community.
This project focuses on methods maximizing the use of water
resources in a planned urban environment, while minimizing their
degradation. Particular attention is being directed towards
determining the biological, chemical, hydrological and physical
characteristics of stormwater runoff and its corresponding role
in the urban water cycle.
Francis T. Mayo
Director
Municipal Environmental Research
Laboratory
m
-------
ABSTRACT
Relative to the project entitled "Maximum Utilization of
Water Resources in a Planned Community" it was essential to study
the effect of wastewater disinfection on aquatic organisms.
Impoundments in The Woodlands will receive reclaimed wastewater
and will provide recreational fishing. To ensure adequate water
quality for aquatic organisms the following experiments were
conducted.
Flow through bioassays vs. standard static bioassays were
run to determine an accurate LCso for chlorine on fingerling
channel catfish, Ictalurus punctatus. The 96 hour LCsg for
chlorine is 0.07 mg/1 (total chlorine) from the flow through vs.
0.45 mg/1 (total chlorine) from the static bioassay.
The 96 hour LC5Q for ozone on fingerling channel catfish, as
determined from flow through bioassay, is 0.03 mg/1 ozone.
Chlorine and ozone exposures had little effect on kidney
functions. Exposure to both chlorine and ozone drastically
reduced the ability of the gills to actively absorb sodium from
the water.
Long term exposure to chlorine drastically reduced both blood
pressure and heart rate while exposure to ozone had little, if any,
effect. Blood pressure and heart rate are very sensitive physio-
logical parameters and changes are indicative of a stressful
environment.
Both chlorine and ozone are extremely toxic to fish at low
levels. If detected in receiving waters by present analytical
techniques, a toxic condition exists.
This report was submitted in fulfillment of USEPA Grant
Number 802433 by Brian R. Hammond and James R. Bishop, Jr. under
the sponsorship of the Environmental Protection Agency. This
report covers a period from July 1973 to May 1976 and work was
completed as of May 1976.
iv
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CONTENTS
Foreword iii
Abstract iv
Figures vi
Tables viii
Acknowledgments ix
1. Introduction 1
Chlorine 3
Ozone 4
2. Conclusions 5
3. Experimental Methods 5
Fish Holding and Preparation 5
4. Results 10
Flow Through Bioassays 10
Physiological Observations 15
References 37
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FIGURES
Number Page
1 A comparison of survival-mortality characteristics of
fingerling catfish to chlorine exposure in a static
bioassay and in a flow through bioassay 12
2 Survival-mortality characteristics of fingerling cat-
fish to ozone exposure in a flow through bioassay . 14
3 Effect of short term chlorine exposure on the glomer-
ular filtration rate and corresponding urine flow
of Ictalurus punctatus 16
4 Effect of short term chlorine exposure on the glomer-
ular filtration rate and corresponding urine flow
of Ictalurus punctatus 17
5 Effect of short term chlorine exposure on the concen-
tration of ions in the urine of Ictalurus punctatus 18
6 Effect of short term chlorine exposure on the rate of
excretion of ions in the urine of Ictalurus punc-
tatus 19
7 Effect of short term ozone exposure on the glomer-
ular filtration rate and corresponding urine flow
of Ictalurus punctatus 20
8 Effect of short term ozone exposure on the rate of
excretion of ions in the urine of Ictalurus punc-
tatus 22
9 Chlorine uptake by five catfish 23
10 Effect of short term chlorine exposure on the uptake
2^Na by the gills of Ictalurus punctatus 25
11 Effect of short term ozone exposure on the uptake of
22Na by the gills of Ictalurus punctatus 26
12 Influence of short term chlorine exposure on the heart
rate and blood pressure of Ictalurus punctatus. . . 27
VI
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FIGURES (Continued)
Number
13 Heart rate and blood pressure of Ictalurus punctatus
immediately following exposure to different concen-
trations of chlorine 28
14 Summary of changes in fish's blood pressure and heart
rate following exposure to chlorine 30
15 Influence of long term chlorine exposure on the blood
pressure and heart rate of Ictalurus punctatus. . . 31
16 Influence of long term chlorine exposure on the blood
pressure and heart rate of Ictalurus punctatus. . . 32
17 Summary of changes in blood pressure following long
term exposure in chlorine 33
18 The influence of chlorine at a dose approximating
the 96 hour LC5Q on heart rate and blood pressure of
Ictalurus punctatus 34
19 The influence of chlorine at a dose less than half of
the 96 hour LC$Q on heart rate and blood pressure
in Ictalurus punctatus 36
vii
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TABLES
Number Page
Summary of Standard Chemical and Physical'Tests
Used in this Research
Ninety-Six Hour LC5Q Bioassay Data of Chlorine for
Ictalurus punctatus: 11
Ninety-Six Hour LC5Q Flowthrough Bioassay Data of
Ozone for Ictalurus punctatus 13
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ACKNOWLEDGMENTS
This project was supported by the U.S. Environmental
Protection Agency, The Woodlands Development Corporation and
Rice University.
Thanks are due the Texas Parks and Wildlife - Sheldon Fish
Hatchery,and Catfish Acres of Winnie, Texas for their donation
of fish used in these experiments.
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SECTION 1
INTRODUCTION
Non-point source pollution has become of greater concern due to
increasingly stringent point source effluent standards and rapid develop-
ment of urban areas. Urban stormwater runoff is one of the major non-
point pollution sources and innovative methods must be developed to
minimize the impact of stormwater runoff on receiving waters. One such
imaginative water resource plan has been developed for The Woodlands,
a planned community in southeast Texas. The U.S. Environmental Protection
Agency, Woodlands Development Corporation and Rice University have com-
pleted a three year research/demonstration project to evaluate the water
resource system at The Woodlands and develop strategies for maximizing
the benefits to the community while minimizing the effect on receiving
waters.
The hydrological characteristics of a natural watershed change with
urbanization. Replacement of flow-retarding vegetation with impervious
surfaces, such as roads and buildings, increases the rate and amount
of stormwater runoff. Removal of the water is traditionally implemented
by the use of an urban drainage system consisting of storm sewers and/or
deep, concrete-lined drainage ditches, designed specifically for rapid
drainage. Increased runoff volumes and peak flow rates result, creating
problems of downstream flooding and channel erosion.
Infiltration of stormwater is a major groundwater recharge source,
however emphasis on surface removal minimizes the infiltration rate,
resulting in a lowered water table and possible urban land subsidence
problems. Water quality deteriorates because natural purification
provided by infiltration in compromised.
i
The urban environment, typified by high population density, provides
a major pollutant source for runoff waters. Recent investigations
recognize the significance and magnitude of pollution problems from
urban stormwater runoff. In terms of specific pollutants, the sediment
yield problem is the most dramatic. Due primarily to urban construction,
urban sediment loads were found to be as much as 75 times greater than
loads in agricultural regions. Other runoff pollutants reported
higher in urban regions include dissolved solids, coliform,
biochemical oxygen demand and chemical oxygen demand (BOD and COD),
polychlorinated biphenyls, heavy metals, pesticides and fertilizers.
An increase in bacterial content is also reported. Claudon
stated that agricultural and urban runoff regularly contribute Salmonella
s£. to recreational waters. High fecal coliform and fecal streptococcus
1
-------
numbers have been found in urban runoff. Snowmelt and related agricultural
runoff in far northern climes in the continental U.S. have been shown to
contribute high densities of indicator bacteria and pathogens to runoff.
The addition of large quantities of inorganic nutrients particularly
nitrogen and phosphorous, to freshwater lakes poses a serious problem in
lake management. Municipal sewage, agricultural drainage, managed forestland
drainage, and fertilization often accelerate the natural process of eutrophi-
cation, thus enhancing the growth of bacteria, algae and aquatic vascular
plants. Population densities of these organisms often reach nuisance propor-
tions and interfere with the aesthetic qualities and recreational values
of lakes. These "blooms" may discolor, impart unsatisfactory tastes and
excrete toxins into the water. They can also clog treatment plant filters,
and upon decomposition, produce foul odors. Late summer "blooms" can create
anoxic conditions and cause the death of fish. Accordingly, values of
lake properties may depreciate and there may be increased burdens on munici-
pal water systems due to added costs of filtration and deodorization of
the water.
Although literature on eutrophication is extensive, most limnological
studies were conducted on lakes that were either eutrophic, or non-eutrophic
at the time of study, and few studies were continued long enough to follow
changes in the trophic status of lakes. Even fewer studies have been initiated
on a drainage system before the construction of lakes. Thus, complete
developmental histories of the water resources of particular areas are
lacking. Also, it is evident that information on more effective methods
of removing mineral nutrients from effluents prior to release into natural
waters is badly needed.
OBJECTIVES
The overall goal of this research project was to evaluate the water
resource plan for The Woodlands and to make recommendations as necessary
to maximize its effective utilization through alternations in design or
management.
One of the major objectives was to modify and expand the capabilities
of the EPA Storm Water Management Model (SWMM) to apply to the "natural
drainage" system designed for The Woodlands. SWMM has been expanded to
include the following additional water quality parameters: total COD,
total Kjeldahl nitrogen (TKN), nitrates and phosphates. The model has
been used to evaluate the effectiveness of the "natural drainage" system
in minimizing changes in storm runoff quantity and quality and to assist
engineers and planners in designing the drainage system for future develop-
ment phases at The Woodlands.
-------
Since urban areas are typified by a variety of land uses, including
residential, commercial and industrial as the broadest categories,
determining effects of land use on stormwater quality was conceived
as another major project objective. The Woodlands development plan
could not provide sufficiently diverse study areas during the project
period. Consequently, urban watersheds of similar physiographic and
drainage characteristics in Houston were studied to further relate
storm runoff water quality to urban land use.
In order to accomplish these two primary objectives, a massive
sampling and monitoring program was established. Rainfall, stream-
flow and over twenty-five water quality parameters were monitored
on a regular basis. Including in the water quality program were
intensive programs concentrating on bacteriological water quality,
chlorinated hydrocarbons and phytoplankton identification and enumeration.
The sampling program included bacteriological tests to evaluate
the traditional relationship between indicator organisms (e.g. fecal
coliform, fecal streptococcus) and pathogens in stormwater runoff.
Disinfection experiments were conducted to determine relative effective-
ness of C12, Br2» and 03 in untreated stormwater runoff. Toxicological
testing determined maximum tolerable concentrations of disinfectant in
the receiving stream for maintenance of fish populations. Algal bio-
assays were conducted to experimentally determine conditions, including
nutrient concentrations, necessary to prevent eutrophication in The
Woodlands' lakes.
Finally, the performance of a porous pavement was compared to a
conventional pavement with regard to runoff amount and quality, wet
skid resistance, hydroplaning and other characteristics related to
driveability.
The present requirements for the disinfection of domestic and industrial
wastes often requires the use of chlorine and/or ozone in their treatment.
In part the study entitled "Maximum Utilization of Water Resources in a
Planned Community" was to determine safe environmental conditions for these
disinfectants using both flow through 96 hour 1X59 bioassays and the
monitoring of physiological parameters as possible indicators under both
static and flow through conditions.
CHLORINE
In a recent review by Brungs (1) entitled, "Effects of Residual Chlorine
on Aquatic Life", it is suggested that continuous exposure to 0.002 mg/1 or
less of residual chlorine would not harm most aquatic organisms, but
continuous exposure to 0.01 mg/1 would harm some species of fish at some
life stages. Present data for trout and salmon would indicate a maximum
chlorine exposure of 30 min/day at a concentration of residual chlorine up
to, but not exceeding, 0.01 mg/1. Brungs (1) also states that, "No
comparable criterion for warm-water fish can be suggested at this time".
Considerable work has been done to evaluate the toxic effects of chlorine
on a wide variety of aquatic species.
-------
Tsai (2) reported that species shifts occurred following the introduction
of chlorine into a Maryland river. Also, it has been reported that the
reproduction of fathead minnows is drastically affected by exposure to
sub-lethal concentrations of chlorine (3).
An extensive review on chlorine toxicity has been made by Becker and
Thatcher (4). However, no acceptable chlorine standard has been established
to date.
OZONE
The flow through system was essential for the bioassay of ozone toxicity
(96 hour LC50) and for the exposure of adult fish to stable ozone concentrations.
The previous early work showed that ozone concentrations of 0.01 mg/1 were
lethal to fish and invertebrates (5). Ozone has been shown to retard meiosis
and cleavage, cause irregular polar bodies and abnormal nuclei cleavage in
the eggs of the American oyster (Crassostrea virginica) (6). Although ozone
is shown to be toxic to aquatic organisms it has been successfully used in
the control of fungal disease in incubating trout eggs (7). It has also
been used for the oxidation of dissolved organics and for the reduction of
microorganisms in aquaria systems (8). However, its use is not wide spread
since following ozone exposure animals show increased disease susceptibility
as a result of surface damage, and death if over exposed (9). With these
advantages and disadvantages in mind, the following experiments were designed
to establish safe environmental conditions for warm water fishes.
-------
SECTION 2
CONCLUSIONS
The environmentally safe surface water concentration for channel
catfish (Ictalurus punctatusj exposure to chlorine is 0.007 mg/1 . This
limit is based on 1/10 the 96 hour LC$n value of 0.07 mg/1 measured in the
water before contacting the fish and the absence of harmful physiological
responses at low chlorine exposures.
The channel catfish is sensitive to chlorine at toxic concentrations
and would probably avoid exposure. This is evident by observable pulmonary
irritation and a significant cardiovascular response to chlorine.
A flow through bioassay compared to a static bioassay produces a lower
LC5Q for chlorine (0.07 mg/1 vs. 0.45 mg/1).
The 96 hour LC^Q flow through bioassay of ozone for catfish finger! ings
is 0.03 mg/1 ozone measured in the water before contacting the fish. Since
this value is the lower limit of analysis it can be concluded that if ozone
can be detected, the receiving waters are toxic to fish.
Short term exposure of catfish to both chlorine and ozone did not
appreciably affect renal function. No physiological changes were noted
in tubular water reabsorption, ion excretion, glomerular filtration rate
(GFR) and their corresponding urine flows.
Short term exposure of catfish to both chlorine and to ozone was shown
to appreciably reduce the ability of gill epithelium to take up sodium from
the external medium. Chlorine is actively taken up by the gills of catfish.
Uptake averaged 0.4 mg Cl2/min/kg.
Long term exposures of catfish to ozone had no significant effect on
blood pressure or heart rate. This suggests that fish might not be aware
of ozone at low concentrations and would exhibit no avoidance reaction to
its presence. This lack of avoidance was actually observed in the bio-
assay tanks.
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SECTION 3
EXPERIMENTAL METHODS
FISH HOLDING AND PREPARATION
Holding
Adult catfish (0.25 - 2.0 kg) were netted and returned to
the laboratory. They were held in 150 gal circulating tanks at
22 ± 0.5C. The circulating water was dechlorinated tap water
with approximately 6.5 mg/1 of dissolved oxygen. The light
regime was 12 hours light and 12 hours dark. The fish were fed
three times weekly on standard catfish pellets (Purina). All
fish were acclimated for two weeks before being used in exper-
iments . Only healthy fish were used for physiological obser-
vation.
Cannulation
A single fish was netted and placed in the anaesthetic,"MS
222" (Cresent Research Chemicals, Inc.), dissolved in water
from which the animal came. Following the loss of all movement
the fish was transferred to a plastic operating box which allow-
ed the gills to be covered and the tail exposed. The procedure
for cannulation was as follows and involved the following custom
made materials:
a. 2 1/2" long 20 gauge thin wall needle with a specially
constructed deflected point,
b. Guide wire with a 4 foot long 0.010" diameter core and
a 1/2" long 0.003" diameter felxible end. Over the core
and flexible end was attached a wound flexible cover
of 0.021" which allowed it to fit down the 20 gauge
thin wall needle.
c. Polyethylene cannula with an inside diameter of
0.023" (Clay Adams Intramedic PE 50).
Cannula insertion was as follows: a small hole was made with
a sewing needle directly lateral to the haemal arch and approx-
imately 1" before the last caudal vertebrae. Following the re-
moval of the sewing needle, the 20 gauge needle was inserted
with the deflection pointed downward. This needle was advanced
at a very flat angle until the point slipped between two haemal
arches and entered either the dorsal aorta or the caudal vein.
-------
Entrance of a vessel was determined by blood entering the syringe.
The needle was then rotated 180° so the deflected opening faced
forward. At this point, the fish was heparinized with approxi-
mately 0.5 cc of 1000 unit sodium heparin and the syringe removed
leaving the needle in place. Immediately the flexible guide wire
was introduced down the needle and into the vessel. The needle
was then removed leaving the end of the wire several inches up
the vessel. The measured cannula was then introduced over the
guide wire and the end placed several inches away from the punc-
ture site. The guide wire was then withdrawn and the cannula
filled with heparized saline and plugged with a headless stain-
less pin. Usually a single suture through a caudal ray held the
tube in place. Visual observation of the blood pressure in the
cannula identified the vessel cannulated. To select the other
vessel,be it the vein or dorsal aorta, it was only necessary to
enter the haemal arch at the last vertebrae. The needle was in-
serted along the hypural plate in line with the haemal arch. If
the aorta was needed, the deflected point was turned dorsally
and ventrally for the vein. The guide wire would then enter the
correct vessel and follow it along since they are both enclosed
in a bony sheath. With this method, no loss of blood occurred
during the operation or following, since the puncture made by
the 20 gauge needle was smaller than the outside diameter of the
cannula.
Catheterization
While the fish was anaesthetized, a bilumen rubber or poly-
ethylene (Intramedic) heart-shaped catheter was inserted into
the bladder through the urogenital papilla. If necessary, the
catheter was secured in place with a purse-string suture around
the papilla and also a suture through an anal fin ray. To assure
complete collection of the urine sample and to remove it as
quickly as possible from bladder influences, saturated air was
introduced a few seconds before the end of the collection period
in order to empty the catheter and collect the complete sample.
Using this method, it was possible to use larger sized catheters
and thus prevent blockage which occurred with smaller tubes.
To prevent evaporation, the tare-weighed collection tubes were
covered as the, fraction collector rotated.
Experimental Holding
Following the completion of the operation, the anaesthetized
fish was placed in a clear plastic box fitted with an inside
partition which was adjusted to prevent the fish from turning
over and tangling or dislodging the catheter and cannula tubes.
These tubes led to the outside through water tight rubber seals
in the end of the box. Water was pumped to the anterior end
of the box at a rate of 3-5 liters per minute. The fish exhib-
ited only slight opercular movements after complete recovery
-------
from the anaesthetic.
Blood Pressure
Following cannulation and catheterization, the fish was
transferred to the holding box and allowed to acclimate for sev-
eral days. Several hours before an experiment, the dorsal aortic
cannula was connected to a pressure transducer and a recording
physiograph (E and M Instrument). The transducer was calibrated
with a mercury manometer and normal blood pressure was recorded.
Gill Sodium Transport
In order to measure sodium uptake by the gills, a fish was
placed in a recirculating system containing 10 liters of deion-
ized water, with a known amount of NaCl labelled as 22Na (Amer-
sham/Searle). Removal of 22Na from the circulation tank was
measured employing a Beckman LS-133 scintillation counter.
Static Bioa.ssays
Using the procedure described in Standard Methods, static
bioassays (96 hour LC^Q) were run on fingerling channel catfish.
Each survival -mortality experiment was repeated three times.
For the tests, fingerlings averaging between 6 and 10 cm in
length were used. Temperature was maintained at 22 ± 0.5 C.
Oxygen was continually monitored and maintained above 4 ppm by
the intermittent introduction of air. Chlorine added to the de-
chlorinated tap water as sodium hypochlorite, was also monitored
and was returned to the initial value every 24 hours. In all
cases, it was observed that chlorine was rapidly removed from
the water of the assay bottles in the presence of respiring
fish.
Chlorine Uptake
A single fish was placed in a circulating tank containing
40 liters of water. A calculated dose of chlorine (as aqueous
solution of sodium hypochlorite) was introduced and the concen-
tration of chlorine in the tank measured at 5-10 minute inter-
vals until chlorine disappeared.
Continuous Flow or Flow Through Bioassay
Using methods described by Sprague (10) and Standard Meth-
ods, 13th Editon (11)Continuous flow or flow through bioassays
(96 hour LC5g) were run on fingerling catfish. Chlorine (sod-
ium hypochlorite) was continually added to the inflow water by
a syringe infusion pump, resulting in a constant chlorine level
in the water. Ozone was added to the water by aeration at the
inflow, using a Sander Ozonisator Model IV and pure oxygen.
Ozone concentrations were controlled by regulating the oxygen
supply using Kontes flowmeters.
8
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TABLE 1. SUMMARY OF STANDARD CHEMICAL AND PHYSICAL TESTS
USED IN THIS RESEARCH
Parameter Reference
(Refer to Refej
Section. )
Calcium
Chloride
Chlorine
Dissolved oxygen
Ifegnesium
Potassium
Sodium 22 (Beckman Liquid Scintillation Counter #133)
Inulin 14C (Beckman Liquid Scintillation Counter #133)
Sodium
Statis bioassay
Flow through Bioassay
Ozone
(12)
(11)
(11)
(12)
(12)
(11)
(13)
(13).
(11)
(11)
(10)
(11)
(11)
pp.
pp.
pp.
pp.
pp.
pp.
pp.
pp.
pp.
pp.
pp.
pp.
pp.
102
377
112
60
112
283
1
1
317
569
5
570
271
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SECTION 4
RESULTS
FLOW THROUGH BIOASSAYS
Chlorine
The survival-mortality characteristics of fingerling chan-
nel catfish to chlorine were examined and results shown in Table
2 and Figure 1. A flow through bioassay compared to a static
bioassay results in a lower 96 hour LC5Q/ 0.07 mg/1 chlorine vs.
0.45 mg/1 chlorine. The chlorine concentration at which 50% of
the fish survived was read directly from Figure 1. A flow
through or continuous flow bioassay is a more accurate measure-
ment for 96 hour LC5Q of a highly reacting toxicant like chlor-
ine (10). The chlorine concentrations in Figure 1 are based on
the measured amounts of chlorine added to the inflow and do not
represent concentrations during tests with fish. In this report
it is shown that the gills of catfish rapidly remove chlorine
from the medium. Thus the chlorine concentrations shown are
levels that would be representative at treatment outfalls. Sam-
ples taken from receiving waters should show chlorine concentra-
tions significantly lower. Based on our 96 hour LCso of 0.07
mg/1 chlorine and using the "Aquatic Life Water Quality Criteria"
(14) of 1/10 the 96 hour LC50/ the final concentration of chlor-
ine in the receiving waters should not exceed 0.007 mg/1. It
should be noted that this concentration is below the limits of
accurate analyses (11).
Ozone
The flow through bioassay was also used for ozone and the
survival-mortality characteristics of fingerling catfish were
examined and results are in Table 3 and Figure 2. Ozone proved
to be more toxic to fingerling channel catfish than chlorine.
The 96 hour LC5Q for ozone, read directly from Figure 2 is 0.03
mg/1. As in the case of chlorine, the ozone concentrations are
based on the amounts of ozone added to the water without fish in
the system. Because ozone is highly reactive, it was rapidly
decomposed with the presence of organics (fish).
In the presence of fish,ozone could not be detected in the
bioassay waters due to the insensitivity of the lower limits of
10
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TABLE 2. NINETY-SIX HOUR LC5Q BIOASSAY DATA FOR ICTALURUS PUNCTATUS
Initial
Chlorine
Concen.
(mg/1)
2.0
1.0
.9
.8
.7
.6
.5
.4
.3
.2
.1
.08
.05
.03
0
STATIC
Average % Kill
for 3 duplicate
experiments
100%
100
100
100
100
100
63
37
30
13
10
-
-
-
0
BIOASSAY
Time Corresponding
to average % Kill
(hours )
2-3
4-8
36
48
60
60
96
96
96
96
96
-
-
-
96
FLOW THROUGH
Average % Kill
for 2 duplicate
experiments
—
100%
100
100
100
100
100
100
-
100
95
75
5
0
0
BIOASSAY
Time Corresponding
to average % Kill
(hours )
-
24
24
24
48
48
72
72
-
72
96
96
96
96
96
-------
ro
% KILL
100,.
D STATIC BIOASSAY
OF LOW THROUGH BIOASSAY
LC 50
0-45 mg /
0
0-2 0-3 0-4 0-5 0-6 0-7 0-8 0-9 1-0 >l-0
CHLORINE, mg/l
Figure 1. A comparison of survival-mortality characteristics
of fingerling catfish to chlorine exposure in a
static bioassay and in a flow through bioassay.
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CO
TABLE 3. NINETY-SIX HOUR LC FLOW THROUGH BIOASSAY
oU
DATA OF OZONE FOR ICTALURUS FUNCTATUS
Initial Ozone
concentration
(mg/1)
.20
.18
.16
.14
.13
.12
.10
.05
.04
.018
0
Percent kill for three
duplicate experiments
expt. 1 expt. 2 expt. 3
100
100
100
100
100
80
60
60
60
0
0
100
100
100
100
100
100
100
80
60
0
0
100
100
100
100
100
100
100
80
80
0
0
Average
100
100
100
100
100
93
86
73
66
0
0
Time corresponding to
average % kill (hours)
24
24
24
24
36-48
96
96
96
96
96
96
-------
% KILL
100
80
60
40
20
02 -04 -06 -08 -10 -12 -14
OZONE, mg/|
•16 -18 -20 >-20
Figure 2. Survival-mortality characteristics of fingerling
catfish to ozone exposure in a flow through
bioassay.
-------
the analytical technique. However, the presence of ozone was
detectable by its characteristic odor. At the lower level ozone
concentrations were estimated from the oxygen flow rates (mg/1
63: 1/min 02). Based on acceptable water quality criteria (1/10,
96 hour LC50),if ozone can be detected either by analytical
techniques or by its odor (detectable odor 0.02-0.05 ppm, Feld-
stein, 15),it can be assumed that its concentration exceeds a
safe environmental limit.
PHYSIOLOGICAL OBSERVATIONS
The results of the bioassay experiments established a refer-
ence point for the physiological evaluation of the effects of
chlorine and ozone exposure on adult fish. Using the LC5Q as a
guide, adult catfish (0.25-2 kg) were exposed to various levels
of chlorine and ozone and monitored for those physiological
changes which were considered most important. Parameters moni-
tored included: kidney functions (ion excretion, tubular water
reabsorption, glomerular filtration rate), gill sodium uptake,
blood pressure and heart rate.
Kidney Function
Chlorine — The use of plasma insulin clearance as a measure
of kidney glomerular filtration rate in mammals and fish is
well established (16,17). All of the calculations performed
on kidney function parameters were done according to Pitts (16).
Figures 3 and 4 show typical glomerular filtration rates and
urine flows for two different fish, both before and after the
addition of chlorine. In these two experiments chlorine was
added to the 10 liter reactor 10 hours after the start of the
experiment. The concentration of chlorine added was 0.4 mg/1 in
the experiment shown in Figure 3, and 0.2 mg/1 in the experiment
shown in Figure 4.
A very high correlation between glomerular filtration rate
and urine flow is evident in Figures 2 and 3. This strong cor-
relation indicates that tubular water reabsorption is highly
regulated. There is also' a significant fluctuation of urine
flow and glomerular filtration rate with time. This suggests
that there is a high variation in the permeability of water
through the gill and body surfaces of the catfish. A correspond-
ing variability in urine ion concentration (Figure 5) and rate
of ion excretion (Figure 6) is also observed through time.
After carefully observing the renal function of catfish,
before and after exposure to chlorine, it is concluded that
exposure of this fish to toxic concentrations of chlorine does
not markedly affect flow rate of urine, urine ion concentration,
or glomerular filtration rate.
15
-------
to
3=
o
_J
u.
U
z
—
o:
•o
c
o
OC
U.
CD
D GFR
URINE FLOW
Figure 3, Effect of short term chlorine exposure on the glomerular filtration
rate (GFR) and corresponding urine flow of Ictalurus punctatus (Fish #5
wt. 1.155 kg, and 0.4 mg/1 of chlorine added 10 hours after start of
experiment).
-------
Figure 4. Effect of short term chlorine exposure on the glom-
erular filtration rate (GFR) and corresponding urine
flow of Ictalurus punctatus (Fish #16, wt. 0.368 kg,
and 0.2 iiig/1 of chlorine added 10 hours after the
start of the experiment).
17
-------
72r
48 -
w
z
o
o e
o
u
8
TIME
12
IN H OUR S
16
Figure 5.
Effect of short term chlorine exposure on the
concentration of ions in the urine of Ictalurus
punctatus (Fish #16, wt. 0.368 kg and 0.2 mg/1
of chlorine added 10 hours after the start of
experiment).
18
-------
560
D No
O K
C I
Figure 6. Effect of short term chlorine exposure on the rate of excretion of ions
in the urine of Ictalurus punctatus (Fish #16, wt. 0.368 kg, and 0.2 mg/1
of chlorine added 10 hours after the start of the experiment).
-------
ro
o
15
12
liJ o> Q
Z •*
E ^
D •«=
^
• E 6
QC
U.
D GFR
O URINE FLOW
pH 7
TKMP 22 i . 5C
Figijre 7. Effect of short term ozone exposure on the
glomerular filtration rate and corresponding
urine flow of j[cta_lurus PHHS^il.!^ (fish //37,
wt. 1.102 kp,,0. 36 mg7T~ozoneT7~
-------
Ozone — In these three experiments ozone was added by aera-
tion to the 10 liter flow through system. Figures 7 and 8 show
the experiment where concentration of ozone added was 0.36 mg/1.
Similar results were obtained using ozone concentrations of 1.8
and 0.9 mg/1. The length of exposure was from 12 to 24 hours at
concentrations well above the 96 hour LC5Q.
A very high correlation between glomerular filtration rate
and urine flow is evident in Figure 7- This strong correlation
indicates that tubular water reabsorption is highly regulated
and that there is also a significant fluctuation of urine flow
and glomerular filtration rate with time. This suggests that
there is a high variation in the permeability of water through
the gills and body surfaces of catfish. A corresponding varia-
bility in urine ion concentration (Figure 8) is also observed.
After carefully observing the renal functions of channel
catfish before and after exposure to ozone, it is concluded
that short term exposure of this fish to toxic concentrations
of ozone does not affect urine volume, urine ion concentration
or glomerular filtration rate. However, these results do not
preclude the possibility of long term ozone exposure affecting
renal function.
Gill Function
Uptake of Chlorine by Gills — Chlorine is rapidly removed
from aqueous solution by the gills of fish. This occurrence was
deduced from the experimental data presented in Figure 9. In
these studies, a different concentration of chlorine (1.5-10
mg/1) was added to the circulating system for each fish. The
concentration of chlorine was measured initially and at 5-10
minute intervals until the chlorine had disappeared. The exper-
imental system contained 40 liters of water, in which only slight
loss of chlorine was noted when no fish were present (Line A in
Figure 9). Similarly, only a small amount of chlorine was re-
moved when an anesthetized fish with no opercular movements was
placed in holding tank (Line B in Figure 9). The near coinci-
dence of Lines A and B in Figure 9 indicates that the body sur-
face of the fish does not react markedly with chlorine. However,
when a normal respiring fish is present in the system and chlor-
ine is introduced, dramatic changes occur. The fish immediately
reacts by significantly increasing opercular pumping rate. Peri-
odically, at this time the fish appears to "cough" in an apparent
attempt to clear the gills. Also, the rate of removal of chlor-
ine in the presence of respiring fish (upper four lines in Fig-
ure 9) is markedly higher than that for an anesthetized fish.
The average rate of uptake of chlorine for a respiring fish is
relatively constant at 0.4 mg chlorine per minute per kg of fish
and is valid over a wide range of fish sizes (0.37-1.65 kg). The
obvious conclusion from the experimental data shown in Figure 9
is that chlorine is rapidly removed by the gills of fish.
21
-------
I80r
ro
ro
pH 7
TEMP 22 ± .5C
TIME
Figure 8. Effect of short term ozone exposure on the rate
of excretion of ions in the urine of Ictalurus
punctatus (fish #37, wt. 1.102 kg, 0.36 mg/1
ozone)
-------
ro
co
TEMP. 22±-5C
1
e
___ • " BOX, NO FISH
_— * — :
i i i
12 16 20
TIME. IN MINUTES
1
24
LINE A
i I
28
Figure 9. Chlorine uptake by five catfish. Four respiring fish had uptake rates
averaging 0.4 mg Cl/min/kg. An anesthetized fish (no opercular move-
ments) had an upatke rate of 0.05 mg Cl/min/kg. Loss of chlorine from
the experimental system with no fish present was negligible.
-------
Gill Sodium Transport
Chlorine — The influence of chlorine exposure on the uptake
of 22Na by gills of fish is shown in Figure 10. The disappear-
ance of 22Na from aqueous phase (ordinate axis in Figure 10) is
a measure of the removal of 22Na by the gills of fish. The addi-
tion of 0.1 mg/1 of chlorine at the start of the experiment mark-
dely reduced the ability of the gills to remove sodium from aque-
ous solution. This reduction in uptake rate following chlorine
exposure suggests an impairment of normal physiological function
in the sodium transport system.
Ozone —The influence of ozone exposure on the uptake of
22Na by the gills of fish is shown in Figure 11. The disappear-
ance of 22Na from the aqueous compartment is a measure of the
removal of 22Na by the gills of the fish. Phase I of the exper-
iment utilized the fish as its own control. A one day recovery
period followed. The addition of 0.1 mg/1 ozone in the second
phase of the experiment markedly reduced the ability of the gills
to remove sodium from the aqueous solution. This reduction in
uptake following ozone exposure suggests an impairment of normal
physiological functions in the sodium transport system of the gill
epithelium.
Heart Rate and Blood Pressure
Single Dose Exposure to Chlorine — The effect of short term
exposure to chlorine on the heart rate and blood pressure of
adult channel catfish was determined. Blood pressure was con-
tinuously recorded prior to/ and following, the introduction of
chlorine into the 10 liter recirculating system. Figure 12
demonstrates the temporal changes in dorsal aortic blood pressure
and heart rate after exposure to 1 mg/1 of chlorine, while Figure
13 illustrates the immediate response in the above two parameters
after exposure to various concentrations of chlorine. In both
experiments, each fish had an initial mean blood pressure of 23
mm Hg, a pulse pressure of 2 mm Hg and a heart rate of 78 to 84
beats per minute. Measureable variations and fluctuations in
blood pressure were recorded prior to the addition of chlorine.
These variations may be due partially to opercular movements and
changes in gill blood vessel resistance. Swimming movements also
significantly alter dorsal aorta blood pressure.
The introduction of a single dose of chlorine to the exper-
imental system caused an immediate drop in blood pressure. The
mean blood pressure dropped from a pretreatment level of 23 mm
Hg to a low of 10 mm Hg between one to five minutes after
chlorine was added. Blood pressure did not return to pretreat-
ment levels in the 30 minute period following initial exposure.
Generally the recovery time and change in blood pressure was
increased with increasing chlorine concentration. It was also
observed that rhythmic changes in mean blood pressure correlated
24
-------
CI2 Added
7
1200
ac.
UJ
Q_
ro
01
CM
CM
800
400
0
pH 7
D.O. 6.5
TEMP. 22±.5C
0
A
22
No + Chlorine
22
Q No
8 12 16
TIME, IN HOURS
20
24
Figure 10.
22,
Effect of short term chlorine exposure on the uptake of ^Na by the gills
of Ictalurus punctatus (Fish #34, wt. 1.119 kg, and 0.1 mg// of chlorine
added at start of experiment).
-------
1000
ro
800
UJ
I-
z
s
v»
CO
z
o
u
o
•z.
OJ
C\J
600
400
200
22Na
22Na + Ozone
pH 7
TEMP 22 ± .5C
•QFISH 48, WT. 0-56 kg.
-DFISH 51, wt. MO kg.
6
TIME
9
HOURS
12
15
Figure 11.
Effect of short term ozone exposure on the uptake
of 22jvja by the gills of Ictalurus punctatus
(0.1 mg/1 ozone).
-------
30r
X
E
6
NORMAL
1-0 PPM
CHLORINE
1-0 MINUTE
5 MIN LATER
10 MIN LATER
ro
30r-
15 MIN LATER
20 MIN LATER
LL
25 MIN LATER
30 MIN LATER
Figure 12. Influence of short term chlorine exposure on the heart rate and blood
pressure of Ictalurus punctatus (fish #26, wt. 1.036 kg, and 1.0 mg/1
of C12 added at start of experiment).
-------
30
x
E
£
10
NORMAL
0-03 PPM
0-07 PPM
0-07 PPM
f
0-07 PPM
sJa
1-0 MINUTE
ro
oo
0-07 PPM
0-2 PPM
0-3 PPM
0-3 PPM 5MIN
LATER
Figure 13. Heart rate and blood pressure of Ictalurus punctatus immediately following
exposure to different concentrations of chlorine (fish #30, wt. 0.985 kg).
-------
with opercular movements. Severe opercular movements were assoc-
iated with drastic fluctuations in mean pressure.
The heart rate of the fish was also affected by short term
exposure to chlorine. Heart rate declined from a normal value
of 78 beats/min to 42 beats/min, 20 minutes after initial expo-
sure.
The most significant physiological effect of single dose
chlorine exposure were changes in dorsal aortic blood pressure.
Figure 14 summarizes the changes in blood pressure and heart
rate following exposure to 1.0 mg/1 of chlorine. The change in
blood pressure is dramatic (vagal inhibition) and the time for
recovery increases with increasing chlorine exposure.
Continuous Exposure to Chlorine — The effect of long term
exposure to low levels of chlorine on the heart rate and blood
pressure of adult channel catfish was determined. Blood pressure
was continually recorded prior to and after introduction of
chlgrine to the flow through system. Figure 15 demonstrates the
change in dorsal aortic pressure and heart rate after a 7.5 hour
exposure to 0.22 mg/1 chlorine. Figure 16 demonstrates the
change in dorsal aortic blood pressure and heart rate after a
5 hour exposure to 0.17 mg/1 chlorine.
Fish #38, Figure 15, had an initial mean blood pressure of
27 mm Hg, a pulse pressure of 3 mm Hg, and a heart rate of 55-
60 beats per minute. The introduction of 0.22 mg/1 chlorine
caused an immediate drop in mean blood pressure, from 27 mm Hg
to 17 mm Hg, an initial decrease in heart rate of 50% but an
increase in the pulse pressure. The initial response is proba-
bly due to vagal inhibition. During the 7.5 hours of chlorine
exposure blood pressure dropped to 16 mm Hg, pulse pressure to
2 mm Hg and heart rate decreased to 40-45 beats per minute.
Fish #39, Figure 16, had an initial mean blood pressure of
27 mm Hg, a pulse pressure of 2 mm Hg,- and a heart rate of 65-
70 beats per minute. The introduction of 0.17 mg/1 chlorine
caused a gradual drop in' blood pressure from 27 mm Hg to 20 mm
Hg. There was no apparent change in pulse pressure or heart
rate. During 5 hours of chlorine exposure, blood pressure
dropped to 21 mm Hg, pulse pressure dropped to 1 mm Hg and heart
rate decreased to 55-60 beats per minute.
Figure 17 is a condensed summary of blood pressure fluctua-
tions (Fish #38 and Fish #39) due to chlorine exposure over an
8 hour period.
Low Level Chlorine Exposure — The exposure of fish to
chlorine at levels approaching the 96 hour LC5Q was immediately
detected by the pronounced drop in blood pressure (Figure 18).
Continued exposure for 5 hours resulted in an increase in heart
29
-------
CO
CO
-------
30
en
X
E
E
10 '-
NORMAL
I MINUTE
0-22 mg/1 CHLORINE
mm
I
5 MIN LATER
CO
10 ("HOUR "LATER
1-5 HOURS LATER
75 HOURS LATER
Figure 15. Influence of long term chlorine exposure op
the blood pressure and heart rate of Ictalurus
punctatus (fish #38, wt. a832 kg).
-------
30
E
E
I01
NORMAL
0-17 mg/l CHLORINE
I MINUTE
co
ro
30
o»
X
E
E
in
•WL* *•«*''• W'M !¥«'"*''
^^uf*^H.»*>**
-------
456
TIME, HOURS
oo
00
CD
456
TIME, HOUR S
Figurfi 17. Summary of chanp,es in blood pressure following
long term exposure to chlorine (A- fish #38,
wt. 0.823 kg, 0.22 mg/1 chlorine, B- fish #39,
wt. 0.85M kg, 0.17 mg/1 chlorine).
-------
30
X1
E
E:
10'
NORMAL
I MINUTE
0-08 mg / I C HLORINE'
oo
.£»
30
E
E
1/2 HOUR LAT ER
'^YWW^^
I HOUR LATER
5 HOURS LATER
Figure 18. The inJ Lu
-------
rate from 22 beats/minute to 45 beats/minute. At an exposure of
0.03 mg/1 chlorine there was no initial reaction shown by blood
pressure or heart rate. After two hours the fish appeared normal
with no apparent disfunction (Figure 19). Thus it can be con-
cluded that on exposure to a chlorine concentration of 0.007 mg/1
chlorine (1/10 of 96 hour LCsg) fish should show no physiological
response and the environment should be safe for their survival.
Ozone
Continuous Exposure to Ozone — Tests to determine the
effects of ozone on blood pressure, pulse pressure and heart rate
were run. Levels of ozone from 0.1 to 0.5 mg/1 for 10 hours had
no apparent effect on these parameters. This leads to the con-
clusion that ozone has no effect on gill vascular resistance.
It was also noted that the pronounced vagal inhibitions associat-
ed with chlorine exposure did not occur with ozone. The absence
of this drop in blood pressure normally attributed to vagal
inhibitions induced by external stimuli leads to the conclusion
that fish may be insensitive to the presence of ozone at low
levels in the water and any avoidance reaction would not take
place. Thus, it can be concluded that since ozone is so extreme-
ly toxic to fish and that they apparently lack the ability to
sense its presence at low levels in their environment, the area
immediately adjacent to treatment outfalls poses a threat. Mea-
sures should therefore be introduced to assure that no ozone
reaches the aquatic environment.
35
-------
30
X ,;
E
E
10
NORMAL
MINUTE
ic'2
0-03 mg I CHLORINE
CO
-------
REFERENCES
1. Brungs, W.A. Effects of residual chlorine on aquatic life. Jour.
Water Poll. Control Fed., 45, 2180-2193, 1973.
2. Tsai, C., "Effects of Chlorinated Sewage Effluents on Fish in
Upper Patuxent River, Maryland." Chesapeake Sci. 9, 2, 83 (1968).
3. Arthur, J.W. and J.G. Eaton, "Chloramine Toxicity to the Amphipod,
Gammarus Pseudolimnaeus, and the Fathead Minnow, Pimephales promelas"
Journ. Fish. Res. Bd. Can., 28, 1841 (1971).
4. Becker, C.D. and T.O. Thatcher. Toxicity of Power Plant Chemicals
to Aquatic Life. U.S. Atomic Energy Commission. Report, Wash-1249,
UC-11, June 1973.
5. Hubbs, C.L. High Toxicity of Nascent Oxygen. Physiological Zoology
3(4):441-460, 1969.
6. MacLean, S.A., et al. The Effects of Ozone-treated Seawater on the
Spawned, Fertilized, Meiotic, and Cleaving Eggs of the Commercial
American Oyster. Mutation Research. 2:283-285, 1973.
7. Benoit, R.F., N.A. Matlin. Control of Saprolegnia on Eggs of Rainbow
Trout (Salmo gairdneri) with ozone. Trans. Amer. Fish. Soc.
95(4):43-432, 1966
8. Spotte, S.H. Fish and Invertebrate Culture. Wiley-Interscience, Inc.,
New York. 1970.
9. Adger, S., Hawaiian Marine Imports, Houston, Texas* Personal communication.
10. Sprague, J.B. The ABC's of Pollution Bioassay using Fish. A paper
presented at The Symposium on Environmental Monitoring, Los Angeles,
California. June 27-28, 1972.
11. Standard Methods, 13th Edition. American Public Health Assoc., New
York, N.Y. 1971.
12. Methods for Chemical Analysis of Water and Wastes. U.S. EPA Report No.
EPA-625-/6-74-003, 1974.
13. Beckman Instruments, Inc. Fullerton, California. Bulletin 800--C.
37
-------
14. Horton, R.K. Aquatic Life Water Quality Criteria. Environ. Sci.
Technol, (11):888,1967.
15. Feldstein, M. "Air Pollution" in Handbook of Analytical Toxocology,
ed. I. Sunshine. The Chemical Rubber Co. 708, 1969.
16. Pitts, R.F. Physiology of the Kidneys and Body Fluids. Amer.
Public Health Asso., New York, N.Y. 1971.
17. Hickman, C.P., Jr., B.F. Trump. The Kidney, in Fish Physiology,
Vol. 1, ed. W.S. Hoar, D.J. Randall, New York Academic Press.
91-239, 1969.
38
-------
TECHNICAL REPORT DATA
(Please read Instructions on the reverse before completing)
1. REPORT NO.
EPA-600/2-79-050e
3. RECIPIENT'S ACCESSION NO.
4. TITLE AND SUBTITLE
MAXIMUM UTILIZATION OF WATER RESOURCES IN A PLANNED
COMMUNITY
Chlorine and Ozone Toxicity Evaluation
5. REPORT DATE
August 1979 (Issuing Date)
6. PERFORMING ORGANIZATION CODE
7. AUTHOR(S)
8. PERFORMING ORGANIZATION REPORT NO.
Brian Hammond and James Bishop, Jr.
9. PERFORMING ORGANIZATION NAME AND ADDRESS
Department of Biology
Rice University
Box 1892
Houston, Texas, 77001
Present address:
Research Secretariat
Alberta Environment
Canada T5T 1B8
10. PROGRAM ELEMENT NO.
1BC822, SOS #2, Task 02
11. CONTRACT/GRANT NO.
802433
12. SPONSORING AGENCY NAME AND ADDRESS
Municipal Environmental Research Laboratory--Cin.,OH
Office of Research and Development
U.S. Environmental Protection Agency
Cincinnati, Ohio
13. TYPE OF REPORT AND PERIOD COVERED
Final 7/73 - 12/76
14. SPONSORING AGENCY CODE
EPA/600/14
15. SUPPLEMENTARY.NOTES
One in a series of volumes of one report. P.O. Anthony N. Tafuri and Richard Field,
Storm and Combined Sewer Section, FTS 340-6675, (201) 321-6675.
16. ABSTRACT
To ensure adequate water quality for impoundments receiving disinfected wastewate :
in The Woodlands, Texas the following experiments were conducted. Using fingerling
channel catfish, Ictalurus punctatus the 96 hour LC^Q for chlorine is 0.07 mg/1
(total chlorine) and 0.03 mg/1 for ozone in flow through bioassays.
Chlorine and ozone exposures had little effect on kidney functions. Exposure
to both chlorine and ozone drastically reduced the ability of the gills to actively
absorb sodium from the water.
Long term exposure to chlorine drastically reduced both blood pressure and heart
rate while exposure to ozone had little,if any, effect. Blood pressure and heart rate
are very sensitive physiological parameters and changes are indicative of a stressful
environment.
Both chlorine and ozone are extremely toxic to fish at low levels. If detected
in receiving waters by present analytical techniques, a toxic condition exists.
17.
KEY WORDS AND DOCUMENT ANALYSIS
DESCRIPTORS
b.IDENTIFIERS/OPEN ENDED TERMS
c. COSATl Field/Group
Tpxicology, Chlorine, Ozone, Fresh water,
Pollution, Physiology, Blood pressure,
Kidney, Fishes, Catfishes
Fish toxicity, Fish
physiology, Gills
13B
18. DISTRIBUTION STATEMENT
RELEASE TO PUBLIC
19. SECURITY CLASS (ThisReport)
UNCLASSIFIED
21. NO. OF PAGES
49
20. SECURITY CLASS (This page)
UNCLASSIFIED
22. PRICE
EPA Form 2220-1 (Rev. 4-77)
39
iLs.anBMen mane oract n» -»57-i«><
------- |