EPA-600/3-76-036
April 1976
Ecological Research Series
EFFECTS OF CHLORINE AND
SULFITE REDUCTION ON
LAKE MICHIGAN INVERTEBRATES
Environmental Research Laboratory
Office of Research and Development
U.S. Environmental Protection Agency
Duluth, Minnesota 55804
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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 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 and 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 ECOLOGICAL RESEARCH series. This series
describes research on the effects of pollution on humans, plant and animal
species, and materials. Problems are assessed for their long- and short-term
influences. Investigations include formation, transport, and pathway studies to
determine the fate of pollutants and their effects. This work provides the technical
basis for setting standards to minimize undesirable changes in living organisms
in the aquatic, terrestrial, and atmospheric environments.
This document is available to the public through the National Technical Informa-
tion Service, Springfield, Virginia 22161.
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EPA-600/3-76-036
April 1976
EFFECTS OF CHLORINE- AND SULFITE
REDUCTION ON LAKE MICHIGAN INVERTEBRATES
by
A. M. Beeton
P. K. Kovacic
A. S. Brooks
Center for Great Lakes Studies
The University of Wisconsin-Milwaukee
Milwaukee, Wisconsin 53201
Grant No. R-801035-01
Project Officer
D. T. Olson
Environmental Research Laboratory
Duluth, Minnesota 55804
U.S. ENVIRONMENTAL PROTECTION AGENCY
OFFICE OF RESEARCH AND DEVELOPMENT
ENVIRONMENTAL RESEARCH LABORATORY
DULUTH, MINNESOTA 55804
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DISCLAIMER
This report has been reviewed by the Environmental Research
Laboratory, U.S. Environmental Protection Agency, and approved
for publication. Approval does not signify that the contents
necessarily reflect the views and policies of the U.S. Environ-
mental Protection Agency, nor does mention of trade names or
commercial products constitute endorsement or recommendation
for use.
XI
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ABSTRACT
The acute toxicity of residual chlorine was determined for
the copepod Cyclops bicuspidatus thomasi and the rotifer
Keratella cochlearis. The 96-hour TL5Q value for Cyclops
was,0.084 mg/1 total residual chlorine added as monochlora-
mine. When Cyclops was exposed to sodium hypochlorite the
96-hour TL50 was 0.069 mg/1 total residual chlorine. The
4-hour TLso value for Keratella was 0.019 mg/1 total res-
idual chlorine added as monochloramine.
Chemical studies determined that sodium sulfite was an
efficient, inexpensive chemical agent for reducing chlorine
residuals which did not produce undesirable by-products.
Complete reduction was accomplished in less than 20 seconds
with a calculated km^n of 43 sec"-'-. Bioassay studies indi-
cated that sodium sulfite added to chlorinated water com-
pletely eliminated the acute toxicity of residual chlorine
to both Cyclops bicuspidatus thomasi and Keratella coch-
learis.
Field studies in Milwaukee Harbor and adjacent Lake Michigan
indicated that measurable chlorine residuals were confined
to a very small area surrounding the effluent from the Jones
Island Sewage Treatment Plant. Significant reductions in
the populations of benthic organisms were observed in the
effluent plume area after the start of chlorination.
This report was submitted in fulfillment of Grant No.
R-801035-01 by The University of Wisconsin-Milwaukee, under
the sponsorship of the Environmental Protection Agency.
Work was completed as of August, 1974.
111
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CONTENTS
Section Page
I Conclusions 1
II Recommendations 2
III Introduction 3
IV Field Studies 5
V Chemical Studies 30
VI Bioassay Studies 47
VII References 64
VIII Publications Resulting from Project 72
IX Appendices 73
v
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FIGURES
Page
1 Sampling Stations in Milwaukee Harbor and 7
Adjacent Lake Michigan.
2 Abundance (#/m2) of Oligochaeta in Milwaukee 12
Harbor and Adjacent Lake Michigan, June 1971
and June 1972.
3 Abundance of Oligochaeta in June 1972 as a 15
Percentage of the June 1971 Abundance.
4 Percentage of Tubifex tubifex in the Oligo- 16
chaete Population of Milwaukee Harbor and
Adjacent Lake Michigan.
5 Percentage of Limnodrilus hoffmeisteri in the 17
Oligochaete Population of Milwaukee Harbor
and Adjacent Lake Michigan.
6 Percentage of Peloscolex multisetosus multi- 18
setosus in the Oligochaete Population of
Milwaukee Harbor and Adjacent Lake Michigan.
7 Percentage of Stylodrilus heringianus and 19
Limnodrilus cervix-claparedeanus in the
Oligochaete Population of Milwaukee Harbor
and Adjacent Lake Michigan.
8 Mean Monthly Temperatures in Milwaukee Harbor, 28
1971 and 1972 (Adapated from Wisconsin Surface
Water Monitoring Data 1969-72).
9 Current-Voltage Curve of HOC1 and Blank. 36
LO Current-Voltage Curve of HOC1 and Blank with 37
Triton X-100 Added.
LI Current-Voltage Curve of NH2Cl and Blank with 38
Triton X-100 Added.
vx
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FIGURES (Continued)
Page
12 Current-Voltage Curve for Sodium Sulfite 40
13 Schematic Representation of the Three-Electrode 42
Cell Used in Obtaining Current-Voltage Curves
for Chlorine and Chloramine.
14 Current-Voltage Curves for Various Concen- 43
trations of NH^Cl. Detection Limit was 1 mg/1.
15 Current-Voltage Curves for NH2C1 Following 44
Electrode Pretreatment. Dectection Limit was
0.5 mg/1.
16 Relationship Between Current and Concentration 45
for NH2C1 at -0.50 Volts.
17 Proportional Diluter Used for Bioassay 48
Experiments
18 Test Aquarium Used in Bioassay Experiments 51
With Insert Jar Used for Rotifer Tests.
19 The Toxicity of Residual Chlorine, as Sodium 53
Hypochlorite, to Cyclops bicuspidatus thomasi
During 96-Hour Exposures at 15°C.
20 The Toxicity of Residual Chlorine, as Mono- 58
chloramine, to Cyclops bicuspidatus thomasi
During 96-Hour Exposures at 15°C.
21 The Toxicity of Residual Chlorine, as Mono- 60
chloramine, to Keratella cochlearis During
4-Hour Exposures at 15°C.
vi i
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TABLES
No. Pag<
1 Physical Characteristics of Benthic Sampling 9
Stations, 1971 and 1972.
2 Comparison of Means of Paired Sets of Data of 14
Oligochaeta From 7 June 1971 and 20 June 1972.
The Level of Significance of the Difference
Between Means According to a Two-Tailed t-Test.
3 Total Residual Chlorine Values Observed in 22
Milwaukee Harbor and Adjacent Lake Michigan.
All Samples Pumped From a Depth of 2 m Unless
Designated as Surface Sample.
4 Average Chemical and Physical Analyses of 56
Untreated Lake Michigan Water From the
Linnwood Avenue Water Purification Plant,
Milwaukee, for 1973.
5 Summary of Effects of Residual Chlorine on 61
Aquatic Life (Adapted From Brungs 1973).
Vlll
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ACKNOWLEDGMENTS
The field study section of this report is based primarily
on the Master's thesis of P. Scott Hausmann ("The benthic
macrofauna of Milwaukee Harbor and adjacent Lake Michigan,"
Department of Zoology, The University of Wisconsin-Milwaukee,
August 1974).
Studies on the chemical reduction of chlorine were conducted
by John W. Strand as Master's thesis research (Part III—
"Reduction of chloramines in chlorinated sewage," Department
of Chemistry, The University of Wisconsin-Milwaukee, Sep-
tember 1972).
Additional chemical studies were undertaken by Drs. Calvin
O. Huber, Roy A. Crochet, F. Ryan Sullivan and Christopher
Daniels of the Department of Chemistry.
Bioassay data on the rotifers is from the Master's thesis of
Nevin E. Grossnickle ("The acute toxicity of residual chlor-
amine to the rotifer Keratella cochlearis (Gosse) and the
effects of dechlorination with sodium sulfite," Department
of Zoology, The University of Wisconsin-Milwaukee, August
1974) .
Crustacean bioassays were completed with the assistance of
R. Scott Burkhardt, Lynn L. Frederick, and David Latimer.
We also wish to acknowledge the assistance provided by the
staff of the City of Milwaukee Linnwood Avenue Water Purif-
ication Plant in obtaining lake water, and the staff of the
Milwaukee Sewage Commission Jones Island Sewage Treatment
Plant for providing information on plant operations.
Figures were prepared by Ratko J. Ristic and the manuscript
was typed and assembled by Irene Berg.
Contribution number 137, Center for Great Lakes Studies,
The University of Wisconsin-Milwaukee, Milwaukee, Wisconsin.
IX
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SECTION I
CONCLUSIONS
1. The chlorinated effluent from the Jones Island Sewage
Treatment Plant appears to have reduced the popula-
tions of benthic organisms in areas of Milwaukee
Harbor influenced by the effluent plume.
2. The rotifer Keratella cochlearis is one of the most
sensitive species to chlorine residuals having a
4-hour TLgg value of 0.019 mg/1 total residual chlorine
added as monochloramine.
3. The copepod Cyclops bicuspidatus thomasi is among the
more sensitive aquatic species to residual chlorine
with a 96-hour TI^Q value of 0.084 mg/1 total residual
chlorine added as monochloramine and 0.069 mg/1 total
residual chlorine in a solution of hypochlorite and
monochloramine.
4. Sodium sulfite is an effective chemical agent for
reducing chlorine residuals. It is not toxic to Cyclops
bicuspidatus thomasi or to Keratella cochlearis at
levels sufficient to reduce chlorine residuals observed
in chlorinated sewage. And, it effectively reduces the
toxicity of residual chlorine to these organisms without
producing undesirable by-products.
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SECTION II
RECOMMENDATIONS
1. A careful evaluation of the use of chlorine as a
disinfectant for sewage treatment should be undertaken
in terms of its effectiveness as a biocide and its
adverse effects on the aquatic environment.
2. The rate of chlorine application to sewage should be
carefully regulated in terms of the chlorine demand
of the sewage and receiving waters to minimize chlorine
residuals in the effluent plume.
3. Total residual chlorine levels should be kept below
0.01 mg/1 when applied continuously in order to protect
all but the most sensitive species. To protect even
the most sensitive species, total residual chlorine
levels should not exceed 0.002 mg/1 when applied on a
continuous basis.
4. Sodium sulfite should be used to reduce chlorine
residuals in situations where residuals cannot be
maintained within acceptable limits by sound operating
procedures and it can be demonstrated that a continu-
ously chlorinated effluent would have adverse effects
on important organisms in the receiving water body.
5. Studies should be undertaken to determine if other
compounds are formed in the process of chlorinating
sewage, such as chlorinated organics, which may not be
detectable as residual chlorine yet are toxic to aquatic
life and humans.
6. Pilot studies should be undertaken to determine the
effects of chlorine reduction by sodium sulfite, and
the reaction products formed, on receiving water bodies
of varying water qualities.
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SECTION III
INTRODUCTION
Chlorine, a strong oxidizing agent, has been, and continues
to be used in the treatment of sewage to control pathogenic
organisms, reduce odor, and to reduce the biochemical
oxygen demand. The Chlorine Institute (1974) estimates
that 187,000 tons of chlorine were used for the treatment
of sewage in the U. S. in 1973. Chlorination is considered
desirable from an operational standpoint because it improves
the physiochemical characteristics of the effluent and
reduces the bacterial load of sewage although its effective-
ness in controlling pathogenic organisms has been questioned
(Durham and Wolf 1973). Chlorinated sewage represents a
hazard to the receiving waters, however, since the residual
chlorine is toxic and chloramine and unknown chlorinated
organic compounds are produced which are toxic to aquatic
life and generally more persistent than free chlorine.
Numerous studies have been conducted which demonstrate the
toxicity of chlorine to various types of organisms. Brungs
(1973) has compiled an excellent review on the effects of
residual chlorine on aquatic life which summarizes this
literature.
This project was undertaken to determine the effect on
invertebrates of Milwaukee Harbor and adjacent Lake Michigan
of the chlorinated effluent from the Jones Island Sewage
Treatment Plant and to determine what additional chemical
treatment could be employed to eliminate or reduce the tox-
icity of the residual chlorine in the effluent.
To accomplish these objectives, three basic approaches were
taken. First, field studies were conducted to sample the
benthic community of the harbor prior to and subsequent to
the start of chlorination. The original research proposal
included sampling of fish and plankton, but at the request
of the Environmental Protection Agency studies of fish were
not included. Data on plankton populations in the harbor
-------
are not included because the frequent exchange of various
water masses into and through the harbor make such data
meaningless in assessing the effects of a single effluent.
The measurement of residual chlorine in Milwaukee Harbor
and adjacent Lake Michigan was also included in the field
program. Secondly, laboratory bioassay experiments were
conducted to determine the toxicity of residual chlorine
to copepods and rotifers, which are normally the most
abundant organisms within the study area. Bioassays on
organisms such as amphipods were not included since the
results of the field studies indicated that they were not
an important portion of the biota in the study area. The
third approach involved laboratory chemical studies which
were undertaken to identify compounds which would reduce
or eliminate the toxicity of residual chlorine in chlor-
inated sewage. Bioassay experiments were conducted using
the most promising compound to determine its toxicity and
effectiveness in reducing chlorine toxicity to the test
organisms.
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SECTION IV
FIELD STUDIES
INTRODUCTION
Many workers have suggested that changes in environmental
quality are inevitably reflected in the composition of the
benthos and that the best way to determine the extent and
kind of change taking place in the environment is by the
study of the benthos (Hynes 1960; Mackenthun 1969). Henson
(1966) has stated that maximum sensitivity in detecting
environmental changes requires investigation at the species
level. Changes in the community structure and abundance of
organisms have provided useful indices of pollution (Hooper
1969; Hynes 1960). In several instances changes in the
benthic biota have been valuable in documenting eutrophi-
cation in the Great Lakes (Beeton 1965, 1966; Howmiller and
Beeton 1970, 1971). While warning that no universal indi-
cator species exists, Brinkhurst (1965) suggests that
analysis of benthos and substrate will give an indication
of the source, nature and extent of pollution.
Fortunately with increasing awareness of the continued
decrease in water quality there has been increased research
on the macrobenthos of the Great Lakes from which an ade-
quate inventory of the fauna is being built (Brinkhurst 1966a,
1966b, 1967, 1969, 1970; Brinkhurst, Hiltunen and Herrington
1968; Carr and Hiltunen 1965; Henson 1966; Hiltunen 1967,
1969a, 1969b, 1969c; Howmiller and Beeton 1970, 1971; Johnson
and Matheson 1968; Robertson and Alley 1966; Schuytema and
Powers 1966; Veal and Osmond 1968). From these studies it
has been shown that of all the benthic organisms the Oligo-
chaeta are assuming the greatest importance in terms of
abundance, especially in areas of organic pollution, e.g.,
harbors. Oligochaetes have been used to assess increases
in pollution CCarr and Hiltunen 1965).
-------
This study was undertaken to document the composition of
the benthic fauna, especially that of the aquatic oligo-
chaetes, of Milwaukee Harbor and adjacent Lake Michigan and
to determine whether chlorination of sewage from the Jones
Island Waste Water Treatment Plant, Sewage Commission of
the City of Milwaukee, has affected the benthic community.
SITE DESCRIPTION
Milwaukee Harbor is an artificial harbor formed by man-
made breakwaters enclosing approximately 5.84 sq Km of a
natural embayment of Lake Michigan (Figure 1). An inland
harbor area encompasses about 0.93 sq Km which includes the
mouths of the Milwaukee, Kinnickinnic, and Menomonee Rivers.
The major tributary of the harbor, the Milwaukee River,
contributes an average of 14.6 m3/sec of water to the system
while the Kinnickinnic and Menomonee Rivers discharge about
0.48 m^/sec each. The dissolved oxygen content of all three
rivers may be less than 1.0 mg/1 in summer months (Depart-
ment of Natural Resources 1969).
The Jones Island Sewage Treatment Plant, operated by the
Sewage Commission of the City of Milwaukee, is an activated
sludge plant which began discharging into Milwaukee Harbor
in 1925. The plant began continuous chlorination of sewage
on 21 June 1971. At present the average flow of sewage
treated by the plant is 8.5 m^/sec. The chlorine, which is
in the liquid state, is vaporized and injected into the
sewage through a perforated mixer. From 1300 to 1500 Kg of
chlorine are used per day at a rate of 25 I/sec. After
chlorination, the effluent flows approximately 225 m before
entering Milwaukee Harbor at station 2 (Figure 1). Flow
time between the points of chlorination and discharge is
10-20 minutes which results in an average chlorination con-
tact time of 15 minutes. The chlorine residual near the
point of chlorination ranges from 0.5 to 1.0 mg/1. Ammonia
concentrations range from zero in the summer to 10 mg/1 in
the winter.
METHODS AND MATERIALS
Benthos samples for this study were taken from 16 sampling
stations in Milwaukee Harbor and adjacent Lake Michigan
(Figure 1) from the R/V NEESKAY on June 7, 1971, two weeks
before chlorination of the Jones Island effluent commenced
and July 6 and 7, December 1, 1971, and April 17 and
June 20, 1972, after chlorination was started.
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AREA LOCATION MAP
Figure 1. Sampling stations in Milwaukee Harbor and
adjacent Lake Michigan.
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On each sampling date three samples were taken at each
station with a 23 cm x 23 cm Ponar grab. The Ponar grab
was selected for this study because of its greater effic-
iency in a wider variety of sediments than the commonly
used Petersen or Ekman grabs (Glannagan 1970; Howmiller
1971; Poers and Robertson 1967). Each grab sample was
placed in a large plastic tub and then screened immediately
through a U. S. Std. No. 30 mesh (mesh opening 0.62 mm)
screening box. Residue remaining after screening was trans-
ferred into glass bottles, and preserved in 10% formalin.
At the time samples were collected, sediments were des-
cribed according to "categories for field evaluation of
soil characteristics" (Roelofs 1944) (Table 1). Depth at
each station was determined using the recording fathometer
on the R/V NEESKAY (Table 1).
In the laboratory the preserved samples were washed in a
U. S. Std. No. 60 mesh screen box to remove the formalin
solution. Samples were then placed in white enamel pans
and all organisms were hand picked and counted using for-
ceps and the aid of a Dazor magnifying lamp.
The abundance of organisms, based on the numbers in the
sample, were calculated by multiplying the numbers in the
sample by 19 to convert from the area of the sampler to a
square meter. Numbers of organisms in this report are, for
each station, based on the arithmetic mean of the three
samples taken at that station.
A volume displacement method (Anderson and Hooper 1956) was
used to determine the abundance of oligochaetes at stations
1, 2, 4 and 16. The volumes displaced by a known number of
worms and by the worms in the sample were determined. A
specifically modified centrifuge tube with a screen basket
insert (Howmiller 1972) was used to rid the worms of ex-
ternally adhering water.
Oligochaete worms picked from each sample at the 16 stations
were examined for species identification. Usually not more
than 30 worms per sample (90 per station), were mounted on
microscope slides in a mixture of Turtox CMC and Turtox CMCS
(this mixture gives the best viscosity for handling and
mounting) and were examined at a magnification of at least
100X.
In cases where only a portion of a sample was examined, the
worms to be examined were selected at random. The collec-
tion was spread out evenly in a pan (in about 1 cm of
water), which had a numbered grid layed out on its bottom.
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Table 1. PHYSICAL CHARACTERISTICS OF BENTHIC SAMPLING
STATIONS, 1971 AND 1972.
Station
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
Water depth (m) Bottom type
10.4
9.7
10.4
10.7
12.8
9.4
15.5
14.6
16.1
14.6
4.3
10.7
7.9
10.7
11.6
7.3
Oily organic silt, much debris
Organic silt, debris, fly ash
Organic silt, sand, crushed
Mollusca shells
Organic silt, clay
Gravel, sand over hard-packed
clay
Sand
Sand
Sand
Sand
1971, Sand, clay; 1972,
Organic silt over sand, clay
Gravel, sand over hard-packed
clay
Gravel over hard-packed clay
Organic silt, Mollusca shells
Gravel, sand over hard-packed
clay
Organic silt, debris, ash
Organic silt, sand, crushed
Mollusca shells
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The grid squares were numbered from 01 to 77. Pairs of
digits were read from a table of random numbers and worms
from the indicated squares taken for examination until
the desired number of specimens were obtained.
The taxonomic works of Brinkhurst (1964, 1965); Brinkhurst
and Cook (1966); and Brinkhurst, Hamilton and Herrington
(1968) were used for identification of the Oligochaeta.
The works of Hiltunen (1967) and Kennedy (1969) were par-
ticularly helpful with the genus Limnodrilus. Nomenclature
used follows that in the recent monograph of Brinkhurst
and Jamieson (1971), except that Limnodrilus cervix-clapare-
deanus is used here to describe worms that seem almost
exactly intermediate between L. claparedeanus and L. cervix.
The form of the penis sheath of these worms was similar to
that described by Brinkhurst and Cook (1966) and Kennedy
(1969) as intermediate between L. claparedeanus and L.
cervix and treated by Hiltunen (1967, 1969b, 1970) and
Howmiller (1972) as a variant of L. cervix.
All the midge larvae from a sample were examined. The
identification of chironomid larvae requires microscopic
examination. This is necessary because the structures
associated with the larval head are characteristic for each
genus. The head was dissected off each larva and mounted
ventral side up in CMCS. The body was mounted under a
separate coverslip. The taxonomic keys of Hilsenhoff and
Narf (1968), Johannsen (1937) and Mason (1968) were used
in identification.
No special methods were employed for the identification of
Mollusca, Hirudinea, or Crustacea. Sphaeriidae were sorted
and counted as a major taxonomic group with no attempt to
speciate them. Nematoda were not sampled quantitatively,
hence not counted, but their presence or absence was noted
for each station. Taxonomic keys in Edmondson (1959) and
Pennak (1953) were used for organisms other than oligo-
chaetes and midges.
Water samples to determine chlorine content of the harbor
and adjacent lake waters were collected at stations 1, 2,
4, 9, 13, 15, and at several other points in close prox-
imity to station 2 at the Jones Island sewage effluent
(Figure 1). Complete surveys of these stations were con-
ducted on April 5, 26, May 31, and August 22, 1973. In
addition, station 2 was sampled June 7, 1973, and February
12 and 28, 1974. Samples were obtained from the circulating
water system of the ship which draws water from a depth of
2 m. Additional samples were collected at the surface with
a plastic bucket and at depths greater than 2 m using a
10
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4 H van Dorn bottle. All samples were analyzed immediately
after collection on board the ship using an amperometric
titration procedure described in the chemical section of
this report.
RESULTS
Organic sediments were found at all stations in the harbor
(Table 1). The dominant component of the bottom in this
area was an organic silt ranging in thickness, from approx-
imately 2.5 cm to greater than the penetration depth of the
grab (10.2 cm). Stations within the inner harbor and near
its entrance (stations 1, 2, 15 and 16) contained abundant
hair, debris and partly burned materials and ash; these
last items probably originated from the City of Milwaukee
incinerator, which had stood on the north side of the inner
harbor entrance.
Hard bottom was present in each of the passages through the
breakwaters. In most cases these hard bottoms were of clay
with varying overlays of course gravel.
Gravel and sand were the dominant sediments of the embayment
(Lake Michigan). In 1972 a fine layer (.5 to 1.0 cm) of
silt appeared at station 10. This layer had not been
present during sampling in 1971. Its distribution in just
this one area and absence of such sediment in the rest of
the embayment suggests that this silt fraction was a
current-carried contribution from the harbor.
The dominant organisms within the harbor were the oligo-
chaete worms, accounting for over 90% of the total benthos
at all stations (Table 1 to 5, Appendix A). Every station
inside the breakwater had a density of worms of 10,000 to
>50,000/m2. Maximum numbers occurred in the mouth of the
Milwaukee River and in the immediate area of the Jones
Island Sewage Treatment Plant outfall. Minimum numbers of
worms were present at the breakwater entrances.
In the embayment proper, worms were present in concentra-
tions usually reported for Lake Michigan. Only in the
extreme north end of the embayment (station 11) did the
number of oligochaetes exceed the usual range of 0 to about
500/m2.
Oligochaetes generally were significantly lower in abun-
dance at harbor stations in 1972 (Figure 2). A standard
two-tailed test (Elliot 1971) was used to test the signif-
icance of difference between means of paired sets of data
11
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JUNE 1971
JUNE 1972
o
<25,000
25,000 - 75,000
75,000 - 125,000
> 125,000
Figure 2. Abundance (#/m2) of Oligochaeta in Milwaukee
Harbor and adjacent Lake Michigan, June 1971
and June 1972.
12
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from June 7, 1971, and June 20, 1972 (Table 2, Figure 3).
Stations in mid-harbor and near the Jones Island outfall
(stations 2, 14, 15 and 16) all had differences in popu-
lations which proved to be significant at the 99% level.
Station 13 in the northern part of the harbor also showed
a significant decrease (99% level) in worm populations in
1972. Populations of worms at the inner harbor station
C#D and in the extreme northern and southern parts of the
outer harbor (stations 4, 5, 12) as well as the embayment
stations (6-11) generally decreased, but the differences
were not significant. Station 3, located near the munici-
pal docks, also followed this pattern of a non-significant
decrease.
The most abundant species throughout the study were Tubifex
tubifex, Limnodrilus hoffmeisteri, and Peloscolex multi-
setosus (Figures 4-6) and (Tables 6-10 Appendix A) . Four
other species were found: Stylodrilus heringianus, Dero
digitata, Limnodrilus cervix-claparedeanus, and Limnodrilus
cervix. Stylodrilus heringianus (Figure 7) was restricted
to the embayment and breakwater entrance stations, while
Dero digitata was restricted to the south and north ends of
the main harbor. Limnodrilus cervix-claparedeanus (Figure
7) was generally distributed at main harbor stations, with
as many as 17,327/m2 found at one station. Limnodrilus
cervix was found at two stations twice during the study
accounting for about 6% of the total worms found at those
stations (Tables 7-9 Appendix A).
Because many common tubificids can be positively identified
only when they are sexually mature and have genital chaetae
and/or penis sheaths, it was impossible to identify many of
the worms in the samples, especially those taken early or
late in the year. These are listed as undetermined im-
matures and placed into two groups: those with hair chaetae
and those without hair chaetae. One can make a reasonable
guess, though, at the identity of immature specimens based
upon this knowledge of which species are present in the
sample from the positive identification of mature specimens.
Thus, the tables in Appendix A list the probable abundance
of Limnodrilus hoffmeisteri and Tubifex tubifex. These
numbers include mature specimens and a portion of or all of
the immature worms which could fit into the category. In
cases where a portion of the immature worms could have
belonged to another species the immature worms were assigned
to the two species on the basis of relative abundance of
positively identified mature specimens.
13
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Table 2. COMPARISON OF MEANS OF PAIRED SETS OF DATA OF
OLIGOCHAETA FROM 7 JUNE 1971 AND 20 JUNE 1972.
THE LEVEL OF SIGNIFICANCE OF THE DIFFERENCE
BETWEEN MEANS ACCORDING TO A TWO-TAILED t-TEST.
Oligochaeta (number/m2 )
Station 7 June 1971 20 June 1972
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
146357
164242
84949
68590
203
70
38
101
57
0
1545
0
138244
70
33662
133285
+ 4660
+ 7037
+ 7766
+ 4200
+ 44
± 17
± 25
+ 28
+ 22
+ 290
+ 5089
± 6
+ 1662
+ 12191
151145
112480
72783
55436
228
19
38
89
158
2261
1995
25
99649
19
13072
31863
+ 8873
+ 3923
+ 7378
+ 5648
+ 94
+ 11
+ 29
+ 50
± 61
+ 378
+ 295
± 17
+ 5090
± 1:L
+ 1889
+ 7422
Level of
t-value significance
0.86
6.42
1.14
1.44
0.24
1.09
0.00
0.21
1.58
1.09
7.58
4.07
8.18
7.11
n.s.
99
n.s.
n.s.
n. s.
n.s.
n.s.
n.s.
n.s.
n.s .
99
98
99
99
14
-------
500 1000 1500 METER
1/2 3/4 I STATUTE MILE
Figure 3. Abundance of Oligochaeta in June 1972 as a
percentage of the June 1971 abundance.
15
-------
Tubifex tubifex
500 1000 1500 METER
\fi 3/4 I STATUTE MILE
Figure 4. Percentage of Tubifex tubifex in the oligochaete
population of Milwaukee Harbor and adjacent
Lake Michigan.
16
-------
Limnodrilus hoffmeisteri
1000 500 1000 1500 METE
i
1/5 3/4 I STATUTE MILE
• 0
t
N
CD
J
O
i
UJ
Figure 5. Percentage of Limnodrilus hoffmeisteri in the
oligochaete population of Milwaukee Harbor and
adjacent Lake Michigan.
17
-------
Pe/osco/ex multisetosus multisetosus
Figure 6. Percentage of Peloscolex multisetosus multisetosus
in the oligochaete population of Milwaukee Harbor
and adjacent Lake Michigan.
18
-------
Limnodrilus cervix - claparedeanus
Styloan /us henngianus ::
Figure 7. Percentages of Stylodrilus heringianus and
Limnodrilus cervix-claparedeanus in the
oligochaete population of Milwaukee Harbor
and adjacent Lake Michigan.
19
-------
The distribution of the tendipedids (chironomids, midge
larvae) was markedly different from that of the oligo-
chaetes. Near the north and south ends of the area inside
the breakwaters and the entrances in the breakwaters, there
were small populations (20-100/m2) of midges (Tables 11-15
Appendix A). Elsewhere few midges were collected. Outside
the breakwaters midge abundance was usually less than 50/m2.
Only in the extreme north end of the embayment (station 11)
did the numbers of tendipedids approach usual Lake Michigan
concentrations, 0 to 500/m2. Three taxa were identified.
The dominant taxon at all stations was Hetrotrissocladius
sp. Procladius and Chironomus species occurred in small
numbers at stations 11 and 13.
No marked changes in the relative abundance of tendipedids
was noticed between 1971 and 1972, primarily because so
few were collected within the harbor.
Leeches and snails were minor components of the benthos
(Tables 11-15 Appendix A). They were found only in the
area inside the breakwater and never in numbers exceeding
63/m2 or about three individuals per grab. Only one species
of leech was found, Helobdella stagnalis. The snails were
represented by Vivaparus and Valvata sp. Many empty shells
of these taxa occurred at many of the stations within the
harbor.
Fingernail clams (Pelecypoda, Sphaeriidae), like snails
and leeches, were not very abundant, and found generally in
the area north of the main breakwater entrance (Tables 11-
15 Appendix A). Maximum numbers (772/m2) occurred at
station 16, just north of the main entrance and near the
west harbor wall. Collections included numbers of Sphaerium
corneum, £5. transversum, and £. lacustre form jayense.
Shells of these taxa were at many stations in the harbor,
but only about one in six were found alive, the rest being
empty shells. No clams were found in the area outside the
breakwater, except at station 11, where one to two indiv-
iduals were found per grab. These clams are believed to
be the naiad clam (Unionidae) Lampsilis siliquoidea.
Isopods (Asellus intermedius) and amphipods occurred only
in the embayment outside the breakwaters and in the break-
water entrances themselves (Tables 11-15 Appendix A).
Usually less than 50/m2 were collected, if there were any
amphipods or isopods at all. Localized areas of higher
populations of both lay north and northeast of the north
harbor entrance (stations 10 and 11). The amphipods,
Gammarus fasciatus, Hyalella azteca and Pontoporeia affinis,
were identified from specimens taken at these stations.
20
-------
The common and abundant benthic nematodes in this study
were about 3-4.5 mm long and 0.15 mm in diameter. An
animal this size cannot be collected quantitatively using
a No. 30 (0,516 mm aperture) screen and thus, nematodes
were not counted in the samples. Nematodes were found at
all harbor stations and at one embayment station (Tables
2-6 Appendix A).
The results of the chlorine determinations in the harbor
and adjacent lake are presented in Table 3. The only area
of the harbor where measurable residual chlorine was
detected was in the immediate vicinity of the Jones Island
Treatment Plant effluent (station 2, Figure 1). The
highest concentration of total residual chlorine observed
at this station was 0.220 mg/1. Samples obtained 200 m
from the effluent at both the surface and the bottom did
not contain measurable residual chlorine.
DISCUSSION
Aquatic oligochaete worms, especially the Tubificidae, are
probably the most common member of the benthos in eutrophic
or organically polluted areas. Pollution biologists have
long associated an abundance of oligochaetes, along with a
scarcity of other benthic invertebrates, with severs organic
pollution.
The abundance and distribution of worms in relation t~>
pollution has been discussed in numerous reports (Brinkhurst
1965, 1966a, 1966b, 1968, 1970; Brinkhurst, e_t al. 1970;
Carr and Hiltunen 1965; Goodnight and Whitley 1960; Howmiller
and Beeton 1971). The view held is that worms can be used as
indicators of organic pollution and that it is possible to
define species regularly associated with organically pol-
luted and eutrophic water and those associated with cleaner
oligotrophic systems.
Stylodrilus heringianus (Lumbriculidae) is apparently an
oligotrophic species, "typical of wide reaches of the Great
Lakes where there is little evidence of eutrophication"
(Brinkhurst 1969). It is abundant in Georgian Bay, Lake
Huron (Brinkhurst, et al. 1968) and oligotrophic Lake
Superior (Beeton and Hausmann, unpublished; Hiltunen 1969).
It is common throughout Lake Ontario, except where the
environment is considered unfavorable (Hiltunen 1969b) and
it is the dominant oligochaete throughout the central basin
of Lake Michigan (Hiltunen 1967; Howmiller 1972). Stylo-
drilus heringianus was the dominant oligochaete at all
21
-------
Table 3. TOTAL RESIDUAL CHLORINE VALUES OBSERVED IN
MILWAUKEE HARBOR AND ADJACENT LAKE MICHIGAN.
ALL SAMPLES PUMPED FROM A DEPTH OF 2 m
UNLESS DESIGNATED AS SURFACE SAMPLE.
Date
Apr.
Apr.
May
5, 1973 1
2a
2b
2c
26, 1973 9
4
15
2a
2b
2c
13
15
1
31, 1973 9
4
15
15
1
2
2
2
2
2
2
2
2
2
2
Residual
chlorine
Station no. (mg/1)
60 m east of outfall
60 m east of outfall
30 m east of outfall
3 mi offshore
south of channel
30 m east of outfall
surface
30 m east of outfall
surface
15 m east of outfall
north of channel
3 mi offshore
south of channel
north of channel
surface
5 m east of effluent
surface
5 m east of effluent
surface
60 m east of effluent
6 m deep 60 m east
of effluent
surface
200 m east of effluent
6 m deep 200 m east of
effluent
surface
200 m S.E. of effluent
6 m deep 200 m S.E. of
effluent
surface 200 m N.E. of
effluent
6 m deep 200 m N.E. of
effluent
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
.000
.000
.000
.000
.000
.000
.000
.020
.020
.220
.000
.000
.000
.000
.000
.000
.000
.000
.136
.131
.030
.000
.000
.000
.000
.000
.000
.000
Temp.
C°C)
10
8
—
—
6
14
14
15
15
15
15
14
15
10.
15
14
14
18
17
17
15
13
16
14.
15
-
17.
14.
5
5
5
5
22
-------
Table 3. (continued) TOTAL RESIDUAL CHLORINE VALUES
OBSERVED IN MILWAUKEE HARBOR AND ADJACENT
LAKE MICHIGAN. ALL SAMPLES PUMPED FROM A
DEPTH OF 2 m UNLESS DESIGNATED AS SURFACE SAMPLE,
Date
Residual
chlorine Temp.
Station no. (mg/1) (°C)
Jun. 7, 1973 2 30 m east of effluent 0.000 21.2
Aug. 22, 1973 1 0.000 23.5
2 at effluent 0.001 20.5
2 30 m east of effluent 0.006 20.5
4 0.000 20
15 south of channel 0.000 19.5
15 north of channel 0.000 19.5
13 0.000 20
Feb. 12, 1974 2 30 m east of effluent 0.000 3.7
Feb. 28, 1974 2 30 m east of effluent 0.000 2.5
stations outside the breakwaters, except station 11, in the
present study. Stylodrilus heringianus was also present in
about equal abundance with other worm species at the break-
water entrances.
It has been shown that there are two species of Tubificidae
that have the least demanding ecological requirements
(Brinkhurst and Kennedy 1962) and consistently occur in
greatest numbers in highly organic sediments and water
highly "polluted" or eutrophic (Brinkhurst 1965, 1966a, 1966b,
1970- Hiltunen 1969; Howmiller and Beeton 1970). These two
species are Tubifex tubifex and Limnodrilus hoffmeisteri.
in great abundance and in the absence of other benthic
organisms, including other tubificid species, indicate gross
organic pollution (Brinkhurst 1970). Where conditions are
less extreme L. hoffmeisteri is often the most abundant
species, generally found together with other Limnodrilus
species and Peloscolex multisetosus. Peloscolex multi-
setosus is the only member of the genus closely associated
with organic pollution and highly organic sediments.
Howmiller (1972) found that the distribution of P. multi-
setosus extended far into the southern polluted area of
Green Bay. Brinkhurst (1969) has found that in the Great
Lakes in general, P. multisetosus appears to be restricted
to polluted area.
23
-------
In Milwaukee Harbor, T. tubifex, L. hoffmeisteri and
p. multisetosus dominated the benthos and were present in
great abundance. T. tubifex dominated the worm population
of the inner harbor (Milwaukee River) and along with
L. hoffmeisteri made up about 97% of the benthos near the
Jones Island outfall. Brinkhurst (1970) has found a similar
distribution pattern in the polluted Toronto Harbor near
the mouth of the Don River, which is the major source of
organic pollution to the harbor. T. tubifex was absent
from all the stations outside the breakwater, except statxon
11 and in 1972 station 10, both areas possibly affected by
dredging spoils.
In the northern and southern areas of the harbor Limnodrilus
cervix-clapredeanus was common. Howmiller (1972) reported
this form common in lower Green Bay and Hiltunen (1967,
1969) has reported it from enriched areas of Lake Michigan
and from western Lake Erie.
Dero digitata, a naidid, was found by Brinkhurst (1967) to
be the commonest naidid in Saginaw Bay, Lake Huron, which is
influenced by the inflow of the polluted Saginaw River.
Howmiller (1972) found D. digitata to be restricted to the
lower bay and western shore of Green Bay. D. digitata was
found in low numbers at only two stations in the northern
and southern ends of Milwaukee Harbor.
The midges, especially the Chironomidae, include many species
with larvae and pupae tolerant of the very low oxygen con-
centrations associated with organic pollution. The aquatic
larvae of midges were the second most widespread members of
the benthic fauna during this study but nowhere did they
reach the abundance reported by Ayers and Huang (1966) in
their 1964 study of Milwaukee Harbor. While Ayers and Huang
found populations both in the harbor and the embayment,
midges were observed only in the extreme north and south ends
of the harbor and in the embayment in this study.
Fingernail clams, like the midges, were less abundant in
1971-72 than in 1964. Distribution patterns did, though,
follow those found by Ayers and Huang, with clams found
everywhere in the harbor except off Jones Island and at the
mouth of the Milwaukee River. Rofritz (1973) reported high
populations of sphaeriids near the west wall of the north
section of the harbor but did not differentiate between
living and relict shells. Emmling (1975) observed a distri-
bution pattern dependent on sediment grain size with a maxi-
mum abundance of sphaeriids occurring in the extreme north
and south portions of the harbor. The lower abundance of
24
-------
sphaeriids observed in this study is probably a function
of an accurate separation of living from relict shells
which was not done in previous studies.
While no naiad clams (Unionidae) were reported by Ayers and
Huang, Lampsilis siliquoidea was found at several Lake
Michigan embayment stations. The presence of naiads gen-
erally indicates clean water (Carr and Hiltunen 1965).
Howmiller (1972) found these naiads in the sandy substrates
of middle and upper Green Bay, and noted that "soft muds
. . . are not a suitable substrate for these large clams."
Leeches and snails were minor components of the benthos in
the harbor and showed little variation in numbers. Neither
leeches nor snails appear able to tolerate decomposition
gases at low oxygen concentrations (Pennak 1953). Carr and
Hiltunen (1965) concluded that, in western Lake Erie, they
were adversely affected by polluted conditions near river
mouths.
Amphipods and isopods occurred only at the stations outside
the breakwater. Amphipods and isopods did occur occasion-
ally at the breakwater entrances, but their presence at
these stations was probably a function of water movements.
Ayers and Huang (1968) believed, based on their measure-
ments of transparency, sulphide levels, and distribution of
benthic organisms, that the long-term mean water movement
was to the northeast in the embayment. Bellaire (1964)
found northward currents along Milwaukee during four days
of October 1963. The FWPCA (1966) reported primary north-
ward currents through the embayment during September
through March and weaker southward currents during the five
months of April through August. The low numbers of benthic
organisms in the southern and central part of the embayment,
and the tendencies toward higher concentrations of benthic
invertebrates to the north and east of the northern harbor
opening observed in this survey is taken to indicate a
long-term tendency of harbor water to move northeast from
the harbor.
The relative abundance of worms, the abundance of certain
worm species themselves and the lack of other benthic
organisms in the inner harbor and central area of the harbor
probably indicate a zone of gross pollution. The appearance
of other worm species and invertebrates in the north and
south areas of the harbor would appear to indicate dilution
of the pollution from the river and sewage effluent with
lake water. The northern and southern ends of the harbor
25
-------
could best be described as mesotrophic areas based on the
benthic invertebrate fauna. While the benthos found in
this area could only be described as pollution-tolerant,
the assemblages and greater species diversity indicate
less extreme conditions, as more and more dilution and
mixing with lake water entering through the breakwater
entrances takes place.
The lower abundance of oligochaetes in the southern end
would also seem to indicate that circulation of lake water
is better in this area and that the currents carrying the
detritus and bacteria-rich harbor water within the harbor
were primarily northward.
Emmling (1975) in his study of sphaeriid-sediment inter-
action in the Milwaukee Harbor found sediment with the
highest organic matter primarily in the central portion of
the main harbor off Jones Island and extending into the
north and south-central regions of the harbor. Both the
extreme north and south ends of the harbor had greater
percentages of sand and lower organic matter than the sedi-
ments of the central region and a greater abundance and
diversity of non-oligochaete macroinvertebrates.
Observations of water color within the harbor would seem to
confirm these conclusions. Dark brown water enters the
harbor from the Milwaukee River and from the Jones Island
Sewage Treatment Plant. In the central area of the harbor
the water progressively becomes a lighter brown moving away
from these sources, as the harbor water is diluted by lake
water entering through the central harbor entrance. In the
south end of the harbor the water takes on a blue-green-
brown color indicating extensive mixing of harbor and lake
water. The north end of the harbor remains a light brown
color probably because of impaired circulation due in part
to the curved internal bulkhead located in the north end
and the general northward movement of harbor water.
The significant decrease of oligochaetes in 1972 and the
subsequent disappearance of midges from Milwaukee Harbor
suggests a major environmental degradation. Investigation
of possible spills of toxicants into the harbor or the
rivers which might have contributed to the dramatic changes
in population levels revealed no reported spills other than
a few minor (less than 15 gallons) oil spills in 1971-72
(Department of Natural Resources 1973a).
Monthly Performance Reports for the Jones Island Sewage
Plant (Sewage Commission of the City of Milwaukee 1971-72),
indicate no periods of time when the 15-minute chlorine
26
-------
residual measured less than 0.31 mg/1 or greater than 1.62
mg/1 for the total plant effluent (includes plant bypass).
Yearly averages for the plant were 1.01 mg/1 in 1971 and
0.65 mg/1 in 1972. Total residual chlorine in the harbor,
however, was not measurable except directly over the efflu-
ent pipe of the plant.
Records of climatological data for the Milwaukee area (U. S.
Department of Commerce, 1971, 1972, 1973) in 1970-72 did not
show any variations in average wind speed and direction or
air temperature great enough to influence any major differ-
ences in the harbor environment for the winter of 1971-72
compared to previous years. Water temperature data (Depart-
ment of Natural Resources 1973b) for Milwaukee Harbor in
1969-72 (Figure 8) shows lower water temperatures in the
winter months of 1971-72 than in previous years. It has
been shown that in many northern European lakes tubificids
are only able to breed during a restricted period in late
summer when the water temperature rises above 15°C. This
influence of temperature has been reported for Limnodrilus
hoffmeisteri (Kennedy 1966a) and L. udekemianus (Kennedy
1966b). Brinkhurst and Jamieson (1971) have stated that
where the habitat is very productive the greatest number of
breeding specimens were found in the winter months, but a
spell of very low temperatures (below 4°C) can cause a
temporary cessation in activity.
The fact that the worm populations decreased only in the
central and north-central area of the harbor near the Jones
Island effluent and not in the rest of the harbor, the
rivers, or in adjacent Lake Michigan, suggests adverse con-
ditions associated with the sewage outfall. Although no
residual chlorine was measured in these areas of the harbor
it is possible that toxic levels of chlorine could have
existed there for short periods of time. Learner and
Edwards (1968) demonstrated that 0.5 mg/1 chlorine was lethal
to Nais (Oligochaeta) within a period of 35 minutes. Hart
(1957) found that 0.5 mg/1 chlorine killed Nais within one
hour.
It is also possible that chlorinated compounds formed in the
sewage that were undetectable by amperometric analysis may
have been responsible for the changes observed.
Natural variations in the benthic community could also have
influenced the results. Sampling over a number of years
would be required to establish natural fluctuations in the
abundance and composition of the benthic community in Mil-
waukee Harbor.
27
-------
22
UJ 20
Q
£'«
UJ
<•> 14
to
UJ IP
UJ I*
QC
a 10
o
IT
| 6
oc.
UJ
0- 4
UJ
H- 2
1971
19720^
F M A M J
SON
Figure 8. Mean monthly temperatures in Milwaukee Harbor,
1971 and 1972 (adapted from Wisconsin Surface
Water Monitoring Data 1969-72).
SUMMARY
Macroscopic benthic invertebrates of Milwaukee Harbor and
surrounding Lake Michigan were studied from grab samples
taken at 16 stations in 1971 and 1972. Comparison of the
present findings with data from studies made in 1964 indi-
cate that many types of invertebrates are less abundant now.
These include Hirudinea, Sphaeriidae, Gastropoda and prob-
ably Amphipoda and Isopoda. Oligochaeta and Chironmidae
were present in similar numbers in 1971, but declined sig-
nificantly in 1972 in the central and north-central area of
the harbor. This change might be indicative of chlorine
toxicity and further studies are needed to determine if this
is really so.
28
-------
Oligochaetes were the most abundant macroinvertebrate in
the harbor, generally composing over 90% of the total
benthos. Seven species were recorded. Species distri-
bution patterns were similar to those found in other studies
of polluted areas of the Great Lakes.
Other benthic invertebrates composed only about 10% of the
total specimens collected with clams probably being the
most abundant, although being concentrated in only certain
areas. Chironomids were found at about half of the stations
and in subnormal lake concentrations (0-50/m2). Taxa
identified included the genera Chironomus, Procladius and
Heterotrissocladius. Subsequent studies of the harbor area
(Emmling 1975) have reported the almost complete absence of
midge larvae.
This study has been adequate to determine distribution
patterns of the dominant elements of the benthic fauna. It
thus may serve as a reference for further bioassessment of
water quality in the harbor area. Further studies are
needed to determine accurately the abundance and distri-
bution patterns of the less numerous benthic species.
29
-------
SECTION V
CHEMICAL STUDIES
INTRODUCTION
The goal of this section of the study was to find a fast,
economical, and safe method for the removal of residual
chlorine from chlorinated sewage. To achieve this goal,
the following objectives were pursued:
1. To screen selected agents, e.g., sodium
sulfite (Na2S03), sodium bisulfite
(NaHSO3), sulfur dioxide (S02), which
are chemically synonymous, and sodium
thiosulfate (Na2S203) as reductants of
residual chlorine as monochloramine.
2. To ascertain the kinetics of the reduc-
tion of monochloramine by the preferred
reducing agent.
Several chemical and physical methods for removal of chlor-
ine and chloramines have been used and reported in the
literature. Watzl (1929) exposed chlorinated water to gran-
ular charcoal which safely removed the excess chlorine. To
remove chloramines from tap water for use in fish tanks,
Conventry, et al. (1935) added thiosulfate.
2NA2S203 + NH2C1 + 2H20 —-»-
Na2S406 + NH40H + NaOH + NaCl
[1]
Coventry, e_t al. (1935) also used several other compounds,
with some degree of success. These included NaHSO3, Fe++
salts and CH2 = CH2.
30
-------
Boiling chloramine solutions for 30 minutes had little
effect until acid was added while aeration alone removed
some chloramine (Adams and Buswell 1933). The use of
activated carbon to remove chlorine has recently been
reviewed by Magee C1956). Complete utilization of the
sorption properties of activated carbon depends on the
concentration of ammonia (Itlina 1958). Elimination of
free chlorine by heating was most effective with an iron
kettle; 0.4-0.8 mg/1 were completely removed by heating
to 30-50° (Kikuchi, et al. 1958). Elimination by stir-
ring was not effective; stirring for 30 minutes at 75-
300 rpm could reduce the level by about one-half, regard-
less of the initial concentration (Kikuchi, et al. 1958).
The addition of Na2S2C>3 or NaHSQs could instantly destroy
free chlorine (Kikuchi, et al. 1958). The amount of
Na2S2C>3 required to neutralize 1 mg of chlorine ranges
from 1.6 to 3 mg over a pH range of 6.2-9 (Al'terman 1958).
The removal of up to 0.5 ppm of free chlorine from tap
water over long periods of time was affected by the addi-
tion of Na2S2C>3 (McCauley and Scott 1960). This compound
had previously been used only on a short-term basis. The
compound was added continuously at a constant flow rate as
an aqueous solution. The amount needed was calculated by
titrating a known volume of tap water with a standard
solution of the reagent. The strength of the reducing
solution was calculated on the basis of a maximum concen-
tration of free chlorine of 0.5 mg/1. At this concentra-
tion, 1.5 g per day of thiosulfate was required for a
water flow of 1.1 liters per minute. No ill effects were
observed on sea lampreys, goldfish, brown bullheads, and
white suckers held in the treated water for several months.
The addition of sodium thiosulfate to effluent removed the
residual chlorine, which was tested in laboratory and field
conditions (Zillich 1972).
The nature of Cl2-NH3reaction depends on temperature, pH,
and concentration of ammonia. At a pH above 8, monochlora-
mine is formed, between pH 3-5, dichloramine is generated,
and below pH 3, trichloramine results (Corbett, et al.
1953). At any one pH only two N-chloramine species can be
present and the points of intersection are isosbestic
(Corbett, et al. 1953).
When chlorine is added to water, it hydrolyzes very quickly
to produce hypochlorous acid and hydrochloric acid equation
[2] (Mark 1963).
C12 + H20 £=± HOC1 + H+ + Cl~, K]_ =
3.94 x 10~4 at 20-25° [2]
31
-------
This reaction is nearly instantaneous (Draley 1972). If_
the [H+] concentration is taken as 1 x 10~7M and the [Cl~]
concentration as 3.81 x 10~^M, the mean values for sewage
effluent, the [HOC1]/[C1] ratio is constant at 1.03 x 107.
The hypochlorous acid partially ionizes according to
equation [3] (Mark 1963).
HOCI ?=± H+ + OC1~, K2 =
3.2 x 10~8 at 20-25°
[3]
This gives a [OC1 ]/[HOCl] ratio of 3.2 x 10 *, also con-
sidered a constant because the rate is instantaneous. Thus
chlorine water at pH 7 is actually approximately 1 part
hypochlorite anion and 3 parts hypochlorous acid with
essentially no free chlorine. Ammonia or ammonium ion forms
chloramine rapidly with a rate constant of 5.1 x 10^ liter
mole"1 sec"1, (Morris 1967) equation [4] (Mark 1963).
NH3 + HOCI £Z± NH2C1 + H20, K3 =
3.6 x 109
[4]
The reaction between ammonia and hypochlorous acid is not
inhibited by the ammonia-ammonium equilibrium which is
rapidly reversible. If the NH3 concentration is 5.9 x
10~4M, the maximum ammonia concentration in the City of
Milwaukee effluent, the [NH2C1]/[HOCI] ratio is 2.1 x 104.
The reaction of monochloramine and hypochlorous acid occurs
much slower, with a rate constant of 3.4 x 102 liter mole"1
sec"1 (Morris 1967). If the [HOCI] concentration is 2 x
10"5M, the [NH2C1]/[NHC12] ratio would be 2.7 x 101 equation
[5] (Draley 1972).
NH2C1 + HOCI £I± NHC12 + H20, K =
1.33 x 106 at 20-25°
Although dichloramine is favored in the Milwaukee effluent
from equilibrium considerations, little is formed due to the
slow rate, and hence monochloramine is the predominant
species.
32
-------
MATERIALS AND METHODS
All chlorine determinations for the field studies and those
made in conjunction with the laboratory bioassay studies
were determined using a modification of the amperometric
titration procedure described in American Public Health
Assn. (1971), with the following exceptions:
(1) a more dilute solution of phenylarsine
oxide titrant with a normality of 0.0028
was used;
(2) a mechanical Metrohm piston-type burette
to introduce the titrant to the titrating
vessel was employed;
(3) a 3-electrode cell (platinum test, platinum
counter, saturated calomel reference)
connected to a Heath model EUA-19-2-4
polarograph for measuring the current in
the titration solution was used; and
(4) a strip-chart recorder was used to record
current changes in the solution being
titrated.
Using the Metrohm burette and keeping the tip of the
delivery tube below the surface of the solution being
titrated, it was possible to accurately introduce as little
as 0.005 ml of titrant at one time. With the normality of
the titrant at 0.0028, 0.005 ml of the titrant was equiva-
lent to 0.005 mg/1 residual chlorine. Through the use of
the strip-chart recorder, and interpolation, it was pos-
sible to determine the endpoint of the titration, to within
0.001 mg/1 residual chlorine.
In studies undertaken to examine the effects of chlorine on
biological species, it is desirable to distinguish between
"free" chlorine and that combined as chloramines. The
amperometric titration procedure developed for this study
was unable to clearly distinguish between "free" and com-
bined chlorine. The o-tolidine test similarly measures a
combination of both "free" and combined chlorine. A method
which measures "free" chlorine, in combination with other
analytical procedures would provide information on the
amount of each chlorine compound present in a solution.
33
-------
Bauer and Rupe (1971) have developed a method for the
analysis of "free" chlorine in the presence of bound forms.
Syringaldazine was found to be a chomophoric agent which is
sensitive to hypochlorite, but insensitive to combined
chlorine such as chloramines. The reagent reacts on a mole-
for-mole basis with chlorine. Unfortunately, this photo-
metric method did not work well at low concentrations such
as those encountered in the lake and in the laboratory bio-
assay experiments. Hence, all values expressed here are as
total residual chlorine.
The concentration of sulfite used in the bioassay experi-
ments was measured spectrophotometrically using the 5,5'
dithiobis-(2-nitrobenzoic acid) method of Humphrey, et al.
(1970). A Hitachi Perkin-Elmer 139 UV-VIS spectrophotometer
with a tungsten lamp was used at a wavelength of 412 nm with
no filter. Five-centimeter pathlength cells were employed.
Absorbance readings were converted to sulfite concentrations
by the Beer's Law data furnished in Table 1 of Humphrey,
et al. (1970). Sulfite concentrations were expressed by
convention as sulfur dioxide (862)•
Standard Preparation of Chloramine for Chemical Studies
To prepare a 1.5 M solution of sodium hypochlorite, 80 g of
sodium hydroxide is dissolved in 200 ml of distilled water,
and the mixture is cooled by Dry Ice-acetone bath. After
the addition of 200 g of cracked ice, chlorine gas is passed
into the solution through an 8 mm glass tube. The chlorine
addition is continued until the desired increase in weight
(142 g) has been attained or until there is no longer a
yellow precipitate of mercuric oxide when a sample of the
solution is added to mercuric chloride. When the reaction
is completed, the pH should be 7. The solution is diluted
to 600 ml and titrated with standard 0.1 N sodium thiosul-
fate. Caution should be exercised to avoid overchlorination.
In a 1-liter beaker is placed 200 ml of 1.5 M ammonia which
is cooled to 0°. After addition of 100 g of cracked ice,
200 ml of 1.5 M sodium hypochlorite is slowly introduced
with stirring. This yields chloramine of 0.2 molarity.
Decomposition of Chloramine
A fresh solution of chloramine was equilibriated in a con-
stant temperature bath at 25+l°C. At 6-minute intervals
during an 80-minute period, 1 ml aliquots were removed and
added to a solution of 40 ml of acetic acid, 10 ml of water,
and 2 g of sodium iodide. The concentration was plotted
34
-------
versus time indicating a decomposition rate of -0.00019
mole per minute at pH 9. This experiment indicated that
chloramine is stable enough to permit a study of the rate
of reduction with various chemical reducing agents.
Reduction of Chloramine
A fresh solution of chloramine (0.208M) was equilibriated
at 25° + 0.1°. Aqueous solutions (0.2M) of ferrous chloride
sodium bisulfite, sodium thiosulfate, sodium sulfite, sodium,
metabisulfite, and sulfur dioxide were also maintained at
the same temperature. Individual experiments were conducted
during which the chloramine solution was added to each of
the reducing agents with magnetic stirring. Aliquots (1 ml)
were withdrawn as rapidly as possible, timed, and quenched
in an acidic KI solution. Titration with thiosulfate was
used to determine the concentration of chloramine versus
time. In this way, the time required for total reduction
of chloramine by each reducing agent was determined.
Kinetic Studies
To study the kinetics of chloramine reduction in the rag/1
range, it was necessary to use a much more sensitive anal-
ytical procedure. Voltammetry with a platinum electrode
was used to obtain current-voltage curves for sodium hypo-
chlorite and monochloramine in the mg/1 range in order to
determine reduction potentials. With hypochlorous acid at
20 mg/1, buffered at a pH of 7, 0.1 M Na2S04 supporting
electrolyte, and with deaeration, two waves resulted with
maxima at -0.11 v and -0.51 v vs. S.C.E. (Figure 9). When
0.02% Triton X-100 (a surface active agent) was added, a
limiting current was obtained with an E1/2=+0-42v vs- S.C.E.
(Figure 10). The double wave could not be eliminated with
gradual drop-wise addition of Triton X-100. R> limiting
curve or maxima were obtained with oxygen free 200 ppm mono-
chloramine, buffered at pH 7, 0.1 M K2SO4. Again when
Triton X-100 was added, a limiting current was obtained with
a half wave value of -0.15v (Figure 11). The difference in
half wave potentials was great enough to distinguish between
the two species (HOC1 and NH2C1).
The current-voltage curves of hypochlorous acid and mono-
chloramine have been obtained using a rotating platinum
electrode (Marks and Bannister 1947), and by the dropping
mercury electrode method (Heller and Jenkins 1946). A set
of double polarographic waves was obtained with the dropping
mercury electrode, which were shown to result from simple
chemical reaction at the mercury surface (Jenkins 1951).
35
-------
-220/xa-f
.0 v +0.6v + 0.2v
20
-0.2v -0.6v -I.Ov
Figure 9. Current-voltage curve of HOC1 and blank.
36
-------
HCIO
-220 yua--
-180AQ --
-I40/AQ--
fl.Ov
0.6v f0.2v
+ 20/AQ
Blank
-0.2v -0.6v -I.Ov
Figure 10. Current-voltage curve of HOC1 and blank with
Triton X-100 added.
37
-------
-0.2v -0.6v -I.Ov
Figure 11.
Current-voltage curve of NI^Cl and blank with
Triton X-100 added.
38
-------
Recently in two papers (Harrison and Khan 1971; Schwarzer
and Landsberg 1968) on the reduction of hypochlorous acid
at a platinum and a carbon electrode, an oxide film was
reported to form in both cases. The oxide on platinum
inhibits reduction strongly in the early stages, followed
by a relaxation in the oxide structure, after which the
inhibition disappears. With the carbon electrode, one part
of the oxide layer is reduced with the anion and the rest
at a more negative potential. It is believed that the
double wave in the current-voltage curve of hypochlorous
acid is due to an oxide film on the platinum surface and
that, with both hypochlorous acid and chloramine, Triton
X-100 prevents formation of the oxide. Since the double
wave was not completely eliminated with drop-wise addition
of Triton X-100, this suggests there are sites on the sur-
face of the platinum for which oxygen and Triton X-100 are
competing.
When a current-voltage curve of Milwaukee sewage effluent
was determined, added chloramine could be detected in the
ppm range. It appears that the metal cations present are
not reduced at this potential. When the current-voltage
curve was taken of sodium sulfite, the reducing agent of
principal interest, no wave was detected that might be mis-
taken for the chloramine wave (Figure 12). Also the oxi-
dized product of S03=, i.e., SC>4=, does not interfere and
is used as a supporting electrolyte.
Voltammetric Limits of Detection
Current-voltage curves were determined for solutions of 20,
10, 5, 1 and 0.5 ppm monochloramine. The plots were used to
determine the lower limit of chloramine detectable. To
accomplish the lower limit of detection, an electrode pre-
treatment process was employed. The three-electrode system
was placed in the solution, deaerated for 20 minutes with
N2, set at a potential of O.Ov for 30 seconds, 2 drops
Triton X-100 (0.02%) added, and deaerated again for 3 minutes
with N2- The current-voltage curves were then determined.
Kinetic studies were done after electrode pretreatment, using
9.6 x 10~5 M (5 mg/1) chloramine solution (0.02% Triton X-100,
0.1 M sodium sulfate, 0.01 M phosphates, pH 7 and under nitro-
gen). The potential of the polarograph was set on -O.SOv on
the limiting plateau. A strip-chart recorder monitored
current versus time as solid sodium sulfite was added. The
recorder followed the chloramine concentration, and resul-
tantly the rate of the reduction reaction. Because the
reaction is so fast the instrumentation response time is the
limiting factor in precisely determining the rate of reduction.
39
-------
Q-
§
O
E
300--
UJ
tr
cc.
3 200
o
Q
O
X
< IOO--
o
1.00
^0.50
200--
-0.50 -1.00
POTENTIAL (volts)
z
LU
CC
o:
^
o
Q
O
Figure 12. Current-voltage curve for sodium sulfite,
40
-------
A new approach was tried to determine fast reaction rates.
Steady-state systems have often been used to measure rapid
rates of reaction. Such a system was constructed (Figure 13)
In this system, chloramine was added at various distances
from the platinum indicator electrode by an infusion pump.
An increase in current showed that chloramine was reaching
the indicator electrode before the reduction reaction was
complete. Knowledge of rate of addition of chloramine, the
stirring rate and distance of the Pt indicator electrode
from the point of chloramine entry should allow calculation
of the rate constant (k). Once an increase in current was
found for a certain rate of addition and distance x, the rate
of addition could be decreased to an equilibrium point where
the current remained constant. In the final case the rate of
addition was proportional to the rate of reaction.
Ninety-five ml of 0.1 M sodium sulfite was placed in the
reaction vessel. A solution of 0.05 M chloramine was then
forced into the system by the infusion pump through a Teflon
tube at various points of entry. The fastest rate of addi-
tion was 46.3 ml/min. A synchronous motor stirred the
solution at 600 rpm.
RESULTS AND DISCUSSION
Reduction of,Chloramine
Ferrous chloride, sodium bisulfite, sodium thiosulfate,
sodium sulfite, sodium metabisulfite, and sulfur dioxide all
reduced chloramine in less than 60 seconds. With sulfur
dioxide, sodium bisulfite, and sodium sulfite, reduction was
complete in less than 20 seconds. Sodium nitrate was elim-
inated as a potential reducing agent since the oxidation
product, nitrate, is a nutrient. Ferrous chloride was elim-
inated because it formed a precipitate. Sodium thiosulfate
and sodium metabisulfite were also excluded because of a
potentially deleterious effect due to the tetrathionate
product. Due to its low cost and efficiency of chloramine
reduction, sodium sulfite was the preferred reducing agent.
Voltammetric Limits of Detection
A detectable current-voltage wave was found down to 1 mg/1
(1.9 x 10~5 M) (Figure 14). The electrode pretreatment
procedure enabled the lower limit to be decreased to 0.5
mg/1 (9.6 x 10~6 M) monochloramine (Figure 15). This
increase in sensitivity is presumably due to renewal of the
electrode surface. Removal of absorbed organics and oxide
41
-------
Direction of
Flow
Magnetic
Stirring Bar
S. C E.
Reference
Electrode
O.I M Na2S03
Pt. Indicator
Electrode
0.05 M NH2CI
Solution Input
(O.I M. Na2S04 Supporting Electrolyte)
Figure 13.
Schematic representation of the three-electrode
cell used in obtaining current-voltage curves
for chlorine and chloraraine.
42
-------
-1.00
POTENTIAL (volts)
Figure 14. Current-voltage curves for various concentra-
tions of NH2C1. Detection limit was 1 mg/1.
formations exposes the active sites on the surface. Figure
16 indicates that at -0.05 volts, on the limiting current
plateau, the current is proportional to the chloramine con-
centration present in solution. On this basis, the moni-
toring of chloramine concentrations can be done efficiently
using voltammetric techniques.
Kinetics of Reduction of Chloramine
Voltammetry was used to monitor the reduction of chloramine
by sodium sulfite. Upon the addition of solid sodium sulfite
to a 5 ppm chloramine solution the current dropped to zero
almost instantaneously. This rapid decrease in current,
i.e., extremely rapid reaction, occurred too fast to permit
measurement of an absolute rate. An attempt to slow the
43
-------
-0.50
-1.00
POTENTIAL (volts)
Figure 15. Current-voltage curves for NH2C1 following
electrode pretreatraent. Detection limit
was 0.5 mg/1.
reaction enough to measure the rate was made by decreasing
the temperature. A kinetic run at 0°C displayed the same
rapid decrease in current. Because of the fast reaction
rate, instrumentation response became the limiting factor.
The response specifications for the recorder at 10 in. per
sec., allowed a minimum rate constant, kmin, to be calcu-
lated, namely kmin = 0.40 sec~l. The final concentration
of sodium sulfite, 9.0 x 10~3 M, was in excess of the
chloramine concentration, thus the reaction can be assumed
to be zero-order with respect to reducing agent. Assuming
the reaction is first-order with respect to chloramine the
reaction should follow equation [6].
44
-------
300r
[NH2CI] (ppm)
Figure 16. Relationship between current and concentration
for NH2C1 at -0.50 volts.
d[NH2Cl]
= -K[NH2C1]
[6]
The steady-state system (Figure 13) was used to determine
the first-order reaction rate constant (k).
When 0.05 M chloramine was added to a solution of supporting
electrolyte without reducing agent, the current increased
linearly with time. The next experiment involved the addi-
tion of chloramine to the reducing agent. A 0.05 M chlor-
amine solution was introduced into a 0.10 M sodium sulfite
solution. With the inlet port as close as 10 mm to the
indicator electrode and an addition rate at 46.3 ml per
second, the maximum rate of the pump, no response was
observed. From this data the minimum rate constant was
-1
This value of k
•mm
calculated to be kmin =43 sec
represents a 100-fold increase over the value determined by
the earlier method.
45
-------
The significance of this value is presumably associated
with the fast nature of the reaction. It is apparently
mass transport limited. Hence mixing is crucial in des-
truction of chloramine in sewage effluent.
Using this value of the rate constant, kmin/ and assuming
a chloramine concentration of 9.6 x 10~5 M (5.0 mg/1), a
half-life value can be calculated using the integrated form
of equation [6J, i.e., equation [7].
InlNH^Cl],- -In [NH^Cl] +_+ = -k t1/2
£. t—U £. 1/2
[7]
The half-life calculates to be t1//2 = 0.016 sec.
Of concern in destruction of chloramine by sodium sulfite
is the stability of the reducing agent. Studies on the
oxidation of sodium sulfite by dissolved oxygen have been
done (Fuller and Crist 1941). The first-order rate constant
of oxidation for [NA2S03] <_ 0.015 M, saturated with 02 at
1 atmosphere was found to be kavcf = 0.013 sec""1 at 25°C
(Fuller and Crist 1941) . The kmj_n = 43 sec"1 for reduction
of chloramine is ~ 3 x 10^ greater, thus added sodium sul-
fite should react with residual chloramine before any
appreciable oxidation by dissolved 02 can take place.
Trace catalysis by Cu2+ was also observed. With [Cu2+]
>^ 10~9 M the rate constant was found to be kavg = 2.5 x 10°
£/mole/sec (Fuller and Crist 1941). However, second-order
kinetics are implied by the units and a direct comparison of
the values should not be made.
SUMMARY
Sodium sulfite was selected as the preferred reducing agent
of chloramine because of its lower relative cost, efficiency
and speed of reduction, and lack of reaction products poten-
tially deleterious to the aquatic environment. The minimum
first-order reaction rate constant (kmin) for the reduction
of chloramine by sulfite was determined voltametrically to
be 43 sec -1. The half-life of a chloramine solution of
5 mg/1 in the presence of sulfite was 0.016 sec. Therefore,
complete reduction of chloramine by sulfite in sewage efflu-
ents would occur almost instantly.
46
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SECTION VI
BIOASSAY STUDIES
INTRODUCTION
Numerous toxicity tests have estimated the tolerance of
certain forms of aquatic life to chlorine residuals (Brungs
1973). Chlorine toxicity tests have been common using fish
but aquatic invertebrates have been largely neglected.
This study involved two common aquatic invertebrate groups,
the rotifers and the cyclopoid copepods, which heretofore
have not been studied in chlorine toxicity tests. The
purpose of this investigation was to determine the acute
toxicity of residual chlorine to these organisms, and to
investigate whether the addition of sodium sulfite, a
reducing agent, could eliminate or reduce the toxicity of
chlorine residuals to the test organisms.
Two test organisms were selected because of their abundance
within the study area and their wide distribution throughout
the Great Lakes. Previous field work conducted at the
Center for Great Lakes Studies indicated that Cyclops bicus-
pidatus thomasi is one of the more common zooplankton
species found in the inshore; areas of Lake Michigan. The
rotifer, Keratella cochlearis, was shown by Stemberger
(1973) to be the most abundant rotifer in the Milwaukee—area
of Lake Michigan. Another consideration in choosing these
test organisms was the fact that both could be maintained
in the laboratory without significant mortalities.
METHODS AND MATERIALS
Bioassay experiments were conducted in a continuous flow
proportional diluter, modified from the design of Mount and
Brungs (1967) (Figure 17). Modifications of the original
design were incorporated to overcome the problems of exces-
sive consumption of dilution water, of starting several
47
-------
DILUTION WATER
FROM STORAGE TANK
CELLS (W)
HEAD TANK
AIR*
'ERFLOW
BOTTLE
vMICROSWITCH
S-l SHUTOFF
FLOAT CONTROLED
STOPCOCK VALVE
UARmrvr CHEMICAL
MARRIOT DELIVERY
VACUUM BOTTLE SYSTEM
CHEMICAL
MIXING
CHAMBER(M)
CHEMICAL
PROPORTIONING
CELLS (C)
MIXING CHAMBERS
TO SOLENOID
''VALVES
REPEAT
CYCLE
TIMER
>^ OVERFLOW
TO WASTE
UNIVERSITY OF WISCONSIN-MILWAUKEE
CENTER FOR GREAT LAKES STUDIES
DRAWN SY R4TKO J. RISTIC
DATE: e.JAN. 1975
Figure 17. Proportional diluter used for bioassay
experiments,
48
-------
siphons from a single vacuum line, of precisely timing flow
rates and the lag time between the synthesis of labile
chlorine compounds and their delivery to the test aquaria.
All of the "fail-safe" features of the system described by
Mount and Brungs were retained in the modified design.
The heart of the modified diluter consisted of a repeat
cycle timer run by a chronometrically governed 12 VDC motor.
This timer sequentially opened and closed a series of
switches, which in turn energized a series of seven 12 VDC
solenoid valves. The entire system operated by gravity flow
and a 12-volt automobile storage battery. The power supply
was totally independent of external power sources and,
hence, was not subject to interruption by external power
failures. A fully charged 60-amp-hour battery was more
than adequate to power the system for the duration of a 96-
hour experiment.
Dilution water was collected from Lake Michigan at the City
of Milwaukee Linnwood Avenue Water Purification Plant prior
to any treatment or chemical additions. The water was
transported to the laboratory in 15-gallon FDA food grade
polyolefin drums. Water in the drums was filtered through
a number 20 Nitex nylon net to remove zooplankton and then
pumped into glass-lined steel storage tanks suspended from
the ceiling of the laboratory. From the storage tanks, the
water flowed into a temperature-controlled environmental
chamber housing the bioassay dilution system. A constant
head tank received the inflowing water where aeration and
temperature equilibration occurred. The temperature of the
environmental chamber was maintained at 15°C for all experi-
ments. Light in the chamber was supplied from cool white
fluorescent tubes on a 16-8 hour light-dark cycle. The
light level at the surface of the test aquaria was between
40 and 60 foot candles.
Once per hour the repeat cycle timer opened a solenoid valve
(S-l Figure 17) allowing water to flow from the head tank
through a 20 mesh Nitex filter into the dilution water
chambers (Wl-6 Figure 17) of the diluter. When the last
chamber filled, water overflowed into a bottle which
depressed and opened a microswitch which closed the sole-
noid valve in the inflow line. Water in the overflow
bottle slowly drained through a capillary tube allowing the
microswitch to close before the next cycle began. The
microswitch did not close, however, until the timer switch
opened, breaking the circuit to the solenoid valve.
49
-------
The next valve to open drained dilution water from the W-l
cell through a vacuum venturi which drew the chemical to
be tested into the chemical-mixing chamber (M Figure 17)
where it was mixed with the dilution water. An overflow
siphon in the mixing chamber started as the last portion
of dilution water entered, drawing the mixed solution into
the chemical cells (Cl-5 Figure 17} and to the test aquaria
having the greatest concentration of test chemical.
The chemicals were measured and delivered to the mixing
chamber via a standpipe siphon tube arrangement described
by McAllister, et al. 1972. When dechlorination required
the addition of two chemical solutions simultaneously, two
such delivery systems were connected to the diluter.
The next timer switch to close opened five valves, simul-
taneously draining dilution water cells W-2 through W-6.
The water flowed through vacuum venturies which siphoned
the chemical solution from the chemical cells into mixing
chambers where dilution and mixing occurred. The volumes of
the dilution water cells and the chemical cells were such
that 400 ml of each test concentration was prepared with a
50% dilution between each concentration. The W-6 cell sup-
plied 400 ml of unadultered dilution water for the control
aquaria. The solutions flowed from the mixing chambers and
were split in two, half the flow going to each of tne two
test aquaria.
The test aquaria were 2000 ml beakers with central overflow
standpipes screened with number 20 Nitex netting (Figure 18).
Incoming water entered the beaker at the bottom and flowed
out at the surface to enhance mixing. The volume of water
in the test aquaria was maintained at 1600 ml for the
Cyclops experiments and at 1200 ml for the rotifer tests.
With 200 ml of fresh water entering each test aquaria per
hour, the flushing time for the test aquaria was once every
eight hours for Cyclops runs and once per 6 hours for the
rotifer experiments.
The entire dilution system was constructed of glass, silicon
rubber, teflon and a small amount of nylon for screening.
Double distilled water, reagent grade 5% sodium hypochlorite
solution, ammonium hydroxide and sodium sulfite were used to
prepare all test solutions. Water chemistry data on the
dilution water from Lake Michigan was monitored by the City
of Milwaukee at the Linnwood Water Purification Plant.
All of the test organisms used in these experiments were
collected from Lake Michigan near Milwaukee. The rotifers
for the bioassay experiments were obtained by filtering them
50
-------
NFLOW
TUBE
ROTIFER
AQUARIUM
MESH
DRAIN
PIPE
2-LITER BEAKER
Figure 18. Test aquarium used in bioassay experiments
with insert jar used for rotifer tests.
51
-------
with a number 20 Nitex net from untreated Lake Michigan
water taken from the Linnwood Avenue Water Purification
Plant. Lake Michigan water entered the purification plant
from an intake crib 17 meters below the surface of the lake
approximately 1.6 km offshore 6 to 7 km north of Milwaukee
Harbor. The filtered water remaining from this filtration
process was used as the dilution water in the rotifer bio-
assay experiments. The rotifers were returned to the lab-
oratory and acclimated overnight in a. 4-liter container at
the same temperature and lighting conditions which were to
be employed during the bioassay exposures.
The copepods used in the bioassay studies were either
collected in a manner similar to that described for the
rotifers or obtained from a vertical plankton tow taken
from the R/V NEESKAY at a point in Lake Michigan 5 km north-
east of Milwaukee Harbor. The net used for vertical tows
was a one-half meter number 20 mesh Nitex net. The organ-
isms collected from both the vertical tow and from the
purification plant were returned to the laboratory and
acclimated in the environmental chamber in a 20-liter
aquarium for at least 24 hours before the start of a bio-
assay run.
Prior to the start of each bioassay experiment, 20 actively
swimming organisms were pipetted into twelve vials, one for
each test aquaria. The distribution of the organisms to
the vials was randomized to avoid the aggregation of easily
caught individuals in any one test chamber. After all the
test organisms were in the vials, they were transferred to
their respective test aquaria.
Test organisms were removed from the aquaria at the end of
each bioassay by one of two methods. Rotifers were removed
and concentrated for observation simply by lifting the small
test chamber shown in Figure 18 from the 2000 ml beaker
allowing the water to drain through the screened opening
near the bottom. The water and organisms remaining in the
container were then transferred to a microscope counting
chamber for mortality assessment. Copepods were removed
from the 2000 ml beakers by gently pouring the contents
through a screened jar similar to that used for the rotifer
exposure chambers. The organisms concentrated in this
manner were then transferred to a microscope counting chamber
for mortality assessment. Test organisms were considered
dead if they did not swim or exhibit any internal or exter-
nal movement when examined under 100 X magnification. Occa-
sionally a few animals would be unaccounted for at the time
52
-------
of examination. It was assumed that these losses occurred
in transferring the organisms to or from the test aquaria.
The percent mortalities reported were calculated on the
basis of the actual number of living and dead organisms
observed at the end of each experiment.
Observations of test organism mortality were only made at
the termination of each experiment. More frequent observa-
tions of mortality would have been desirable but since mor-
tality was determined under the microscope, considerable
handling was involved in transferring the organisms from the
test aquaria to the observation chamber. It was felt that
once the test organisms were removed from the test aquaria
they would no longer be fit for further study due to the
rigors of handling.
Measurements of total residual chlorine, pH and temperature
were made at the start and finish of each 4-hour bioassay
and at 24-hour intervals for the duration of the 96-hour
bioassay exposures. The dissolved oxygen in the test
aquaria was measured periodically to assure that the aerated
dilution water was at or near the saturation valve.
Bioassay experiments with the rotifer Keratella cochlearis
were terminated at 1, 4, and 24 hours. The 4-hour exposure
period was chosen as the best compromise between sufficient
exposure time and minimal control mortality which increased
with longer exposures. All experiments with Cyclops bicus-
pidatus thomasi were conducted over a 96-hour exposure
period.
The results of the bioassay experiments are reported as the
median tolerance limit, or TLgQ. This value is the estimated
concentration at which 50 percent of the test organisms sur-
vived for the exposure period indicated. The TL^Q values for
each experiment where mortality was observed were determined
by plotting the toxicant concentration on a log scale versus
percent mortality on a log-probit scale (American Public
Health Assn. 1971). A regression line was then drawn through
the points on the graph. The toxicant concentration at which
the regression line intersected the 50 percent mortality level
was taken to be the TL50 value for the experiment.
Chlorine concentrations cited are expressed as total resid-
ual chlorine. This includes both free and combined chlorine
species which were detectable using an amperometric titra-
tion procedure described in the chemistry section of this
report. It was not possible to accurately differentiate
between free chlorine and combined forms using this tech-
nique. Based on the calculated equilibrium concentrations
of Draley (1972), it was assumed that monochloramine was the
53
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dominant chlorine constituent present when monochloramine
was added as the test chemical. Because of the presence of
ammonia in the dilution water, some chloramines were undoubt-
edly formed when sodium hypochlorite was added as the test
chemical. Hence, experiments run adding hypochlorite reflect
the effects of the hypochlorite ion and monochloramine. This
would be the situation in most natural waters where added
chlorine would react with ammonia to yield chloramines. The
relative amounts of chlorine and chloramine present would be
determined by the pH, ammonia content of the water and the
amount of chlorine added (Draley 1972). Since the bioassay
experiments were designed to simulate the natural chemical
conditions in Milwaukee Harbor and Lake Michigan, it was
deemed unsuitable to remove ammonia from the dilution water
or to substitute ammonia free water for dilution water
obtained from Lake Michigan.
RESULTS
Seventeen duplicate bioassay experiments were performed
during the course of this study. Nine 96-hour bioassay
experiments were conducted with the copepod Cyclops bicus-
pidatus thomasi. Three of these were undertaken using
sodium monochloramine. One Cyclops bioassay was conducted
using sodium sulfite alone, and two experiments usiiig
sodium sulfite to reduce monochloramine were completed.
Seven 4-hour bioassay experiments were performed with the
rotifer Keratella cochlearis. Three of these experiments
were run exposing the organism to monochloramine. In
addition, one bioassay was undertaken adding sodium sulfite
alone, while three dechlorination bioassays were completed
using sodium sulfite to reduce monochloramine.
As discussed previously, chlorine concentrations measured
in conjunction with the bioassay experiments will be
presented as total residual chlorine. The dominant com-
bined chlorine consituent present in the test solutions
was monochloramine. No dichloramine or trichloramine was
ever detected. Although free chlorine could not be measured
accurately, the hypochlorite ion was undoubtedly present in
the test aquaria when sodium hypochlorite was added as the
test chemical. No attempt was made to calculate the amount
of free chlorine present using published equilibrium con-
stants. The uncertainties involved in applying laboratory-
derived equilibrium conditions to natural waters over-
shadowed the value of making such determinations.
Other parameters which could have influenced the toxicity
tests, namely dissolved oxygen, pH and temperature, were
held relatively constant for all bioassay experiments. The
54
-------
temperature of the test aquaria was maintained at 15 +
0.5°C for all runs. The pH ranged between 8.1 and 8.5" over
the course of all experiments but did not vary more than
0.2 pH unit during any one individual test. Dissolved
oxygen was maintained at or near saturation by constant
aeration of the incoming dilution water. The saturated
value for dissolved oxygen at 15°C is 10.2 mg/1. Frequent
measurements of dissolved oxygen in the test aquaria were
always within a few tenths of this value. Specific values
for pH measurements are presented in Appendix C in conjunc-
tion with the results of the individual bioassay experiments,
A summary of the water quality characteristics of the dilu-
tion water obtained from the Linnwood Avenue Water Purifi-
cation Plant is contained in Table 4.
The results of the three bioassay experiments exposing
Cyclops bicuspidatus thomasi to residual chlorine added as
sodium hypochlorite are contained in Tables 1-3 Appendix C.
The 96-hour TL5p value derived by pooling the results of
the three experiments was 0.069 mg/1 total residual chlorine
(Figure 19). No mortalities were observed after exposures
of 96 hours at total residual chlorine concentrations below
0.01 mg/1. Nearly complete mortality was observed at con-
centrations above 0-20 mg/1 total residual chlorine after
96 hours of exposure. The scatter of points observed on
Figure 19 may be due in part to changing chlorine equil-
ibrium conditions between individual bioassay experiments.
Test solutions with varying proportions of free and com-
bined chlorine species could possibly account for the vari-
ability observed. Under more defined chemical conditions
where monochloramine predominated, much less variability
was seen.
Tables 4-7 Appendix C contain the results of the four
Cyclops bioassay experiments run adding monochloramine as
the test chemical. The 96-hour TL5Q value calculated from
the pooled data from the three experiments with monochlor-
amine is 0.084 mg/1 total residual chlorine (Figure 20). No
mortality was observed below 0.01 mg/1 total residual
chlorine while nearly complete mortality would be expected
to occur at concentrations above 0.30 mg/1 total residual
chlorine following exposures of 96-hour duration.
Bioassay experiments to determine the toxicity of sodium
sulfite to Cyclops bicuspidatus thomasi indicated that
sodium sulfite was not toxic at maximal concentrations
ranging between 0.510 and 0.637 mg/1 for a 96-hour exposure
period. Mortalities observed within this concentration
range and lower were equal to or less than mortalities
55
-------
Table 4. AVERAGE CHEMICAL AND PHYSICAL ANALYSES OF
UNTREATED LAKE MICHIGAN WATER FROM THE
LINNWOOD AVENUE WATER PURIFICATION PLANT,
MILWAUKEE, FOR 1973.
Concentration
Parameter (mg/1)
Dissolved Solids 160
Suspended Solids 12
Total Solids 172
Turbidity - J. T. U. 5.5
Bicarbonate Alkalinity - as CaCO., 108
Carbonate Alkalinity - as CaC03 0
Non-carbonate Hardness - as CaCO., 28
Total Hardness - as CaCO- 136
Chlorides (Cl~) 9.1
Free Ammonia (as N) 0.029
Sulfate (S04) 21.1
Chemical Oxygen Demand 7
PH 8.17
56
-------
98
95
90
80
570
| 60
°50
1-40
S 30
o
2l 20
a.
10
2 11
0.001
96 hour TLgQ = 0.069 mg/l
* .
0.010 0.100
TOTAL RESIDUAL CHLORINE mg/L
1.000
Figure 19. The toxicity of residual chlorine, as soiium
hypochlorite, to Cyclops bicuspidatus thomasi
during 96-hour exposures at 15°C.
observed in control organisms (Table 8 Appendix C). The
highest concentration of sulfite used for this bioassay
was adequate to reduce chlorine residual levels commonly
observed at sewage plant outfalls.
Preliminary Cyclops bioassay experiments using sodium sul-
fite to reduce chlorine residuals failed to protect the
organisms over the entire 96-hour exposure period although
some reduction in mortality was noted. It was later found
that the strength of stock sulfite solution added to the
bioassay system degraded quite rapidly, thereby losing its
ability to reduce chlorine. If fresh sulfite solutions
57
-------
98
95
90
>- 80
I70
5: 60
§ 50
Z 40
$30
cr
£20
10
•
5 r- / ~ 96hour TL50 = 0.084 mg/l
f J. .
2 ll
o.ooi o.oio o.ioo i.ooo
TOTAL RESIDUAL CHLORINE mg/l
Figure 20. The toxicity of residual chlorine, as mono-
chloramine, to Cyclops bicuspidatus thomasi
during 96-hour exposures at 15°C.
were prepared daily, chlorine residuals were reduced or
eliminated and the test organisms survived the 96-hour
exposure.
Tables 9 and 10 Appendix C contain the results of the suc-
cessful sodium sulfite dechlorination bioassay experiments.
Monochloramine was added to the chemical-mixing chamber, as
was done for experiments previously described, at levels
sufficient to produce a range of concentrations in the test
aquaria between 0 and 1.0 mg/l had dechlorination not been
employed. Simultaneously, an equimolar concentration of
sodium sulfite was added to the chemical-mixing chamber
which completely reduced the monochloramine. After an
exposure of 96 hours to the sulfite dechlorinated water
the mortalities of Cyclops observed in the test aquaria
were equal to or less than those experienced by the control
organisms.
58
-------
Results of the three 4-hour Keratella cochlearis bioassay
experiments with monochloramine are contained in Tables 11-
13 Appendix C. The 4-hour TL5g value derived by pooling
the results of the three experiments was 0.019 mg/1 total
residual chlorine (Figure 21). Residual chlorine concen-
trations above 0.05 mg/1 produced nearly complete mortality
after a 4-hour exposure. No mortalities were recorded
below 0.003 mg/1 total residual chlorine following a 4-hour
exposure period.
Bioassay experiments exposing Keratella to sodium sulfite
concentrations as high as 0.821 mg/1 for 4 hours did not
result in any significant mortalities in the test aquaria
in comparison to the controls (Table 14 Appendix C).
Having established that sodium sulfite by itself was not
toxic to Keratella at levels sufficient to reduce chlorine
residuals observed at sewage outfalls, three 4-hour dechlor-
ination bioassays were undertaken. Monochloramine was added
to the chemical-mixing chamber of the diluter at a level
sufficient to produce a range of concentrations in the test
aquaria between 0 and 1.0 mg/1 total residual chlorine had
dechlorination not been employed. An equimolar amount of
sodium sulfite was added to the mixing chamber simultan-
eously with the monochloramine to reduce the chlorine
residual. The results of the dechlorination bioassay exper-
iments with Keratella are contained in Tables 15-17 Appendix
C. Sodium sulfite effectively eliminated the residual
chlorine toxicity to Keratella. The highest mortality
observed in any test aquaria over the entire series of exper-
iments was only 11.8%.
DISCUSSION
The 96-hour TL50 values calculated for Cyclops bicuspidatus
thomasi and Keratella cochlearis indicate that these inver-
tebrates are among the more sensitive organisms with respect
to the acute toxicity of chlorine residuals. Table 5 com-
pares these tolerance limits with those for several other
aquatic species. The 0.084 mg/1 96-hour TL50 value reported
here for Cyclops exposed to monochloramine is within the
lower range of TLsg values reported by Arthur (1972) as
cited in Brungs 1973 for the scud Gammarus pseudolimnaeus
and the stonefly Acronearia lycorias exposed to chlorinated
sewage for seven days. The 0.019 mg/1 4-hour TL50 value
reported here for Keratella exposed to monochloramine is
well below any TL50 values reported in the literature for
aquatic organisms.
59
-------
98
95
90
60
40
O
g
Q.
10
5
2'
0.001
4-hour TLgQ = 0.019 mg/l
* L
_i
0.010 0.100
TOTAL RESIDUAL CHLORINE mg/l
1.000
Figure 21. The toxicity of residual chlorine, as mono-
chloramine, to Keratella cochlearis during
4-hour exposures at 15°C.
The lower 96-hour TL5Q value of 0.069 mg/l calculated for
Cyclops exposed to hypochlorite together with monochlor-
amine suggests that free chlorine may be somewhat more toxic
to this species than monochloramine alone. It is difficult
to make a more definitive statement in this situation with-
out knowing the exact quantities of the various active
chlorine species which were present during the tests. This
observation is consistent with findings summarized by Brungs
(1973). Brungs (1973) notes that "the toxicities of the
principal components of residual chlorine (free chlorine,
dichloramine and monochloramine) are not sufficiently dif-
ferent to preclude using a measure of residual chlorine to
define acute toxicity." He goes on to state: "When a high
percentage of residual chlorine exists as free chlorine,
toxicity will be greater and its effect will occur more
quickly."
60
-------
Table 5. SUMMARY OF EFFECTS OF RESIDUAL CHLORINE ON
AQUATIC LIFE (ADAPTED FROM BRUNGS 1973).
Species
Brook trout
Rainbow trout
Coho salmon
Fathead minnow
Fathead minnow
Fathead minnow
Largemouth bass
Largemouth bass
Largemouth bass
Yellow perch
Yellow perch
Yellow perch
Daphnia magna
Gammarus
ps eudo 1 imnaeus
Cyclops
bicuspidatus
thomasi
Keratella
Effect
endpoint
TL5Q
TL50
TL50
TL50
TL50
TL50
TL50
TL50
TL50
TL50
TL50
TL50
Safe Con-
centration
TL50
TL50
TL50
Residual
chlorine
Time cone. (mg/1) Reference
7-day 0.083
7-day 0.08
7-day 0.083
1-hour 0.79
12-hour 0.26
7-day 0.082-
0.115
1-hour 0.74
12-hour 0.365
7-day 0.261
1-hour 0.88
12-hour 0.494
7-day 0.205
0.003
96-hour 0.220
96-hour 0.069-
0.084
4-hour 0.019
Arthur, 1972
Merkens, 1958
Arthur, 1972
n n
"
II II
II It
,,
n n
n n
n n
n «
Arthur, 1972
Arthur and
Eaton, 1971
This report
cochlearis
61
-------
The criteria proposed by Brungs (1973) and The National
Academy of Sciences (1972) for continuous chlorine appli-
cation are supported by the results of the bioassay exper-
iments reported here. A total residual chlorine concen-
tration of 0.01 mg/1 would appear to protect most adult
Cyclops bicuspidatus thomasi while producing some mortal-
ities of Keratella cochlearis. A 0.002 mg/1 total residual
chlorine concentration applied continuously would appear to
be adequate to protect the adults of both species tested
although the safety factor for Keratella would be minimal.
Additional research will be required to determine if the
TL50 values calculated for the adults of these species would
permit reproduction to occur and the subsequent development
of pre-adult life stages.
The reduction of chlorine residuals with sodium sulfite
appears to be a viable means of protecting aquatic life
from the adverse effects of chlorinated effluents. Sodium
sulfite itself was not toxic to either of the test organisms
used in the bioassay experiments reported here when applied
at levels sufficient to reduce chlorine residuals observed
in the field. Sodium sulfite preferentially reacts with
residual chlorine constituents over dissolved oxygen and
hence would not increase the oxygen demand of an effluent
if added in concentrations equivalent to the residual
chlorine present. The reaction products resulting from
residual chlorine reduction are common constituents in most
freshwaters which should not pose a serious threat to the
aquatic environment. Additional research should be under-
taken to document this under varying field conditions.
SUMMARY
The 96-hour TL5Q values calculated for Cyclops bicuspidatus
thomasi exposed to chlorine residuals added as monochlora-
mine was 0.084 mg/1 total residual chlorine. The 96-hour
TLso value for this species exposed to a combination of free
chlorine and monochloramine was 0.069 mg/1 total residual
chlorine.
The 4-hour TL50 value calculated for Keratella cochlearis
exposed to chlorine residuals added as monochloramine was
0.019 mg/1 total residual chlorine.
Sodium sulfite was not toxic to either Cyclops bicuspidatus
thomasi when exposed to concentrations as high as 0.637 mg/1
for 96 hours or to Keratella cochlearis exposed to levels up
to 0.821 mg/1 for four hours.
62
-------
The addition of sodium sulfite to solutions containing up
to 1.0 mg/1 total residual chlorine effectively reduced
the chlorine residual and eliminated its toxicity to both
test organisms.
63
-------
SECTION VII
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aluminum chloride. J. Amer. Chem. Soc. 95C9): 2977-2982.
70
-------
United States Department of Commerce. 1971-73. Wisconsin
climatological data. U. S. Weather Bureau, Wisconsin Crop
Reporting Service, Madison, Wis.
United States Environmental Protection Agency. 1973. Data
update for 1966 Federal Water Pollution Control Administra-
tion Report, "A Comprehensive Water Pollution Control Program,
Lake Michigan Basin, Milwaukee, Wisconsin, area." Region V.
Chicago, 111. 4 p.
Veal, D. M. and D. S. Osmond. 1968. Bottom fauna of the
western and nearshore Canadian waters of Lake Erie. In;
Proc. llth Conf. Great Lakes Res., Internat. Assoc. Great
Lakes Res. War Memorial Center on the Lakeshore of Mil-
waukee, Wis. April, pp. 151-160.
Watzl, E. 1929. Superchlorination and subsequent dechlor-
ination over carbon of water for municipal supply. Ind.
Eng. Chem. 21: 156-158.
Zillich, J. A. 1972. Toxicity of combined chlorine res-
iduals to freshwater fish. J. Water Poll. Con. Fed. 44:
212-219.
71
-------
SECTION VIII
PUBLICATIONS RESULTING FROM THIS PROJECT
1. Strand, J. W. and P. Kovacic. 1972. Preferred route
for vic-dichloride formation from alkanes. Comparison
of cyclahexene chlorination with chlorine, sulfuryl
chloride and trichloramine. Synthetic communications
2(3): 129-137.
2. Strand, J. W. and P. Kovacic. 1973. Amination of
toluene, adamantane, and tert-butyl chloride with
monochloramine-aluminum chloride. Journal of the
American Chemical Society 95: 2977-2982.
3. Brooks, A. S., N. E. Grossnickle and A. M. Beeton.
1975. The acute toxicity of residual chlorine to the
copepod Cyclops bicuspidatus thomasi and the rotifer
Keratella cochlearis. Submitted to Journal of the
Fisheries Research Board of Canada.
4. Grossnickle, N. E., A. S. Brooks and A. M. Beeton.
1975. Reduction of residual chlorine toxicity to the
copepod Cyclops bicuspidatus thomasi and the rotifer
Keratella cochlearis with sodium sulfite. Submitted
to Journal of the Water Pollution Control Federation.
72
-------
SECTION IX
APPENDICES
Page
A. Field Studies 76
Table A-l. Abundance of Benthic Invertebrates, 77
7 June 1971
Table A-2. Abundance of Benthic Invertebrates, 78
6 and 7 July 1971
Table A-3. Abundance of Benthic Invertebrates, 79
1 December 1971
Table A-4. Abundance of Benthic Invertebrates, 80
17 April 1972
Table A-5. Abundance of Benthic Invertebrates, 81
20 June 1972
Table A-6. Abundance of Oligochaete Taxa, 82
7 June 1971
Table A-7. Abundance of Oligochaete Taxa, 85
6 and 7 June 1971
Table A-8. Abundance of Oligochaete Taxa, 88
1 December 1971
Table A-9. Abundance of Oligochaete Taxa, 91
17 April 1972
Table A-10. Abundance of Oligochaete Taxa, 94
20 June 1972
Table A-ll. Abundance of Invertebrate Taxa, 97
7 June 1971
73
-------
Table A-12. Abundance of Invertebrate Taxa,
6 and 7 July 1971
Table A-13. Abundance of Invertebrate Taxa,
1 December 1971
Table A-14. Abundance of Invertebrate Taxa,
17 April 1972
Table A-15. Abundance of Invertebrate Taxa,
20 June 1972
B. Chemistry of N-Chloramines
C. Bioassay Tests
Table C-l. Toxicity of Residual Chlorine Added
as Sodium Hypochlorite to Cyclops
bicuspidatus thomasi.
May 19-23, 1973
Table C-2. Toxicity of Residual Chlorine Added
as Sodium Hypochlorite to Cyclops
bicuspidatus thomasi .
June 1-5, 1973
Table C-3. Toxicity of Residual Chlorine Added
as Sodium Hypochlorite to Cyclops
bicuspidatus thomasi.
June 14-18, 1973
Table C-4. Toxicity of Residual Chlorine Added
as Monochloramine to Cyclops bicus-
pidatus thomasi.
July 13-17, 1973
Table C-5. Toxicity of Residual Chlorine Added
as Monochloramine to Cyclops
bicuspidatus thomasi.
July 19-23, 1973
Table C-6. Toxicity of Residual Chlorine Added
as Monochloramine to Cyclops
bicuspidatus thomasi.
August 9-13, 1973
Table C-7. Toxicity of Residual Chlorine Added
as Monochloramine to Cyclops
bicuspidatus thomasi.
August 16-20, 1973
98
99
100
101
102
104
105
106
107
108
109
no
74
-------
Table C-8. Toxicity of Sodium Sulfite to
Cyclops bicuspidatus thomasi.
January 11-15, 1974
Table C-9. Toxicity of Residual Chlorine to
Cyclops bicuspidatus thomasi.
Dechlorinating with Sulfite.
April 16-20, 1974
Table C-10. Toxicity of Residual Chlorine to
Cyclops bicuspidatus thomasi.
Dechlorinating with Sodium Sulfite.
June 3-7, 1974
Table C-ll.
Table C-12.
Table C-13.
Toxicity of Residual Chlorine Added
as Monochloramine to Keratella
cochlearis. September 27, 1973
Toxicity of Residual Chlorine Added
as Monochloramine to Keratella
cochlearis. October 11, 1973
Toxicity of Residual Chlorine Added
as Monochloramine to Keratella
cochlearis. October 18, 1973
Table C-14. Toxicity of Sodium Sulfite Alone on
Keratella cochlearis.
November 29, 1973
Table C-15. Toxicity of Residual Chlorine Added
as Monochloramine to Keratella coch-
learis. Dechlorinating with Sodium
Sulfite. January 26, 1974
Table C-16. Toxicity of Residual Chlorine Added
as Monochloramine to Keratella coch-
learis. Dechlorinating with Sodium
Sulfite. January 31, 1974
Table C-17. Toxicity of Residual Chlorine Added
as Monochloramine to Keratella coch-
learis. Dechlorinating with Sodium
Sulfite. February 9, 1974
112
113
114
115
116
117
118
119
120
121
75
-------
APPENDIX A
FIELD STUDIES
76
-------
Table A-l.
ABUNDANCE OF BENTHIC INVERTEBRATES, INDIVIDUALS/m2, AT STATIONS SAMPLED
7 JUNE 1971, BASED ON THE MEAN OF COUNTS FROM THREE 23 cm x 23 cm PONAR
GRAB SAMPLES, FOLLOWED BY THE STANDARD ERROR OF THE MEAN. Presence (+)
or absence (0) of nematodes is indicated.
Station
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
Hirun- Gastro-
Oligochaeta dinea poda
146357+4660
164242±7037
84949±7766 6±6
68590±4200 6±6 13±13
203±44
70±17
38±25
101±28
57±22
0
1545±290 6±6
0
138244±5089 63±39
70 + 6
33662±1662 6±6
133285±12191
Amphi- Chirono-
Pelycpoda poda Isopoda midae Nematoda
+
+
+
1621±83 82±35 +
57±22 +
0
0
6 + 6 0
0
0
19±11 114±105 424±89 +
0
38±29 95±22 +
32±32 +
281±45 +
722±140 +
-------
Table A-2.
ABUNDANCE OF BENTHIC INVERTEBRATES, INDIVIDUALS/m2, AT STATIONS SAMPLED
6 AND 7 JULY 1971, BASED ON THE MEAN OF COUNTS FROM THREE 23 cm x 23 cm
PONAR GRAB SAMPLES, FOLLOWED BY THE STANDARD ERROR OF THE MEAN. Presence
(+) or absence (0) of nematodes is indicated.
00
Station
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
Oligochaeta
150258±18950
160233±15134
101732±7487
68400±6349
329±63
89±63
25±25
76±11
1697±199
1672148
139±10
127186±14776
89±42
34953±11166
136813±9789
Hirun- Gastro-
dinea poda Pelycpoda
19±11 51±28
19±11 1336±158
19±11
6±6 13±6
44±25
19±11
25±6 5131134
25117 19111
4501125
25111 773H21
Amphi-
poda
25117
108190
32H3
32117
38111
616
Chirono-
Isopoda midae
13±13
139±61
38119 25H7
127172 82±13
203+102 323129
6i6 462177
72±146
82123
Nematoda
+
+
+
+
0
0
0
0
0
0
+
0
+
+
+
+
-------
Table A-3. ABUNDANCE OF BENTHIC INVERTEBRATES, INDIVIDUALS/m2, AT STATIONS SAMPLED
1 DECEMBER 1971, BASED ON THE MEAN OF COUNTS FROM THREE 23 cm x 23 cm
PONAR GRAB SAMPLES, FOLLOWED BY THE STANDARD ERROR OF THE MEAN.
Presence (+) or absence (0) of nematodes is indicated.
Station
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
Hirun- Gastro- Amphi-
Oligochaeta dinea poda Pelycpoda poda
165414+9589
173008+4748
86178±10519
7325116974 2014±177
215±94 6±6
19±11
196±45
616
1596+900 616
14181130 44128
63135
143874±4749 13+13 184134
89139
3181213078 329196
14528016418 1316 773145
Chirono-
Isopoda midae Nematoda
+
+
+
38H1 +
+
0
0
616 0
0
+
139+28 +
0
42+19 +
+
+
+
-------
Table A-4.
ABUNDANCE OF BENTHIC INVERTEBRATES, INDIVIDUALS/m2, AT STATIONS SAMPLED
17 APRIL 1972, BASED ON THE MEAN OF COUNTS FROM THREE 23 cm X 23 cm PONAR
GRAB SAMPLES, FOLLOWED BY THE STANDARD ERROR OF THE MEAN. Presence ( + )
or absence (0) of nematodes is indicated.
00
o
Station
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
Hirun- Gastro- Amphi-
Oligochaeta dinea poda Pelycpoda poda
14773115665
119314±4880
83726±13321
52022±5194 32±32 1659±221
127±63
51±17
6±6
19 + 19
101±35
2128±320
1393±139 13±13
11046±8031 6±6 386±121
12103±2141 6±6 108±56
33662±1662 6±6 609±134
Chirono-
Isopoda midae
6±6
171±58
19 + 19
19±11
51±28
6±6
9±6 432±116
51±6
Nematoda
+
+
+
+
0
0
0
0
0
0
+
0
+
0
+
+
-------
Table A-5.
ABUNDANCE OF BENTHIC INVERTEBRATES, INDIVIDUALS/m2, AT STATIONS SAMPLED
20 JUNE 1972, BASED ON THE MEAN OF COUNTS FROM THREE 23 cm x 23 cm PONAR
GRAB SAMPLES, FOLLOWED BY THE STANDARD ERROR OF THE MEAN. Presence (+)
or absence (0) of nematodes is indicated.
CO
Station
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
Hirun-
Oligochaeta dinea
151145+8873
112480±3923
72783+7378
55436±5648
228±94
19±11
38±29
89±50
158±61
2261±378
1995±295
25±17
99649±5090
19±11 6±6
13072±1889 19±11
31863±7422
Gastro- Amphi- Chirono-
poda Pelycpoda poda Isopoda midae
1545±204 203±110
6±6
6±6 44±44 13±13
6+6 6+6
13±13 25±19 6±6
44±44 95±22
6±6 32117 19±19
241±84
6±6
6±6 336±92 19±11
25±17
19±19 133+58
558+155
Nematoda
+
+
+
+
0
0
0
0
0
0
+
+
+
+
+
+
-------
Table A-6.
ABUNDANCE OF OLIGOCHAETE TAXA, INDIVIDUALS/m2, AT STATIONS SAMPLED
7 JUNE 1971, BASED ON THE MEAN OF COUNTS FROM THREE 23 cm x 23 cm
PONAR GRABS; AND IN PARENTHESES, THE RELATIVE ABUNDANCE OF EACH TAXON
AS A PERCENTAGE OF THE TOTAL. The lower portion of the table lists
probable number of each species in each collection, probable number (%)
of Limnodrilus hoffmeisteri, Tubifex tubifex, and the total number of
specimens examined.
00
to
Taxon 1
Stylodrilus heringianus
Station
2 3
4 5
89 (44)
Dero digitata
Limnodrilus cervix
L. cervix-claparedeanus
L. hoffmeisteri
Feloscolex multisetosus
multisetosus
Tubifex tubifex
Undetermined immature
With hair chaetae
Without hair chaetae
Total
23417 (16)
4927 (3)
52557 (32)
54152 (37) 26279 (16)
54152 (37)
13172 (9)
146357±4660
11497 (7)
68982 (42)
16424217037
5097 (6)
22936 (27)
38159 (45)
5946 (7)
1757 (2)
14441 (17)
84949±7766
686 (1)
10289 (15)
15090 (22)
20577 (30)
4801 (7)
4116 (2)
13032 (19)
68590±4200
51 (25)
63 (31)
203+44
Probable L. hoffmeisteri 25%
Probable T. tubifex 74%
Probable No. species 2
No. specimens examined 86
74%
23%
3
74
9%
4
86
41%
9%
5
114
56%
2
32
-------
Table A-6. (continued) ABUNDANCE OF OLIGOCHAETE TAXA, INDIVIDUALS/m2, AT STATIONS
SAMPLED 7 JUNE 1971, BASED ON THE MEAN OF COUNTS FROM THREE 23 cm X 23 cm
PONAR GRABS; AND IN PARENTHESES, THE RELATIVE ABUNDANCE OF EACH TAXON AS
A PERCENTAGE OF THE TOTAL. The lower portion of the table lists probable
number of each species in each collection, probable number (%) of Limno-
drilus hoffmeisteri, Tubifex tubifex, and the total number of specimens
examined.
00
CO
S tation
Taxon 678 9 10
Stylodrilus heringianus 57 (82) 38 (100) 88 (87) 57 (100)
Dero digitata
Limnodrilus cervix
L. cervix-claparedeanus
L. hoffmeisteri 13 (18)
Peloscolex multisetosus
multisetosus
Tubifex tubifex
Undetermined immature
With hair chaetae
Without hair chaetae 13 (13)
Total 70±17 38±25 101±28 57±22
Probable L. hoffmeisteri 13%
Probable T. tubifex
Probable No. species 21 21
No. specimens examined 11 6 15 9
11
325 (11)
371 (24)
201 (13)
77 (5)
572 (37)
1545±290
61%
18%
3
62
-------
Table A-6.
(continued) ABUNDANCE OF OLIGOCHAETE TAXA, INDIVIDUALS/m2, AT STATIONS
SAMPLED 7 JUNE 1971, BASED ON THE MEAN OF COUNTS FROM THREE 23 cm x 23 cm
PONAR GRABS; AND IN PARENTHESES, THE RELATIVE ABUNDANCE OF EACH TAXON AS
A PERCENTAGE OF THE TOTAL. The lower portion of the table lists probable
number of each species in each collection, probable number (%) of Limno-
drilus hoffmeisteri, Tubifex tubifex, and the total number of specimens
examined.
00
Taxon
12
Station
13 14
15
16
Stylodrilus heringianus
Dero digitata
Limnodrilus cervix
L. cervix-claparedeanus
L. hoffmeisteri
Peloscolex multisetosus
multisetosus
Tubifex tubifex
Undetermined immature
With hair chaetae
Without hair chaetae
Total
Probable L. hoffmeisteri
Probable T. tubifex
Probable No. species
No. specimens examined
12442 (9)
20091 (29)
42860 (31)
9677 (7)
6908 (5)
27649 (20)
138244±5089
43%
12%
4
123
15 (22)
31 (44)
32 (33)
70±6
77%
2
9
2693 (8)
12792 (38)
6745 (20)
1347 (4)
1333 (4)
8752 (26)
33662±1162
58%
8%
4
91
17327 (13)
49316 (37)
17327 (13)
6664 (5)
1533 (1)
42651 (32)
133285112191
58%
6%
4
126
-------
Table A-7.
ABUNDANCE OF OLIGOCHAETE TAXA, INDIVIDUALS/m2, AT STATIONS SAMPLED 6 AND
7 JULY 1971, BASED ON THE MEAN OF COUNTS FROM THREE 23 cm x 23 cm PONAR
GRABS; AND IN PARENTHESES, THE RELATIVE ABUNDANCE OF EACH TAXON AS A
PERCENTAGE OF THE TOTAL. The lower portion of the table lists probable
number of each species in each collection, probable number (%) of Limno-
drilus hoffmeisteri, Tubifex tubifex, and the total number of specimens
examined.
CO
Taxon
Stylodrilus heringianus
Dero dicjitata
Limnodrilus cervix
L. cervix-claparedeanus
L. hoffmeisteri
Peloscolex multisetosus
multisetosus
Tubifex tubifex
Undetermined immature
With hair chaetae
Without hair chaetae
Total
Probable L. hoffmeisteri
Probable T. tubifex
Probable No. species
No . specimens examined
1
22539 (15)
60103 (40)
40570 (27)
27046 (18)
150258±18950
33%
67%
2
95
2
Station
3
1602 (1)
43263 (27)
40058 (25)
25637 (16)
49672 (31)
160233±15134
57%
41%
3
77
7121 (7)
28485 (28)
35586 (35)
9156 (9)
6124 (6)
14243 (14)
10173217487
38%
15%
4
116
4
10944
14364
27360
1368
1368
12996
68400
32%
4%
4
128
5
(16)
(21)
(40)
(2)
(2)
(19)
+ 6349
145 (44)
82 (25)
69 (21)
329±179
46%
2
52
-------
Table A-7.
CO
(continued) ABUNDANCE OF OLIGOCHAETE TAXA, INDIVIDUALS/m2, AT STATIONS
SAMPLED 6 AND 7 JULY 1971, BASED ON THE MEAN OF COUNTS FROM THREE 23 cm
x 23 cm PONAR GRABS; AND IN PARENTHESES, THE RELATIVE ABUNDANCE OF EACH
TAXON AS A PERCENTAGE OF THE TOTAL. The lower portion of the table
lists probable number of each species in each collection, probable
number (%) of Limnodrilus hoffmeisteri, Tubifex tubifex, and the total
number of specimens examined.
Taxon 6 7
Stylodrilus heringianus 82 (92) 25 (100)
Dero digitata
LTrnnodrilus cervix
L. cervix-claparedeanus
L. hof fmeisteri
Peloscolex multisetosus
multisetosus
Tubifex tubifex
Undetermined immature
With hair chaetae
Without hair chaetae 7 (8)
Total 89±63 25±25
Probable L. hof fmeisteri 8%
Probable T. tubifex
Probable No. species 2 1
No. specimens examined 13 4
Station
8 9 10
76 (100) 458 (27)
102 (6)
526 (31)
34 (2)
560 (33)
76±11 16971199
58%
1 3-4
12 51
11
318 (19)
518 (31)
217 (13)
184 (11)
451 (27)
1672±48
58%
24%
3
75
-------
Table A-7.
00
(continued) ABUNDANCE OF OLIGOCHAETE TAXA, INDIVIDUALS/m2, AT STATIONS
SAMPLED 6 AND 7 JULY 1971, BASED ON THE MEAN OF COUNTS FROM THREE 23 cm X
23 cm PONAR GRABS; AND IN PARENTHESES, THE RELATIVE ABUNDANCE OF EACH
TAXON AS A PERCENTAGE OF THE TOTAL. The lower portion of the table lists
probable number of each species in each collection, probable number (%)
of Limnodrilus hoffmeisteri, Tubifex tubifex, and the total number of
specimens examined.
Taxon 12
Stylodrilus heringianus 70 (50)
Dero digitata
Limnodrilus cervix
L. cervix-claparedeanus
L. hof fmeisteri 20 (14)
Peloscolex multisetosus
multisetosus
Tubifex tubifex
Undetermined immature
With hair chaetae
Without hair chaetae 50 (36)
Total 139±10
Probable L. hoffmeisteri 50%
Probable T. tubifex
Probable No. species 2
No. specimens examined 22
13
Station
14
13 (14)
3816 (3)
12719 (10)
30525 (24) 63 (71)
35612 (28)
8903 (7)
6360 (5)
29253 (23) 13 (14)
127186±14776 89±42
38%
12%
5
92
85%
2
14
15
2796 (8)
11884 (34)
9437 (27)
2097 (6)
1049 (3)
8039 (23)
3495211166
50%
9%
4
113
16
10945
47885
36940
2736
4104
34203
13681:
58%
4%
4
121
(8)
(35)
(27)
(2) f
(3)
(25)
3±9789
-------
Table A-8.
ABUNDANCE OF OLIGOCHAETE TAXA, INDIVIDUALS/m2, AT STATIONS SAMPLED
1 DECEMBER 1971 BASED ON THE MEAN OF COUNTS FROM THREE 23 cm x 23 cm
PONAR GRABS; AND IN PARENTHESES, THE RELATIVE ABUNDANCE OF EACH TAXON
AS A PERCENTAGE OF THE TOTAL. The lower portion of the table lists
probable number of each species in each collection, probable number (%)
of Limnodrilus hoffmeisteri, Tubifex tubifex, and the total number of
specimens examined.
oo
00
Taxon
Stylodrilus heringianus
Dero digitata
Limnodrilus cervix
L. cervix-claparedeanus
L. hoffmeisteri
Peloscolex multisetosus
multisetosus
Tubifex tubifex
Undetermined immature
With hair chaetae
Without hair chaetae
Total
Probable L. hoffmeisteri
Probable T. tubifex
Probable No. species
No. specimens examined
1
13233 (8)
43008 (26)
81053 (49)
28120 (17)
165414±9589
25%
75%
2
104
2
Station
3
32872 (19)
20761 (12)
39792 (23)
77854 (45)
173008±4748
65%
35%
2
98
862 (1)
15512 (18)
29300 (34)
4308 (5)
22406 (26)
31024 (36)
86178±10519
52%
8%
4
90
4
5
733 (10)
16848 (23)
28001 (38)
1300 (2)
13918 (19)
19778 (27)
7325116974
38%
3
121
-------
Table A-8.
(continued) ABUNDANCE OF OLIGOCHAETE TAXA, INDIVIDUALS/m2, AT STATIONS
SAMPLED 1 DECEMBER 1971 BASED ON THE MEAN OF COUNTS FROM THREE 23 cm X
23 cm PONAR GRABS; AND IN PARENTHESES, THE RELATIVE ABUNDANCE OF EACH
TAXON AS A PERCENTAGE OF THE TOTAL. The lower portion of the table lists
probable number of each species in each collection, probable number (%)
of Limnodrilus hoffmeisteri, Tubifex tubifex, and the total number of
specimens examined.
Taxon
Station
8 9
10
11
Stylodrilus heringianus
Dero digitata
2 Limnodrilus cervix
L_. cervix-claparedeanus
L. hoffmeisteri
Peloscolex multisetosus
multisetosus
Tubifex tubifex
Undetermined immature
With hair chaetae
Without hair chaetae
Total
163 (76) 19 (100) 129 (66) 6 (100)
26 (12)
10 (5)
26 (12)
215±94 19±11
57 (29)
196±45
6±6
798 (50)
32 (2)
192 (12)
199 (14)
355 (25)
80 (5) 326 (23)
511 (32) 539 (38)
1596±900 1418±130
Probable L. hoffmeisteri 24%
Probable T_. tubifex
Probable No. species 2
No. specimens examined 34
1
3
34%
2
41
1
1
40%
3
66
63%
2
108
-------
Table A-8.
(continued) ABUNDANCE OF OLIGOCHAETE TAXA, INDIVIDUALS/m2, AT STATIONS
SAMPLED 1 DECEMBER 1971 BASED ON THE MEAN OF COUNTS FROM THREE 23 cm x
23 cm PONAR GRABS; AND IN PARENTHESES, THE RELATIVE ABUNDANCE OF EACH
TAXON AS A PERCENTAGE OF THE TOTAL. The lower portion of the table
lists probable number of each species in each collection, probable
number (%) of Limnodrilus hoffmeisteri, Tubifex tubifex, and the total
VO
o
number of specimens examined.
Taxon 12
Stylodrilus heringianus 45 (73)
Dero digitata
Limnodrilus cervix
L. cervix-claparedeanus
L. hoffmeisteri
Peloscolex multisetosus
multisetosus
Tubifex tubitex
Undetermined immature
With hair chaetae
Without hair chaetae 17 (27)
Total 6 3 ±35
Probable L. hoffmeisteri 27%
Probable T. tubifex
Probable No. species 2
No. specimens examined 11
Station
13 14
32 (36)
12949 (9)
33091 (23) 32 (36)
35970 (25)
5755 (4)
5753 (4)
48917 (34) 26 (29)
143874±4749 89±39
44%
8%
4
112
65%
2
14
15
954 (3)
8589 (27)
8589 (27)
1591 (5)
2227 (7)
10182 (32)
3181213078
55%
11%
4
107
16
1453 (1)
34867 (24)
40678 (28)
5811 (4)
3075 (2)
49395 (34)
14528016418
57%
13%
4
145
-------
Table A-9.
ABUNDANCE OF OLIGOCHAETE TAXA, INDIVIDUALS/m2, AT STATIONS SAMPLED
17 APRIL 1972 BASED ON THE MEAN OF COUNTS FROM THREE 23 cm x 23 cm
PONAR GRABS; AND IN PARENTHESES, THE RELATIVE ABUNDANCE OF EACH
TAXON AS A PERCENTAGE OF THE TOTAL. The lower portion of the table
lists probable number of each species in each collection, probable
number (%) of Limnodrilus hoffmeisteri, Tubifex tubifex, and the
total number of specimens examined.
Taxon
Stylodrilus heringianus
Dero digitata
Limnodrilus cervix
L. cervix-claparedeanus
L. hof fmeisteri
Peloscolex multisetosus
multisetosus
Tubifex tub if ex
Undetermined immature
With hair chaetae
Without hair chaetae
Total
Probable L. hof fmeisteri
Probable T. tubifex
Probable No. species
No. specimens examined
1
20682 (14)
39887 (27)
56138 (38)
31024 (21)
14773115665
35%
65%
2
117
2
S tation
3
1193 CD
29829 (25)
10738 (9)
26249 (22)
51305 (43)
11931414880
67%
31%
3
103
12559 (15)
36002 (43)
2512 (3)
8374 (10)
24281 (29)
8372613321
44%
13%
3
91
4
1040 (2)
4162 (8)
9884 (19)
2601 (5)
10404 (20)
52022+5194
31%
10%
5
122
5
70 (55)
19 (15)
19 (15)
19 (15)
127163
20%
2
20
-------
Table A-9. (continued) ABUNDANCE OF OLIGOCHAETE TAXA, INDIVIDUALS/m2, AT STATIONS
SAMPLED 17 APRIL 1972 BASED ON THE. MEAN OF COUNTS FROM THREE 23 cm x
23 cm PONAR GRABS; AND IN PARENTHESES, THE RELATIVE ABUNDANCE OF EACH
TAXON AS A PERCENTAGE OF THE TOTAL. The lower portion of the table
lists probable number of each species in each collection, probable
number (%) of Limnodrilus hoffmeisteri, Tubifex tubifex, and the total
number of specimens examined.
to
T axon 6 7
Stylodrilus heringianus 51 (100) 6 (100)
Dero digitata
Limnodrilus cervix
L. cervix-claparedeanus
L. hoffmeisteri
Peloscolex multisetosus
multisetosus
Tubifex tubifex
Undetermined immature
With hair chaetae
Without hair chaetae
Total 51+17 6 + 6
Probable L. hoffmeisteri
Probable T. tubifex
Probable No. species 1
No. specimens examined 1
S tation
8 9 10
19 (100) 101 (5)
511 (24)
277 (13)
213 (10)
106 (5)
979 (46)
19±19 101±35 2128±320
70%
15%
114
3 16 63
11
320 (23)
223 (16)
56 (4)
153 (11)
655 (47)
13931139
63%
15%
3
75
-------
Table A-9.
(continued) ABUNDANCE OF OLIGOCHAETE TAXA, INDIVIDUALS/m2, AT STATIONS
SAMPLED 17 APRIL 1972 BASED ON THE MEAN OF COUNTS FROM THREE 23 cm X
23 cm PONAR GRABS; AND IN PARENTHESES, THE RELATIVE ABUNDANCE OF EACH
TAXON AS A PERCENTAGE OF THE TOTAL. The lower portion of the table
lists probable number of each species in each collection, probable
number (%) of Limnodrilus hoffmeisteri, Tubifex tubifex, and the total
number of specimens examined.
to
S tation
Taxon
12
13
14
15
16
Stylodrilus heringianus
Dero digitata
Limnodrilus cervix
L. cervix-claparedeanus
L. hoffmeisteri
Peloscolex multisetosus
multisetosus
Tubifex tubifex
Undetermined immature
With hair chaetae
Without hair chaetae
Total
Probable L. hoffmeisteri
Probable T. tubifex
Probable No. species
No. specimens examined
5523 (2)
20987 (19)
36452 (33)
6628 (6)
8636 (16)
32033 (29)
110460±8031
45%
14%
4
118
605 (5)
5026 (25)
3147 (26)
726 (6)
969 (8)
3752 (31)
1210312141
45%
14%
4
81
2356 (7)
8752 (26)
5050 (15)
1683 (5)
2692 (8)
12792 (38)
33662±1662
54%
13%
4
84
-------
Table A-10. ABUNDANCE OF OLIGOCHAETE TAXA, INDIVIDUALS/m2, AT STATIONS SAMPLED
20 JUNE 1972, BASED ON THE MEAN OF COUNTS FROM THREE 23 cm x 23 cm
PONAR GRABS; AND IN PARENTHESES, THE RELATIVE ABUNDANCE OF EACH
TAXON AS A PERCENTAGE OF THE TOTAL. The lower portion of the table
lists probable number of each species in each collection, probable
number (%) of Limnodrilus hoffmeisteri, Tubifex tubifex, and the
total number of specimens
T axon
Stylodrilus heringianus
Dero digitata
Limnodrilus cervix
L. cervix-claparedeanus
L. hoffmeisteri
Peloscolex multisetosus
multisetosus
Tubifex tubifex
Undetermined immature
With hair chaetae
Without hair chaetae
Total
Probable L. hoffmeisteri
Probable T. tubifex
Probable No. species
No. specimens examined
1
30229 (20)
54412 (36)
43832 (29)
22672 (15)
151145±8873
35%
65%
2
116
examined .
2
S tation
3
37118 (33)
23621(21)
22496 (20)
30370 (27)
112480±3923
60%
40%
2
101
18924 (26)
32024 (44)
3639 (5)
4267 (6)
13829 (19)
7278317378
45%
11%
3
93
4
554 (1)
554 (1)
2772 (5)
15522 (28)
19402 (35)
3098 (11)
3326 (6)
7207 (13)
55436±5648
39%
17%
6
128
5
139 (61)
7 (3)
50 (22)
7 (3)
25 (11)
228±94
32%
4
36
-------
Table A-10.
(continued) ABUNDANCE OF OLIGOCHAETE TAXA, INDIVIDUALS/m2, AT STATIONS
SAMPLED 20 JUNE 1972, BASED ON THE MEAN OF COUNTS FROM THREE 23 cm x
23 cm PONAR GRABS; AND IN PARENTHESES, THE RELATIVE ABUNDANCE OF EACH
TAXON AS A PERCENTAGE OF THE TOTAL. The lower portion of the table
lists probable number of each species in each collection, probable
number (%) of Limnodrilus hoffmeisteri, Tubifex tubifex, and the total
number of specimens examined.
Taxon 6 7
Stylodrilus heringianus 19 (100) 32 (83)
Dero digitata
Limnodrilus cervix
L. cervix-claparedeanus
L. hoffmeisteri
Peloscolex multisetosus
multisetosus
Tubifex tubifex
Undetermined immature
With hair chaetae
Without hair chaetae 16 (17)
Total 19±11 48±29
Probable L. hoffmeisteri
Probable T. tubifex
Probable No. species 1 2
Station
8 9 10
89 (100) 158 (100) 68 (3)
588 (26)
588 (26)
68 (3)
68 (3)
904 (40)
89±50 158±61 22611378
66%
6%
1 1
11
519 (26)
778 (39)
219 (11)
459 (23)
1995±295
62%
3
No. specimens examined
14
25
99
-------
Table A-10.
(continued) ABUNDANCE OF OLIGOCHAETE TAXA, INDIVIDUALS/m2, AT STATIONS
SAMPLED 20 JUNE 1972, BASED ON THE MEAN OF COUNTS FROM THREE 23 cm x
23 cm PONAR GRABS; AND IN PARENTHESES, THE RELATIVE ABUNDANCE OF EACH
TAXON AS A PERCENTAGE OF THE TOTAL. The lower portion of the table
lists probable number of each species in each collection, probable
number (%) of Limnodrilus hoffmeisteri, Tubifex tubifex, and the total
number of specimens examined.
VD
(Tl
Taxon 12
Stylodrilus heringianus 9 (75)
Dero digitata
Limnodrilus cervix
L. cervix-claparedeanus
L. hof fmeisteri
Peloscolex multisetosus
multisetosus
Tubifex tubifex
Undetermined immature
With hair chaetae
Without hair chaetae 6 (25)
Total 27±17
Probable L. hoffmeisteri 25%
Probable T. tubifex
Probable No. species 2
No. specimens examined 4
Station
13 14
8968 (9)
26905 (27)
28898 (29)
7972 (8)
6975 (7)
20926 (21) 19 (100)
99649±5090 19±11
41% 100%
25%
4 1
116 3
15
1176 (9)
3529 (27)
4445 (34)
392 (3)
261 (3)
3399 (26)
13072+1889
47%
6%
4
90
16
956 (3)
9878 (31)
6691 (21)
2868 (9)
2231 (7)
9240 (29)
3186317422
57%
16%
4
98
-------
Table A-ll. ABUNDANCE OF INVERTEBRATE TAXA, INDIVIDUALS/m2, AT STATIONS SAMPLED
7 JUNE 1971, BASED ON THE MEAN COUNTS FROM THREE 23 cm x 23 cm
PONAR GRABS.
123
Chironomids
Heterotrissocladius sp.
Procladius sp.
Chironomus sp.
Station
456789
82
10 11
399
25
12 13
89
6
14 15 16
Hirudinea
Helobdella stagnalis
Gastropoda
Vivaparus sp.
Valvata sp.
Isopods
Asellus intermedius
Amphipods
Gammarus
fasciatus
Hyalella azteca
Pontoporeia affinis
6 6
13
114
6
57
6
-------
Table A-12. ABUNDANCE OF INVERTEBRATE TAXA, INDIVIDUALS/m2, AT STATIONS SAMPLED
6 AND 7 JULY 1971, BASED ON THE MEAN COUNTS FROM THREE 23 cm x 23 cm
PONAR GRABS.
Station
6 7 8 9 10 11 12 13 14 15 16
vo
oo
Chironomids
Heterotrissocladius sp.
Procladius sp.
Chironomus sp.
Hirudinea
Helobdella. stagnalis
Gastropoda
Vivaparus sp.
Valvata sp.
Isopods
Asellus intermedius
13 139
19 19
25 82 323 253
127
82
19
21 82
51
25
25
25
38 127 203
Amphipods
Gammarus fasciatus
Hyalella azteca
Pontoporeia affinis
13
25 95
11
28 21 38
-------
Table A-13. ABUNDANCE OF INVERTEBRATE TAXA, INDIVIDUALS/m2, AT STATIONS SAMPLED
1 DECEMBER 1971, BASED ON THE MEAN COUNTS FROM THREE 23 cm x 23 cm
PONAR GRABS.
Station
6789
10 11 12 13 14 15 16
vo
Chironomids
Heterotrissocladius sp.
Procladius spl
Chironomus sp.
Hirudinea
Helobdella stagnalis
Gastropoda
Vivaparus sp.
Valvata sp.
Isopods
Asellus intermedius
Amphipods
Gammarus fasciatus
Hyalella azteca
Pontpppreia affinis
38
139
42
13
13
-------
Table A-14. ABUNDANCE OF INVERTEBRATE TAXA, INDIVIDUALS/m2, AT STATIONS SAMPLED
17 APRIL 1972, BASED ON THE MEAN COUNTS FROM THREE 23 cm X 23 cm
PONAR GRABS.
Station
6789
10 11 12 13 14 15 16
o
o
Chironomids
Heterotrissocladius sjp.
Procladius spT
Chironomus sp.
Hirudinea
Helobdella stagnalis
Gastropoda
Vivaparus sp.
Valvata sp.
Isopods
Asellus intermedius
Amphipods
Gammarus fasciatus
Hyalella azteca
Pontoporeia affinis
6 171 19 19
26
6
51 6 142 51
108
82
3
10
6 6
-------
Table A-15. ABUNDANCE OF INVERTEBRATE TAXA, INDIVIDUALS/m2, AT STATIONS SAMPLED
20 JUNE 1972, BASED ON THE MEAN COUNTS FROM THREE 23 cm x 23 cm
PONAR GRABS.
Station
789
10 11 12 13 14 15 16
Chironomids
Heterotrissocladius sp.
Procladius sp.
Chironomus sp.
Hirudinea
Helobdella stagnalis
Gastropoda
Vivaparus sp.
Valvata sp.
Isopods
Asellus intermedius
Amphipods
Gammarus fasciatus
Hyalella azteca
Pontoporeia affinis
203
13 6 6 95 19 154
87
19 25
6 19
13
6
13
-------
APPENDIX B
CHEMISTRY OF N-CHLORAMINES
1. Amination of Toluene, Adamantane, and tert-Butyl
Chloride with Monochloramine-Aluminum Chloride
The reaction of monochloramine with toluene in the presence
of aluminum chloride at -35°C yielded 13-15% of m-toluidine,
whereas trichloramine gave 39-43% yields. To rationalize
the meta substitution, a mechanistic scheme entailing addi-
tion-elimination is proposed. Amination of adamantane with
monochloramine under Friedel-Crafts conditions gave 1-
aminoadamantane in 40% yield. An analogous reaction with
tri-chloramine-aluminum chloride provided 1-aminoadamantane
in 85% yield with no detectable 2-aminoadamantane. The
reaction pathway presumably involves formation of the 1-
adamantyl cation followed by attack by the nitrogen-contain-
ing nucleophile. Reaction of tert-butyl chloric monochlor-
amine, and aluminum chloride yielded tert-butylamine (7-20%).
Similarly, trichloramine generated tert-butylamine in 50-56%
yield and 2, 2-dimethylaziridine in 7-12% yield. Mechanis-
tically, the tert-butyl cation is thought to participate as
an intermediate. Possible reasons are discussed for the
lower yields in all cases with monochloramine, as compared
to trichloramine.
2. Preferred Route for vic-Dichloride Formation from
Alkenes. Comparison of Cyclohexene Chlorination with
Chlorine, Sulfuryl Chloride, and Trichloramine
The best method for generating vic-dichlorides was investi-
gated by comparing chlorination of cyclohexene with chlorine,
sulfuryl chloride, and trichloramine. In relation to clean-
ness of reaction, trichloramine is the reagent of choice
(97% yield of trans-l;2-dichlorocyclohexane). If some
sacrifice in yieTd is'not critical, sulguryl chloride or
chlorine would be preferred on the basis of simpler oper-
ating procedures.
102
-------
3. Rearrangement of o-Hydroxyaldehydes and Ketones to
o-Hydroxyanxlides by Monochloramxne
o-Hydroxyaldehydes and ketones are converted in good yield
to o-hydroxy anil ides by reaction with monochloramine in
base. The reaction was carried out with benzene nuclei
containing alkyl, methoxyl, chlorine, and nitro substit-
uents, as well as with a naphthalene nucleus. The overall
transformation is similar to the Beckmann, Schmidt, Theil-
acker, and Pearson rearrangement. There appears to be
mechanistic similarity to the Dakin oxidation.
4. A Novel, Directed Synthesis of Unsymmetrical Azoxyal-
kanes and Azoxyaralkanes from N,N-Dihaloamine and
Nitroso Precursors
A novel, directed synthesis of unsymmetrical azoxyalkanes
and azoxyaralkanes from nitroso compounds (RNO) and N,N-
dichloramines (R'NC^) in the presence of methanolic caustic
is described. An investigation of the scope of the reaction
revealed that the highest yields of azoxy compounds were
produced when R is tert-alkyl or aryl and R1 is tert-alkyl.
This method possesses advantages not offered by prior tech-
niques. Possible mechanistic pathways are also discussed.
5. Dealkylation of N,N-Dichloroalkylamines with Silver
Salts
Reaction of N,N-dichloro-t-butylamine in refluxing ac^toni-
trile with silver (I) fluoride gave isobutylene in 80-35%
yield (Sharts 1969). Investigation of N,N-dichloro-l-araino-
1-methylcyclohexane in dimethyl sulfoxide with silver ace-
tate at room temperature afforded 50-70% alkene (45% exo:
55% endo). This reaction is being studied further.
103
-------
APPENDIX C
BIOASSAY TESTS
104
-------
Table C-l. TOXICITY OF RESIDUAL CHLORINE ADDED AS SODIUM
HYPOCHLORITE TO CYCLOPS BICUSPIDATUS THOMAS I.
Date: May 19-23, 1973.
Exposure Period: 96 hrs.
Temperature: 15 °C
Average
No. of total residual Mortality
Aquarium organisms chlorine (mg/1) %
1A
IB
2A
2B
3A
3B
4A
4B
5A
5B
6A
6B
20
20
20
20
20
20
20
20
20
20
20
20
0.000
0.000
0.011
0.010
0.024
0.021
0.081
0.072
0.170
0.178
0.264
0.269
15.8
10.0
25.0 .
25.0
22.0
36.8
61.1
50.0
90.0
95.0
100.0
100.0
Average
PH
8.15
8.15
8.10
8.05
8.10
8.10
8.15
8.10
8.10
8.10
8.15
8.15
105
-------
Table C-2. TOXICITY OF RESIDUAL CHLORINE ADDED AS SODIUM
HYPOCHLORITE TO CYCLOPS BICUSPIDATUS THOMASI.
Date: June 1-5, 1973. Exposure Period: 96 hrs .
Temperature: 15 °C
Aquarium
1A
IB
2A
2B
3A
3B
4A
4B
5A
5B
6A
6B
No. of
organisms
20
20
20
20
20
20
20
20
20
20
20
20
Average
total residual
chlorine (mg/1)
0.000
0.000
0.020
0.021
0.046
0.047
0.083
0.109
0.185
0.191
0.401
0.376
Mortality
%
0.0
10.0
25.0
10.0
33.3
21.1
22.2
42.1
45.5
38.5
90.5
85.0
Average
pH
8.20
8.15
8.18
8.18
8.20
8.18
8.20
8.18
8.23
8.28
8.25
8.25
106
-------
Table C-3. TOXICITY OF RESIDUAL CHLORINE ADDED AS SODIUM
HYPOCHLORITE TO CYCLOPS BICUSPIDATUS THOMASI.
Date: June 14-18, 1973. Exposure Period: 96 hrs .
Temperature: 15°C
Aquarium
1A
IB
2A
2B
3A
3B
4A
4B
5A
5B
6A
6B
No. of
organisms
20
20
20
20
20
20
20
20
20
20
20
20
Average
total residual
chlorine (mg/1)
0.000
0.000
0.029
0.027
0.075
0.081
0.184
0.178
0.234
0.238
0.630
0.660
Mortality
%
20.0
65.0
30.0
55.0
25.0
14.3
88.2
73.0
89.5
90.0
94.7
100.0
Average
PH
8.20
-
8.20
8.20
8.22
8.21
8.23
8.20
8.22
8.18
8.20
8.21
107
-------
Table C-4. TOXICITY OF RESIDUAL CHLORINE ADDED AS MONO-
CHLORAMINE TO CYCLOPS BICUSPIDATUS THOMASI.
Date: July 13-17, 1973. Exposure Period: 96 hrs.
Temperature 15 °C
Aquarium
1A
IB
2A
2B
3A
3B
4A
4B
5A
5B
6A
6B
No. of
organisms
20
20
20
20
20
20
20
20
20
20
20
20
Average
total residual
chlorine (mg/1)
0.000
0.000
0.018
0.015
0.042
0.036
0.136
0.165
0.236
0.312
1.025
0.982
Mortality
%
5.6
6.7
5.3
5.9
20.0
5.9
84.2
87.5
100.0
100.0
100.0
100.0
Average
PH
-
-
8.15
8.20
8.21
8.22
8.19
8.19
8.21
8.25
8.29
8.20
108
-------
Tabe C-5. TOXICITY OF RESIDUAL CHLORINE ADDED AS MONO-
CHLORAMINE TO CYCLOPS BICUSPIDATUS TKOMASI.
Date: July 19-23, 1973.
Exposure Period: 96 hrs.
Temperature: 15 °C
Aquarium
1A
IB
2A
2B
3A
3B
4A
4B
5A
5B
6A
6B
No. of
organisms
20
20
20
20
20
20
20
20
20
20
20
20
Average
total residual
chlorine (mg/1)
0.000
0.000
0.012
0.024
0.067
0.041
0.169
0.168
0.308
0.398
0.979
0.990
Mortality
%
5.0
5.9
0.0
0.0
10.5
10.5
73.7
100.0
100.0
100.0
100.0
100.0
Average
pH
8.37
8.39
8.39
8.36
8.37
8.36
8.35
8.36
8.34
8.35
8.37
8.37
109
-------
Table C-6. TOXICITY OF RESIDUAL CHLORINE ADDED AS MONO-
CHLORAMINE TO CYCLOPS BICUSPIDATUS THOMASI.
Date: Aug. 9-13, 1973.
Aquarium
1A
IB
2A
2B
3A
3B
4A
4B
5A
5B
6A
6B
No. of
organisms
20
20
20
20
^_20
20
20
20
20
20
20
20
Average
total residual
chlorine (mg/1)
0.000
0.000
0.014
0.018
0.067
0.061
0.147
0.156
0.323
0.359
0.838
0.868
Exposure Period: 96 hrs.
Temperature: 15°C
Mortality
%
15.0
0.0
10.5
17.6
41.2
70.0
90.0
90.0
100.0
100.0
100.0
100.0
Average
PH
8.33
8.30
8.40
8.34
8.45
8.37
8.49
8.42
8.50
8.45
8.53
8.48
110
-------
Table C-7.
TOXICITY OF RESIDUAL CHLORINE ADDED AS MONO-
CHLORAMINE TO CYCLOPS BICUSPIDATUS THOMASI.
Date: Aug. 16-20, 1973.
Exposure Period: 96 hrs.
Temperature: 15°C
Aquarium
1A
IB
2A
2B
3A
3B
4A
4B
5A
5B
6A
6B
No. of
organisms
20
20
20
20
20
20
20
20
20
20
20
20
Average
total residual
chlorine (mg/1)
0.000
0.000
0.022
0.031
0.089
0.083
0.227
0.195
0.447
0.507
1.110
1.194
Mortality
%
0.0
0.0
5.0
10.0
38.9
31.6
85.7
88.9
100.0
100.0
100.0
100.0
Average
PH
8.39
8.42
8.32
8.39
8.38
8.45
8.39
8.40
8.42
8.40
8.40
8.32
111
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Table C-8. TOXICITY OF SODIUM SULFITE TO CYCLOPS
BICUSPIDATUS THOMAS I.
Date: Jan. 11-15, 1974. Exposure Period: 96 hrs.
Temperature 15°C
Aquarium
1A
IB
2A
2B
3A
3B
4A
4B
5A
5B
6A
6B
No. of
organisms
20
20
20
20
20
20
20
20
20
20
20
20
Average
SO^ as S02
Cmg/1)
0.000
0.000
0.011
0.015
0.030
0.035
0.050
0.062
0.130
0.137
0.477
0.563
Mortality
25.0
12.5
20.0
17.6
10.5
23.5
5.9
20.0
22.2
11.1
12.5
15.8
Average
PH
8.27
8.32
8.32
8.30
8.30
8.30
8.30
8.32
8.30
8.33
8.32
8.29
112
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Table C-9.
TOXICITY OF RESIDUAL CHLORINE TO CYCLOPS
BICUSPIDATUS THOMASI. DECHLORINATING
WITH SULFITE.
Date: April 16-20, 1974. Exposure Period: 96 hrs.
Temperature: 15 ° C
Average Average
No. of total residual SOo as S02 Mortality Average
Aquarium organisms chlorine (mg/1) (mg/1) % pH
1A 20
IB 20
2A 20
2B 20
3A 20
3B 20
4A 20
4B 20
5A 20
5B 20
6A 20
6B 20
0.000
0.000
0.000
0.000
0.000
0.000
0.000
0.000
0.000
0.000
0.000
0.000
0-000
0.000
0.000
0.005
0.024
0.018
0.028
0.026
0.000
0.019
0.014
0.069
5.0
11.1
5.0
10.5
5.0
5.0
10.0
0.0
5-0
0.0
10.5
5.0
8.08
8.20
8.19
8.20
8.24
8.19
8.24
8.20
3.29
8:26
8.30
8.32
113
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Table C-10. TOXICITY OF RESIDUAL CHLORINE TO CYCLOPS
BICUSPIDATUS THOMASI. DECHLORINATING
WITH SODIUM SULFITE.
Date: June 3-7, 1974. Exposure Period: 96 hrs,
Temperature: 15°C
Average Average
No. of total residual SO^ as SO2 Mortality
Aquarium organisms chlorine Cmg/1) (mg/1) %
1A
IB
2A
2B
3A
3B
4A
4B
5A
5B
6A
6B
20
20
20
20
20
20
20
20
20
20
20
20
0.000
0.000
0.000
0.000
0.000
0.000
0.000
0.000
0.000
0.000
0.000
0.000
0.000
0.000
0.006
0.004
0.006
0.011
0.026
0.022
0.042
0.039
0.099
0.098
22.2
5.0
14.3
10.0
5.0
5.0
11.1
18.8
5.3
11.8
15.0
15.8
Average
PH
8.08
8.16
8.35
8.18
8.31
8.39
8.15
8.27
8.32
8.30
8.36
8.38
114
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Table C-ll. TOXICITY OF RESIDUAL CHLORINE ADDED AS
MONOCHLORAMINE TO KERATELLA COCHLEARIS.
Date:
Sept. 21, 1973. Exposure Period: 4 hrs.
Temperature: 15 °C
Aquarium
1A
IB
2A
2B
3A
3B
4A
4B
5A
5B
6A
6B
No. of
organisms
20
20
20
20
20
20
20
20
20
20
20
20
Average
total residual
chlorine (mg/1)
0.000
0.000
0.000
0.003
0.006
0.004
0.039
0.019
0.055
0.062
0.135
0.156
Mortality
%
5.0
19.0
12.5
21.1
31.6
30.0
47.4
41.2
94.4
100.0
100.0
100.0
Average
pH
8.10
8.10
8.10
8.10
8.10
8.10
8.10
8.10
8.10
8.10
8.15
8.15
115
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Table C-12. TOXICITY OF RESIDUAL CHLORINE ADDED AS
MONOCHLORAMINE TO KERATELLA COCHLEARIS.
Aquarium
1A
IB
2A
2B
3A
3B
4A
4B
5A
5B
6A
6B
Date:
No. of
organisms
20
20
20
20
20
20
20
20
20
20
20
20
Oct. 11, 1973.
Average
total residual
chlorine (mg/1)
0.000
0.000
0.000
0.000
0.000
0.000
0.008
0.005
0.026
0.037
0.077
0.107
Exposure Period: 4 hrs.
Temperature: 15 °C
Mortality
%
5.0
0.0
29.4
5.9
16.7
15.8
35.0
27.8
45.0
52.4
95.0
94.7
Average
PH
8.20
8.20
8.20
8.20
8.20
8.20
8.20
8.20
8.20
8.20
8.20
8.25
116
-------
Table C-13.
TOXICITY OF RESIDUAL CHLORINE ADDED AS
MONOCHLORAMINE TO KERATELLA COCHLEARIS.
Date:
Oct. 18, 1973.
Exposure Period: 4 hrs.
Temperature: 15 °C
Aquarium
1A
IB
2A
2B
3A
3B
4A
4B
5A
5B
6A
6B
No. of
organisms
20
20
20
20
20
20
20
20
20
20
20
20
Average
total residual
chlorine (mg/1)
0.000
0.000
0.000
0.000
0.000
0.000
0.008
0.005
0.029
0.025
0.103
0.124
Mortality
%
5.0
14.3
16.7
5.9
15.8
11.1
25.0
16.7
50.0
42.1
94.7
100.0
Average
PH
8.15
8.15
8.15
8.15
8.15
8.15
8.15
8.15
8.15
8.15
8.20
8.20
117
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Table C-14. TOXICITY OF SODIUM SULFITE ALONE
ON KERATELLA COCHLEARIS.
Date: Nov. 29, 1973. Exposure Period: 4 hrs,
Temperature: 15°C
303 as S02
No. of (mg/1) Mortality Average
Aquarium organisms Start Finish % pH
1A 20 0.000 0.000 14.3 8.20
IB 20 0.000 0.000 5.0 8.20
2A 20 0.692 0.750 5.0 8.20
2B 20 0.750 0.861 9.5 8.20
118
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Table C-15. TOXICITY OF RESIDUAL CHLORINE ADDED AS
MONOCHLORAMINE TO KERATELLA COCHLEARIS.
DECHLORINATING WITH SODIUM SULFITE.
Date: Jan. 26, 1974. Exposure Period: 4 hrs.
Temperature: 15°C
Average Average
No. of total residual 803 as S02 Mortality Average
Aquarium organisms chlorine (mg/1) (mg/1) % pH
1A
IB
2A
2B
3A
3B
4A
4B
5A
5B
6A
6B
20
20
20
20
20
20
20
20
20
20
20
20
0
0
0
0
0
0
0
0
2.5
3-0
14.0
13.5
0.000
0.000
0.000
0.000
0.000
0.000
0.000
0.000
0.000
0.000
0.000
0.000
11.1
0.0
10.5
6.3
0.0
5.9
0.0
11.1
5.9
0.0
11.1
10.5
8.40
8.40
8.40
8.40
8.40
8.40
8.40
8.40
8.40
8.40
8.40
8.40
119
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Table C-16. TOXICITY OF RESIDUAL CHLORINE ADDED AS
MONOCHLORAMINE TO KERATELLA COCHLEARIS.
DECHLORINATING WITH SODIUM SULFITE.
Date: Jan. 31, 1974. Exposure Period: 4 hrs
Temperature: 15°C
Average Average
No. of total residual SO^ as S02
Aquarium organisms chlorine (mg/1) (mg/1)
1A
IB
2A
2B
3A
3B
4A
4B
5A
5B
6A
6B
20
20
20
20
20
20
20
20
20
20
20
20
0
0
0
0
0
0
0
0
0
0
6
6
0.000
0.000
0.000
0.069
0.023
0.000
0.000
0.000
0.000
0.000
0.000
0.000
Mortality Average
% pH
0.0
0.0
0.0
5.9
10.0
11.8
0.0
6.7
6.3
0.0
5.9
0.0
8.40
8.40
8.40
8.40
8.40
8.40
8.40
8.40
8.40
8.40
8.40
8.40
120
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Table C-17. TOXICITY OF RESIDUAL CHLORINE ADDED AS
MONOCHLORAMINE TO KERATELLA COCHLEARIS.
DECHLORINATING WITH SODIUM SULFITE.
Date: Feb. 9, 1974. Exposure Period: 4 hrs.
Temperature: 15°C
Aquarium
1A
IB
2A
2B
3A
3B
4A
4B
5A
5B
6A
6B
Average Average
No. of total residual 303 as S02
organisms chlorine Crag/1 ) tmg/1)
20
20
20
20
20
20
20
20
20
20
20
20
0
0
0
0
0
0
0
0
0
0
0
0
0.000
0.000
0.000
0.034
0.000
0.080
0-069
0.242
0.196
0.161
0.161
0.253
Mortality
%
6.3
0.0
0.0
0.0
5.0
0.0
6.3
6.7
0.0
5.3
5.9
5.3
Average
PH
8.40
8.40
8.40
8.40
8.40
8.40
8.40
8.40
3.40
8.40
8.40
8.40
121
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TECHNICAL REPORT DATA
(Please read Instructions on the reverse before completing)
1. REPORT NO.
EPA-600/3-76-036
3. RECIPIENT'S ACCESSION NO.
4. TITLE ANDSUBTITLE
EFFECTS OF CHLORINE AND SULFITE REDUCTION
ON LAKE MICHIGAN INVERTEBRATES
5. REPORT DATE
April 1976
(Issuing Date)
6. PERFORMING ORGANIZATION CODE
7. AUTHOR(S)
A. M. Beeton, P. K. Kovacic, and
A. S. Brooks
8. PERFORMING ORGANIZATION REPORT NO
Center for Great Lakes
Studies Contrib. No. 137
9. PERFORMING ORGANIZATION NAME AND ADDRESS
The University of Wisconsin-Milwaukee
Center for Great Lakes Studies
Milwaukee, Wisconsin 53201
10. PROGRAM ELEMENT NO.
1BA608
11.R3NE8XKT/GRANT NO.
R-801035
12. SPONSORING AGENCY NAME AND ADDRESS
13. TYPE OF REPORT AND PERIOD COVERED
Environmental Research Laboratory
Office of Research and Development
U.S. Environmental Protection Agency
Duluth, Minnesota 55804
Final
14. SPONSORING AGENCY CODE
EPA-ORD
15. SUPPLEMENTARY NOTES
16. ABSTRACT
The acute toxicity of residual chlorine was determined for the copepoc
Cyclops bicuspidatus thomasi and the rotifer Keratella cochlearis. The
96-hour TLso value for Cyclops was 0.084 mg/1 total residual chlorine
added as monochloramine. When Cyclops was exposed to sodium hypochlo-
rite the 96-hour TLso was 0.069 mg/1 total residual chlorine. The
4-hour TLso value for Keratella was 0.019 mg/1 total residual chlorine
added as monochloramine.
Chemical studies determined that sodium sulfite was an efficient, in-
expensive chemical agent for reducing chlorine residuals which did not
produce undesirable by-products. Complete reduction was accomplished
in less than 20 seconds with a calculated km£n of 43 sec"1. Bioassay
studies indicated that sodium sulfite added to chlorinated water com-
pletely eliminated the acute toxicity of residual chlorine to both
Cyclops bicuspidatus thomasi and Keratella cochlearis.
Field studies in Milwaukee Harbor and adjacent Lake Michigan indicated
that measurable chlorine residuals were confined to a very small area
surrounding the effluent from the Jones Island Sewage Treatment Plant.
Significant reductions in the populations of benthic organisms were
observed in the effluent plume area after the start of chlorination.
17.
KEY WORDS AND DOCUMENT ANALYSIS
DESCRIPTORS
b.IDENTIFIERS/OPEN ENDED TERMS
c. cos AT I Field/Group
Water pollution
Chlorination
Sewage treatment
Bioassay
Toxicity
Sodium sulfites
Lake Michigan
Municipal wastes
Sewage effluents
Benthic fauna
Oligochaetes
Copepods
Rotifers
Chlorine toxicitv
6T
8. DISTRIBUTION STATEMENT
RELEASE UNLIMITED
19. SECURITY CLASS (ThisReport)
UNCLASSIFIED
21. NO. OF PAGES
132
20. SECURITY CLASS (Thispage)
UNCLASSIFIED
22. PRICE
EPA Form 2220-1 (9-73)
122
«USGPO: 1976 — 657-695/5405 Region 5-1
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