EPA-600/3-76-105
October 1976
EFFECTS OF EXPOSURE TO HEAVY METALS ON
SELECTED FRESHWATER FISH
Toxicity of Copper, Cadmium, Chromium and Lead to
Eggs and Fry of Seven Fish Species
by
Scott Sauter
Kenneth S. Buxton
Kenneth J. Macek
Sam R. Petrocelli
EG&G, Bionomics
Aquatic Toxicology Laboratory
Wareham, Massachusetts 02571
Contract 68-01-07^0
Project Officer
Duane A. Benoit
Environmental Research Laboratory
Duluth, Minnesota 55804
ENVIRONMENTAL RESEARCH LABORATORY
OFFICE OF RESEARCH AND DEVELOPMENT
U.S. ENVIRONMENTAL PROTECTION AGENCY
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.
ii
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FOREWORD
Our nation's freshwaters are vital for all animals and plants, yet our
diverse uses of water for recreation, food, energy, transportation, and
industry physically and chemically alter lakes, rivers, and streams. Such
alterations threaten terrestrial organisms, as well as those living in water.
The Environmental Research Laboratory in Duluth, Minnesota develops methods,
conducts laboratory and field studies, and extrapolates research findings
—to determine how physical and chemical pollution affects aquatic
life
—to assess the effects of ecosystems on pollutants
—to predict effects of pollutants on large lakes through use of models
—to measure bioaccumulation of pollutants in aquatic organisms that
are consumed by other animals, including man
This report demonstrates the effects of lead, chromium, copper, and
cadmium on the early life stages of rainbow trout, lake trout, brook trout,
channel catfish, bluegill, white sucker, northern pike, and walleye. The
results could be used in establishing water quality criteria for aquatic life.
Donald I. Mount, Ph.D.
Director
Environmental Research Laboratory
Duluth, Minnesota
iii
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ABSTRACT
Eggs and fry of rainbow trout, lake trout, channel catfish,
bluegill, white sucker, northern pike and walleye were con-
tinuously exposed for a maximum of 60 days after hatching,
to a series of concentrations of lead and chromium in soft
water (32.6-40.? mg/1 as CaCC>3). Similarly, eggs and fry
of "brook trout, channel catfish and walleyes were exposed to
copper and cadmium in "both soft (35-0-37.5 mg/1 as CaCOo)
and hard (185.0-189.0 mg/1 as CaCO^) water. Observations
of the hatchability of eggs and the survival and growth,
quantitated as total length and wet weight, of fry were made
after 30 and 60 days continuous exposure of the fry of these
species to metals.
Results of exposures were used to estimate the range of
metal concentrations which bracket the maximum acceptable
toxicant concentration (MATC) for a given metal and fish
species. The MATC's generated indicated that copper and
cadmium were similar in toxicity and were toxic at signifi-
cantly lower concentrations than lead and chromium. Water
hardness did not appear to have a significant effect on the
observed toxicity in most cases. Most importantly, MATC's
estimated from these relatively short duration egg and fry
tests were generally similar and in some cases, nearly
identical to those estimated from chronic studies of much
greater duration. This indicates that egg and fry studies
are an effective, reliable means of assessing chronic toxicity
of certain compounds to selected freshwater fish species in
relatively short-term studies.
iv
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CONTENTS
SECTION
I Introduction
II Conclusions 3
III Recommendations 4
IV Materials and Methods 5
Exposure Systems 5
Chemical Methods 5
Biological Methods 8
Statistics 9
V Results 11
Water Chemistry 11
Exposure of Fish to Lead 11
Rainbow trout 11
Lake trout 21
Channel catfish 21
Bluegill 24
White sucker 24
Northern pike 2?
Walleye 29
Exposure of Fish to Chromium 30
- Rainbow trout 30
Lake trout 33
Channel catfish 36
Bluegill 36
White sucker 39
Northern pike 39
Walleye 42
Exposure of Fish to Copper 43
Brook trout (soft water) 43
Brook trout (hard water) 43
Channel catfish (soft water) 46
Channel catfish (hard water) 46
Walleye (soft and hard waters) 49
Exposure of Fish to Cadmium 50
Brook trout (soft water) 50
Brook trout (hard water) 52
Channel catfish (soft water) 52
Channel catfish (hard water) 55
Walleye (soft and hard waters) 55
VI Discussion 59
Exposure of Fish to Lead 59
Exposure of Fish to Chromium 63
Exposure of Fish to Copper 65
Exposure of Fish to Cadmium 6?
VII Literature Cited 71
V
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TABLES
1. Source, age of eggs at beginning of exposure,
incubation time and temperature, and food utilized
in the exposure of eggs and fry of selected
freshwater fish species to metals . ]_Q
2. Means and standard deviations (x ± S.D.), ranges and
number of observations (N) of measured water quality
parameters during exposure of eggs and fry of selected
freshwater fish species to lead. 12
3. Means and standard deviations (x ± S.D.), ranges and
number of observations (N) of measured water quality
parameters during exposure of eggs and fry of selected
freshwater fish species to chromium. 13
4. Means and standard deviations (x ± S.D.), ranges and
number of observations (N) of measured water quality
parameters during exposure of eggs and fry of selected
freshwater fish species to copper in "soft" and "hard"
water. Ik
5. Means and standard deviations (x ± S.D.), ranges and
number of observations (N) of measured water quality
parameters during exposure of eggs and fry of selected
freshwater fish species to cadmium in "soft" and "hard"
water. 15
6. Means and standard deviations (x ± S.D.), ranges and
number (N) of measured concentrations of lead during
continuous exposure of eggs and fry of selected fresh-
water fish species. 16
7. Means and standard deviations (x ± S.D.), ranges and
number (N) of measured concentrations of chromium during
continuous exposure of eggs and fry of selected fresh-
water fish species. 17
8. Means and standard deviations (x ± S.D.), ranges and
number (N) of measured concentrations of copper during
continuous exposure of eggs and fry of selected fresh-
water fish species in "soft" and "hard" waters. 18
9. Means and standard deviations (x ± S.D.), ranges and
number (N) of measured concentrations of cadmium during
continuous exposure of eggs and fry of selected fresh-
water fish species in "soft" and "hard" waters. 19
vi
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(10.. PAGE
10. Mean percentage hatch of eggs, mean survival, total
lengths and wet weight of rainbow trout (Salmo gairdneri )
fry continuously exposed to lead in soft water
± 3.2 mg/1 as CaC0). 20
11. Mean percentage hatch of eggs, mean survival,
total lengths and wet weight of lake trout
(Salvelinus namaycush) fry continuously exposed
to lead in soft water (32.6 ±3.9 mg/1 as CaCOo) 22
12. Mean percentage hatch of eggs, mean survival, total
lengths and wet weight of channel catfish (Ictalurus
punctatus) fry continuously exposed to lead in soft
water (36.0 ± 1.0 mg/1 as CaCOo). 23
13. Mean percentage hatch of eggs, mean survival, total
lengths and wet weight of "bluegill (Lepomis macrochirus)
fry continuously exposed to lead in soft water
(*K>.7 ± 7-3 mg/1 as CaC03). 25
1^. Mean percentage hatch of eggs, mean survival, total
lengths and wet weight of white sucker (Catostomus
commersoni) fry continuously exposed to lead in soft
water (37.6 ±5.9 mg/1 as CaCC^). 26
15. Mean percentage hatch of eggs, mean survival, total
lengths and wet weight of northern pike (Esox lucius)
fry continuously exposed to lead in soft water (35«^
±4.8 mg/1 as CaCO^). 28
16. Mean percentage hatch of eggs, mean survival and
total lengths of walleye (Stizostedion vitreum) fry
continuously exposed to lead in soft water (37«6 ±5-2
mg/1 as CaCOo). 29
17. Mean percentage hatch of eggs, mean survival, total
lengths and wet weights of rainbow trout (Salmo gairdneri )
fry continuously exposed to chromium in soft water
(3^.0 ± 3.2 mg/1 as CaC03). 31
18. Mean percentage hatch of eggs, mean survival, total
lengths and wet weight of rainbow trout (Salmo gairdneri )
fry continuously exposed to chromium in soft water
(33.4- ± ^.5 mg/1 as CaC03). 32
19. Mean percentage hatch of eggs, mean survival, total
lengths and wet weight of lake trout (Salvelinus namaycush )
fry continuously exposed to chromium in soft water
(33.0 ± 3.9 mg/1 as CaCO^). 3^
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NO.
PAGE
20. Mean percentage hatch of eggs, mean survival, total
lengths and wet weight of lake trout (Salvelinus namaycusM
fry continuously exposed to chromium in soft water
(3^.0 ± 4.8 mg/1 as CaCOo). 35
21. Mean percentage hatch of eggs, mean survival, total
lengths and wet weight of channel catfish (Ictalurus
punctatus) fry continuously exposed to chromium in
soft water (36.2 ±1.2 mg/1 as CaCO^). 37
22. Mean percentage hatch of eggs, mean survival, total
lengths and wet weight of bluegill (Lepomis macrochirus)
fry continuously exposed to chromium in soft water
(38.3 ± 4.6 mg/1 as CaCO-). 38
23. Mean percentage hatch of eggs, mean survival, total
lengths and wet weight of white sucker (Catostomus
commersoni) fry continuously exposed to chromium in
soft water (38.8 ± 5.3 mg/1 as CaCOo). 40
24. Mean percentage hatch of eggs, mean survival, total
lengths and wet weight of northern pike (Esox lucius)
fry continuously exposed to chromium in soft water
(37-8 ± 5-6 mg/1 as CaC03). 41
25. Mean percentage hatch of eggs, mean survival and total
lengths of walleye (Stizostedion vitreum) fry continuously
exposed to chromium in soft water (38.5 ± 4.2 mg/1 as
CaCOo). 42
26. Mean percentage hatch of eggs, mean survival, total
lengths and wet weight of brook trout (Salvelinus
fontinalis) fry continuously exposed to copper in
soft water (37-5 ± 7-3 mg/1 as CaCOo). 44
27. Mean percentage hatch of eggs, mean survival, total
lengths and wet weight of brook trout (Salvelinus
fontinalis) fry continuously exposed to copper in hard
water (187.0 ± 22.0 mg/1 as CaCO-J. 45
28. Mean percentage hatch of eggs, mean survival, total
lengths and wet weight of channel catfish (Ictalurus
punctatus) fry continuously exposed to copper in soft
water (36.0 ± 1.1 mg/1 as CaCOo). 47
29. Mean percentage hatch of eggs, mean survival, total
lengths and wet weight of channel catfish (Ictalurus
punctatus) fry continuously exposed to copper in hard
water (186.3 ±38.7 mg/1 as CaCO,,). 48
viii
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NO.
PAGE
30. Mean percentage hatch of eggs, mean survival and total
lengths of walleye (Stizostedion vitreum) fry continu-
ously exposed to copper in soft water" (35-0 ± 1.8 mg/1
as CaCOo). 49
31. Mean percentage hatch of eggs, mean survival, total
lengths and wet weight of walleye (Stizostedion vitreum)
fry continuously exposed to copper in hard water (189.0
±33.0 mg/1 as CaCO ). 50
32. Mean percentage hatch of eggs, mean survival, total
lengths and wet weight of brook trout (Salvelinus
fontinalis) eggs and fry continuously exposed to various
concentrations of cadmium in soft water (37.0 ± 7.2
mg/1 as CaCOo). 51
33. Mean percentage hatch of eggs, mean survival, total
lengths and wet weight of brook trout (Salvelinus
fontinalis) fry continuously exposed to cadmium in
hard water (188.0 ± 27.0 mg/1 as CaCO.,). 53
34. Mean percentage hatch of eggs, mean survival, total
lengths and wet weight of channel catfish (Ictalurus
punctatus) fry continuously exposed to cadmium in soft
water (37.0 ±1.3 mg/1 as CaCOo). 5^
35• Mean percentage hatch of eggs, mean survival, total
lengths and wet weight of channel catfish (Ictalurus
punctatus) fry continuously exposed to cadmium in hard
water (185.0 ±35.0 mg/1 as CaCOo). 56
36. Mean percentage hatch of eggs, mean survival and total
lengths of walleye (Stizostedion vitreum) fry continuously
exposed to cadmium in soft water (35.0 ± 1.2 mg/1 as
CaC03). 57
37. Mean percentage hatch of eggs and survival of walleye
(Stizostedion vitreum) fry continuously exposed to cadmium
in hard water (187.0 ± 36.0 mg/1 as CaCO.,). 58
38. Summary of the maximum acceptable toxicant concentration
(MATC) of lead and chromium for selected freshwater
fish species in soft water (32.6-^0.7 mg/1 as CaCOo). 60
39. Summary of the maximum acceptable toxicant concentration
(MATC) of copper and cadmium for selected freshwater
fish species in soft (35.0-37.5 mg/1 as CaCOo) and hard
(185.0-189.0 mg/1 as CaCOo) water. * 61
^0. Summary of the actual duration of exposure to lead and
chromium for selected freshwater fish species. 61
IX
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ACKNOWLEDGEMENTS
During the course of this study we were fortunate to have
laboratory assistance from Jerry Dean, Mark Lindberg, Steve
Derr, Pat Costa, Gerry LeBlanc, Emily Dionne and Stephan Ells
who performed many of the tasks necessary in conducting these
exposures. We also extend our appreciation to Mr. D. Curtis
Hutchinson for constructing the proportional dilution apparatus.
We are indebted to Mr. Richard Seamans, chief of the Division
of Inland and Marine Fisheries and to Mr. Arthur Newell, Mr.
Peter Brezosky and Mr. Charles Thoits of the New Hampshire
Fish and Game Department for providing the lake trout and white
sucker eggs needed in this study. We are further grateful to
Mr. John Spinner of the Iowa State Fish Hatchery for providing
northern pike eggs and Mr. Wally Jorgensen of the Spirit Lake
Fish Hatchery for providing eggs of the walleye.
Our sincere appreciation is extended to Mr. Duane Benoit,
Project Officer, NWQL (now the Environmental Research Laboratory),
Duluth, Minnesota for his patience and constructive advice during
the performance of these studies.
X
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SECTION I
INTRODUCTION
[Che establishment of a realistic and valid water quality
criteria for chemicals entering surface waters requires an
understanding of both the acute and chronic effects of these
chemicals on aquatic organisms. Although the former is
relatively easy to estimate, definition of the latter requires
a time consuming and expensive effort. Data on estimates of
acute toxicity of a myriad of chemicals to aquatic forms have
been collected and are available (McKee and Wolf, 1963; Battelle-
Columbus Laboratories, 1971). However, most of these data report
the results of acute static bioassays and provide only a relative
index of acute toxicity. Mount and Stephan (1967) and Brungs
(1969) have described a method for estimating the maximum accept-
able toxicant concentration (MATC) based on chronic toxicity
studies. However, time consuming studies of this type have
been performed for only a relatively small number of chemicals,
primarily heavy metals and pesticides. Chronic studies have
been completed using fathead minnows (Pimephales promelas) and
copper (Mount, 1968; Mount and Stephan"i1969), zinc (Brungs,
1969) and cadmium (Pickering and Gast, 1972); with brook trout
(Salyelinus fontinalis) and copper (McKim and Benoit, 1971»
197*0; with rainbow trout (Salmo gairdneri) and lead (Davies
and Everhart, 1973); and with brown bullheads (Ictalurus nebulosus)
and copper (Christensen et al., 1972, Brungs et al., 1973T-
Similar studies have been performed with a number of pesticides
(Mount and Stephan, 1967; Eaton,1970; Hermanutz et al., 1973;
Macek et al., 1975, 1976, 1976).
These data suggest that the differences between acutely safe
and chronically safe concentrations can be much greater for
heavy metals than for pesticides. In addition, these data
suggest that eggs and/or fry of fish may be the most sensitive
life stages to chemical exposure. If this is a valid general-
ization, then it is possible that one could develop a realistic
estimate of the MATC of a chemical for a fish species by
defining the effects of that chemical on the eggs and fry.
The successful development of such an approach would obviously
result in a great saving of time and effort and/or reduce
the need for the use of arbitrary application factors in the
development of water quality criteria.
More recently, significant attention has been directed
towards the evaluation of water chemistry (i.e., hardness,
alkalinity and pH), and its influence on the toxicity of
particular metal species to aquatic forms. Complexing
properties of a series of chelating agents were evaluated
on a number of heavy metals in order to determine mobilization
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and accumulation dynamics in "biological tissue (Erickson
1972). Laboratory studies with copper and fathead minnows
brook trout and rainbow trout indicated that this metal is'
highly complexed by carbonate and hydroxide ions as in natur 1
waters and these phenomena determine the concentration of
copper in solution. Further investigations suggested that
alkalinity is the controlling factor in copper complexation
(Pagenkopf et al., 197^). The ability of water hardness
and pH to alter the toxicity of metals to fishes has also
been investigated (Cairns and Scheier, 1957; Pickering and
Henderson, 1965; Mount, 1966; Tabata, 1969).
The primary objective of this study was to empirically esti-
mate the "no effect" levels of four heavy metals, lead,
chromium, copper and cadmium, during continuous exposure of
eggs and fry of eight species of fish in two types of water
(^0 and 200 mg/1 hardness as CaCO^). The fish species
selected for this investigation were:
1. Rainbow trout (Salmo gairdneri)
2. Lake trout (Salvelinus namaycush)
3. Channel catfish (Ictalurus punctatus)
^. White sucker (Catostomus commersoni)
5. Bluegill (Lepomis macrochirus)
6. Northern pike (Esox lucius)
7. Walleye (Stizostedion vitreum)
8. Brook trout (Salvelinus fontinalis)
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SECTION II
CONCLUSIONS
1. Cadmium and copper were similar in toxicity to the
freshwater fish species tested and were significantly
more toxic than were lead or chromium.
2. The sensitivity of all seven species of freshwater fish
exposed to lead and chromium appears to be similar when
MATC's are considered relative to the actual duration
of exposure.
3. Fish eggs appeared to be the most resistant of the life
cycle stages tested in this study. Hatchability of eggs
was the parameter least affected by exposure to metals.
4. Water hardness did not appear to exert a significant
influence on the effect of the metals tested on fry
survival and growth over the long-term.
5. Scoliosis was an abnormality observed to have a signifi-
cant incidence in fish exposed to lead.
6. The effects of both chromium and cadmium appeared to be
cumulative during the entire exposure period.
7. Significant feeding problems experienced with northern
pike and walleye may indicate that these species are
unsuitable for this type of test.
8. Egg and fry tests appear to be effective and reliable
short-term means of assessing potential hazards associated
with environmental contaminants.
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SECTION III
RECOMMENDATIONS
1. Egg and fry tests should be utilized for preliminary
screening of all compounds whose potential hazard as
environmental contaminants are unknown. If results of
egg and fry tests are ambiguous then full-chronic assays
should be performed.
2. The "Proposed Recommended Bioassay Procedure for Egg and
Fry Stages of Freshwater Fish" should be revised on the
basis of information generated in this study.
3. The recommended test temperature of 1?° - 18°C for
northern pike is too high. Temperatures of 10° - 12°C
are more appropriate during egg incubation and hatching.
During swim-up, exposure temperature should be increased
gradually to 14° - 15°C.
^•. Forage fish should be reared in large numbers concurrently
with growing northern pike on a time scale to insure
that forage fish size is 1/2 to 2/3 that of northern
pike fry. A minimum of 5-10 appropriately-sized forage
fish per northern pike fry is necessary to prevent
cannibalism.
5. The recommended test temperature of 15°C for walleye is
adequate for egg incubation and hatching but should be
increased gradually to 18° - 21°C during swim-up to
stimulate feeding.
6. Walleye fry test chambers should be entirely blackened
on the sides, ends and bottom so that illumination is
entirely from above. This results in more even distribu-
tion of fry in test tanks and improved feeding activity.
?. To improve survival and growth, fewer fry should be held
in each growth chamber during chronic exposures. Reduction
of the recommendation of 50 fry initially, thinned to 25
after 30 days should be reduced to 10-20 fry in each
chamber during the entire 60 day exposure.
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SECTION IV
MATERIALS AND METHODS
methodology for these tests generally followed the
proposed Recommended Bioassay Procedure for Egg and Fry
Stages of Freshwater Fish, issued by the National Water
Quality Laboratory, Duluth, Minnesota (U.S. EPA, 1972 a).
EXPOSURE SYSTEMS
proportional diluters (Mount and Brungs, 196?) with a dilution
factor of 0.5 were utilized for the delivery of six toxicant
concentrations and a control to two replicate test aquaria.
Chambers to promote mixing of toxicant and diluent water were
utilized prior to the delivery of water to the test aquaria
which measured 35 x 30 x 30 cm with a water depth of 20 cm.
Each test chamber was divided to provide space for two growth
chambers which measured 25 x 30 x 12.5 cm and had stainless
steel screening (^-0 mesh) attached to one end to allow water
to drain to a depth of 2.5 cm upon removal from the aquaria.
This design allowed for the transfer of growth chambers to a
fluorescent light box for photographing the fish. Test
aquaria were maintained in circulating water baths at the
recommended temperature for the species being tested. Egg
incubation cups, made from k-oz., 2-in. OD round glass jars
'with the bottoms cut off and replaced with nylon screen (A-0
mesh), were oscillated in the test water by means of a rocker
arm apparatus driven by a 2 rpm motor (Mount, 1968). Eggs and
fry were shielded from all sources of light by means of black
polyethylene curtains.
Diluent water obtained from a ^00 ft. bedrock well had a
measured total hardness of 35 mg/1 (as CaC03). Well water
was delivered to test units through PVC pipes in the soft
water exposures. In the hard water exposures, well water
was passed through a water hardner as described by Lemke
(1969). The hardner utilized the principle of carbonate/
bicarbonate ion buffering system and supplied water with a
nominal hardness of 180 mg/1 (as CaCO^).
CHEMICAL METHODS
Water quality parameters such as acidity, alkalinity, hardness
and pH of diluent water were measured according to the methods
described in APHA et al. (1971). Dissolved oxygen concentration
of test water was measured with a YSI Oxygen Meter and oxygen/
temperature probe.
Stock solutions of the appropriate metal salts were dissolved
in distilled water and. delivered to diluters from a Mariotte
bottle via a volumetric delivery system. Analytical grade
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chemicals utilized to obtain respective metal stock solutions
were cadmium chloride (CdCl2), sodium dichromate (Na2Cr07),
copper sulfate (CuSO^.^HgO) and lead nitrate [Pb(N03J2]0
The analyses of metals in hard and soft water were performed
by atomic absorption spectroscopy using conventional acetylene-
air flame aspiration of the sample „ In the case of several of
the lower concentrations of lead, copper and cadmium in water
samples, residues were concentrated by solvent extraction
prior to flame atomization and analysis.
Distilled water was obtained from a Corning AG-1 all-glass still
and passed through a Barnstead mixed-bed ion exchange cartridge
before use. An analytical standard of each of the four metals
was prepared separately in deionized distilled water at a
concentration of 1,000 mg/1. These four standard solutions
were used throughout the study, however, dilutions of the
analytical standards were made at the time of analysis and
discarded after use. Metal standards were: lead nitrate,
#2322, Lot ^-5008, J.T. Baker; chromium trioxide, #A-99, Lot
731023, Fisher Scientific; copper metal, #1736, Lot ^3671,
J.T. Baker; and cadmium metal, #CX13-CBl-52, Lot 16, Matheson,
Coleman and Bell.
Organometallic compounds used to standardize instrument
response prior to analyzing water extracts were: lead
cyclohexanebutyrate, #10395. Eastman; copper cyclohexanebutyrate,
#10389, Eastman; and cyclohexanebutyric acid, cadmium salt,
#10386, Eastman. No standardization of this type was necessary
for chromium as it was always measured directly and never
required solvent extraction.
Lead, chromium and copper standards were prepared and diluted
in deionized distilled water according to U.S. EPA (1971). The
cadmium standard was prepared similarly according to Perkin-
Elmer (1971).
Organometallic compounds were dried for ^-8 hours in a desiccator
over fresh phosphorus pentoxide, then weighed and diluted with
methyl isobutyl ketone (MIBK) (#^53, Burdick & Jackson) to
concentrations of 100 mg of metal per liter of MIBK. Further
dilutions of all metal standard solutions used to calibrate
instrument response, were prepared on the day of analysis and
discarded after use.
All analyses of metals were performed with a Perkin-Elmer
Model 305-A atomic absorption spectrophotometer, using an
acetylene-air flame. The instrument contains a factory-
installed deuterium background corrector which was used in
these analyses. Instrument response to light absorption by
the metal of interest was measured with a Perkin-Elmer Model
56 strip chart recorder.
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Water samples (unfiltered) were collected in high-density
polyethylene "bottles which were cleansed and prepared for
use according to U.S. EPA (1972 b).
Concentrated nitric acid was added to each water sample (one
percent by volume) and the sample was stored at room tempera-
ture prior to analysis (2-3 weeks). As a check for contamina-
tion, a solution of one percent nitric acid in deionized
distilled water was analyzed for lead, chromium, copper, and
cadmium. No detectable concentrations of these metals were
present in the solution. No suppression or enhancement of
signal response was observed when known concentrations of
these metals were added to the solution and the signal response
obtained was compared to the response of analytical standards
containing from 0.1 to 5 percent nitric acid.
Concentrations of lead in water to 60 (j.g/1, copper to 25 |J.g/l
and cadmium to 25 |J-g/l were analyzed by direct sample atomiza-
tion. Metal concentrations less than those stated above were
measured by extracting the metal solution, according to U.S.
EPA (1971), and aspirating the MIBK extract. The percentage
metal recovery of the extraction procedure was accounted for
in the procedure since the absorbance of samples was compared
to the absorbance of inorganic metal standards which were
also extracted in the same manner.
The extraction procedure concentrates the metal by a factor
of ten in the MIBK solvent. In addition, the absorbance of
lead, copper and cadmium in MIBK solvent was ca four times
the absorbance of these metals in aqueous solution. Therefore,
the effective concentration achieved by extracting the metal
was 40X.
All concentrations of chromium were determined by direct
atomization of the sample.
AA operating conditions; acetylene-air flame with three-slot
burner atomizing 6 ml of water or k ml of MIBK per minute.
Lead: Analytical wavelength - 2170 Angstroms
Bandpass - 0.7 Angstroms
Recorder - 0.060 to 0.200 mg/1 range; 27 and 87 mm
peak heights, respectively.
Chromium: Analytical wavelength - 3579 Angstroms
Response - a. 0.015 to 0.500 mg/1 range; 6 and 212 mm
peak heights, respectively.
Response - b. 0.015 to 0.200 mg/1 range: 1^ and 1?6 mm
peak heights, respectively.
All other conditions were the same as for "lead" above.
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Copper: Analytical wavelength - 23^7 Angstroms
Response - 0.025 to 0.100 mg/1 range; 40 a
peak heights, respectively.
All other conditions were the same as for "lead" above.
Cadmium: Analytical wavelength - 2288 Angstroms
Response - 0.025 to 0.200 mg/1 range; 30 and 225 mm
heights, respectively.
All other conditions were the same as for "lead" above.
Standard deviation of the measurement: Measurement involves
placing a nebulizer tube into the sample or standard solution,
allowing 20-90 seconds for the recorder pen response to
stablize and then returning the nebulizer tube to deionized
distilled water to obtain a baseline reading. The pen response
to standard solutions was the same when aspiration occurred
continuously for two minutes or for five separate trials,
therefore the standard deviation is essentially the error
involved in measuring the peak heights with a millimeter
ruler (i.e., very small).
BIOLOGICAL METHODS
Egg exposures were initiated by randomly selecting two groups
of 100 eggs each for each replicate tank, i.e. 4-00 eggs per
concentration. Egg mortality was recorded daily and dead eggs
were removed to prevent fungus growth. When hatching commenced,
the number of eggs hatching in each group was recorded daily
until hatching was completed. Surviving fry were impartially
reduced to two groups of 50 per replicate tank, (200 fry per
concentration) and placed in the duplicate growth chambers.
Survival was recorded daily and after 30 days, fish were
transferred to a box with a translucent millimeter grid for
photographic determination of total (tip of snout to end of
caudal fin) length (McKim and Benoit, 1971). At this time, the
remaining fry were impartially reduced to 25 per growth chamber
(100 per concentration) and returned to the appropriate test
chambers. After 60 days post-hatch, photographs for growth
measurements were repeated, total group wet weights were deter-
mined and the experiment was terminated.
Specific information on source of test organisms, incubation
data and feeding for each species is presented in Table 1.
All fry were fed a minimum of four times daily. All aquaria
were siphoned daily to remove fecal material, excess food and
detritus and aquaria were brushed when algal growth became
excessive.
-------
STATISTICS
Measured biological parameters from duplicate containers during
continuous exposure were averaged and subjected to analysis of
variance according to Steele and Torrie (i960). When treatment
effects were indicated, the means of these effects were subjected
to Duncan's Multiple Range Test to determine which treatments
were statistically different from the controls. All differences
were considered significant at a probability of 0.05.
-------
TABLE 1 - SOURCE, AGE OF EGGS AT BEGINNING OF EXPOSURE, INCUBATION TIME AND TEMPERATURE,
AND FOOD UTILIZED IN THE EXPOSURE OF EGGS AND FRY OF SELECTED FRESHWATER FISH SPECIES
TO METALS
Species
Rainbow trout
(Salmo gairdneri)
Lake trout
(Salvelinus namaycush)
Channel catfish
(Ictalurus punctatus)
White sucker
(Catostomus commersoni)
Bluegill
(Lepomis macrochirus)
Northern pike
(Esox lucius)
Walleye
(Stizostedion vitreum)
Brook trout
(Salvelinus fontinalis)
Source
Wareham,
Mas So
Laconia,
N.H.
Lonoke,
Ark,
Laconia,
N.H.
Wareham,
Mass.
Lansing,
Iowa
Spirit Lake
Iowa
Wareham ,
Mass =
Age
0-1 day
0-1 day
2-3 days
0-1 day
<12 hours
5 days
(eyed)
2 days
0-1 day
Incubation
time
35-37 days
51-55 days
6-8 days
10-13 days
2 days
4 days
9-12 days
35 days
Temp.
(°C)
x ± S.D.
10 ± 1
10 ± 1
22 + 1
17 + 1
25 + 1
17 ± 1
15 + 1
10 + 1
Food
Oregon moist
trout starter
Oregon moist
trout starter
ground liver,
trout starter
daphnids, brin
shrimp nauplii
trout starter
mixed zooplank
daphnids, brin
shrimp nauplii
daphni ds , whi t
sucker fry
brine shrimp
nauplii, daphn
Oregon moist
trout starter
-------
SECTION V
RESULTS
WATER CHEMISTRY
Results of the chemical analysis of water samples indicated
that chemical characteristics were not significantly variable
among treatments within a test. Therefore, only means (and
standard deviations) and ranges for the various parameters
in each test are presented (Tables 2, 3» ^» 5)-
The results of the atomic absorption analysis of water samples
taken periodically during continuous exposure to materials
indicated that mean measured concentrations closely approxi-
mated nominal concentrations and were relatively constant
throughout exposure (Tables 6, 7, 8, 9)« The notable excep-
tions to this were the two instances where we attempted to
continuously expose rainbow trout and lake trout to 1000 p.g/1
dissolved lead. In both cases we were able to measure only
^80-672 p.g/1 with the remainder occurring as a precipitate at
the bottom of the aquaria.
EXPOSURE OF FISH TO LEAD
Rainbow trout
The percentage of rainbow trout eggs successfully hatching
when continuously exposed to 672 [ig/1 lead during incubation
was significantly lower than the percentage hatch observed
among control eggs and those exposed to lesser concentrations
of lead (Table 10). Survival of trout fry continuously
exposed to 672, ^43 and 250 |ig/l lead for 30 days was
significantly lower than fry survival in the other three
concentrations and the controls. Further evidence of the
effect of exposure to lead concentrations of 250 [ig/1 and
higher on trout fry was the significantly smaller total
lengths of survivors in these concentrations after 30 days
continuous exposure when compared with controls.
After 60 days exposure to 250 |ig/l lead only two juvenile
rainbow trout had survived and no fish survived 60 days
continuous exposure to W-3 and 672 p.g/1. Survival among
rainbow trout continuously exposed to 1*4-6 p.g/1 lead was
slightly but not significantly reduced after 30 days.
However, survival had decreased significantly during the 30-60
day exposure interval indicating a cumulative effect of lead
at this concentration.
Total lengths and wet weights of rainbow trout at the
termination of exposure (oO days post-hatch) were similar for
controls and all lead concentrations with surviving fish.
11
-------
TABLE 2 - MEANS
(N) OF MEASURED
FRESHWATER FISH
AND STANDARD DEVIATIONS (x ± S.D.), RANGES AND NUMBER OF OBSERVATIONS
WATER QUALITY PARAMETERS DURING EXPOSURE OF EGGS AND FRY OF SELECTED
SPECIES TO LEAD
Parameter
Species
Rainbow trout
x ± S.D.
Range
(N)
Lake trout
x ± S.D.
Range
(N)
Channel catfish
x ± S.D.
Range
(N)
White sucker
x ± S.D.
Range
(N)
Bluegill
x ± S.D.
Range
(N)
Northern pike
x ± S.D.
Range
(N)
Walleye
x ± S.D.
Range
(N)
Acidity
(mg/1)
3.5±1.1
2.9-4.8
(14)
3. 4±1.1
1.9-4.0
(16)
3 . 4±0 . 8
2.9-4.8
(6)
3.5±0.9
1.9-4.8
(8)
5-3±o.9
4.0-6.2
(6)
3.6±1.2
2.9-6.0
(8)
3-7±l.o
2.9-4.8
(4)
Alkalinity
(mg/1 as CaC03)
30.2±3.8
24.0-33.0
(14)
29.6±4.1
23.1-34.1
(16)
33-6±1.3
31.9-35-2
(6)
34.8±4.5
31.9-^2.0
(8)
33-3±3.8
28.0-36.0
(6)
33 . 1±3 • 8
30.0-42.0
(8)
33.8±4.0
31.9-^2.0
(4)
Hardness
(mg/1 as CaC03)
3^- . 6±3 . 2
32.0-^2.0
(1^)
32.6±3.9
25.0-38.0
(16)
36.o±l.o
2^.5-37.0
(6)
37-6±5.9
32.0-48.0
(8)
4o.7±7.3
35.0-53-0
(6)
35.4+4.8
31.0-45.0
(8)
37.6±5.2
32.0-45.0
(4)
Dissolved D£
(mg/1)
lo.3±o.9
8.6-11.3
(118)
9.7±1.4
8.6-11.3
(182)
8.5±0.7
6.5-9.2
(99)
8.8±0.6
7-3-9.8
(96)
6.9±2.o
5.0-8.7
(203)
8.7±o.6
7.6-9.3
(84)
10.3±1.1
9.2-10.4
(68)
pH
6.9-7.4
(14)
7.0-7.3
(16)
6.8-7.3
(6)
6.7-7.1
(8)
6.7-7.2
(8)
6.7-7-3
(8)
6.7-7.1
(b)
-------
TABLE 3 - MEANS
(N) OF MEASURED
FRESHWATER FISH
AND STANDARD DEVIATIONS (x ± S.D.), RANGES AND NUMBER OF OBSERVATIONS
WATER QUALITY PARAMETERS DURING EXPOSURE OF EGGS AND FRY OF SELECTED
SPECIES TO CHROMIUM
Parameter
Species
Rainbow trout
x ± S.D.
Range
(N)
Lake trout
x 1 S.D.
Range
(N)
Channel catfish
x ± S.D.
Range
(N)
White sucker
x ± S.D.
Range
(N)
Bluegill
x ± S.D.
Range
(N)
Northern pike
x ± S.D.
Range
(N)
Walleye
x ± S.D.
Range
(N)
Acidity
(mg/1)
3.3±1.2
1.0-4. 8
(16)
3.6±1.1
1.9-4.8
(14)
4.0+0.7
2.9-^.8
(6)
3-2±o.?
2.9-4.0
(8)
6.6±1.1
5.0-7.8
(6)
3.3±i.o
1.9-4.8
(8)
3.4±0.8
2.9-4.6
(4)
Alkalinity
(mg/1 as CaC03)
30.3+4.0
23.0-34.1
(16)
31.5±2.8
24.2-33.0
(14)
33.7±l-3
31-9-35.2
(6)
34.6±5.6
30.0-44.0
(8)
33.0±5.3
25.0-39.0
(6)
35.6±4.9
31.9-43.0
(8)
33.8±1.8
31.9-36.0
(4)
Hardness
(mg/1 as CaC03)
33-4±4.5
30.0-42.0
(16)
34 . 0±4 . 8
31.0-42.0
(14)
36.2±1.2
35.0-38.0
(6)
38.815.3
32.0-46.0
(8)
38.3±4.6
33.0-45.0
(6)
37.8±5.6
31.0-46.0
(8)
38.5±4.2
32.0-46.0
(4)
Dissolved Op
(mg/1)
9 . l±o . 6
6.7-12.2
(288)
9.5±0.9
6.7-12.2
(340)
8.1±1.5
5.6-9.4
(87)
8.9±0.6
7.5-10.4
(96)
6.611.3
3.3-9.7
(195)
9.010.6
7-5-10.4
(84)
9.510.6
8.5-10.9
(68)
PH
5.7-7.0
(14)
5.8-7.1
(14)
7.0-7.4
(6)
5.9-7.2
(8)
5.7-7-1
(6)
S. 7-7-0
1 (8)
6.8-7-2
(4)
-------
TABLE 4 - MEANS
(N) OF MEASURED
FRESHWATER FISH
AND STANDARD DEVIATIONS (x ± S.D.), RANGES AND NUMBER OF OBSERVATIONS
WATER QUALITY PARAMETERS DURING EXPOSURE OF EGGS AND FRY OF SELECTED
SPECIES TO COPPER IN "SOFT" AND "HARD" WATER
Parameter
Species and
type water
Brook trout
soft water
x ± S.D.
Range
(N)
hard water
x ± S.D.
Range
(N)
Channel catfish
soft water
x ± S.D.
Range
(N)
hard water
x ± S.D.
Range
(N)
Walleye
soft water
x ± S.D.
Range
(N)
hard water
x ± S.D.
Range
(N)
Acidity
(mg/1)
3-3±1.7
1.9-7.0
(8)
8.3±2.1
7.0-11.0
(8)
3.2±o.5
2.9-3-8
(6)
4.8+1.6
2.9-6.7
(6)
3.3±0.7
2.9-4.0
(4)
4.6H.7
2.9-6.8
(4)
Alkalinity
(mg/1 as CaCO-^)
27.8±3.8
22.0-31.9
(8)
177.6i30.4
150.7-204.0
(8)
34.1+1.8
31.9-36.3
(6)
172.9±34.6
130.9-210.0
(6)
34.011.9
31.0-39.0
(4)
176.0130.0
151.0-209.0
CO
Hardness
(mg/1 as CaC03)
37.5±7.3
32.0-51.0
(8)
187.0±22.0
167.0-208.0
(8)
36.o±l.l
35-0-37.0
(6)
186.3±38.7
136.0-229.0
(6)
35.011.8
30.0-39.0
CO
189.0+33.0
170.0-238.0
(4)
Dissolved D£
(mg/1)
10.010.7
87.0-11.1
(24)
11.011.2
9.5-13.2
(28)
7.3±2.4
6.3-10.6
(100)
8.7±0.9
6.2-10.6
(48)
9.9+0.7
7.8-11.2
(68)
pH
_
6.6-7.1
(8)
-
6.7-7.1
(8)
_
7.4-7.6
(6)
_
7-5-7.8
(6)
_
6.8-7-3
(4)
,
8.9±0.8
7.9-10.1 7.0-?. 3
(33) I (b)
-------
TABLE 5 - MEANS
(N) OF MEASURED
FRESHWATER FISH
AND STANDARD DEVIATIONS (x ± S.D.), RANGES AND NUMBER OF OBSERVATIONS
WATER QUALITY PARAMETERS DURING EXPOSURE OF EGGS AND FRY OF SELECTED
SPECIES TO CADMIUM IN "SOFT" AND "HARD" WATER _
Parameter
Species and
type water
Brook trout
soft water
x ± S.D.
Range
(N)
hard water
x ± S.D.
Range
(N)
Channel catfish
soft water
x ± S.D.
Range
(N)
hard water
x ± S.D.
Range
(N)
Walleye
soft water
x ± S.D.
Range
(N)
hard water
x ± S.D.
Range
(N)
Acidity
(mg/1)
3.5±1.8
1.9-7.0
(8)
8.1±?.0
6.7-9-5
(6)
3.o±o.7
1.9-3-8
(6)
4-.811.0
3.8-5-7
(6)
3.8±1.6
2.9-5-8
CO
4-. 8+1. 9
1.9-7.0
(4-)
Alkalinity
(mg/1 as CaC03)
30.011.4-
28.0-32.0
(8)
I77.o±32.o
14-8.0-205.0
(6)
34-. oil. 6
33.0-36.0
(6)
172.0133.4-
132.0-207.0
(6)
33.0±1.3
31.9-35.0
CO
172.0±33.1
154-. 0-210.0
(4-)
Hardness
(mg/1 as CaC03)
37-0±7.2
33-0-51.0
(8)
188.0127-0
164-. 0-213.0
(6)
37.0±1.3
35-0-38.0
(6)
185.0±35.0
14-2.0-223.0
(6)
35.o±l.2
32.0-39.0
(4-)
187.0±36.0
164-. 0-24-0.0
CO
Dissolved 02
(mg/1)
10.0±0.8
8.6-11.0
(37)
10.6±1.5
8.3-12.7
(33)
7-6±0.9
5.6-9.8
(91)
8.6±0.5
7.4-9-3
(4-8)
io.5±i.l
7.7-10.6
(68)
8.8±0.9
7.6-10.3
(34-)
PH
6.5-7.2
(10)
6.7-7.1
(6)
7.5-7.6
(6)
7-7-7.8
(6)
6.8-7.3
(4-)
6.9-7-3
(4-)
-------
TABLE 6 - MEANS AND STANDARD DEVIATIONS (x ± S.D.), RANGES AND NUMBER (N) OF MEASURED
CONCENTRATIONS OF LEAD DURING CONTINUOUS EXPOSURE OF EGGS AND FRY OF SELECTED FRESH-
WATER FISH SPECIES
Species and
nominal cone.
-------
TABLE 7 - MEANS AND STANDARD DEVIATIONS (x ± S.D.), RANGES AND NUMBER (N) OF MEASURED
CONCENTRATIONS OF CHROMIUM DURING CONTINUOUS EXPOSURE OF EGGS AND FRY OF SELECTED FRESH-
WATER FISH SPECIES
Species and
nominal cone.
(ng/D
Rainbow trout
Lake trouta
750
375
187
9^
^7
Northern pike
White suckera
2000
1000
500
250
125
Walleye
2000
1000
500
250
125
62
Measured cone, (p.g/1)
x ± S.D.
822±87
38^±42
19^±32
105±12
51±13
1975±125
963±125
538±133
290± 57
123+ 9
2l67±153
1125±189
558± 7^
288± i»4
133± 35
80± 24-
Range
700-9*4-0
330-^10
1^0-220
90-120
30- 60
1800-2100
800-1100
ij-10- 710
2^-0- 370
no- 130
2000-2300
1000-1*4-00
J4-90- 660
250- 350
no- 170
59- no
(N)
5
5
5
5
5
5
5
5
5
5
Li-
it
4
4
i*
*f
Species and
nominal cone.
(ns/D
Channel catfish
1250
620
310
155
77
39
Bluegill
1000
500
250
125
62
31
Measured cone. (p,g/l)
x ± S.D.
1290±1^
570±28
305±35
150±16
73± 3
39± 2
1122±97
522±38
265±13
1^0±14-
70± l
57±35
Range
1280-1300
550- 590
280- 330
137- 179
71- 76
36- ^2
980-1200
^70- 560
250- 280
120- 150
68- 70
20- 100
(N)
1*
1*
^
b
b
^
^
^
4
4
it-
4
a
Tests run concurrently from same if- liter dilution apparatus
-------
TABLE 8 - MEANS AND STANDARD DEVIATIONS (x ± S.D.), RANGES AND NUMBER (N) OF MEASURED
CONCENTRATIONS OF COPPER DURING CONTINUOUS EXPOSURE OF EGGS AND FRY OF SELECTED FRESH-
WATER FISH SPECIES IN "SOFT" AND "HARD" WATERS
Soft Water
Species and
nominal cone.
WD
Brook trout
100
50
25
12
6
3
Channel catfish
25.0
12.5
6.2
3.1
1.6
0.8
Walleye
100
50
25
12
• 6
3
Measured cone. (|ig/l)
x ± S.D.
95±7.4
51±5.1
2?±3.2
13±2.1
7±1.6
5±1.2
21±4.5
15+3-6
9±2.1
4±1.5
3±0.7
o.5±o.3
9l±7.l
47
21±2.1
13+4.5
8±2.6
3±1.1
Range
73-107
2k- 55
13- 28
7- 14
5- 8
3-5.5
16-27
10-18
6-11
2- 6
2- 3
0.2-0.7
86-96
-
19-22
10-16
6-10
2- 4
(N)
7
7
7
7
7
7
4
4
4
4
4
4
2
2
2
2
2
2
Hard Water
Species and
nominal cone.
(usA)
Brook trout
100
50
25
12
6
3
Channel catfish
100
50
25
12
6
3
Walleye
100
50
25
12
6
3
Measured cone. (p,g/l)
x ± S.D.
74±9.4
49±7.0
21±5.l
13±3-6
8±2.3
5±1.6
66±8.7
34±6.4
19±6.5
13±5.1
10±9.4
7±1.9
71±3.5
38±7.8
24±2.8
17
14±1.4
9±0.7
Range
60-81
43-58
15-27
8-18
4-10
3- 7
53-73
28-43
11-26
8-17
6-28
6-11
68-73
32-43
22-26
_
13-15
8.6-9.6
(N)
7
7
7
7
7
7
4
4
4
4
4
4
2
2
2
2
2
2
00
-------
TABLE 9 - MEANS AND STANDARD DEVIATIONS ( x ± S.D.), RANGES AND NUMBER (N) OF MEASURED
CONCENTRATIONS OF CADMIUM DURING CONTINUOUS EXPOSURE OF EGGS AND FRY OF SELECTED FRESH-
WATER FISH SPECIES IN "SOFT" AND "HARD" WATERS
Soft Water
Species and
nominal cone.
(^g/1)
Brook trout
50.0
25-0
12.5
6.2
3-1
1.5
Channel catfish
60.0
30.0
15.0
7.5
3.8
1.9
Walleye
50.0
25.0
12.5
6.2
3-1
1.5
Measured cone.
x ± S.D,
47±4 . 8
24±2 . 6
10±1.2
6 . ifcfcO . 1
3 . 2±0 . 5
2.0±0.1
54±7 . 0
32±5.1
20±5.9
17±6.7
11±4.7
6±2.3
55±2.6
24.7±3-2
8.6±2.1
3.7±1.2
l.8±0.7
0.9
Range
41-52
20-26
9-12
6o4-6.6
2.5-3.6
1.8-2.2
M1.-63
20-33
7-21
4-19
2-13
2.6-6.9
52-57
20-26
7.^-11
2.3-4.7
1.3-2.6
lUfi'l)
(N)
4
4
4
4
4
4
5
5
5
5
4
4
3
3
3
3
3
3
Hard Water
Species and
nominal cone.
(Jig/1)
Brook trout
100
50
25
12
6
3
Channel catfish
100
50
25
12
6
3
Walleye
100
50
25
12
6
3
Measured cone, (us/1)
x ± S.D.
91±12
50±11
21± 3
12± 3
7± 1
3±0.4
59±18
33± 8
17± 2
12± 3
5± 2
2±0.5
86.7±10.7
44. 3± 6.8
19. 0± 2.6
8.4±. 2.3
3.4± 2.4
1.3± 0.2
Ranges
73-101
37- 59
20- 25
10- 16
6- 9
2.6-3.6
52-96
30-50
16-21
7-14
2- 7
1.1-2.5
75-96
39-52
16-21
6.6-11
1.6-6.1
1.1-1.5
(N)
4
4
4
4
4
4
5
5
5
5
5
5
3
3
3
3
3
3
-------
TABLE 10 - MEAN PERCENTAGE HATCH OF EGGS, MEAN SURVIVAL, TOTAL LENGTHS AND WET WEIGHT
OF RAINBOW TROUT (Salmo gairdneri) FRY CONTINUOUSLY EXPOSED TO LEAD IN SOFT WATER
(34 ±3.2 mg/1 as
Mean
measured
lead cone.
(ne/D
6?2 A
B
443 A
B
250 A
B
146 A
B
71 A
B
49 A
B
Control A
B
Mean
hatch
w
28
29a
96
87
88
87
91
92
90
87
93
94
87
82
1-30 Days
Survival
w
18
26a
2
I4a
24
12a
86
82
98
98
90
90
96
96
Mean total
length (mm)
20 (1.0)
20 (0.9)a
21 (1.1)
20 (1.5)a
23 (3-0)
24 (3.Da
28 (2.0)
28 (1.0)
29 (2.2)
27 (1.6)
28 (1.6)
28 (1.5)
29 (1.7)
29 (1.7)
31-60 Days
Survival
(*)
0
"0
0
0
4
0
56
52a
84
96
96
100
96
92
Mean total
length (mm)
_
-
38
39 (3-D
38 (3.0)
40 (4.1)
37 (3.2)
38 (3-9)
37 (4.9)
40 (4.8)
38 (3-8)
Mean total
wet wt. (nig)
_
_
700
710
690
820
650
680
660
750
670
N3
O
Denotes values significantly lower than the controls (Duncan's Multiple Range P=0.05)
-------
Approximately 2Q% and 1.2% of rainbow trout fry exposed to
672 and 443 M-g/1 lead, respectively, exhibited scoliosis
(a lateral curvature of the spine) during the initial 30 day
post-hatch exposure.
Based on these data, the MATC for lead and rainbow trout
eggs and fry is estimated to be "between 71 and 146 p.g/1.
Lake trout
No significant effect on the percentage of lake trout eggs
successfully hatching was observed during continuous exposure
to lead concentrations as high as 483 p.g/1 (Table 11). The
reduced hatch observed among eggs exposed to 83 M-g/1 and among
the A replicate of control eggs was apparently due to fungus.
Survival of lake trout fry exposed to 483 and 404 n.g/1 lead
for 30 days was significantly lower than survival among control
fry and those exposed to lesser concentrations of lead. In
addition, growth among fry exposed to 438 and 404 (ig/1 lead
was also significantly reduced when compared to controls.
No lake trout fry survived 60 days exposure to 483 and 4o4 (ig/1
lead and survival was significantly reduced among fry exposed
for 60 days to 198, 120 and 83 fig/1 lead. Survival in the
latter three concentrations had not been affected during the
initial 30 days exposure indicating a cumulative toxic effect
of lead at these concentrations. No statistically significant
reduction in total length or wet weight was observed among
surviving fish although fry exposed to 198 (ig/1 lead were
smaller than all other fry.
The scoliosis condition observed in rainbow trout exposed to
672 and 44-3 (J-g/1 lead was not observed among lake trout
exposed to a similar (483 M-g/1) concentration.
Based on these data,the MATC for lead and lake trout eggs and
fry is estimated to be between 48 and 83 (J.g/1.
Channel catfish
Percentage hatch among catfish eggs in duplicajes continuously
exposed to 460 [ig/l were 93 and QQfo indicating no significant
effects of exposure on the hatching process (Table 12).
Hatching success was variable among other treatments and
poor among controls presumably due to an obvious fungus
infection. None of the catfish fry continuously exposed
to 460 and 280 [ig/1 lead survived 30 days continuous exposure.
In addition, 30 day exposure to 136 p.g/1 lead significnatly
21
-------
TABLE 11 - MEAN PERCENTAGE HATCH OF EGGS, MEAN SURVIVAL, TOTAL LENGTHS AND WET WEIGHT
OF LAKE TROUT (Salvelinus namaycush) FRY CONTINUOUSLY EXPOSED TO LEAD IN SOFT WATER
(32.6 ± 3-9 ~~
Mean
measured
lead cone.
(ne/D
483 A
B
404 A
B
198 A
B
120 A
B
83 A
B
48 A
B
Control A
B
Mean
hatch
(*)
78
83
80
82
92
84
66
66
34
22
71
81
38
82
1-30 Days
Sur vi val
(*)
22
36a
16
I6a
68
86
88
76
90
86
86
88
64
68
Mean total
length (mm)
22 (1.1)
20 (l.l)a
21 (1.6)
21 (1.3)a
24 (1.8)
25 (1.3)
25 (1.8)
25 (1.8)
26 (1.5)
24 (1.6)
26 (2.2)
26 (2.5)
25 (1.5)
27 (1.4)
31-60 Days
Survival
(*)
0
oa
Oa
Oa
58
54a
56
58a
72
80a
96
96
96
92
Mean total
length (mm)
-
—
27 (2.8)
26 (1.7)
28 (2.4)
27 (2.2)
29 (2.0)
28 (2.6)
31 (2.5)
30 (2.8)
30 (2.9)
30 (2.8)
Mean total
wet wt. (mg)
-
-
120
130
180
130
170
150
200
190
190
170
a
Denotes values significantly lower than the controls (Duncan's Multiple Range P=0.05)
-------
TABLE 12 - MEAN PERCENTAGE HATCH OF EGGS, MEAN SURVIVAL, TOTAL LENGTHS AND WET WEIGHT
OF CHANNEL CATFISH (Ictalurus punctatus) FRY CONTINUOUSLY EXPOSED TO LEAD IN SOFT WATER
(36.0 ± 1.0 mg/1 as CaC03)
Mean
measured
lead cone.
. WD
1+60 A
B
280 A
B
136 A
B
75 A
B
33 A
B
17 A
B
Control A
B
Mean
hatch
<*)
93
88
97
59
97
66
96
68
5^
6^
^8
71
21
20
1-30 Days
Survival
(*)
0
oa
0
oa
12 V
ya, D
53
50
f3
62
39
^8
70
60
Mean total
length (mm)
-
-
18 (l.*0
18 (l.*0
20 (2.2)
19 (3-D
20 (2.3)
19 (2.8)
21 (2.7)
21 (2.5)
19 (2.3)
19 (2.2)
31-60 Days
Survival
<*)
-
-
79
90
92
84-
76
9I
96
8*J.
76
Mean total
length (mm)
_
-
2^ (3.3)a
30 (2.9)
27 (^.3)
29 (3-5)
28 (i+.b)
31 (5-3)
32 (*K3)
29 (^.5)
29 (*K7)
Mean total
wet wt. (mg)
^
—
150a
250
210
250
230
230
230
2^-0
2*4-0
a
'Denotes values significantly lower than the controls (Duncan's Multiple Range P=0.05)
All survivors pooled into one experimental unit after 30 days exposure to maintain
comparability of numbers of fry/chamber
-------
reduced survival of catfish fry. Continuous exposure to lead
for 30 days had no significant effect on total length of
surviving catfish.
During days 31-60 of exposure to lead, survival of remaining
catfish was similar in all treatments. However, continuous
exposure to 136 (ig/1 of lead for 60 days significantly
reduced total length and wet weight of catfish when compared
to controls and fish in lower treatments.
Based on these data, the MATC for channel catfish and lead is
estimated to be between 75 and 136 |ig/l.
Bluegill
Continuous exposure to lead concentrations as high as W-7 p.g/1
had no significant effect on the percentage of bluegill eggs
successfully hatching (Table 13). Survival of bluegill fry
during the first 30 days exposure to lead was generally poor
but obvious differences among treatments were observed. None
of the bluegill fry exposed to 447 and 277 (ig/1 of lead
survived for the duration of this period and survival was
significantly lower among fry exposed to 120 [ig/1 than among
controls and bluegill exposed to lower concentrations of lead.
Prior to death, bluegill exposed to 4^7 and 277 M-g/l exhibited
some incidence of scoliosis. In addition, mean total length
of surviving bluegill exposed to 120 |ig/l of lead was signifi-
cantly less than that observed for all other groups where
bluegill survived.
During the period of 31-60 days exposure, survival of bluegill
fry exposed to 120 |ig/l was again significantly reduced when
compared to other treatments and controls. Also, after 60 days
exposure to this concentration, total length and weight of
bluegill fry was significantly lower than all other treatments
and controls.
Based on these data, the MATC of lead for bluegill eggs and fry
is estimated to be between 70 and 120 |ig/l.
White sucker
The percentage of white sucker eggs successfully hatching
was similar for eggs incubated in controls and in lead
concentrations as high as ^83 (ig/1 (Table 14-). Only one
sucker fry survived 30 days exposure to ^-83 |ig/l lead and
survival in one of the replicates of 253 M-g/1 was lower than
generally observed in other treatments. More than half
24
-------
TABLE 13 - MEAN PERCENTAGE HATCH OF EGGS, MEAN SURVIVAL, TOTAL LENGTHS AND WET WEIGHT
OF BLUEGILL (Lepomis macrochirus) FRY CONTINUOUSLY EXPOSED TO LEAD IN SOFT WATER
(40.7 ± 7.3 mg/1 as
Mean
measured
lead cone.
(ng/D
447 A
B
277 A
B
120 A
B
70 A
B
33 A
B
12 A
B
Control A
B
Mean
hatch
(*)
97
924.
85
94
95
91
78
91
71
87
65
95
90
65
1-30 Days
Survival
<*)
0
oa
0
Oa
6
6a
12
16
24
24
14
16
16
24
Mean total
length (mm)
_
-
19 (3.5)
17a(3.D
23 (4.2)
23 (5.D
21 (2.1)
21 (4.2)
23 (3-4)
25 (3-D
24 (2.5)
21 (2.8)
31-60 Days
Survival
(*)
_
-
5°a
75a
100
70
100
88
100
100
100
100
Mean total
length (mm)
_
-
23a (3.6)
23 (3.6)
29 (1.4)
27 (4.3)
25 (3.6)
28 (2.3)
31 (5.6)
27 (4.5)
30 (1.7)
25 (4.4)
Mean total
wet wt. (mg)
-
-
250
250a
510
470
360
420
460
380
410
360
IsJ
01
lDenotes values significantly lower than the controls (Duncan's Multiple Range P=0.05)
-------
TABLE 14 - MEAN PERCENTAGE HATCH OF EGGS, MEAN SURVIVAL, TOTAL LENGTHS AND WET WEIGHT
OF WHITE SUCKER (Catostomus commersoni) FRY CONTINUOUSLY EXPOSED TO LEAD IN SOFT WATER
(37.6 ± 5-9 mg/1 as CaC03)
Mean
measured
lead cone.
(ns/i)
483 A
B
253 A
B
119 A
B
6? A
B
33 A
B
Control A
B
Mean
hatch
(*)
65
70
70
68
66
72
80
78
84
64
63
65
1-30 Days
Survival
w
2
Oa
6a
20
32
30
40
24
22
24
46
30
Mean total
length (mm)
15fl
_a
15 (1°4)
17 U.9)a
18 (1.2)
19 (1.7)
19 (1.4)
20 (1.6)
20 (2.1)
20 (1.2)
22 (1.5)
21 (2.8)
31-60 Days
Survival
w
0
0
2
20
32
24
36
24
20
20
32
22
Mean total
length (mm)
-
18
22 (4.4)a
28 (2.7)
30 (3-8)
29 (2.7)
32 (4.4)
33 (2.9)
32 (2.6)
30 (3.5)
31 (3-3)
Mean total
wet wt. (mg)
-
60
80a
140
230
160
220
250
280
160
220
Denotes values significantly lower than the controls (Duncan's Multiple Range P=0.05)
-------
of the sac fry of white sucker exposed to ^83 and 253 P-g/1
lead exhibited severe scoliosis shortly after hatching and
expired "before reaching the swim-up stage. Survival was
generally low during the initial 30 days due to difficulties
encountered in feeding sucker fry with both live food and
trout starter . Total length of surviving sucker fry after
30 days exposure to ^83 and 253 P-g/1 lead was significantly
less than that observed for control fry. Survival of fry after
60 days, except for one of the duplicates at 253 M-g/1 previously
mentioned, was not significantly different, statistically,
between controls and lead exposed fry.
Total lengths and wet weights of sucker fry exposed to 253
lead for 60 days were significantly less than lengths and
weights of control fry and those exposed to lesser concentrations
of lead.
Based on these data, the MATC for white sucker and lead is
estimated to be between 119 and 253 M.g/1-
Northern pike
The percentage of successful hatching was similar for northern
pike eggs incubated in controls and in lead concentrations as
high as ^4-83 p.g/1 (Table 15). After 20 days exposure, survival
was significantly reduced among pike fry exposed to 483 p.g/1
lead when compared with controls and lesser concentrations of
lead. Approximately 20$ of fry exposed to ^83 M-g/1 lead
exhibited severe scoliosis after one week of exposure.
All data recorded after 20 days exposure are highly variable
due to difficulty in controlling cannibalism among pike fry.
Northern pike hatched and fed in water of 18° C were found to
accept invertebrates as food for only one week after which
we observed cannibalism despite the presence of large quantities
of Daphnia in the growth chamber. During the initial 20 days
after hatching, cannibalism was somewhat controlled by the
constant maintenance of a supply of white sucker or fathead
minnow fry in the growth chamber with young pike. After 20
days, the food size preference of young pike apparently differed
from the supply of forage fish which was available and canni-
balism masked any toxicant induced reduction in survival or
growth.
Based on the reduced survival and incidence of scoliosis in
hatchery pike exposed for 20 days to ^-83 Hg/1 lead, the
MATC for this species is estimated to be between 253 and
27
-------
TABLE 15 - MEAN PERCENTAGE HATCH OF EGGS, MEAN SURVIVAL, TOTAL LENGTHS AND WET WEIGHT
OF NORTHERN PIKE (Esox lucius) FRY CONTINUOUSLY EXPOSED TO LEAD IN SOFT WATER
±4.8 mg/1 as CaCOp
Mean
'measured
lead cone.
(ne/D
483 A
B
253 A
B
119 A
B
6? A
B
33 A
B
Control A
B
Mean
hatch
(*)
72
76
73
88
72
73
80
83
70
83
80
79
1-20 Days
Survival
(*)
18
24a
40
46
48
40
60
4o
56
48
3^
38
Mean total
length (mm)
19 (2.1)
30 (4.0)
33 (6.5)
38 (5.0)
36 (5.D
39 (6.1)
33 (7.7)
42 (6.5)
35 (6.9)
37 (6.0)
31 (8.1)
37 (6.6)
21-50 Days
Survival
(*)
0
8
6
6
10
16
18
14
8
14
16
10
Mean total
length (mm)
_
57 (12.2)
77 (10.3)
81 (14.0)
67 (11.3)
61 ( 6.5)
65 (10.6)
64 (10.1)
74 ( 9.2)
62 (15.0)
65 ( 9.7)
74 (10.2)
Mean total
wet wt. (mg)
_
1310
2960
3310
1840
1200
1610
I46o
2790
1700
1760
2680
Ni
00
Denotes values significantly lower than the controls (Duncan's Multiple Range P=0.05)
-------
Walleye
Exposure to lead concentrations as high as 397 M-g/1 had no
significant effect on the percentage of walleye eggs which
hatched successfully (Table 16). Percentage survival of
walleye fry after 30 days exposure to 397 P-g/1 lead appeared
reduced when compared to controls and lower lead concentra-
tions, however, variability "between replicates and the generally
poor success in feeding walleye fry precluded ascribing statis-
tical significance to this observation. Many of the walleye
fry exposed to 397 (J.g/1 lead exhibited the scoliosis which
was symptomatic of lead toxicity for other species discussed
previously.
TABLE 16 - MEAN PERCENTAGE HATCH OF EGGS, MEAN SURVIVAL AND
TOTAL LENGTHS OF WALLEYE (Stizostedion vitreum) FRY CONTINUOUSLY
EXPOSED TO LEAD IN SOFT WATER (37.6 ± 5.2 mg/1 as
Mean measured
lead cone.
WD
397 A
3
237 A
B
108 A
B
1+9 A
B
29 A
B
22 A
B
Control A
B
Mean hatch
(*)
70
69
64
65
68
61
67
66
58
73
66
59
48
62
1-30 Days
Survival
(*)
0
12
16
16
26
26
18
32
24
14
24
46
38
28
Mean total
length (mm)
10 (2.4)
11 (2.2)
11 (2.6)
8 (1.8)
11 (3.6)
11 (2.9)
12 (1.8)
11 (3.1)
10 (2.0)
12 (3.0)
10 (2.5)
13 (1.7)
13 (1.3)
Based on these data,the MATC of lead for walleye is estimated
to be between 237 and 397 P-g/1.
29
-------
EXPOSURE OF FISH TO CHROMIUM
Rainbow trout
Two separate exposures of rainbow trout eggs and fry to
chromium were completed. During the first exposure, the mean
measured chromium concentrations ranged from 1.6 to 49-7 mg/1
(Table 1?). None of the trout eggs exposed to 26.7 and 49.7
mg/1 hatched successfully. The percentage of eggs success-
fully hatching among groups exposed to 12.2 and 6.1 mg/1 was
significantly lower than controls and eggs exposed to lower
concentrations of chromium. None of the fry survived 30
days exposure to 12.2 mg/1 chromium, and 30 days exposure to
6.1 and 3-2 mg/1 significantly reduced survival of trout fry
when compared to controls. Mean total lengths of trout fry
were significantly less than controls after 30 days exposure to
to all chromium concentrations including 1.6 mg/1 (the lowest
concentration tested). During the period of 31-60 days we
observed that the exposure to 1.6 mg/1 chromium significantly
reduced survival, total length, and total wet weight of
rainbow trout fry when compared to controls.
In view of the fact that we could not estimate limits on the
MATC based on these data, a second exposure of rainbow trout
eggs and fry to chromium was conducted with mean measured
chromium concentrations ranging from 51 to 822 p.g/1 • These
concentrations provided a continuum of the 0.5 dilution
factor from the first test. As would be expected from the
first test, exposure to concentrations of chromium as high as
822 p.g/1 had no significant effect on the percentage of rain-
bow trout eggs successfully hatching or on survival of fry
during the first 30 days post-hatch exposure (Table 18).
However, after 30 days exposure, the total length of trout fry
exposed to 822 p.g/1 was significantly reduced compared with
controls and lesser chromium concentrations.
Continuous exposure to 822 p.g/1 chromium for 60 days signifi-
cantly reduced survival of trout fry when compared to controls
and all other treatments. In addition, total length of fry
was significantly less than controls after 60 days exposure
to 822 and 384 p.g/1 chromium. Finally, total weight of fry
was significantly lower than controls after 60 days exposure
to chromium concentrations _ 105 (-ig/1-
Based on these data, the MATC for rainbow trout and chromium
is estimated to be between 51 and 105 M-g/1-
30
-------
TABLE 1? - MEAN PERCENTAGE HATCH OF EGGS, MEAN SURVIVAL, TOTAL LENGTHS AND WET WEIGHTS
OF RAINBOW TROUT (Salmo gairdrieri) FRY CONTINUOUSLY EXPOSED TO CHROMIUM IN SOFT WATER
(3^.0 ± 3.2 mg/1 as
Mean
measured
chromium cone.
(mg/1)
ij-9.7 A '
B
26.7 A
B
12.2 A
B
6.1 A
B
3.2 A
B
1.6 A
B
Control A
B
Mean
hatch
w
0
oa
0
oa
0
3ia
56
62a
82
85
77
87
76
77
1-30 Days
Survival
(*)
_
-
oa
7
Oa
17
i^a
84
85
96
99
Mean total
length (mm)
-
_
—
20 (l.*0
_a
20 (1.0)
19 (1.3)a
21 (lA)
21 (1.5)a
26 (1.7)
26 (1.6)
31-60 Days
Survival
(*)
-
_
-
0
0
0
0
l^a
2
96
88
Mean total
length (mm)
_
_
-
-
-
21 (1.2)a
28 -
3^ (3-8)
33 (3-3)
Mean total
wet wt . (ing)
_
-
—
-
-
250a
250
^80
ij-60
Denotes values significantly lower than the controls (Duncan's Multiple Range P=0.05)
-------
TABLE 18 - MEAN PERCENTAGE HATCH OF EGGS, MEAN SURVIVAL, TOTAL LENGTHS AND WET WEIGHT
OF RAINBOW TROUT (Salmo gairdneri) FRY CONTINUOUSLY EXPOSED TO CHROMIUM IN SOFT WATER
^ ± 4--5 mg/1 as
Mean
measured
chromium conct
(ns/i)
822 A
B
384 A
B
194 A
B
105 A
B
51 A
B
Control A
B
Mean
hatch
(*)
72
70
68
79
66
68
70
68
66
72
69
76
1-30 Days
Survival
(*)
94
95
96
99
100
97
100
99
99
98
98
91
Mean total
length (mm)
23 (1-5)
23 (2.0)a
26 (1.4)
26 (1.3)
27 (1.4)
28 (1.4)
27 (1.5)
27 (1.6)
28 (1.8)
26 (1.7)
28 (2.3)
26 (2.1)
31-60 Days
Survival
(*)
20
22a
78
86
96
96
88
80
88
86
88
92
Mean total
length (mm)
25 (3.6)
25 (3.8)a
32 (3-3)
33 (2.9)a
35 (4.1)
37 (3.5)
37 (3.8)
35 (3.3)
38 (2.8)
37 (3-3)
36 (4.6)
38 (4.4)
Mean total
wet wt. (mg)
170
170a
270
300a
350
380a
400
370a
450
440
450
550
OJ
ho
''Denotes values significantly lower than the controls (Duncan's Multiple Range P=0.05)
-------
Lake trout
As with the rainbow trout, two separate exposures of lake
trout eggs and fry to chromium were conducted. During the
first exposure, mean measured concentrations of chromium
ranged from 1.4 to 50.7 mg/1 (Table 19)° No lake trout
were observed to hatch successfully during exposure to
50.7 mg/1 and the percentage of lake trout eggs successfully
hatching was severely reduced by exposure to 24.4 mg/1
chromium. None of the lake trout survived 30 days exposure
to 24.4 mg/1 chromium, and survival of lake trout was
significantly reduced as a result of 30 days exposure to
11.6 and 6.0 mg/1 chromium when compared to controls. After
30 days exposure, mean lengths of lake trout were significantly
lower in all chromium concentrations, including 1.4 mg/1,
when compared with controls.
After 60 days exposure to chromium, survival of lake trout
fry appeared to be reduced in all chromium concentrations
when compared with controls. Analysis of variance and
Duncan's Multiple Range Tests indicated that survival was
significantly reduced among lake trout fry exposed to 11.6
and 6.0 mg/1 chromium but that variability between replicates
precluded ascribing statistical significance to the apparent
reduced survival among fry exposed to 1.4 and 2.9 rag/1
chromium. Total lengths and total wet weights of lake trout
after -60 days exposure were significantly lower in all
chromium concentrations when compared with controls.
The second exposure of lake trout eggs and fry to chromium
was conducted concurrently with the second rainbow trout
exposure using mean measured concentrations ranging from
51 to 822 (J.g/1. Percentage hatch of lake trout eggs was
sub-standard in all chromium concentrations and controls,
but generally indicated no adverse effects occurred during
exposure to concentrations as high as 822 (ig/1 (Table 20).
After 30 days exposure, percentage survival and total lengths
of lake trout fry were similar for all test concentrations
of chromium and controls.
After 60 days exposure,survival and total lengths of lake
trout were again similar for controls and chromium concentra-
tions tested. At termination the mean total weights of lake
trout indicated significant differences due to treatment and
a Duncan's Multiple Range Test indicated that total weights
of lake trout exposed to 822, 384- and 194 p.g/1 chromium were
significantly lower than those from controls and other
treatments.
33
-------
TABLE 19 - MEAN PERCENTAGE HATCH OF EGGS, MEAN SURVIVAL, TOTAL LENGTHS AND WET WEIGHT
OF LAKE TROUT (Salvelinus namaycush) FRY CONTINUOUSLY EXPOSED TO CHROMIUM IN SOFT WATER
(33-0 ± 3.9 mg/1 as
Mean
measured
chromium cone.
(mg/1)
50.7 A
B
24.4 A
B
11.6 A
B
6.0 A
B
2.9 A
B
1.4 A
B
Control A
B
Mean
hatch
(*)
0
oa
2
7a
46
39
60
53
64
51
4?
39
40
38
1-30 Days
Survival
(%}
_
0
oa
28
38a
35
49a
39
60
38
60
72
63
Mean total
length (mm)
„
-
21 (0.8)
21 (l.O)a
21 (1.0)
22 (0.9)a
23 (1.4)
22 (1.3)a
22 (1.2)
22 (1.3)a
26 (2.0)
27 (1.9)
31-60 Days
Survival
(*)
-
-
3
13a
0
56a
43
52
54
70
94
84
Mean total
length (mm)
-
—
22
22 (1.6)a
22 (l.l)a
22 (1.6)
22 (1.8)a
22 (1.5)
24 (2.2)a
32 (2.6)
31 (2.5)
Mean total
wet wt. (mg)
-
—
90
90a
6oa
90
iooa
90
90a
200
220
U)
.Denotes values significantly lower than the controls (Duncan's Multiple Range P=0.05)
-------
TABLE 20 - MEAN PERCENTAGE HATCH OF EGGS, MEAN SURVIVAL, TOTAL LENGTHS AND WET WEIGHT
OF LAKE TROUT (Salvelinus namaycush) FRY CONTINUOUSLY EXPOSED TO CHROMIUM IN SOFT WATER
(34.0 ± 4.8 mg/1 as CaCC>3)
Mean
measured
chromium cone.
(p-g/D
822 A
B
384 A
B
194 A
B
105 A
B
51 A
B
Control A
B
Mean
hatch
(*)
23
37
37
37
38
34
35
30
36
34
20-K
0*
30 Days
Survival
W
71
97
95
92
97
85
90
90
83
87
76
Mean total
length (mm)
26 (0.9)
26 (1.8)
26 (1.6)
26 (2.1)
26 (2.5)
27 (2.4)
27 (0.9)
27 (2.0)
27 (1.6)
27 (2.2)
26 (1.1)
60 Days
Survival
(*)
63
84
90
92
96
88
92
88
92
80
88
Mean total
length (mm)
28 (1.8)
27 (6.6)
30 (2.5)
30 (4.3)
30 (1.8)
30 (2.6)
31 (1.6)
30 (2.3)
33 (2.0)
29 (2.3)
31 (1.9)
Mean total
wet wt. (mg)
124
126a
151
135a
155
I59a
173
177
178
175
173
u>
a
Denotes values significantly lower than the controls (Duncan's Multiple Range P=0.05)
A malfunction of the thermoregulatory apparatus was responsible for the mortality
experienced in this duplicate
-------
Based on these data, the MATC for lake trout eggs and fry and
chromium is estimated to "be between 105 and 19^ |ig/l-
Channel catfish
Exposure to chromium concentrations as high as 1290 p.g/1 did
not significantly reduce the percentage of channel catfish
eggs which hatched successfully (Table 21). The reduced
hatch observed in controls and among eggs exposed to 39 M-g/1
chromium was a result of a fungus infection. After 30 days
post-hatch exposure, the percentage survival and total
lengths were significantly lower among channel catfish fry
exposed to concentrations of chromium ^ 305 |ig/l when
compared with controls and lesser chromium concentrations.
During the 31-60 days post-hatch exposure to 305 M-g/1»
survival of remaining catfish fry was similar to survival in
controls and lesser chromium concentrations, presumably
due to the "vigor" of fish which survived the 0-30 day
exposure. None of the channel catfish fry survived 60
days exposure to 1290 |ig/l chromium and the survival, total
length and wet weight of channel catfish exposed for 60 days
to 570 |ig/l chromium was significantly lower when compared
with controls.
Based on these data, the MATC of chromium for egg and fry
stages of channel catfish is estimated to be between 150 and
305 Ug/1.
Bluegill
The percentage of bluegill which hatched successfully was
similar in controls and among eggs exposed to chromium
concentrations as high as 1122 p.g/1 (Table 22). Percentage
survival of bluegill fry 30 days post-hatch was highly
variable and reflected the difficulty encountered in starting
bluegill fry on food. Total lengths of bluegill fry appeared
reduced after 30 days exposure to 1122, 522 and 265 M-g/1
chromium, however variability between duplicates precluded
ascribing statistical significance to these observations.
During the period of 31-60 days exposure, survival of
remaining fry was excellent and indicated no significant
effect due to exposure to chromium concentrations as high
as 1122 (is/1. Total lengths of bluegill after 60 days
exposure again appeared to be reduced as a result of exposure
to the higher chromium concentrations but differences were
not statistically significant due to variability between
duplicates.
36
-------
TABLE 21 - MEAN PERCENTAGE HATCH OF EGGS, MEAN SURVIVAL, TOTAL LENGTHS AND WET WEIGHT
OF CHANNEL CATFISH (Ictalurus punctatus) FRY CONTINUOUSLY EXPOSED TO CHROMIUM IN SOFT
WATER (36.2 ±1.2 mg/1 as
Mean
measured
chromium cone .
(ne/D
1290 A
B
570 A
B
305 A
B
150 A
B
73 A
B
39 A
B
Control A
B
Mean
hatch
(#)
75
95
80
93
83
84
78
74
71
59
33
56
31
37
1-30 Days
Survival
(*)
0
5a
4
12a
24
13a
23
51
74
60
68
70
44
62
Mean total
length (mm)
_
14 (0.4)a
16 (1.7)
16 (1.7)a
18 (2.4)
18 (2.4)a
22 (2.2)
19 (2.1)
23 (2.5)
24 (3.1)
24 (3.4)
21 (3-0)
22 (3.1)
20 (2.8)
31-60 Days
Survival
(#)
0
0
7
10a
72
88
77
87
98
92
96
100
85
86
Mean total
length (mm)
_
-
21 (3.0)
21 (3.0)a
31 (5.0)
24 (4.2)
34 (5.D
28 (4.6)
35 (2.8)
36 (3-5)
38 (5.4)
34 (3.7)
33 (6.8)
31 (4.4)
Mean total
wet wt. (mg)
_
-
120
120a
290
170
320
220
340
350
380
280
360
300
^Denotes values significantly lower than the controls (Duncan's Multiple Range P=0.05)
-------
TABLE 22 - MEAN PERCENTAGE HATCH OF EGGS, MEAN SURVIVAL,- TOTAL LENGTHS AND WET WEIGHT
OF BLUEGILL (Lepomis macrochirus) FRY CONTINUOUSLY EXPOSED TO CHROMIUM IN SOFT WATER
(38.3 ± 4.6 mg/1 as CaC03)
Mean
measured
chromium cone.
(ne/D
1122 A
B
522 A
B
265 A
B
140 A
B
70 A
B
57 A
B
Control A
B
Mean
hatch
(*)
85
62
85
70
88
83
81
83
84
88
89
86
90
85
1-30 Days
Survival
(*)
38
28
32
16
62
42
18
24
50
10
12
18
18
30
Mean total
length (mm)
14 (1.7)
14 (1.9)
16 (3.7)
20 (3.1)
15 (2.2)
18 (3.6)
23 (1.9)
20 (3.6)
17 (3.4)
24 (2.3)
22 (2.5)
22 (2.5)
25 (4.0)
22 (3.0)
31-60 Days
Survival
(*)
94
94
80
100
90
55
100
90
84
100
100
92
83
100
Mean total
length (mm)
20 (2.0)
18 (2.1)
22 (4.6)
24 (3.9)
20 (4.0)
24 (2.9)
27 (3-5)
26 (1.5)
20 (5.0)
31 (4,4)
23 (6.8)
26 (3.8)
27 (4.4)
24 (4,0)
Mean total
wet wt. (mg)
140
120a
230
250
180
220
290
290
180
320
290
300
320
290
UJ
oo
lDenotes values significantly lower than the controls (Duncan's Multiple Range P=0.05)
-------
Total wet weight of bluegill fry exposed for 60 days to
1122 (ig/1 chromium was significantly lower than that of
controls and fry exposed to lesser chromium concentrations.
Although this was the only statistically valid effect
observed in the test it is supported by trends in total
length and the fact that total weight appeared to be the most
sensitive indicator of chromium effects on fry of rainbow
trout and lake trout.
Based on these data, the MATC for chromium and eggs and fry
of bluegill is estimated to be between 522 and 1122 [ig/1,
however, it is possible that growth effects due to exposure
to lesser concentrations were masked by the poor success in
early feeding of these fry.
White sucker
\
The percentage of white sucker eggs which hatched successfully
was similar among controls and eggs exposed to chromium
concentrations as high as 1975 Mg/1 (Table 23). After 30 days
post-hatch exposure, the percentage survival of white sucker
fry was also unaffected by exposure to the range of chromium
concentrations tested. However, total length of white sucker
fry after 30 days exposure to 1975 M-g/1 chromium was signifi-
cantly reduced when compared with controls.
Survival of white sucker fry exposed to any of the chromium
concentrations tested was not significantly affected after
60 days. Both total length and total wet weight of white
suckers after 60 days exposure to 1975 • 963 and 538 |j.g/l
chromium were significantly reduced when compared with
controls. Again, the reduction in total weight of fry
after 60 days exposure appears to be the most sensitive
indicator of sub-lethal effects of chromium.
Based on these data,the MATC of chromium for eggs and fry
of white suckers is estimated to be between 290 and 538 p.g/1.
Northern pike
Exposure to chromium concentrations as high as 1975 M-g/1 had
no significant effect on the percentage of northern pike eggs
which hatched successfully when incubated from the eyed
stage (Table 2^). After 20 days, a significant reduction in
survival of northern pike fry exposed to 1975 and 963 MS/1
chromium was observed.
Data on survival and growth of northern pike after 20 days
exposure is not considered reliable due to the effect of
39
-------
TABLE 23 - MEAN PERCENTAGE HATCH OF EGGS, MEAN SURVIVAL, TOTAL LENGTHS AND WET WEIGHT
OF WHITE SUCKER (Catostomus commersoni) FRY CONTINUOUSLY EXPOSED TO CHROMIUM IN SOFT
WATER ( 38.8 ± 5.3 mg/1 as
Mean
measured
chromium cone.
(ns/i)
1975 A
B
963 A
B
538 A
B
290 A
B
123 A
B
Control A
B
Mean
hatch
(*)
94
97
98
96
100
96
94
97
93
93
88
97
1-30 Days
Survival
(*)
86
54
50
76
64
44
36
54
60
70
74
42
Mean total
length (mm)
16 (2.1)
18 (1.4)a
20 (2.5)
20 (2.6)
21 (1.9)
21 (2.2)
23 (2.0)
22 (2.2)
21 (1.8)
21 (2.5)
20 (1.9)
21 (1.8)
31-60 Days
Survival
<*)
46
10
46
52
48
34
36
50
46
52
48
42
Mean total
length (mm)
18 (2.6)
18 (1.8)a
25 (3.6)
25 (3.7)a
27 (3.4)
28 (1.7)
33 (3.2)
28 (2.9)
32 (2.3)
30 (3.7)
32 (2.5)
31 (4.1)
Mean total
wet wt. (ing)
40
40a
130.
100a
170
170a
260
160
210
180
260
230
^Denotes values significantly lower than the controls (Duncan's Multiple Range P=0.05)
-------
TABLE 24 - MEAN PERCENTAGE HATCH OF EGGS, MEAN SURVIVAL, TOTAL LENGTHS AND WET WEIGHT
OF NORTHERN PIKE (Esox lucius)FRY CONTINUOUSLY EXPOSED TO CHROMIUM IN SOFT WATER
(37.8 ± 5.6 mg/1 as CaCO^T
Mean
measured
chromium cone.
(ns/i)
1975 A
B
963 A
B
538 A
B
290 A
B
123 A
B
Control A
B
Mean
hatch
(*)
83
88
81
83
83
90
76
80
74
87
73
83
1-20 Days
Survival
(*)
14
24a
32
20a
38
38
34
34
42
34
48
48
Mean total
length (mm)
23 (4.6)
26 (3.6)
-
27 (7.5)
27 (4.5)
32 (5.9)
29 (3.6)
31 (3.8)
34 (4.9)
36 (5.D
29 (4.8)
21-50 Days
Survival
(*)
0
8
12
0
8
12
2
8
18
10
14
14
Mean total
length (mm)
40 (9.8)
44 (3.6)
-
57 (6.5)
50 (5.7)
70
58 (5.2)
56 (10.2)
54 (5.4)
54 (7.0)
56 (10.2)
Mean total
wet wt. (mg)
340
440
-
890
640
2000
940
990
780
880
1180
^Denotes values significantly lower than the controls (Duncan's Multiple Range P=0.05)
-------
cannibalism which could not be controlled after pike were
20 days old.
The MATC of chromium for northern pike eggs and fry is esti-
mated to be between 538 and 963 p.g/1 although effects of
lower concentrations on survival and growth may have been
masked by the cannibalism among pike fry in the growth
chambers.
Walleye
Exposure to chromium concentrations as high as 216? M-g/1 had
no significant effect on the percentage of walleye eggs which
hatched successfully (Table 25). Survival of walleye fry after
30 days exposure was low in all treatments but indicated no
significant effects of exposure to the range of chromium
concentrations which were tested. Again, the generally poor
success in feeding walleye fry is responsible for the lack
of more conclusive data on the toxicity of chromium to this
species. Based on the data available, the MATC for chromium
and walleyes is estimated to be >2l6l (j.g/1.
TABLE 25 - MEAN PERCENTAGE HATCH OF EGGS, MEAN SURVIVAL AND
TOTAL LENGTHS OF WALLEYE (Stizostedion vitreum) FRY CONTINUOUSLY
EXPOSED TO CHROMIUM IN SOFT WATER (38.5 ± 4.2 mg/1 as CaCOo)
Mean measured
chromium cone.
(ng/i)
216? A
B
1125 A
B
558 A
B
288 A
B
133 AA
B
80 A
B
Control A
B
Mean hatch
(*)
63
65
71
73
73
65
73
65
58
63
57
67
62
69
1-30 Days
Survival
(*)
18
20
32
30
20
28
28
24
10
14
14
24
10
14
Mean total
length (mm)
10 (3-7)
8 (2.2)
12 (2.9)
11 (2.6)
10 (2.7)
10 (2.5)
11 (3-3)
11 (3.5)
8 (3.8)
10 (1.4)
10 (4.2)
9 (2.1)
10 (2.5)
10 (2.0)
a
Denotes values
Multiple Range
si gni fi cantly
P=0.05)
lower than the controls (Duncan's
42
-------
EXPOSURE OF FISH TO COPPER
Brook trout (soft water)
None of the brook trout eggs exposed to 95 P-g/1 of copper in
soft water (37.5 mg/1 as CaCO-O hatched successfully (Table 26)
Exposure to 51. 27 and 13 M-g/1 copper, significantly reduced
the percentage hatch of brook trout eggs when compared to
controls and eggs exposed to lower concentrations of copper.
Concentrations of 51 and 27 p.g/1 copper significantly reduced
survival of brook trout fry when compared to other treatments
and controls after 30 days exposure. Finally, total lengths
among fry exposed to 51. 27 and 13 ^g/1 copper for 30 days
were significantly less than controls and those at lower
concentrations.
Continuous exposure for 60 days to concentrations of copper
as high as 13 p-g/1 had no significant effect on survival
of brook trout fry. However, exposure to copper concentrations
— 5 M-g/1 significantly reduced total length and wet weight
of fry when compared to controls.
The mean concentration of copper in the diluent water (control)
was measured to be 3 P-g/l« Thus, the MATC of copper for brook
trout in soft water (37.5 mg/1 as CaCOo) is estimated to be
between 3 and 5 P-g/1-
Brook trout (hard water)
The percentage hatch of brook trout eggs was significantly
reduced by exposure to 7^ M-g/1 copper in hard water (187 mg/1
as CaCO-^) when compared to controls (Table 27). None of the
brook trout fry survived 30 days exposure to 7^ P-g/1 copper
and both survival and total length were reduced by 30 days
exposure to ^9 Mg/1 when compared with controls.
None of the brook trout fry survived 60 days exposure to
ij-9 Hg/1 copper while survival among fry exposed to lower
copper concentrations continued to be comparable with
controls. Total length and wet weight of brook trout at the
end of exposure (60 days) were similar for controls and fry
exposed to 5 M-g/1 copper but were significantly reduced by
exposure to 21, 13 and 8 p.g/1 where survival was generally
excellent.
Based on these data, the MATC of copper for brook trout eggs
and fry exposed in hard water (187 mg/1 as CaCO^) is estimated
to be between 5 and 8 |a.g/l.
43
-------
TABLE 26 - MEAN PERCENTAGE HATCH OF EGGS, MEAN SURVIVAL, TOTAL LENGTHS AND WET WEIGHT
OF BROOK TROUT (Salvelinus fontinalis) FRY CONTINUOUSLY EXPOSED TO COPPER IN SOFT WATER
( 37.5 ± 7-3 mg/1 as
Mean
measured
copper cone.
WD
95 A
B
51 A
B
27 A
B
13 A
B
7 A
B
5 A
B
Control A
B
Mean
hatch
(*)
0
Oa
2a
6
25a
63
72
72
77
79
73
1-30 Days
Survival
(*)
-
0
2a
10
6a
96
74
100
98
100
98
100
100
Mean total
length (mm)
-
18 (1.5)a
19 (1.8)
18 (1.9)a
19 (1.8)
18 (1.5)a
22 (2.0)
21 (1.7)
22 (2.3)
22 (1.9)
22 (2.2)
23 (2.3)
31-60 Days
Survival
—
0
8
6a
96
72
98
98
98
90
96
98
Mean total
length (mm)
-
_
, 18 (1.2)
17 (3.8)a
. 22 (7.1)
23 (2.4)a
24 (3.0)
26 (3.3)
25 (4.0)a
28 (5.3)
29 (4.2)
Mean total
wet wt. (mg)
_
_
80
120
101a
126
127a
151a
192
240
3.
Denotes values significantly lower than the controls (Duncan's Multiple Range P=0.05)
-------
TABLE 27 - MEAN PERCENTAGE HATCH OF EGGS, MEAN SURVIVAL, TOTAL LENGTHS AND WET WEIGHT
OF -BROOK TROUT (Salvelinus fontinalis) FRY CONTINUOUSLY EXPOSED TO COPPER IN HARD WATER
( 187.0 ±22.0 mg/1 as CaCOp
Mean
measured
copper cone.
(ne/D
74 A
B
49 A
B
21 A
B
13 A
B
8 A
B
5' A
B
Control A
B
Mean
hatch
(*)
19
12a
35
35
^5
44
48
49
4,9
53
^5
51
56
39
1-30 Days
Survival
(*)
0
oa
52
70a
100
90
100
98
100
100
100
100
100
100
Mean total
length (mm)
—
-
17 (1.6)
17 (1.0)a
19 (1.8)
19 (1.7)
20 (1.2)
20 (1.8)
19 (1.3)
20 (1.4)
20 (1.5)
20 (2.5)
20 (1.7)
20 (1.4)
31-60 Days
Survival
W
_
-
0
oa
76
58
76
90
74
86
68
80
58
100
Mean total
length (mm)
_
-
_
-
19 (2.6)
19 (1.6)a
22 (1.3)
20 (2.3)a
21 (2.2)
19 (2.1)a
24 (2.2)
22 (2.0)
27 (1.8)
24 (3.2)
Mean total
wet wt. (mg)
_
-
V
-
47
48a
81
78a
79
75a
110
117
117
128
^Denotes values significantly lower than controls (Duncan's Multiple Range P=0.05)
-------
Channel catfish (soft water)
Exposure to concentrations of copper as high as 2k \ig/l in
water of 36 mg/1 total hardness appeared to have no effect
on the percentage of channel catfish eggs which hatched
successfully (Table 28). Exposure of catfish fry for 30
days to 2k and 18 [ig/1 copper significantly reduced "both
percentage survival and total length when compared with fry
in controls and those exposed to lower concentrations of
copper.
No catfish fry survived 60 days exposure to 2k p.g/1, however,
survival of fish exposed to 18 fig/1 (during the 31-60 day
period of exposure) was not significantly different from
controls. This is possibly due to the vigor of individuals
which survived the initial 30 day exposure. Total length of
catfish exposed for 60 days to 18 (ig/1 was significantly
reduced compared to controls. Total weight of these fry
appeared reduced at this time but data were too variable to
ascribe statistical significance to this observation.
Based on these data,the MATC of copper for eggs and fry of
channel catfish in water of 36 mg/1 total hardness is
estimated to be between 12 and 18 p.g/1.
Channel catfish (hard water)
Percentage hatchability of channel catfish eggs appeared
reduced by exposure to 66 (ig/1 copper in water with a total
hardness of 186 mg/1 (Table 29), however, variability
between duplicates in other treatments precludes ascribing
statistical significance to this observation. None of the
catfish fry survived 30 days exposure to 66 and 3^ M-g/1
copper, and survival was reduced by exposure to 19 Hg/1
when compared with controls.
After 60 days exposure, the percentage of channel catfish fry
surviving exposure to 19 p.g/1 copper was significantly less
than controls. Total length and wet weight of catfish exposed
to this concentration were also significantly less at this
time when compared with control fish.
Based on these data,the MATC of copper for channel catfish
in hard water (186 mg/1 as CaCOo) is estimated to be between
13 and 19 |ig/l.
-------
TABLE 28 - MEAN PERCENTAGE HATCH OF EGGS, MEAN SURVIVAL, TOTAL LENGTHS AND WET WEIGHT
OF CHANNEL CATFISH (Ictalurus punctatus) FRY CONTINUOUSLY EXPOSED TO COPPER IN SOFT
WATER (36.0 ± 1.1 mg/1 as
Mean
measured
copper cone.
(jxg/1)
24 A
B
18 A
• B
12 A
B
7 A
B
6 A
B
3 A
B
Control A
B
Mean
hatch
(*)
98
86
92
90
76
71
100
80
80
78
55
67
84
6k
1-30 Days
Survival
(*)
1
5a
26
20a
36
52
65
52
49
32
77
50
49
59
Mean total
length (mm)
16 (0.8)
16 (0.8)a
17 (0.8)
17 (2.1)a
21 (2.2)
19 (2.3)
21 (2.5)
21 (2.4)
19 (1.6)
21 (2.5)
20 (2.1)
21 (2.2)
21 (2.3)
22 (2.8)
31-60 Days
Survival
(#)
0
Oa
61
39
69
65
88
92
47
84
96
96
66
86
Mean total
length (mm)
_
-
26 (3.2)
29 (4.7)a
34 (4.3)
32 (4.3)
33 (4.7)
33 (3.6)
31 (4.5)
33 (5.7)
33 (4.5)
32 (4.6)
35 (4.1)
35 (4.5)
Mean total
wet wt. (mg)
_
-
180
230
360
290
330
350
300
34o
310
280
340
400
Denotes values significantly lower than the controls (Duncan's Multiple Range P=0.05)
-------
TABLE 29 - MEAN PERCENTAGE HATCH OF EGGS, MEAN SURVIVAL, TOTAL LENGTHS AND WET WEIGHT
OF CHANNEL CATFISH (Ictalurus punctatus) FRY CONTINUOUSLY EXPOSED TO COPPER IN HARD
WATER (186.3 ±38.7 mg/1 as CaC03)
Mean
measured
copper cone.
(p-g/D
66 A
B
34 A
B
19 A
B
13 A
B
10 A
B
7 A
B
Control A
B
Mean
hatch
(*)
2k
23
35
V?
40
28
51
29
46
50
52
72
83
46
1-30 Days
Survival
(*)
0
oa
0
o.a
14
9a
54
48
53
64
34
72
69
68
Mean total
length (mm)
-
_
21 (3.3)
21 (3.3)
24 (2.5)
24 (3.4)
27 (3-D
24 (2.7)
23 (2.7)
23 (1.9)
23 (1.9)
24 (2.0)
31-60 Days
Survival
(*)
-
—
80
80a
100
95
100
98
98
96
100
100
Mean total
length (mm)
—
—
25 (3.4)
25 (3.4)a
29 (4.6)
30 (5.0)
30 (4.9)
27 (4.4)
34 (4.2)
33 (4.9)
32 (4.2)
34 (3-7)
Mean total
wet wt. (mg)
-
-
170
I70a
270
320
280
230
340
340
320
370
oo
a
Denotes values significantly lower than the controls (Duncan's Multiple Range P=0.05)
-------
Walleye (soft and hard waters)
As mentioned previously, exposures of walleye eggs and fry
were generally unsuccessful due to difficulty in early feeding
of fry. The following observations were made during the early
stages of the copper exposures. In soft water 35 mg/1 as
CaCO-}), no walleye eggs hatched successfully during exposure
to 9i P-g/1 copper and hatchability was significantly reduced
by exposure to ^7 ^g/1 (Table 30)- In hard water (189 mg/1
as CaCC>3) hatchability of walleye was similar for controls
and eggs exposed to copper concentrations as high as 71 p.g/1
(Table 31)- Percentage hatch for eggs in all experimental
units ranged from 4^-69$. In soft water, none of the walleye
fry survived 30 days exposure to ^7 or 21 p.g/1 copper,
although survival was generally poor even in controls. In
hard water, all walleyes including controls died very early
in the expo.sure. Based on the limited available evidence, the
MATC for copper and walleye is estimated to be between 13 and
21 p.g/1 in soft water, and greater than 71 P-g/1 in hard water.
TABLE 30 - MEAN PERCENTAGE HATCH OF EGGS, MEAN SURVIVAL AND
TOTAL LENGTHS OF WALLEYE (Stizostedion vitreum) FRY CONTINUOUSLY
EXPOSED TO COPPER IN SOFT WATER (35.0 ± 1.8 mg/1 as
Mean measured
copper cone.
(ns/i)
91 A
B
^7 A
B
21 A
B
13 A
B
8 A
B
3 A
B
Control A
B
Mean hatch
(*)
0
oa
2*J-
10a
57
56
11
62
71
53
^4-6
60
1-30 Days
Survival
(*)
—
0
0
0
0
12
0
38
16
20
18
18
Mean total
length (mm)
-
_
-
_
10
11
12
9
11
11
9
Denotes values significantly lower than the controls (Duncan's
Multiple Range P=0.05)
-------
TABLE 31 - MEAN PERCENTAGE HATCH OF EGGS, MEAN SURVIVAL
LENGTHS AND WET WEIGHT OF WALLEYE (Stizostedion vitreum) FRY
CONTINUOUSLY EXPOSED TO COPPER IN HARD WATER (189.0 ±33.0
mg/1 as CaCO-})
Mean measured
copper cone.
(US/1)
71 A
B
38 A
B
24 A
B
1? A
B
14 A
B
9 A
B
Control A
B
Mean hatch
(*)
52
44
59
63
69
61
56
45
62
61
58
68
61
54
1-30 Days
Survival
(*)
0
0
0
0
0
0
0
0
0
0
0
0
0
0
EXPOSURE OF FISH TO CADMIUM
Brook trout (soft water)
Hatchability of "brook trout eggs in soft water (37 mg/1 as
CaC03) was similar for controls and eggs exposed to cadmium
concentrations as high as 47 (J-g/1 (Table 32). Survival of
brook trout fry through 30 days post-hatch exposure was also
not affected by exposure to cadmium concentrations as high
as 47 (J-g/1. Total lengths of brook trout were significantly
reduced by JO days exposure to cadmium concentrations > 10 p.g/1.
Survival of brook trout was significantly reduced during 60
days exposure to test concentrations of cadmium > 6 (ig/1. Total
length of brook trout exposed to these same concentrations was
also significantly lower than that of control fish at this
time. Total wet weight of brook trout after 60 days exposure
to concentrations > 3 l-ig/1 cadmium was significantly less
than controls.
50
-------
TABLE 32 - MEAN PERCENTAGE HATCH OF EGGS, MEAN SURVIVAL, TOTAL LENGTHS AND WET WEIGHT
OF BROOK TROUT (Salvelinus fontinalis) EGGS AND FRY CONTINUOUSLY EXPOSED TO VARIOUS
CONCENTRATIONS OF CADMIUM IN SOFT WATER (37.0 ± 7.2 mg/1 as
Mean measured
cadmium cone.
(Hfi/D
4-7 A
B
2k A
B
10 A
B
6 A
B
3 A
B
1 A
B
Control A
B
Mean
hatch
(*)
61
70
7^
74
16
74
65
76
58
73
78
75
75
71
1-30 Days
Survival Mean total
(%") length (nun)
96
89
72
96
48
100
99
96
99
100
100
99
100
100
17 (2.9)
17 (1.7)a
19 (l.7)a
19 (l.9)a
18 (1.5).
18 (1.7)a
20 (1.5)
20 (1.5)
21 (1.6)
22 (1.7)
22 (1.8)
22 (1.5)
22 (1.7)
22 (1.4)
31-60 Days
Survival
W
22a
^a
22a
4
38a
5°a
30a
82
78
90
58
100
100
Mean total
length (mm)
25 (3-8)
27 (1.4)a
25 (i.*0a
24 (3-D
25 (l.*0_
24 (3.0)a
25 (3.8)
25 (2.5)a
27 (3-3)
28 (3.0)
29 (3-7)
30 (3-D
29 (3.8)
30 (3.8)
Mean total
wet wt. (ing)
115
136a
135a
I46a
135a
123a
Iif3a
I48a
191
183
232
226
240
236
^Denotes values significantly lower than the controls (Duncan's Multiple Range P=0.05)
-------
Based on this exposure,the MATC of cadmium for brook trout
eggs and fry exposed in soft water is estimated to be between
1 and 3 (ig/1.
Brook trout (hard water)
Hatchability of brook trout eggs in hard water (188 mg/1
as CaCO-j) was generally lower than hatchability in soft
water but indicated no significant effects resulted from
exposure to cadmium concentrations as high as 91 M-g/1 (Table 33),
Percentage survival of brook trout fry during 30 days post-
hatch exposure was excellent and also indicated no effects of
exposure to the cadmium concentrations tested. Total lengths
of brook trout fry after 30 days exposure to 91. 50 and 21 [ig/I
cadmium were significantly lower than total lengths of control
fishc
None of the brook trout fry survived 60 days exposure to 91
and 50 (J-g/1 and survival was significantly reduced by 60 days
exposure to 21 and 12 p.g/1 indicating an extremely cumulative
effect during the 30-60 day exposure period. In addition,
total length and total weight of surviving brook trout exposed
to 21 and 12 p.g/1 cadmium were also reduced when compared with
controls.
Based on these data, the MATC of cadmium for brook trout exposed
in hard water (188 mg/1 as CaCOo) is estimated to be between
7 and 12 (ig/1.
Channel catfish (soft water)
Exposure to cadmium concentrations as high as 5^ M-S/1 had no
significant effect on the percentage of channel catfish eggs
which hatched successfully (Table 3^)- The survival of channel
catfish fry was significantly reduced by 30 days exposure to
cadmium concentrations > 1? f-ig/1 when compared with controls.
Total lengths of catfish fry after 30 days were slightly less
than that of controls in all cadmium concentrations but
differences were not statistically significant.
During the 31-60 days post-hatch exposure, the survival of
catfish fry which had survived the initial 30 day exposure
was significantly reduced by exposure to 5^ and 32 p.g/1
cadmium. However, survival was not significantly reduced by
exposure to cadmium concentrations of 20 and 1? (J.g/1. which
did significantly reduce survival during the initial 30 day
exposure. Total lengths and wet weights of catfish fry
after 60 days exposure to cadmium were highly variable and
52
-------
TABLE 33 - MEAN PERCENTAGE HATCH OF EGGS, MEAN SURVIVAL, TOTAL LENGTHS AND WET WEIGHT
OF BROOK TROUT (Salvelinus fontinalis) FRY CONTINUOUSLY EXPOSED TO CADMIUM IN HARD
WATER (188.0 ± 27.0 mg/1 as
Mean measured
cadmium cone.
(ns/i)
91 A
B
50 A
B
21 A
B
12 A
B
7 A
B
3 A
B
Control A
B
Mean
hatch
(*)
51
36
53
54
48
45
42
56
46
49
46
43
50
58
1-30 Days
Survival
(*)
100
96
100
100
100
100
100
100
100
100
100
98
100
100
Mean total
length (mm)
16 (1.7)
15 U.7)a
17 !1<5)a
17 U.9)a
18 (1.9).
18 (1.4)a
19 (1.5)
19 (1.4)
20 (1.4)
20 (1.6)
21 (1.5)
20 (1.5)
21 (1.4)
21 (1.0)
31-60 Days
Survival
(*)
0
oa
0°a
^
4o
50a
84
78
78
90
88
74
Mean total
length (mm)
-
-
21 U.9L
21 (2.3)a
22 (2.3).
23 (1.9)a
24 (1.6)
24 (2.4)
24 (2.5)
24 (2.6)
25 (2.0)
25 (2.0)
Mean total
wet wt. (mg)
_
-
85a
85
95a
84a
109
115
114
120
125
136
Ul
OJ
^Denotes values significantly lower than the controls (Duncan's Multiple Range P=0.05)
-------
TABLE 34 - MEAN PERCENTAGE HATCH OF EGGS,- MEAN SURVIVAL, TOTAL LENGTHS AND WET WEIGHT
OF CHANNEL CATFISH (Ictalurus punctatus) FRY CONTINUOUSLY EXPOSED TO CADMIUM IN SOFT
WATER ( 37.0 ± 1.3 mg/1 as CaCO?)
Mean measured
cadmium cone.
(fig/D
54 A
B
32 A
B
20 A
B
17 A
B
11 A
B
6 A
B
Control A
B
Mean
hatch
(*)
93
82
97
98
90
86
79
51
75
55
63
63
77
90
1-30 Days
Survival
(*)
;•
I*
^
17a
30a
46
40
60
61
63
30
Mean total
length (mm)
19
17 (1.2)
17 (1.2)
19 (1.9)
19 (1.9)
17 (1.2)
19 (1.9)
19 (1.5)
19 (1.7)
20 (1.7)
20 (1.8)
21 (2.6)
22 (3.3)
31-60 Days
Survival
(*)
0°*
0
5oa
52
100
58
96
92
85
86
98
84
69
Mean total
length (mm)
_
24~(5.7)a
32 (6.4)
35 (5.0)
25 (5.1)
29 (3.4)
31 (2.6)
30 (4.7)
31 (3.8)
36 (3.7)
35 (4.7)
36 (5.2)
Mean total
wet wt. (mg)
-
170
300
400
170
270
240
240
270
380
380
380
Denotes values significantly lower than the controls (Duncan's Multiple Range P=0.05)
-------
differences could not "be shown to be statistically significant.
Based on these data, the MATC for cadmium and channel catfish
exposed in soft water is estimated to "be "between 11 and 1? |ig/l.
Channel catfish (hard water)
The percentage of channel catfish eggs which hatched successfully
in hard water (185 mg/1 as CaCO^) was not significantly affected
by exposure to cadmium concentrations as high as 59 p.g/1 (Table 35)-
After 30 days post-hatch exposure, survival of channel catfish
was highly variable and no treatment related effects could be
statistically verified. Total length of channel catfish after
30 days exposure to 59 (J-g/1 cadmium was significantly reduced
when compared with controls.
After 60 days post-hatch exposure, survival of channel catfish
was excellent in all concentrations of cadmium which were
tested and in controls. Total length and total wet weight of
catfish exposed to 59. 33 and 17 p.g/1 cadmium for 60 days was
reduced when compared with controls, indicating cumulative
growth effects at these concentrations in hard water. Nearly
identical concentrations of cadmium in soft water had affected
survival of catfish after only 30 days exposure indicating
that increased water hardness delayed or reduced the toxic
effects of cadmium at these concentrations.
Based on these data, the MATC of cadmium for channel catfish
eggs and fry exposed in hard water (185 mg/1 as CaCO^) is
estimated to be between 12 and 17 l-Lg/l-
Walleye(soft and hard waters)
Exposure to concentrations of cadmium as high as 55-0 P-g/1 in
soft water (35.0 mg/1 as CaCOo) (Table 36), and as high as
86.7 iig/1 in hard water (187.0 mg/1 as CaCOo) (Table 37) did
not significantly affect the percentage of walleye eggs which
hatched successfully. Difficulty in feeding newly hatched
walleyes (previously discussed) resulted in extremely poor
survival and the only significant observation was the complete
mortality of walleye fry observed during the initial 10 days
of exposure to 55 and 25 |ig/l in soft water.
Based on these data, the MATC for walleye is estimated to be
between 9 and 25 |ig/l in soft water (35.0 mg/1 as CaCC^) and
>86.7 p.g/1 in hard water (187.0 mg/1 as
-------
TABLE 35 - MEAN PERCENTAGE HATCH OF EGGS, MEAN SURVIVAL, TOTAL LENGTHS AND WET WEIGHT
OF CHANNEL CATFISH (Ictalurus punctatus) FRY CONTINUOUSLY EXPOSED TO CADMIUM IN HARD
WATER (185.0 ±35-0 mg/1 as
Mean measured
cadmium cone.
(ns/D
' 59 A
B
33 A
B
17 A
B
12 A
B
5 A
B
2 A
B
Control A
B
Mean
hatch
(*)
81
81
71
86
57
90
75
61
86
83
41
81
56
^3
1-30 Days
Survival
(*)
38
10
46
81
41
60
22
^7
53
58
59
78
72
53
Mean total
length (mm)
21 (2.6)
21 (2.6)a
24 (2.8)
23 (2.0)
24 (2.3)
23 (1.9)
23 (2.2)
24 (2.2)
25 (2.6)
24 (2.4)
26 (2.3)
25 (2.5)
24 (2.0)
26 (2.2)
31-60 Days
Survival
(*)
88
100
92
98
98
100
100
98
94
96
96
100
98
98
Mean total
length (mm)
29 (3-6)
31 (3-6)a
30 (4.5)a
31 (3.9)a
29 (4.9)_
29 (3.*0a
3^ (3-D
33 (3-5)
33 (4.6)
34 (4.0)
3^ (3-2)
32 (3-D
3^ (3-3)
33 (3-D
Mean total
wet wt. (mg)
230_
270
280
250a
300
290a
360
340
320
330
380
34o
350
340
Denotes values significantly lower than the controls (Duncan's Multiple Range P=0.05)
-------
TABLE 36 - MEAN PERCENTAGE HATCH OF EGGS, MEAN SURVIVAL AND
TOTAL LENGTHS OF WALLEYE (Stizostedion vitreum) FRY CONTINUOUSLY
EXPOSED TO CADMIUM IN SOFT WATER (35-0 ±1.2 mg/1 as
Mean measured
cadmium cone.
(ne/i)
55 A
B
2k A
B
9 A
B
4 A
B
2 A
B
0.9 A
B
Control A
B
Mean
hatch
(*)
64
65
68
57
63
61
61
58
59
69
64
71
64
65
1-30 Days
Survival
(*)
oa
0
oa
0
8
2
12
10
10
18
14
8
2
16
Mean total
length (mm)
-
_
-
10 (1.0)
10 (0.9)
12 (1.3)
10 (1.7)
12 (1.9)
12 (3.2)
12 (2.4)
9 (1.5)
13 (2.3)
12 (0.6)
aDenotes values significantly lower than the controls (Duncan's
Multiple Range P=0.05)
57
-------
TABLE 37 - MEAN PERCENTAGE HATCH OF EGGS AND SURVIVAL OF
WALLEYE (Stizostedion vitreum) FRY CONTINUOUSLY EXPOSED TO
CADMIUM IN HARD WATER (187.0 ± 36.0 mg/1 as CaCO-)
Mean measured
cadmium cone.
(lifi/1)
86.7 A
B
44.3 A
B
19.0 A
B
8.4 A
B
3.4 A
B
1.3 A
' B
Control A
B
Mean Hatch
(*)
60
52
48
66
46
61
48
54
48
48
49
57
37
56
1-30 Days
Survival
(*)
0
0
0
0
0
0
0
0
0
0
0
0
0
0
58
-------
SECTION VI
DISCUSSION
The criteria utilized to estimate the maximum acceptable
toxicant concentrations (MATC's) for metals tested in this
study were generally survival and/or growth (determined "by
measurements of length and weight) of fry. In all cases,
these parameters were affected at metal concentrations
significantly lower than those which were observed to affect
egg hatching. These data support the growing "body of evidence
that the eggs of fishes are generally more resistant to inimical
chemicals than are other life stages.
Analysis of the MATC values generated from the results of this
study indicated that measured concentrations of copper and
cadmium which were effective in significantly reducing survival
and/or growth of fish fry were similar and were generally an
order of magnitude lower than those derived for lead and
chromium with the same fish species. Lead ranked third in
relative toxicity and chromium was the least toxic of the four
metals tested, producing comparable effects only at concentra-
tions 10-100 times greater than those for copper and cadmium
(Table 38 and 39).
MATC's estimated for copper and cadmium from tests conducted
in soft (35-0 - 37.5 mg/1 as CaC03) and hard (185.0 - 189-0
mg/1 as CaCO^) water indicated that there were no significant
differences related to water hardness for brook trout and
channel catfish. In the case of walleyes, however, an increase
in the hardness of exposure water apparently resulted in a
significant increase in the estimated MATC's for both copper
and cadmium.
A superficial analysis of the MATC's estimated for fish exposed
to lead and chromium might indicate a wide range in sensitivity
among the seven fish species tested. However, if the actual
duration of exposure is considered (Table ^0), it appears that
the sensitivities of these species are not significantly
different from one another. That is, the species with the
lowest MATC's are those which were exposed for the longest
period of time. Therefore, the actual duration of exposure
should be considered in interpretation of data generated from
chronic assays.
EXPOSURE OF FISH TO LEAD
While there are now many data in the literature describing the
acute toxicity, generally under static test conditions, of
metals to freshwater fish (e.g., see reviews by McKim et al.,
59
-------
TABLE 38 - SUMMARY OF THE MAXIMUM ACCEPTABLE TOXICANT
CONCENTRATION (MATC) OF LEAD AND CHROMIUM FOR SELECTED
FRESHWATER FISH SPECIES IN SOFT WATER (32.6 - 40.? mg/1 as
Fish Species
MATC
Lead
Chromium
Rainbow trout
(Salmo gairdneri)
Lake trout
(Salvelinus namaycush)
Channel catfish
(Ictalurus punctatus)
Bluegill
(Lepomis macrochirus)
White sucker
(Catostomus commersoni)
Northern pike
(Esox lucius)
Walleye
(Stizostedion vitreum)
>51<105
>75<136
>70<120
>119<253
>237<397
>150<305
>522<1122
>290<538
>538<963
>2l6?
60
-------
TABLE 39 - SUMMARY OF THE MAXIMUM ACCEPTABLE TOXICANT
CONCENTRATION (MATC) OF COPPER AND CADMIUM FOR SELECTED
FRESHWATER FISH SPECIES IN SOFT (35-0 - 37-5 mg/1 as CaC03)
AND HARD (185.0 - 189-0 mg/1 as CaCOo) WATER
Fish Species
Water
hardness
MATC (ug/1)
Copper
Cadmium
Brook trout
(Salvelinus fontinalis)
Channel catfish
(Ictalurus punctatus)
Walleye
(Stizostedion vitreum)
soft
hard
soft
hard
soft
hard
>3<5
>5<8
>7<12
>9<25
>87
TABLE ^0 - SUMMARY OF THE ACTUAL DURATION OF EXPOSURE TO LEAD
AND CHROMIUM FOR SELECTED FRESHWATER FISH SPECIES
Fish Species
Duration of Exposure
(days)
Rainbow trout
(Salmo gairdneri)
Lake trout
(Salvelinus namaycush)
Channel catfish
(Ictalurus punctatus)
Bluegill
(Lepomis macrochirus)
White sucker
(Catostomus commersoni)
Northern pike
(Esox lucius)
Walleye
(Stizostedion vitreum)
95 - 97
111 - 115
66 - 68
62
70 - 73
39 -
61
-------
1973* 197^; Leland et al., 1975). there are still relatively
few data describing sublethal effects of this metal on the
growth and development of early life stages of fish. Davies
and Everhart (1973) report the results of a study in which the
acute and chronic toxicity of lead [as lead nitrate, Pb(N03)2]
to rainbow trout in soft and hard water was determined. Data
regarding hatchability of rainbow trout eggs exposed to lead
concentrations ranging from 5-3 to 101.8 |_ig/l in soft water
(28.1-28.7 mg/1 as CaCO3) were inconclusive since 100% egg
mortality occurred in one replicate of controls and in at
least one replicate of all lead treatments except at 50.9 (ig/1.
In our study (soft water, 3^.6 mg/1 as CaCC^), hatchability
was excellent (>82$) in controls and all lead treatments to
443 (ig/1. However, at 672 (ig/1 lead, only 28 and.29$ of the
exposed eggs hatched successfully.
Davies and Everhart observed significant differences in growth,
determined as mean total length, among fish at the various
lead treatments but concluded that these differences were not
lead related but were due to the use of eggs of significantly
different sizes, with the possibility of significant genetic
differences in brood stock, to initiate this study. We observed
significant differences in mean total lengths of fry exposed to
lead concentrations >250 [ig/1 as compared with controls and those
at lower treatments after 30 days of continuous exposure. Survival
to 60 days post-hatch was near zero at all concentrations >250 |ig/l
lead and was severely reduced at 1^6 (j.g/1 lead. Davies and Everhart
similarly observed severely reduced survival (33$) after 6 months
of exposure to 95-2 (ig/1 lead. Growth measurements for fry
surviving 60 days exposure at all lead treatments were not
significantly different from those of controls.
Physical abnormalities such as "black tail", scpliosis and
caudal fin erosion were observed during the Davies and Everhart
study. Black tail was the most common abnormality observed
and its incidence was ^2$, 91-3$ and 100$ for fish exposed to
lead concentrations of 23.8, ^-7-6 and 95-2 (ig/1, respectively.
Based on black tail, Davies and Everhart estimated an MATC
of >6.0 <11.9 l^g/1 for rainbow trout exposed to lead in soft
water. Scoliosis in fry was also observed in our study but to
a lesser degree. Rainbow trout fry exposed to 4^3 and 672 |ig/l
lead for 30 days post-hatch had a scoliosis incidence of 12
and 28$, respectively. Black tails were never observed on our
fish. Based on our data, the MATC for rainbow trout exposed to
lead is estimated to be >71 <1^6 (ig/1. Discrepancies between
our results and those of Davies and Everhart may be due to a
number of factors. Nevertheless, these approximations of
MATC's are comparable considering factors such as the shorter
62
-------
duration of our study, differences in fish populations and
others.
Similar concentrations of lead, as determined for rainbow
trout, were effective in significantly decreasing survival
and growth of fry of lake trout, channel catfish and bluegill
but among these, only bluegill were observed to be affected
by scoliosis. White sucker, northern pike and walleye appeared
to be more resistant to lead exposure and MATC's estimated for
these species were 2-3 times higher than the others. Scoliosis
was a very common and severe problem with white sucker exposed
to the higher lead concentrations (253 and ^83 p.g/1) .
Northern pike were difficult to study due to their cannibalistic
behavior. Walleye were also difficult to feed adequately under
these conditions. Continuous exposure to a lead concentration
of 39? p-g/1 was associated with a high incidence of scoliosis
not observed in controls or other lead treatments.
Based on the MATC values estimated for 7 species of freshwater
fish chronically exposed to lead in soft water, the recommendation
of a maximum lead in water concentration of 0.03 mg/1 (National
Academy of Sciences, 1973) appears to be adequate for the protection
of most fishes in the aquatic environment.
EXPOSURE OP FISH TO CHROMIUM
There is a paucity of data regarding the effect of chromium
on freshwater fish as a result of chronic exposure. Olson
(1958) and Olson and Foster (1956, 1957). as reported in National
Academy of Sciences (1973) and in McKim and Benoit (1971) t exposed
eggs and fry of rainbow trout and chinook salmon to chromium as
sodium dichromate and determined that fry were more sensitive than
eggs to chromium and that the effect of this metal was cumulative.
Pickering (1971» unpublished data, in National Academy of Sciences,
1973) reported a safe concentration of 1.0 mg/1 for chromium and
fathead minnows in hard water. Benoit (1976, personal communica-
tion) determined that the MATC's for both rainbow and brook trout
chronically exposed to chromium were between 0.2 and 0.^ mg/1.
Based on these data, the maximum permissible concentration of
chromium in the aquatic environment is recommended to be 0.05 rag/1
(National Academy of Sciences, 1973) •
A review of the MATC values generated in the present study indicate
that safe concentrations vary from 51 to >2l67 |ig/l for the 7
freshwater fish species tested.
Chromium concentrations <19^ |ig/l were apparently safe over
63
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the 60 day exposure for the survival and growth in total
length of rainbow trout but wet weights of fry were
significantly lower than those of controls at concentrations
>105
The survival and total length of lake trout exposed to
concentrations as high as 822 p.g/1 for 60 days were apparently
unaffected.
However, as with rainbow trout, deleterious effects on wet
weight of fry were observed at chromium concentrations
>19^ ng/1.
Channel catfish survival and mean total length were significantly
affected by the end of 30 days of exposure to chromium concen-
trations >305 |ig/l. However, at the end of 60 days of exposure
to 305 p.g/l» survival, total length and wet weight of fry were
no longer significantly different from controls. Possibly
this was due to the hardiness of the fish which survived the
initial JO day exposure. Therefore, the MATC estimated for
channel catfish is based on survival and total length data
from the first 30 days of exposure.
There was great variability, some of which was due to feeding
problems, between replicates during the exposure of blue6ill
to chromium. Variabilities were such that statistical
significance could not be ascribed to most observations.
However, wet weights of fry after 60 days exposure to 1122 p.g/1
chromium was significantly different from controls and was the
criterion for estimation of the MATC.
The MATC for white sucker exposed to chromium in soft water
was estimated on the basis of wet weight measurements of fry,
since weights again seemed to be the most sensitive of the
parameters monitored.
Generally, estimates of MATC values on the basis of northern
pike and walleye data are questionable. Severe feeding
problems with both of these species greatly influenced
observations and precluded adequate statistical treatment
of the data.
Despite the feeding problems experienced with northern pike
and walleye, the majority of the data generated from these
chromium exposures indicated a very significant cumulative
effect on fry. This cumulative effect was especially
obvious in the results of exposure of rainbow and lake trout
to this metal. In each of these cases, initial exposures at
64
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mg/1 concentrations resulted in significant inimical effects
at the end of 60 days at all chromium concentrations to which
fry were exposed. Each test was repeated with fry exposed to
a lower range of concentrations and again, in each case,
significant effects of chromium on fry were not detected until
the end of 60 days of exposure.
From the data presented, it appears that the current recommenda-
tion is not stringent enough and the maximum permissible concen-
tration of chromium in the aquatic environment should be reduced.
Consideration of the magnitude of this reduction should obviously
include an analysis of the chronic effects of chromium on aquatic
invertebrates such as crustaceans as well as recognition of the
cumulative toxicity of this metal.
EXPOSURE OF FISH TO COPPER
Many data considering the chronic toxicity of copper to fresh-
water fish are currently available. Mount (1968) reported that
the concentration of copper (as CuSOjj,) in hard water (200 mg/1
as CaCO-^) which did not affect growth and reproduction of
fathead minnows was between 3 and 8$ of the 96 hour median
tolerance limit, that is, between 15 and 33 M-g/l» for that
species. Similar tests with fathead minnows exposed to copper
in soft water (31.^ mg/1 as CaGO^) indicated the MATC was
between 10.6 and 18.^4- |ig/l (Mount and Stephan, 1969). Among
the parameters measured and found to be affected by exposure
to 18.^- p.g/1 copper were survival and length through 30-120
days continuous exposure. McKim and Benoit (1971) exposed
brook trout to Cu(II) in soft water (45 mg/1 as CaCO-^) for
22 months and from the results estimated a MATC between 9-5
and 17.^4- p.g/1. They found no significant difference in sensi-
tivity between fry from parents previously expo'sed to copper
and those from unexposed parents. A copper concentration of
32.5 M-g/1 significantly affected the hatchability of trout
eggs but lower test concentrations did not. The growth of
fry was reduced by all copper concentrations tested during
the first 23 weeks of exposure. However, after 23 weeks,
fish at concentrations of <9«5 M-g/1 grew as well as controls.
These limits for the MATC were apparently confirmed for brook
trout in a subsequent study by McKim and Benoit (197^) in
which they attempted to determine whether or not shorter
duration exposures of fish would yield comparable and meaning-
ful data relative to their earlier study. Results of the
latter study indicated that at copper concentrations <9.^ p.g/1
there were no effects on survival, growth, number of eggs
produced, hatchability and survival of fry subsequently.
65
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Drummond et al., 1973 attempted to identify parameters which
would be useful and reliable in predicting long-term effects
of copper on brook trout based on short-term exposures. Copper
concentrations selected for this study were based on the effect-
no effect concentrations reported by McKim and Benoit (1971)
and parameters observed included cough frequency, locomotion
activity and feeding behavior. Based on their results, Drummond
et al. concluded that selected parameters were significantly
affected at copper concentrations >6 |ig/l, with greatest
distinctions obvious at concentrations >9 M-g/1.
Studies in which blood characteristics, such as red blood
cell count, hematocrit, hemoglobin and associated enzymes, of
freshwater fish exposed to copper were measured in an attempt
to detect effect due to this exposure yielded results compar-
able to those from studies in which reproduction or other
parameters were measured.
McKim et al., (1970) determined an MATC between 9.5 and 17.4
Hg/1 copper based on blood plasma glutamic oxalacetic transaminase
(PGOT) concentrations for brook trout exposed for 6 to 21 days
to cupric sulfate in soft water (46 mg/1 as CaCO^). Christensen
et al. (1972) exposed brown bullheads to copper (as CuSOi(,.5H20)
for~l>, 30 and 600 days in hard water (202 mg/1 as CaCOo) and
estimated an MATC between 11 and 16 jig/1.
The MATC values estimated for brook trout and channel catfish
exposed to copper agree very closely with those previously
reported. Brook trout egg hatchability was significantly
reduced by exposure to concentrations >13 |ig/l copper and total
length and wet weight were reduced after 60 days exposure to
5 (ig/1 in soft water (37-5 mg/1 as CaCO^). Hard water (187.0
mg/1 as CaCC^) appeared to prolong the time to observed effect
while not materially affecting the MATC value. That is, in
hard water, effects appeared later in the exposure (31-60 days),
but at similar concentrations (i.e., 8 vs. 5 M-g/1) • Channel
catfish data followed a pattern similar to that of brook trout
but at slightly higher concentrations, resulting in a higher
range defining the estimated MATC. Survival of newly-hatched
fry through the first 30 days of exposure was comparable in
terms of both effective concentration and relative percentages
in soft and hard water. MATC's estimated for channel catfish
exposed to copper in soft and hard water are virtually identical.
Interpretation of data from walleye exposures is difficult due
to significant problems in feeding the fry. For example, test
concentrations of copper >47 (^g/1 significantly reduced hatching
66
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sTiccess of walleye eggs exposed in soft water while roughly
comparable concentrations in hard water had no effect on
hatchability of eggs, however, no fry (including controls)'
survived to day 30 in hard water while concentrations <8 |ig/l
copper in soft water had no significant effect.
In any event,current water quality criteria (National Academy
of Sciences, 1973) recommend that the concentration of copper
in the aquatic environment not exceed 0.1 of the 96-hour LC50 for
the species of interest in any particular receiving water. Based
on the data in this study, the application factor should probably
be changed.
EXPOSURE OF FISH TO CADMIUM
Cadmium has been shown to be an extremely potent and cumulative
toxicant. Pickering and Gast (1972) reported an estimated MATC
between 37 and 57 M-g/1 cadmium for fathead minnows exposed to
cadmium sulfate in hard water (200 mg/1 as CaCO^). Cadmium
concentrations >57 p.g/1 significantly decreased survival of
developing embryos, apparently the most sensitive stage in the
life cycle of this fish. Concentrations <37 l^g/1 cadmium did
not affect survival, growth or reproduction of these fish.
Hatchability of fathead minnow eggs exposed to 57 l-ig/1 cadmium
§as not significantly different from that of controls. These
nvestigators observed a slow, cumulative mortality occurring
during exposures.
Eaton (197*0 estimated on MATC between 31 and 80 p.g/1 cadmium
for bluegill chronically exposed to cadmium sulfate in hard
water (200 mg/1 as CaCOo) under conditions replicating those
of Pickering and Gast (1972). As in this earlier study, Eaton
observed that fish mortalities generally occurred late into the
exposure periods (i.e», after l5l days), thus corroborating
the cumulative effects of cadmium on freshwater fishes.
In the present study, we observed that exposure to cadmium
concentrations as high as 91 p.g/1 had no significant effect
on hatchability of brook trout, channel catfish and walleye
eggs in soft (35.0-37.0 mg/1 as CaCCH) and hard (185.0-187.0
mg/1 as CaCO^) water. Effects of cadmium on survival of brook
trout fry were not manifested at any concentration during the
first 30 days exposure in both soft (37.0 mg/1 as CaC03) and
hard (188.0 mg/1 as CaCO^) water. However, after 60 days,
survival in soft water (37.0 mg/1 as CaCOo) was affected at
cadmium concentrations of >6 [ig/1. Similarly, at 60 days,
these concentrations significantly affected total length and
wet weight of brook trout fry. Wet weight of fry was also
67
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significantly reduced after 60 days exposure to 3 P-g/l cadmium.
In hard water (188.0 mg/1 as CaCOo), significant effects on
brook trout fry survival, length and weight were observed in
cadmium concentrations >12 p.g/1 after 60 days. Fry exposed to
7 (J.g/1 of cadmium were not apparently affected.
The survival of channel catfish fry exposed to cadmium concen-
trations >1? |ig/l for 30 days in soft water (37.0 mg/1 as CaC03),
was significantly reduced as compared to that of controls. The
survival of catfish fry in hard water (185.0 mg/1 as CaCO-^) was
too variable for statistical treatment. After 60 days exposure
to cadmium in soft water (37.0 mg/1 as CaC03), significant
effects on survival, length and weight were difficult to
demonstrate due to the variability among replicates. In hard
water (185.0 mg/1 as CaCO^), significant effects of cadmium
on the latter two parameters could be established for concen-
trations >17 (J-g/1.
Therefore, MATC's for cadmium and channel catfish eggs and fry
were estimated based on percentage survival at the end of 30 days
exposure in soft water (37.0 mg/1 as CaCCH) and on total length
and wet weight after 60 days exposure in hard water (185.0 mg/1
as
Previously mentioned difficulties in the feeding of walleye
fry resulted in our inability to ascribe statistical significance
to most of these data. However, exposure to cadmium concentrations
>2^ |ig/l resulted in 100$ mortality of walleye fry within 30 days
exposure in soft water (35.0 mg/1 as CaCCh). There was some
survival (2-18$) of fry in control and cadmium treatments <9 M-g/l«
In hard water (187.0 mg/1 as CaCO^) there was 100$ mortality of
all fry in control and all cadmium treatments.
No physical abnormalities were observed in any of the fish
exposed to cadmium. Pickering and Cast (1972; reported observ-
ing many blood clots in the vascular system of cadmium-exposed
fathead minnows. Sangalang and O'Halloran (1972,1973) observed
damage to the testes of brook trout exposed to 25 p.g/1 cadmium
(as- cadmium chloride), for 2^ hours and to 10 jig/1 cadmium
for 21 days in soft water (20.0 mg/1 as CaC03). Gross damage
included general discoloration ("dark purple-brown patches")
of the testes. A histological examination of tissues revealed
distended blood vessels with collapsed walls in some places
and local accumulations of erythrocytes. These investigators
also suggested that steroid (androgen) synthesis was affected
in fish with damaged testicular tissues. None of this testicular
damage was observed after exposure of brook trout to cadmium
concentrations of from 0-5 (J-g/1. These data indicate an MATC
68
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^tween 5 and 10 |ig/l based on testicular damage. These values
agree quite well with the results of the present study.
Based on these data, it appears that the recommendation for a
maximum cadmium concentration of 0.00^ mg/1 in water with a
hardness <100 mg/1 as CaC03 (National Academy of Sciences, 1973)
may just be "barely adequate and possibly should be decreased
further. This same maximum concentration should be extended to
species extant in water with a hardness of >100 mg/1 as CaGO^
since the currently recommended limit of 0«03 mg/1 (National
Academy of Sciences, 1973) is clearly inadequate to protect
aquatic life.
The one significant problem in protocol encountered during this
study was the rearing of both northern pike and walleye fry
under test conditions. Both of these species are highly active,
voracious feeders which require a stenothermal medium for
optimum development during specific portions of their life cycle.
Studies of pond culture of northern pike (White, 1968) and
relationship of water temperature to egg hatching (Walker, 1968)
indicate that temperatures of 10°-12°C are more favorable for
hatching than the 17°-18°C temperatures recommended in the egg
and fry protocol. Furthermore, a gradual increase in water
temperature from 10°-12°C to 1^°-15°C (rather than to 17°-18°C)
would have been adequate to induce feeding without encouraging
kbhe excessive cannibalism experienced. Walker (1968) recommended
Ithe stocking of tanks of swim-up muskellunge fry with white
sucker fry to provide an adequate food source. In the present
study, we found that during the first few weeks of exposure of
northern pike fry, we could provide adequate numbers of white
sucker fry as food. Northern pike fry appeared to feed most
readily on forage fish which were 1/2 to 2/3 their own size.
When offered fish food significantly smaller or larger than
this range, northern pike fry became cannibalistic. Possibly,
rearing these fry at lower temperatures (i.e. 1^°-15°C) would
help to minimize cannibalism.
For walleye, it has been suggested (Lloyd L. Smith, Jr.,
personal communications) that to maximize feeding, fry should
be maintained in aquaria with illumination from above only.
The use of this procedure apparently results in a more even
distribution of fish in the aquarium and improved feeding
activity. A gradual increase in water temperature from 15° C
to 18-21° C during swim-up may also favor increased feeding.
In general, the recommendations of maximum permissible concen-
trations of 0.05 mg/1 for chromium, 0.1 of the 96-hour LC50
for copper and 0.000^-0.004 mg/1 in soft water and 0.0003-0.03 mg/1
in hard water for cadmium in the aquatic environment (National
69
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Academy of Sciences, 1973) appear to be inadequate based on the
results of the present study. In order to suggest concentrations
which may be more appropirate for the protection of the aquatic
environment, a rationale which related acute and chronic toxicity,
the bioconcentration potential of these metals and a margin of
safety was employed. Acute toxicity data for freshwater fishes
exposed to these metals were compiled and then compared with chronic
toxicity data from this study to produce a factor relating the two.
The factor calculated was 0.001. Multiplying the lowest concen-
trations reported for 96-hour LC50 values of each metal and
appropriate freshwater fishes by this factor and then applying
a safety factor resulted in the following recommendations of
maximum permissible concentrations of each metal in the aquatic
environment: 0.03 mg/1 for chromium, 0.1 p.g/1 for copper and
0.01 p.g/1 for cadmium. These concentrations, based on the
results of the present study, appear to be more appropriate
than are others presently available.
In summary, the good correlation between MATC values estimated
from the results of these relatively short-term exposures and
those from much longer-term, full-chronic studies indicates
that egg and fry studies are an cost-effective (in terms of
both time and money) and reliable means of assessing the
potential hazard to selected freshwater fish species of certain
compounds which may impact the aquatic environment.
70
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SECTION VII
LITERATURE CITED
APHA, AWWA and WPCF, 19?1. Standard Methods for the Examination
of Water and Wastewater, 13th edition. Amer. Public Health
Assoc., Washington, DC: 8?5- pp.
Battelle-Columbus Laboratories, 1971. Effects of chemicals on
aquatic life. Water Quality Criteria Data Book, Vol. 3«
U.S. Environmental Protection Agency, Water Pollut. Control
Res. Series No. 18050GWV05/71.
Brungs, W.A., 1969. Chronic toxicity of zinc to the fathead
minnow, Pimephales promelas Rafinesque. Trans. Amer.
Fish. Soc. 98:272-279.
Brungs, W.A., E.N. Leonard and J.M. McKim, 1973* Acute and
long-term accumulation of copper by the brown bullhead,
Ictalurus nebulosus. J. Fish. Res. Bd. Can. 30: 583-586.
Cairns, J., Jr. and A. Scheier, 1957. The effects of temperature
and hardness of water upon the toxicity of zinc to the
common bluegill (Lepomis macrochirus Raf.) Notulae Natur.
(Philadelphia) no. 299: 1-12.
Ohristensen, G.M., J.M. McKim, W.A. Brungs and E.P. Hunt, 1972.
Changes in the blood of the brown bullhead (Ictalurus
nebulosus Leseur) following short and long term exposure
to copper (II). Toxicol. Appl. Pharmacol. 23: ^17-^27.
Davies, P.H. and W.H. Everhart, 1973. Effects of chemical
variations in aquatic environments: Volume III Lead
toxicity to rainbow trout and testing application factor s
concept. U.S. Environmental Protection Agency, Ecological
Res. Series No. EPA-R3-73-011C: 80 pp.
Drummond, R.A., W.A. Spoor and G.F. Olson, 1973. Some short-
term indicators of sublethal effects of copper on brook
trout, Salvelinus fontinali_s. J. Fish. Res. Bd. Can.
30: 698-701.
Eaton, J.G., 1970. Chronic malathion toxicity to the bluegill
(Lepomis macrochirus Rafinesque). Water Res. ^: 673-68^.
Eaton, J.G., 197^. Chronic cadmium toxicity to the bluegill
(Lepomis macrochirus Rafinesque). Trans. Amer. Fish. Soc.
*M 729-735.
Erickson, S.J., 1972. Toxicity of copper to Thallasiosira
pseudonana in unenriched inshore seawater. J. Phycol.
8: 318-323.
71
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Hermanutz, R.O., L.H. Mueller and K.D. Kempfert, 1973. Captan
toxicity to fathead minnows (Pimephales promelas) ,
bluegills (Lepomis macrochirus) and brook trout (Salvelinng
fontinalis). J. Fish. Res. Bd. Can. 30: 1811-18177
Leland, H.V. , E.D. Copenhaver and D.J. Wilkes, 1975« Heavy
metals and other trace elements. J. Water Pollut. Control
Fed. 4-7: 1635-1656.
Lemke, A.E. , 1969* A water hardener for experimental use.
J. Amer. Water Works Assoc. 61:
Macek, K.J., M.E. Barrows, R.F. Krasny, and B.H. Sleight,
1975> Bioconcentration of 1^-C-pesticides by bluegill
sunfish during continuous aqueous exposure. Proc. of
Symposium on Structure Activity Correlation in Studies
of Toxicity and Bioconcentration with Aquatic Organisms.
Canada Centre for Inland Waters, Burlington, Ont.
Macek, K.J. , K.S. Buxton, S.K. Derr, J.W. Dean and S. Sauter,
1976. Chronic toxicity of lindane to selected aquatic
invertebrates and fishes. U.S. Environmental Protection
Agency, Ecological Res. Series No. EPA-600/3-76-0^6 : 55 pp.
Macek, K.J. , K.S. Buxton, S. Sauter, S. Gnilka, and J.W. Dean,
1976. Chronic toxicity of atrazine to selected aquatic
invertebrates and fishes. U.S. Environmental Protection
Agency, Ecological Res. Series No. EPA-600/3-76-0^-7 : 55 pp.
McKee, J.E. and H.W. Wolf, eds., 1963. Water quality criteria.
2nd edit. State Water Quality Control Board, Sacramento,
Calif: 5^8 pp.
McKim, J.M. and D.A. Benoit, 1971. Effects of long-term exposures
to copper on survival, growth, and reproduction of brook
trout (Salvelinus fontinalis) . J. Fish. Res. Bd. Can.
28 : 655-662.
McKim, J.M. and D.A. Benoit, 197^» Duration of toxicity tests
for establishing "no effect" concentrations for copper with
brook trout (Salvelinus fontinalis). J. Fish. Res. Bd. Can.
31: 449-^52.
McKim, J.M,, G.M. Christensen and E.P. Hunt, 1970. Changes
in the blood of brook trout (Salvelinus fontinalis) after
short-term and long-term exposure to copper. J. Fish Res.
Bd. Can. 27: 1883-1889.
McKim, J.M. , G.M. Christensen, J.H. Tucker, D.A. Benoit and
M.J. Lewis, 1973. Effects of pollution on freshwater
fish. J. Water Pollut. Control Fed. k$i 1370-1^07.
72
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McKim, J.M. , G.M. Christensen, J.H. Tucker, D.A. Benoit and M.J.
Lewis, 197^. Effects of pollution on freshwater fish.
J. Water Pollut. Control Fed. ^6: 15^0-1591.
Mount, D.I., 1966. The effect of total hardness and pH on
acute toxicity of zinc to fish. Air Water Pollut. 10: ^9-56.
Mount, D.I., 1968. Chronic toxicity of copper to fathead
minnows (Pimephales promelas, Rafinesque) . Water Res.
2: 215-223.
Mount, D.I. and W.A. Brungs, 196?. A simplified dosing
apparatus for fish toxicology studies. Water Res. 1: 21-29.
Mount, D.I. and C.E. Stephan, 196?. A method for establishing
acceptable toxicant limits for fish-malathion and the
butoxyethanol ester of 2,*J— D. Trans. Amer. Fish. Soc.
96: 185-193.
Mount, D.I. and C.E. Stephan, 1969. Chronic toxicity of copper
to fathead minnow (Pimephales promelas) in soft water.
J. Fish. Res. Bd. Can. 26: 2^9-2^57.
National Academy of Sciences, 1973» Water Quality Criteria,
1972. U.S. Environmental Protection Agency, Ecological
Res. Series No. EPA-R3-73-033: 59^ PP.
Olson, P. A., 1958. Comparative toxicity of Cr (VI) and Cr (III)
in salmon. Hanford biology research-annual report for 1957
(HW 53500) Hanford Atomic Products Operation, Richland,
WA: 215-218 (see National Academy of Sciences, 1973).
Olson, P. A. and R.F. Foster, 1956. Effect of chronic exposure
to sodium dichromate on young Chinook salmon and rainbow
trout. Hanford biology research-annual report for 1955
(HW ^1500) Hanford Atomic Products Operation, Richland,
WA: 35-^7 (see National Academy of Sciences, 1973).
Olson, P. A. and R. F. Foster, 1957. Further studies on the
effect of sodium dichromate on juvenile chinook salmon.
Hanford biology research-annual report for 1956 (HW ^7500)
Hanford Atomic Products Operation, Richland, WA: 21^-224-
(see National Academy of Sciences, 1973).
Pagenkopf, G.K., R.C. Russo and R.V. Thurston, 197^. Effects
of complexation on toxicity of copper to fishes. J. Fish.
Res. Bd, Can. 31:
Perkin-Elmer, 1973* Analytical Methods for Atomic Absorption
Spectrophotometry. Perkin-Elmer Corp., Norwalk, Connecticut.
Pickering, Q.H. and M.H. Cast, 1972. Acute and chronic toxicity
of cadmium to the fathead minnow (Pimephales promelas) . '
J. Fish. Res. Bd. Can. 29: 1099-1106.
73
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Pickering, Q.H. and C. Henderson, 19&5. The acute toxicity of
some heavy metals to different species of warm water fishes.
Proc. 19th Indust. Waste Conf . , Purdue Univ.: 578-591.
Sangalang, G.B. and M.J. O'Halloran, 1972. Cadmium-induced
testicular injury and alterations of androgen synthesis
in brook trouto Nature 2*4-0:
Sangalang, G.B. and M.J. O'Halloran, 1973. Adverse effects of
cadmium on "brook trout testis and on in vitro testicular
androgen synthesis. Biol. Reprod0 9'
Steele, R.G.D. and J.H. Torrie, I960. Principles and Procedures
of Statistics. McGraw-Hill, New York: 481 pp. ~
Tabata, K, , 1969. Studies on the toxicity of heavy metals to
aquatic animals and the factors to decrease the toxicity.
IV. On the relationship between the toxicity of heavy
metals and the quality of environmental water. Bull.
Tokai Fish. Res. Lab. 58: 2^-3-253.
U.S. EPA, 1971. Methods for chemical analysis of water and
wastes. Analytical Quality Control Laboratory, Cincinnati, OH.,
U.S. Environmental Protection Agency, Water Pollut. Control
Res. Series No. 1805GWV05/71.
U.S. EPA, 1972a. Proposed recommended bioassay procedure for
egg and fry stages of freshwater fish. U.S. Environmental
Protection Agency, National Water Quality Laboratory, Duluth, MN,
U.S. EPA, 1972bo Handbook for analytical quality control in
water and wastewater laboratories, Analytical Quality Control
Laboratory, Cincinnati, OH.
Walker, K.W. , 1968. Temperature control in northern pike and
muskellunge egg hatching, p 13-17. In: Proc. of North Central
Warm Water Fish Culture Workshop, Feb. 15-16, Ames, 10,
White, Bo, 1968. Northern pike pond culture, p 21-23. In: Proc.
of North Central Warm Water Fish Culture Workshop, Feb.
15-16, Ames, 10.
74
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TECHNICAL REPORT DATA
(Please read Instructions on the reverse before completing)
; REPORT NO.
EPA-600/3-76-105
3. RECIPIENT'S ACCESSION NO.
4.TITLE ANDSUBTITLE
EFFECTS OF EXPOSURE TO HEAVY METALS ON SELECTED
FRESHWATER FISH
5. REPORT DATE
npfnhpr 1Q76 (Issuing date)
6. PERFORMING ORGANIZATION CODE
7. AUTHOR(S)
Scott Sauter, Kenneth S. Buxton, Kenneth J,
Macek and Sam R. Petrocelli
8. PERFORMING ORGANIZATION REPORT NO
9. PERFORMING ORGANIZATION NAME AND ADDRESS
EG&G, Bionomics
Aquatic Toxicology Laboratory
Wareham, Massachusetts 02571
10. PROGRAM ELEMENT NO.
11. CONTRACT/GRANT NO.
Contract
68-01-0740
12. SPONSORING AGENCY NAME AND ADDRESS
Environmental Research Laboratory - Duluth, Minn.
Office of Research and Development
U.S. Environmental Protection Agency
13. TYPE OF REPORT AND PERIOD COVERED
Final
Diiliit-h.
SSftflA
14. SPONSORING AGENCY CODE
EPA/600/03
5. SUPPLEMENTARY NOTES
Subtitled: Toxicity of Copper, Cadmium, Chromium and Lead to Eggs and Fry
of Seven Fish Species
6. ABSTRACT
Embryo and larvae of rainbow trout, lake trout, channel catfish, bluegill, white
sucker, northern pike, and walleye were exposed for 60 days after hatch to lead
and chromium in soft water. Brook trout, channel catfish, and walleyes were also
exposed for 60 days after hatch to copper and cadmium in soft and hard water. The
effects on survival and growth indicated that copper and cadmium were toxic at
much lower concentrations than lead and chromium. Water hardness did not appear
to have a significant effect on the observed toxicity in most cases.
KEY WORDS AND DOCUMENT ANALYSIS
DESCRIPTORS
b.lDENTIFIERS/OPEN ENDED TERMS C. COS AT I Field/Group
Bioassay
Freshwater fishes
Metals
Toxicity
Water pollution
Lead, Chromium, Cadmium,
Copper, Embryo-larvae,
Brook trout, Rainbow
trout, Lake trout, Channep.
catfish, Bluegill, White
sucker, Northern pike,
Walleye
06F
06A
06S
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RELEASE TO PUBLIC
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21. NO. OF PAGES
85
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UNCLASSIFIED
22. PRICE
EPA Form 2220-1 (9-73)
75
a U.S. GOVERNMENTPftlXnNGOmCfclOT-757-056/5503
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