Document ID: EPA-HQ-OW-2003-0079-0036
Agency: epa
Document Type: Supporting & Related Material
Title: 
Posted Date: 2003-11-18T05:00Z

H­
2
Evaluation
of
the
Chronic
Data
Available
for
Freshwater
Species
Following
is
a
species­
by­
species
discussion
of
each
chronic
test
on
copper
evaluated
for
this
document.
Also
presented
are
the
results
of
regression
analysis
and
probability
distribution
analysis
of
each
dataset
that
was
from
an
acceptable
chronic
test
and
contained
sufficient
acceptable
data.
For
each
such
dataset,
this
appendix
contains
a
figure
that
presents
the
data
and
regression/
probability
distribution
line.

Brachionus
calyciflorus.
The
chronic
toxicity
of
copper
was
ascertained
in
4­
day
renewal
tests
conducted
at
regular
intervals
throughout
the
life
of
the
freshwater
rotifer,
B.
calyciflorus
(
Janssen
et
al.
1994).
The
goal
of
this
study
was
to
develop
and
examine
the
use
of
this
rotifer
as
a
viable
test
organism.
The
effect
of
copper
on
the
age­
specific
survivorship
and
fertility
of
B.
calyciflorus
was
determined,
but
no
individual
replicate
data
were
provided
and
only
three
copper
concentrations
were
tested,
which
precludes
these
data
from
further
regression
analysis.
Chronic
limits
based
on
the
intrinsic
rate
of
natural
increase
were
2.5

g/
L
total
copper
(
NOAEC)
and
5.0

g/
L
total
copper
(
LOAEC).
The
chronic
value
determined
via
traditional
hypothesis
testing
is
3.54

g/
L
total
copper
(
Table
2a).

Campeloma
decisum.
Adult
C.
campeloma
were
exposed
to
five
concentrations
of
total
copper
and
a
control
(
Lake
Superior
water)
under
flow­
through
conditions
in
two
6­
week
studies
conducted
by
Arthur
and
Leonard
(
1970).
Adult
survival
in
the
two
separate
chronic
copper
toxicity
test
trials
was
markedly
reduced
in
the
two
highest
copper
concentrations,
14.8
and
28.0

g/
L,
respectively.
The
authors
reported
that
growth,
as
determined
from
cast
exoskeleton,
was
not
measurable
for
this
test
species,
although
the
authors
did
observe
that
the
adult
snails
would
not
consume
food
at
the
two
highest
copper
concentrations.
Control
survival
was
80
percent
or
greater.
Chronic
values
of
10.88

g/
L
total
copper
were
obtained
for
survival
based
on
the
geometric
mean
of
the
NOAEC
and
LOAEC
of
8.0
and
14.8

g/
L,
respectively,
in
both
tests.
The
corresponding
EC20s
were
8.73
and
10.94

g/
L
(
Table
2a).

Ceriodaphnia
dubia.
The
chronic
toxicity
of
copper
to
C.
dubia
was
determined
in
ambient
river
water
collected
upstream
of
known
point­
source
discharges
of
domestic
and
industrial
wastes
as
part
of
a
water
effect
ratio
study
(
Carlson
et
al.
1986).
In
this
study,
survival
and
young
production
of
C.
dubia
were
assessed
using
a
7­
day
life­
cycle
test.
Organisms
were
not
affected
at
total
copper
concentrations
ranging
from
3
to
12

g/
L
(
5
to
10

g/
L
dissolved
copper).
There
was
a
62.7
percent
reduction
in
survival
and
97
percent
reduction
in
the
mean
number
of
young
produced
per
female
at
32

g/
L
total
copper
(
27

g/
L
dissolved
copper).
No
daphnids
survived
to
produce
young
at
91

g/
L
total
copper.
Control
survival
during
the
study
was
80
percent,
which
included
one
male.
The
chronic
value
EC20
selected
for
C.
dubia
in
this
study,
9.17

g/
L
derived
from
a
nonlinear
regression
evaluation,
was
based
on
mean
number
of
young
produced
(
reproduction).

The
effects
of
water
hardness
on
the
chronic
toxicity
of
copper
to
C.
dubia
were
assessed
by
Belanger
et
al.
(
1989)
using
7­
day
life­
cycle
tests.
C.
dubia
2
to
8
hours
old
were
exposed
to
copper
in
ambient
surface
water
from
the
New
and
Clinch
Rivers,
Virginia.
Mean
water
hardness
levels
were
179
and
94
mg/
L
as
CaCO3,
respectively.
Test
water
was
renewed
on
days
3
and
5.
The
corresponding
chronic
values
for
reproduction
based
on
the
NOAEC
and
LOAEC
approach
were
7.9
and
<
19.3

g/
L
dissolved
copper,
respectively.
The
EC20
value
for
number
of
young
(
neonates)
produced
in
Clinch
River
water
(
water
hardness
of
94
mg/
L
as
CaCO3)
was
19.36

g/
L
dissolved
copper.
The
EC20
for
young
produced
in
New
River
water
was
not
calculated.
The
chronic
values
were
converted
to
total
copper
using
the
freshwater
conversion
factor
for
copper
0.96
(
e.
g.,
7.897/
0.96).
The
resulting
total
chronic
values
for
the
New
and
Clinch
rivers
are
8.23
and
20.17

g/
L,
respectively.

Copper
was
one
of
12
toxicants
examined
by
Oris
et
al.
(
1991)
in
their
comparisons
between
a
4­
day
survival
and
reproduction
toxicity
test
utilizing
C.
dubia
and
a
standard
7­
day
life­
cycle
test
for
the
H­
3
species.
The
reported
7­
day
chronic
values
for
survival
and
reproduction
(
mean
total
young
per
living
female)
in
two
tests
based
on
the
traditional
hypothesis
testing
techniques
were
24.5
and
34.6

g/
L
total
copper.
Comparable
point
estimates
for
these
7­
day
tests
could
not
be
calculated
using
regression
analysis.

Daphnia
magna.
Blaylock
et
al.
(
1985)
reported
the
average
numbers
of
young
produced
for
six
broods
of
D.
magna
in
a
14­
day
chronic
exposure
to
copper.
A
significant
reduction
was
observed
in
the
mean
number
of
young
per
female
at
a
concentration
of
30

g/
L
total
copper,
the
highest
copper
concentration
tested.
At
this
concentration,
young
were
not
produced
at
brood
intervals
5
and
6.
Reproduction
was
not
affected
at
10

g/
L
total
copper.
The
chronic
value
determined
for
this
study
(
17.32

g/
L
total
copper)
was
based
on
the
geometric
mean
of
the
NOAEC,
10

g/
L,
and
LOAEC,
30

g/
L.

Van
Leeuwen
et
al.
(
1988)
conducted
a
standard
21­
day
life­
cycle
test
with
D.
magna.
The
water
hardness
was
225
mg/
L
as
CaCO3.
Carapace
length
was
significantly
reduced
at
36.8

g/
L
total
copper,
although
survival
was
100
percent
at
this
concentration.
Carapace
length
was
not
affected
at
12.6

g/
L
total
copper.
No
daphnids
survived
at
110

g/
L
concentration.
The
highest
concentration
not
significantly
different
from
the
control
for
survival
was
36.8

g/
L.
The
lowest
concentration
significantly
different
from
the
control
based
on
survival
was
110

g/
L,
resulting
in
a
chronic
value
of
63.6

g/
L
for
survival.
The
chronic
value
based
on
carapace
length
was
21.50

g/
L.
The
21­
day
EC10
as
reported
by
the
author
was
5.9

g/
L
total
copper.

Chronic
(
21­
day)
renewal
toxicity
tests
were
conducted
using
D.
magna
to
determine
the
relationship
between
water
hardness
(
nominal
values
of
50,
100,
and
200
mg/
L
as
CaCO3,
respectively)
and
the
toxicity
of
total
copper
(
Chapman
et
al.
unpublished
manuscript).
All
test
daphnids
were
<
1
day
old
at
the
start
of
the
tests.
The
dilution
water
was
well
water
from
the
Western
Fish
Toxicology
Station
(
WFTS),
Corvallis,
Oregon.
Test
endpoints
were
reproduction
(
total
and
live
young
produced
per
female)
and
adult
survival.
The
survival
of
control
animals
was
100
percent
at
nominal
water
hardness
levels
of
50
and
200
mg/
L
as
CaCO3,
and
80
percent
at
a
hardness
of
100
mg/
L
as
CaCO3.
The
chronic
values
for
total
young
produced
per
female
(
fecundity)
based
on
the
geometric
mean
of
the
NOAEC
and
LOAEC
were
13.63,
29.33,
and
9.53

g/
L
at
the
nominal
hardness
levels
of
50,
100,
and
200
mg/
L
as
CaCO3,
respectively.
The
corresponding
EC20
values
for
reproduction
calculated
using
nonlinear
regression
analysis
were
12.58,
19.89,
and
6.06

g/
L
total
copper.
The
chronic
toxicity
of
copper
to
D.
magna
was
somewhat
ameliorated
from
an
increase
in
water
hardness
from
50
to
100
mg/
L
as
CaCO3,
but
slightly
increased
from
100
to
200
mg/
L
as
CaCO3.

Daphnia
pulex.
Winner
(
1985)
evaluated
the
effects
of
water
hardness
and
humic
acid
on
the
chronic
toxicity
(
42­
day)
of
copper
to
D.
pulex.
Contrary
to
the
expectation
that
sublethal
endpoints
are
more
sensitive
indicators
of
chronic
toxicity,
reproduction
was
not
a
sensitive
indicator
of
copper
stress
in
this
species.
Water
hardness
also
had
little
effect
on
the
chronic
toxicity
of
copper
(
similar
to
D.
magna
trends),
but
humic
acid
significantly
reduced
chronic
toxicity
of
copper
when
added
to
the
varying
water
types.
The
survival
chronic
values
based
on
the
NOAEC
and
LOAEC
values
for
the
three
low
to
no
humic
acid
studies
were
4.90,
7.07,
and
12.25

g/
L
total
copper
at
hardnesses
of
57.5,
115,
and
230
(
0.15
mg/
L
HA)

g/
L
as
CaCO3,
respectively.
The
EC20
values
calculated
for
the
low
and
high
hardness
studies
using
nonlinear
regression
techniques
were
2.83
and
9.16

g/
L
at
hardness
values
of
57.5
and
230
(
0.15
mg/
L
HA)

g/
L
as
CaCO3,
respectively.

Clistoronia
magnifica.
The
effects
of
copper
on
the
lifecycle
of
the
caddisfly,
C.
magnifica,
were
examined
in
Nebeker
et
al.
(
1984b).
The
test
included
continuous
exposure
of
first­
generation
aquatic
larvae
and
pupae
through
to
a
third
generation
of
larvae.
A
significant
reduction
in
adult
emergence
occurred
at
13.0

g/
L
total
copper
from
first­
generation
larvae.
No
observed
adverse
effect
to
adult
emergence
occurred
at
8.3

g/
L
total
copper.
Percent
larval
survival
was
close
to
the
control
value
of
80
H­
4
percent.
The
chronic
value
based
on
hypothesis
testing
was
10.39

g/
L
total
copper.
The
corresponding
EC20
value
for
adult
emergence
was
7.67

g/
L
total
copper.

Oncorhynchus
mykiss.
The
growth
and
survival
of
developing
O.
mykiss
embryos
continuously
and
intermittently
exposed
to
copper
for
up
to
85
days
post­
fertilization
was
examined
by
Seim
et
al.
(
1984).
Results
only
from
the
continuous
exposure
study
are
considered
here
for
deriving
a
chronic
value.
A
flow­
through
apparatus
was
used
to
deliver
six
concentrations
and
a
control
(
untreated
well
water;
average
of
3

g/
L
copper)
to
a
single
incubation
chamber.
Continuous
copper
exposure
of
steelhead
embryos
in
the
incubation
chambers
was
begun
6
days
post­
fertilization.
At
7
weeks
post­
fertilization,
when
all
control
fish
had
hatched
and
reached
swim­
up
stage,
subsamples
of
approximately
100
alevins
were
transferred
to
aquaria
and
the
same
exposure
pattern
continued.
Dissolved
oxygen
remained
near
saturation
throughout
the
study.
Water
hardness
averaged
120
mg/
L
as
CaCO3.
Survival
of
steelhead
embryos
and
alevins
exposed
continuously
to
total
copper
concentrations
in
the
range
of
3
(
controls)
to
30

g/
L
was
greater
than
90
percent
or
greater.
Survival
was
reduced
at
57

g/
L
and
completely
inhibited
at
121

g/
L.
A
similar
effect
on
survival
was
observed
for
embryos
and
alevins
exposed
to
a
mean
of
51
(
peak
263)
and
109
(
peak
465)

g/
L
of
copper
in
the
intermittent
exposure,
respectively.
The
adverse
effect
of
continuous
copper
exposure
on
growth
(
measured
on
a
dry
weight
basis)
was
observed
at
concentrations
as
low
as
30

g/
L.
(
There
was
a
30
percent
reduction
in
growth
during
the
intermittent
exposure
at
16

g/
L.)
The
chronic
limits
for
survival
of
embryos
and
alevin
steelhead
trout
exposed
continuously
to
copper
were
16
and
31

g/
L,
respectively
(
geometric
mean
=
22.27

g/
L).
The
EC20
for
biomass
for
the
continuous
exposure
was
27.77

g/
L.

Besser
et
al.
(
2001)
conducted
an
ELS
toxicity
test
with
copper
and
the
rainbow
trout,
O.
mykiss,
starting
with
eyed
embryos
and
continuing
for
30
days
after
the
fish
reached
the
swim­
up
stage.
The
total
test
period
was
58
days.
The
test
was
conducted
in
ASTM
moderately
hard
reconstituted
water
with
a
hardness
of
approximately
160
to
180
mg/
L
as
CaCO3.
Twenty­
five
eyed
embryos
were
held
in
each
of
four
replicate
egg
cups
at
each
concentration.
Survival
was
monitored
daily.
At
the
end
of
the
test,
surviving
fish
in
each
replicate
chamber
were
weighed
(
dry
weight).
Dry
weights
were
used
to
determine
growth
and
biomass
of
surviving
fish.
The
no
observed
effect
concentrations
(
NOECs)
for
survival
and
biomass
were
both
12

g/
L
and
the
lowest
observed
effect
concentrations
(
LOECs)
for
survival
and
biomass
was
also
the
same
for
both
endpoints,
22

g/
L.
The
chronic
values
for
biomass
and
survival
based
on
the
geometric
mean
of
the
NOEC
and
LOEC
were
16.25

g/
L.
The
corresponding
EC20
for
biomass
was
20.32

g/
L.

Oncorhynchus
tshawytscha.
The
draft
manuscript
prepared
by
Chapman
(
1975/
1982)
provides
the
results
from
a
4­
month
egg
through
fry
partial
chronic
test
conducted
to
determine
the
effects
of
copper
on
survival
and
growth
of
O.
tshawytscha.
Continuous
exposure
occurred
from
several
hours
post­
fertilization
through
hatch,
swim­
up,
and
feeding
fry
stages.
The
test
was
terminated
after
14
weeks
post­
hatch.
The
dilution
water
was
WFTS
well
water.
Because
of
the
influence
of
the
nearby
Willamette
River
on
the
hardness
of
this
well
water,
reverse
osmosis
water
was
mixed
periodically
with
ambient
well
water
to
attain
a
consistent
hardness.
The
typical
hardness
of
this
well
water
was
approximately
23
mg/
L
as
CaCO3.
Control
survival
exceeded
90
percent
for
the
test.
The
measured
total
copper
concentrations
during
the
test
were
1.2
(
control),
7.4,
9.4,
11.7,
15.5,
and
20.2

g/
L,
respectively.
Copper
adversely
affected
survival
at
11.7

g/
L
copper
and
higher,
and
growth
was
reduced
at
all
copper
concentrations
tested
compared
with
the
growth
of
control
fish.
The
chronic
limits
for
copper
in
this
study
were
estimated
to
be
less
than
7.4

g/
L.
The
EC20
value
estimated
for
biomass
is
5.92

g/
L
total
copper
based
on
a
logistic
nonlinear
regression
model.

Salmo
trutta.
McKim
et
al.
(
1978)
examined
the
survival
and
growth
(
expressed
as
standing
crop)
of
embryo­
larval
and
early
juvenile
brown
trout
to
copper.
The
most
sensitive
exposure
was
with
embryos
H­
5
exposed
for
72
days.
The
NOAEC
and
LOAEC,
as
obtained
from
the
figure,
were
20.8
and
43.8

g/
L
total
copper,
respectively.
Data
were
not
available
to
calculate
point
estimates
at
the
20
percent
effect
level
using
regression
analysis.
The
chronic
value
selected
for
this
species
was
29.91

g/
L
total
copper
(
geometric
mean
of
20.8
and
43.8

g/
L
total
copper).

Salvelinus
fontinalis.
Sauter
et
al.
(
1976)
examined
the
effects
of
copper
on
selected
freshwater
fish
species
at
different
hardness
levels
(
softwater
at
37.5
mg/
L
as
CaCO3;
hardwater
at
187
mg/
L
as
CaCO3)
during
a
series
of
partial
life­
cycle
(
PLC)
tests.
The
species
tested
were
brook
trout
(
Salvelinus
fontinalis),
channel
catfish
(
Ictalurus
punctatus),
and
walleye
(
Stizostedion
vitreum).
Because
of
the
poor
embryo
and
larval
survival
of
control
animals
(
in
all
cases
less
than
70
percent),
results
from
tests
with
channel
catfish
and
walleye
were
not
included
in
Table
2a.
One
of
the
replicate
control
chambers
from
the
PLC
tests
conducted
with
brook
trout
in
hard
water
also
exhibited
poor
hatchability
(
48
percent)
and
survival
(
58
percent)
between
31
and
60
days
of
exposure.
Therefore,
the
data
for
brook
trout
in
hard
water
were
not
included
in
the
subsequent
EC20
(
regression)
analysis
either.

The
softwater
test
with
brook
trout
was
conducted
using
untreated
well
water
with
an
average
water
hardness
of
35
mg/
L
as
CaCO3.
This
PLC
exposure
consisted
of
six
copper
concentrations
and
a
control.
Hatchability
was
determined
by
examining
randomly
selected
groups
of
100
eggs
from
each
replicate
exposure
tank.
Growth
and
survival
of
fry
were
determined
by
impartially
reducing
the
total
sample
size
to
50
fry
per
tank
and
assessing
their
progress
over
30
day
intervals
up
to
60
days
post­
hatch.
The
chronic
limits
based
on
the
growth
(
wet
weight
and
total
length)
of
larval
brook
trout
after
60
days
of
exposure
to
copper
in
soft
water
were
<
5
and
5

g/
L.
The
resultant
chronic
value
for
soft
water
based
on
hypothesis
testing
was
<
5

g/
L.
The
corresponding
EC20
values
based
on
total
length,
wet
weight,
and
biomass
(
the
product
of
wet
weight
and
survival)
for
brook
trout
in
the
soft­
water
exposures
after
60
days
were
not
amenable
to
nonlinear
regression
analysis.

McKim
et
al.
(
1978)
examined
survival
and
growth
(
expressed
as
standing
crop)
of
embryo­
larval
and
early
juvenile
brook
trout
exposed
to
copper.
The
embryo
exposure
was
for
16
days,
and
the
larvalearly
juveniles
exposure
lasted
60
days.
The
NOAEC
and
LOAEC
were
22.3
and
43.5

g/
L
total
copper,
respectively.
Data
were
not
available
to
calculate
point
estimates
at
the
20
percent
effect
level
using
regression
analysis.
The
chronic
value
for
this
species
was
31.15

g/
L
total
copper
(
geometric
mean
of
22.3
and
43.5

g/
L
total
copper).

Salvelinus
namaycush.
McKim
et
al.
(
1978)
examined
the
survival
and
growth
(
expressed
as
standing
crop)
of
embryo­
larval
and
early
juvenile
lake
trout
exposed
to
copper.
The
embryo
exposure
was
for
27
days,
and
the
larval­
early­
juveniles
exposure
lasted
66
days.
The
NOAEC
and
LOAEC
were
22.0
and
43.5

g/
L
total
copper,
respectively.
Data
were
not
available
to
calculate
point
estimates
at
the
20
percent
effect
level
using
regression
analysis.
The
chronic
value
for
this
species
was
30.94

g/
L
total
copper
(
geometric
mean
of
22.0
and
43.5

g/
L
total
copper).

Esox
lucius.
McKim
et
al.
(
1978)
examined
the
survival
and
growth
(
expressed
as
standing
crop)
of
embryo­
larval
and
early
juvenile
northern
pike
exposed
to
copper.
The
embryo
exposure
was
for
6
days,
and
the
larval­
early­
juveniles
exposure
lasted
34
days.
The
NOAEC
and
LOAEC
were
34.9
and
104.4

g/
L
total
copper,
respectively.
The
authors
attributed
the
higher
tolerance
of
E.
lucius
to
copper
to
the
very
short
embryonic
exposure
period
compared
with
salmonids
and
white
sucker,
Catostomus
commersoni.
Data
were
not
available
to
calculate
point
estimates
at
the
20
percent
effect
level
using
regression
analysis.
The
chronic
value
for
this
species
was
60.36

g/
L
total
copper
(
geometric
mean
of
34.9
and
104.4

g/
L
total
copper).
H­
6
Pimephales
notatus.
An
experimental
design
similar
to
that
described
by
Mount
and
Stephan
(
1967)
and
Mount
(
1968)
was
used
to
examine
the
chronic
effect
of
copper
on
the
bluntnose
minnow,
P.
notatus
(
Horning
and
Neiheisel
1979).
Measured
total
copper
concentrations
were
4.3
(
control),
18.0,
29.9,
44.1,
71.8,
and
119.4

g/
L,
respectively.
The
experimental
dilution
water
was
a
mixture
of
spring
water
and
demineralized
City
of
Cincinnati
tap
water.
Dissolved
oxygen
was
kept
at
5.9
mg/
L
or
greater
throughout
the
test.
Total
water
hardness
ranged
from
172
to
230
mg/
L
as
CaCO3.
The
test
was
initiated
with
22
6­
week­
old
fry.
The
fish
were
later
separated
according
to
sex
and
thinned
to
a
sex
ratio
of
5
males
and
10
females
per
duplicated
test
chamber.
Growth
(
total
length)
was
significantly
reduced
in
parental
and
first
(
F1)
generation
P.
notatus
after
60
days
of
exposure
to
the
highest
concentration
of
copper
tested
(
119.4

g/
L).
Survival
of
parental
P.
notatus
exposed
to
this
same
high
test
concentration
was
also
lower
(
87
percent)
at
the
end
of
the
test
compared
with
the
other
concentrations
(
range
of
93
to
100
percent).
Copper
at
concentrations
of
18

g/
L
and
greater
significantly
reduced
the
number
of
eggs
produced
per
female.
The
number
of
females
available
to
reproduce
was
generally
the
same
up
to
about
29.9

g/
L
of
copper.
The
chronic
limits
were
based
on
an
NOAEC
and
LOAEC
of
<
18
and
18

g/
L
for
number
of
eggs
produced
per
female.
An
EC20
was
not
estimated
by
nonlinear
regression;
nevertheless,
in
this
case
an
EC20
is
likely
to
be
substantially
below
18
µ
g/
L.

Pimephales
promelas.
The
results
from
a
30­
day
ELS
toxicity
test
to
determine
the
chronic
toxicity
of
copper
to
P.
promelas
using
dilution
water
from
Lake
Superior
(
hardness
ranging
from
40
to
50
mg/
L
as
CaCO3)
was
included
in
Table
2a
from
a
manuscript
prepared
by
Lind
et
al.
in
1978.
In
this
experiment,
five
test
concentrations
and
a
control
were
supplied
by
a
continuous­
flow
diluter.
The
exposure
began
with
embryos
1
day
post­
fertilization.
Pooled
results
from
fish
dosed
in
replicate
exposure
chambers
were
given
for
mean
percentage
embryo
survival
to
hatch,
mean
percentage
fish
survival
after
hatch,
and
mean
fish
wet
weight
after
30
days.
The
percentage
of
embryo
survival
to
hatch
was
not
affected
by
total
copper
concentrations
as
high
as
52.1

g/
L
total
copper.
Survival
after
hatch,
however,
was
compromised
at
26.2

g/
L,
and
mean
wet
weight
of
juvenile
fathead
minnows
was
significantly
reduced
at
13.1

g/
L
of
copper.
The
estimated
EC20
value
for
biomass
was
9.376

g/
L
total
copper.

Catastomus
commersoni.
McKim
et
al.
(
1978)
examined
the
survival
and
growth
(
expressed
as
standing
crop)
of
embryo­
larval
and
early
juvenile
white
sucker
exposed
to
copper.
The
embryo
exposure
was
for
13
days,
and
the
larval­
early­
juvenile
exposure
lasted
27
days.
The
NOAEC
and
LOAEC
were
12.9
and
33.8

g/
L
total
copper,
respectively.
The
resulting
chronic
value
based
on
hypothesis
testing
for
this
species
was
20.88

g/
L
total
copper
(
geometric
mean
of
12.9
and
33.8

g/
L
total
copper).

Lepomis
macrochirus.
Results
from
a
22­
month
copper
life­
cycle
toxicity
test
with
bluegill
(
L.
macrochirus)
were
reported
by
Benoit
(
1975).
The
study
included
a
90­
day
embryo­
larval
survival
and
growth
component.
The
tests
were
conducted
at
the
U.
S.
EPA
National
Water
Quality
Laboratory
in
Duluth,
Minnesota,
using
Lake
Superior
water
as
the
dilution
water
(
average
water
hardness
=
45
mg/
L
as
CaCO3).
The
test
was
initiated
in
December
1969
with
2­
year­
old
juvenile
L.
macrochirus.
In
May
1971,
the
fish
were
sexed
and
randomly
reduced
to
three
males
and
seven
females
per
tank.
Spawning
commenced
on
10
June
1971.
The
90­
day
embryo­
larval
exposure
was
initiated
when
12
lots
of
50
newly
hatched
larvae
from
one
of
the
two
control
groups
were
randomly
selected
and
transferred
to
duplicate
grow­
out
chambers
at
1
of
6
total
copper
concentrations:
3
(
control),
12,
21,
40,
77,
and
162

g/
L,
respectively.
In
the
22­
month
juvenile
through
adult
exposure,
survival,
growth,
and
reproduction
were
unaffected
at
77

g/
L
of
copper
and
below.
No
spawning
occurred
at
162

g/
L.
Embryo
hatchability
and
survival
of
4­
dayold
larvae
at
77

g/
L
did
not
differ
significantly
from
those
of
controls.
However,
after
90
days
of
exposure,
survival
of
larval
L.
macrochirus
at
40
and
77

g/
L
was
significantly
lower
than
for
controls,
and
no
larvae
survived
at
162

g/
L.
Growth
remained
unaffected
at
77

g/
L.
Based
on
the
90­
day
survival
of
bluegill
larvae,
the
chronic
limits
were
estimated
to
be
21
and
40

g/
L
(
geometric
mean
=
28.98

g/
L).
The
corresponding
EC20
for
embryo­
larval
survival
was
27.15

g/
L.
H­
7
H­
8
Log
Cu
(
ug/
L
)
Fracti
on
Surv
iv
al
0
.2
.4
.6
.8
1.0
1.2
1.4
1.6
0
.2
.4
.6
.8
1.0
1.2
Log
Cu
(
ug/
L
)
Fracti
on
Surv
iv
al
0
.2
.4
.6
.8
1.0
1.2
1.4
1.6
0
.2
.4
.6
.8
1.0
1.2
Log
Cu
(
ug/
L
)
Y
oung
per
Female
.2
.4
.6
.8
1.0
1.2
1.4
1.6
1.8
2.0
2.2
2.4
0
5
10
15
20
Campeloma
decisum
(
Test
1),
Life­
cycle,
Arthur
and
Leonard
1970
Campeloma
decisum
(
Test
2),
Life­
cycle,
Arthur
and
Leonard
1970
Ceriodaphnia
dubia
(
Clinch
River),
Life­
cycle,
Belanger
et
al.
1989
EC20
=
8.73

g/
L
EC20
=
10.94

g/
L
EC20
=
19.36

g/
L
H­
9
Log
Cu
(
ug/
L
)
Biomass
0
.2
.4
.6
.8
1.0
1.2
1.4
1.6
1.8
0
200
400
600
800
1000
1200
1400
Log
Cu
(
ug/
L
)
Young
per
Original
Female
0
.2
.4
.6
.8
1.0
1.2
1.4
1.6
1.8
2.0
2.2
0
2
4
6
8
10
12
14
16
Log
Cu
(
ug/
L
)
Fracti
on
Surv
iv
al
.2
.4
.6
.8
1.0
1.2
1.4
1.6
1.8
2.0
2.2
2.4
0
.2
.4
.6
.8
1.0
1.2
Lepomis
macrochirus,
Early
Life­
stage,
Benoit
1975
Oncorhynchus
mykiss,
Early
Life­
Stage,
Besser
et
al.
2001
Ceriodaphnia
dubia,
Life­
cycle,
Carlson
et
al.
1986
EC20
=
27.15

g/
L
EC20
=
20.32

g/
L
EC20
=
9.17

g/
L
H­
10
Log
Cu
(
ug/
L
)
Young
per
Original
Female
0
.2
.4
.6
.8
1.0
1.2
1.4
1.6
1.8
2.0
0
50
100
150
200
250
300
Log
Cu
(
ug/
L
)
Young
per
Original
Female
0
.2
.4
.6
.8
1.0
1.2
1.4
1.6
1.8
2.0
2.2
0
50
100
150
200
Log
Cu
(
ug/
L
)
Young
per
Original
Female
0
.2
.4
.6
.8
1.0
1.2
1.4
1.6
1.8
0
50
100
150
200
250
Daphnia
magna
(
Hardness
104),
Life­
cycle,
Chapman
et
al.
Manuscript
Daphnia
magna
(
Hardness
211),
Life­
cycle,
Chapman
et
al.
Manuscript
Daphnia
magna
(
Hardness
51),
Life­
cycle,
Chapman
et
al.
Manuscript
EC20
=
19.89

g/
L
EC20
=
6.06

g/
L
EC20
=
12.58

g/
L
H­
11
Log
Cu
(
ug/
L
)
Fraction
Adult
Emergence
0
.2
.4
.6
.8
1.0
1.2
1.4
1.6
1.8
2.0
2.2
0
.1
.2
.3
.4
.5
.6
.7
.8
.9
EC20
=
7.67

g/
L
Log
Cu
(
ug/
L
)
Biomass
.2
.4
.6
.8
1.0
1.2
1.4
1.6
0
.1
.2
.3
.4
.5
.6
.7
.8
.9
1.0
Log
Cu
(
ug/
L
)
Biomass
0
.2
.4
.6
.8
1.0
1.2
1.4
1.6
1.8
2.0
0
20
40
60
80
100
120
140
Oncorhynchus
tshawytscha,
Early
Life­
Stage,
Chapman
1975
&
1982
Pimephales
promelas,
Early
Life­
stage,
Lind
et
al.
1978
Clistoronia
magnifica,
Life­
cycle,
Nebeker
et
al.
1984a
EC20
=
5.92

g/
L
EC20
=
9.38

g/
L
H­
12
Log
Cu
(
ug/
L
)
Biomass
.2
.4
.6
.8
1.0
1.2
1.4
1.6
1.8
2.0
0
20
40
60
80
100
120
140
160
180
Log
Cu
(
ug/
L
)
Fracti
on
Surv
iv
al
0
.2
.4
.6
.8
1.0
1.2
1.4
1.6
0
.2
.4
.6
.8
1.0
1.2
Log
Cu
(
ug/
L
)
Fracti
on
Surv
iv
al
­
1.0
­.
5
0
.5
1.0
1.5
0
.2
.4
.6
.8
1.0
1.2
Oncorhynchus
mykiss,
Early
Life­
stage,
Seim
et
al.
1984
Daphnia
pulex
(
Hardness
230
HA
0.15),
Life­
cycle,
Winner
1985
Daphnia
pulex
(
Hardness
57),
Life­
cycle,
Winner
1985
EC20
=
27.77

g/
L
EC20
=
9.16

g/
L
EC20
=
2.83

g/
L