Appendix
H.
Hardness
Slopes
As
discussed
in
Section
XX,
EPA's
earlier
freshwater
copper
criteria
recommendations
were
hardness­
dependent
values.
Although
characterized
as
`
hardness­
dependent'
EPA
recognized
that
these
adjusted
criteria
not
only
reflected
the
influence
of
hardness
on
copper
toxicity,
but
also,
hardness
was
a
surrogate
for
other
co­
varying
water
quality
parameters.
In
order
to
compare
the
new
BLM­
based
criteria
with
an
updated
hardness­
dependent
criteria
an
overall
or
`
pooled
slope'
was
needed
to
normalize
the
acute
toxicity
data
to
a
standard
hardness
for
calculating
criteria.
A
pooled
hardness
slope
was
derived
using
all
appropriate
acute
toxicity
data,
regardless
of
the
quality
rating
assigned,
according
to
the
procedures
in
the
1985
Guidelines.

To
account
for
the
apparent
relationship
of
copper
acute
toxicity
to
hardness,
an
analysis
of
covariance
(
Dixon
and
Brown
1979;
Neter
and
Wasserman
1974)
was
performed
using
WINKS
statistical
software
(
WINKS
ETC)
to
calculated
the
pooled
slope
for
hardness
using
the
natural
logarithm
of
the
acute
value
as
the
dependent
variable,
species
as
the
treatment
or
grouping
variable,
and
the
natural
logarithm
of
hardness
as
the
covariate
or
independent
variable.
The
pooled
slope
is
a
regression
slope
from
a
pooled
data
set,
where
every
variable
is
adjusted
relative
to
its
mean.
The
species
are
adjusted
separately,
then
pooled
for
a
single
conventional
least
squares
regression
analysis.
The
slope
of
the
regression
line
is
the
best
estimate
of
the
allspecies
relationship
between
toxicity
and
hardness.

This
analysis
of
covariance
model
was
fit
to
the
data
contained
in
this
appendix
for
the
seven
species
for
which
definitive
acute
values
are
available
over
a
range
of
hardness
such
that
the
highest
hardness
is
at
least
three
time
the
lowest,
and
the
highest
is
also
at
least
100
mg/
L
higher
than
the
lowest.
Other
species
either
did
not
meet
these
criteria,
the
organisms
were
fed
or
as
with
D.
pulex,
D.
pulicaria
and
H.
azteca
did
not
show
any
hardness­
toxicity
trend
possibly
due
to
differences
in
exposure
methods
such
as
unusual
chemical
composition
of
the
dilution
water.

A
list
of
the
species,
acute
toxicity
and
hardness
values
and
the
slopes
used
to
estimate
the
pooled
hardness
slope
are
included
in
this
appendix.
The
slopes
for
the
seven
species
ranged
from
0.4349
to
0.8963,
and
the
pooled
slope
for
these
seven
species
was
0.9584.
An
F­
test
was
used
to
test
whether
a
model
with
separate
species
slopes
for
each
species
gives
significantly
better
fit
to
the
data
than
the
model
with
parallel
slopes.
This
test
showed
that
the
separate
slopes
model
is
not
significantly
better,
and
therefore
the
slopes
are
not
significantly
different
than
the
overall
pooled
slope
(
P=
0.39).
Appendix
G.
Hardness
Slopes
Species
n
Slope
R2
Value
Degrees
of
Freedom
Ceriodaphnia
dubia
27
0.8821
0.6063
0.5893
1.1749
25
Daphnia
magna
46
0.7495
0.6174
0.5702
0.9288
44
Oncorhynchus
clarki
11
0.6461
0.4184
0.0717
1.2204
9
Oncorhynchus
mykiss
56
0.6245
0.6557
0.5010
0.7480
54
Oncorhynchus
tshawytscha
12
0.8963
0.6064
0.3875
1.4051
10
Pimephales
promelas
159
0.4349
0.4447
0.3583
0.5116
157
Lepomis
macrochirus
6
0.7282
0.8499
0.3033
1.1531
4
All
of
the
above
317
0.9584
0.5098
0.8542
1.0625
303
(
p
=
0.389)
95%
Confidence
Limits
Results
of
Covariance
Analysis
of
Freshwater
Acute
Toxicity
Versus
Hardness
G­
2
Appendix
G.
Hardness
Slopes
Species
Lifestage
Method
Hardness
(
mg/
L
as
CaCO3)
LC50
or
EC50
Total
(
ug/
L)
Reference
Ceriodaphnia
dubia
<
4
h
S,
M,
T
52
19.00
Carlson
et
al.
1986
Ceriodaphnia
dubia
<
4
h
S,
M,
T
52
17.00
Carlson
et
al.
1986
Ceriodaphnia
dubia
<
4
h
S,
M,
T
36
20.00
Carlson
et
al.
1986
Ceriodaphnia
dubia
<
4
h
S,
M,
T
36
18.00
Carlson
et
al.
1986
Ceriodaphnia
dubia
<
12
h
S,
M,
D
45
26.04
Belanger
et
al.
1989
Ceriodaphnia
dubia
<
12
h
S,
M,
D
45
17.71
Belanger
et
al.
1989
Ceriodaphnia
dubia
<
12
h
S,
M,
D
45
31.25
Belanger
et
al.
1989
Ceriodaphnia
dubia
<
12
h
S,
M,
D
45
25.00
Belanger
et
al.
1989
Ceriodaphnia
dubia
<
12
h
S,
M,
D
45
29.17
Belanger
et
al.
1989
Ceriodaphnia
dubia
<
12
h
S,
M,
D
45
33.33
Belanger
et
al.
1989
Ceriodaphnia
dubia
<
12
h
S,
M,
D
45
23.96
Belanger
et
al.
1989
Ceriodaphnia
dubia
<
12
h
S,
M,
D
45
20.83
Belanger
et
al.
1989
Ceriodaphnia
dubia
<
12
h
S,
M,
D
45
19.79
Belanger
et
al.
1989
Ceriodaphnia
dubia
<
12
h
S,
M,
D
94.1
27.08
Belanger
et
al.
1989
Ceriodaphnia
dubia
<
12
h
S,
M,
D
94.1
21.88
Belanger
et
al.
1989
Ceriodaphnia
dubia
<
12
h
S,
M,
D
94.1
28.13
Belanger
et
al.
1989
Ceriodaphnia
dubia
<
12
h
S,
M,
D
94.1
38.54
Belanger
et
al.
1989
Ceriodaphnia
dubia
<
12
h
S,
M,
D
94.1
35.42
Belanger
et
al.
1989
Ceriodaphnia
dubia
<
12
h
S,
M,
D
179
69.79
Belanger
et
al.
1989
Ceriodaphnia
dubia
<
12
h
S,
M,
D
179
39.58
Belanger
et
al.
1989
Ceriodaphnia
dubia
<
12
h
S,
M,
D
179
81.25
Belanger
et
al.
1989
Ceriodaphnia
dubia
<
12
h
S,
M,
D
179
84.38
Belanger
et
al.
1989
Ceriodaphnia
dubia
<
12
h
S,
M,
D
97.6
14.58
Belanger
&
Cherry
1990
Ceriodaphnia
dubia
<
12
h
S,
M,
D
97.6
29.17
Belanger
&
Cherry
1990
Ceriodaphnia
dubia
<
12
h
S,
M,
D
97.6
32.29
Belanger
&
Cherry
1990
Ceriodaphnia
dubia
<
12
h
S,
M,
D
182
58.33
Belanger
&
Cherry
1990
Ceriodaphnia
dubia
<
12
h
S,
M,
D
182
87.50
Belanger
&
Cherry
1990
Daphnia
magna
<
24
h
S,
M,
T,
I
100
31.80
Borgmann
&
Ralph
1983
Daphnia
magna
<
24
h
S,
M,
I
100
35.60
Borgmann
&
Charlton
1984
Daphnia
magna
1
d
S,
M,
T
39
9.10
Nebeker
et
al.
1986a
Daphnia
magna
1
d
S,
M,
T
39
11.70
Nebeker
et
al.
1986a
Daphnia
magna
<
2
h
S,
M,
T
38
6.60
Nebeker
et
al.
1986a
Daphnia
magna
<
2
h
S,
M,
T
38
9.90
Nebeker
et
al.
1986a
Daphnia
magna
1
d
S,
M,
T
39
11.70
Nebeker
et
al.
1986a
Daphnia
magna
<
4
h
S,
M,
T
39
6.70
Nebeker
et
al.
1986a
Daphnia
magna
1
d
S,
M,
T
26
9.10
Nebeker
et
al.
1986a
Daphnia
magna
<
2
h
S,
M,
T
27
5.20
Nebeker
et
al.
1986a
Daphnia
magna
<
24
h
S,
M,
T
170
41.20
Baird
et
al.
1991
Daphnia
magna
<
24
h
S,
M,
T
170
10.50
Baird
et
al.
1991
Daphnia
magna
<
24
h
S,
M,
T
170
20.60
Baird
et
al.
1991
Daphnia
magna
<
24
h
S,
M,
T
170
17.30
Baird
et
al.
1991
Daphnia
magna
<
24
h
S,
M,
T
170
70.70
Baird
et
al.
1991
Daphnia
magna
<
24
h
S,
M,
T
170
31.30
Baird
et
al.
1991
Daphnia
magna
<
24
h
S,
M,
I
109.9
7.10
Meador
1991
Daphnia
magna
<
24
h
S,
M,
I
109.9
16.40
Meador
1991
Daphnia
magna
<
24
h
S,
M,
I
109.9
39.90
Meador
1991
Daphnia
magna
<
24
h
S,
M,
I
109.9
18.70
Meador
1991
Daphnia
magna
<
24
h
S,
M,
I
109.9
18.90
Meador
1991
Daphnia
magna
<
24
h
S,
M,
I
109.9
39.70
Meador
1991
Daphnia
magna
<
24
h
S,
M,
I
109.9
46.00
Meador
1991
Daphnia
magna
<
24
h
S,
M,
I
109.9
71.90
Meador
1991
Daphnia
magna
<
24
h
S,
M,
I
109.9
57.20
Meador
1991
Daphnia
magna
<
24
h
S,
M,
I
109.9
67.80
Meador
1991
Daphnia
magna
<
24
h
R,
M,
T
170
31.00
Lazorchak
&
Waller
1993
Daphnia
magna
<
24
h
R,
M,
T
170
38.00
Lazorchak
&
Waller
1993
Daphnia
magna
<
24
h
R,
M,
T
170
35.00
Lazorchak
&
Waller
1993
Daphnia
magna
<
24
h
R,
M,
T
170
58.00
Lazorchak
&
Waller
1993
Daphnia
magna
<
24
h
R,
M,
T
170
37.00
Lazorchak
&
Waller
1993
Daphnia
magna
<
24
h
R,
M,
T
170
51.00
Lazorchak
&
Waller
1993
Daphnia
magna
<
24
h
R,
M,
T
170
39.00
Lazorchak
&
Waller
1993
Daphnia
magna
<
24
h
R,
M,
T
170
50.00
Lazorchak
&
Waller
1993
Daphnia
magna
<
24
h
R,
M,
T
170
52.00
Lazorchak
&
Waller
1993
Daphnia
magna
<
24
h
R,
M,
T
170
31.00
Lazorchak
&
Waller
1993
Daphnia
magna
<
24
h
R,
M,
T
170
30.00
Lazorchak
&
Waller
1993
Daphnia
magna
<
24
h
R,
M,
T
170
46.00
Lazorchak
&
Waller
1993
Daphnia
magna
<
24
h
R,
M,
T
170
63.00
Lazorchak
&
Waller
1993
Daphnia
magna
<
24
h
S,
M,
T
52
26.00
Chapman
et
al.
Manuscript
Daphnia
magna
<
24
h
S,
M,
T
105
30.00
Chapman
et
al.
Manuscript
G­
3
Appendix
G.
Hardness
Slopes
Species
Lifestage
Method
Hardness
(
mg/
L
as
CaCO3)
LC50
or
EC50
Total
(
ug/
L)
Reference
Daphnia
magna
<
24
h
S,
M,
T
106
38.00
Chapman
et
al.
Manuscript
Daphnia
magna
<
24
h
S,
M,
T
207
69.00
Chapman
et
al.
Manuscript
Daphnia
magna
<
24
h
S,
M,
T,
D
7.1
4.80
Long's
MS
Thesis
Daphnia
magna
<
24
h
S,
M,
T,
D
20.6
7.40
Long's
MS
Thesis
Daphnia
magna
<
24
h
S,
M,
T,
D
23
6.50
Long's
MS
Thesis
Oncorhynchus
clarki
larval,
0.34
g
S,
M,
T
169
80.00
Dwyer
et
al.
1995
Oncorhynchus
clarki
larval,
0.57
g
S,
M,
T
169
60.00
Dwyer
et
al.
1995
Oncorhynchus
clarki
7.4
cm,
4.2
g
F,
M,
T,
D
205
398.91
Chakoumakos
et
al.
1979
Oncorhynchus
clarki
6.9
cm,
3.2
g
F,
M,
T,
D
69.9
197.87
Chakoumakos
et
al.
1979
Oncorhynchus
clarki
8.8
cm,
9.7
g
F,
M,
T,
D
18
41.35
Chakoumakos
et
al.
1979
Oncorhynchus
clarki
8.1
cm,
4.4
g
F,
M,
T,
D
204
282.93
Chakoumakos
et
al.
1979
Oncorhynchus
clarki
6.8
cm,
2.7
g
F,
M,
T,
D
83
186.21
Chakoumakos
et
al.
1979
Oncorhynchus
clarki
7.0
cm,
3.2
g
F,
M,
T,
D
31.4
85.58
Chakoumakos
et
al.
1979
Oncorhynchus
clarki
8.5
cm,
5.2
g
F,
M,
T,
D
160
116.67
Chakoumakos
et
al.
1979
Oncorhynchus
clarki
7.7
cm,
4.4
g
F,
M,
T,
D
74.3
56.20
Chakoumakos
et
al.
1979
Oncorhynchus
clarki
8.9
cm,
5.7
g
F,
M,
T,
D
26.4
21.22
Chakoumakos
et
al.
1979
Oncorhynchus
mykiss
larval,
0.67
g
S,
M,
T
169
110.00
Dwyer
et
al.
1995
Oncorhynchus
mykiss
larval,
0.48
g
S,
M,
T
169
50.00
Dwyer
et
al.
1995
Oncorhynchus
mykiss
larval,
0.50
g
S,
M,
T
169
60.00
Dwyer
et
al.
1995
Oncorhynchus
mykiss
swim­
up,
0.25
g
R,
M,
T,
D
44.1
46.70
Cacela
et
al.
1996
Oncorhynchus
mykiss
swim­
up,
0.25
g
R,
M,
T,
D
44.6
24.20
Cacela
et
al.
1996
Oncorhynchus
mykiss
swim­
up,
0.20­
0.24
g
R,
M,
T,
D
38.7
3.54
Welsh
et
al.
2000
Oncorhynchus
mykiss
swim­
up,
0.20­
0.24
g
R,
M,
T,
D
39.3
8.44
Welsh
et
al.
2000
Oncorhynchus
mykiss
swim­
up,
0.20­
0.24
g
R,
M,
T,
D
89.5
17.92
Welsh
et
al.
2000
Oncorhynchus
mykiss
swim­
up,
0.20­
0.24
g
R,
M,
T,
D
89.67
33.33
Welsh
et
al.
2000
Oncorhynchus
mykiss
12­
16
cm
F,
M
300
890.00
Calamari
&
Marchetti
1973
Oncorhynchus
mykiss
alevin
F,
M,
T
23
28.00
Chapman
1975,
1978
Oncorhynchus
mykiss
swim­
up,
0.17
g
F,
M,
T
23
17.00
Chapman
1975,
1978
Oncorhynchus
mykiss
parr,
8.6
cm,
6.96
g
F,
M,
T
23
18.00
Chapman
1975,
1978
Oncorhynchus
mykiss
smolt,
18.8
cm,
68.19
g
F,
M,
T
23
29.00
Chapman
1975,
1978
Oncorhynchus
mykiss
1.2­
7.9
g
F,
M,
T,
D
335
106.25
Fogels
&
Sprague
1977
Oncorhynchus
mykiss
juvenile,
3.9
g
F,
M,
T
125
200.00
Spear
1977,
Anderson
&
Spear
1980b
Oncorhynchus
mykiss
juvenile,
29.1
g
F,
M,
T
125
190.00
Spear
1977,
Anderson
&
Spear
1980b
Oncorhynchus
mykiss
adult,
176
g
F,
M,
T
125
210.00
Spear
1977,
Anderson
&
Spear
1980b
Oncorhynchus
mykiss
1.1
g
F,
M,
T,
D
32
23.33
Howarth
&
Sprague
1978
Oncorhynchus
mykiss
2.2
g
F,
M,
T,
D
31
30.10
Howarth
&
Sprague
1978
Oncorhynchus
mykiss
1.4
g
F,
M,
T,
D
31
31.25
Howarth
&
Sprague
1978
Oncorhynchus
mykiss
2.7
g
F,
M,
T,
D
30
31.25
Howarth
&
Sprague
1978
Oncorhynchus
mykiss
3.2
g
F,
M,
T,
D
101
41.67
Howarth
&
Sprague
1978
Oncorhynchus
mykiss
0.71
g
F,
M,
T,
D
99
34.48
Howarth
&
Sprague
1978
Oncorhynchus
mykiss
0.80
g
F,
M,
T,
D
102
31.98
Howarth
&
Sprague
1978
Oncorhynchus
mykiss
1.5
g
F,
M,
T,
D
101
48.23
Howarth
&
Sprague
1978
Oncorhynchus
mykiss
1.6
g
F,
M,
T,
D
99
49.90
Howarth
&
Sprague
1978
Oncorhynchus
mykiss
1.5
g
F,
M,
T,
D
100
50.10
Howarth
&
Sprague
1978
Oncorhynchus
mykiss
10
g
F,
M,
T,
D
100
84.48
Howarth
&
Sprague
1978
Oncorhynchus
mykiss
1.0
g
F,
M,
T,
D
98
89.48
Howarth
&
Sprague
1978
Oncorhynchus
mykiss
1.0
g
F,
M,
T,
D
366
72.92
Howarth
&
Sprague
1978
Oncorhynchus
mykiss
1.7
g
F,
M,
T,
D
371
85.63
Howarth
&
Sprague
1978
Oncorhynchus
mykiss
6.6
g
F,
M,
T,
D
361
310.42
Howarth
&
Sprague
1978
Oncorhynchus
mykiss
1.8
g
F,
M,
T,
D
371
537.50
Howarth
&
Sprague
1978
Oncorhynchus
mykiss
0.90
g
F,
M,
T,
D
360
321.88
Howarth
&
Sprague
1978
Oncorhynchus
mykiss
3.1
g
F,
M,
T,
D
364
115.63
Howarth
&
Sprague
1978
Oncorhynchus
mykiss
1
g
F,
M,
T,
D
194
176.04
Chakoumakos
et
al.
1979
Oncorhynchus
mykiss
4.9
cm
F,
M,
T,
D
194
88.85
Chakoumakos
et
al.
1979
Oncorhynchus
mykiss
6.0
cm,
2.1
g
F,
M,
T,
D
194
86.77
Chakoumakos
et
al.
1979
Oncorhynchus
mykiss
6.1
cm,
2.5
g
F,
M,
T,
D
194
107.29
Chakoumakos
et
al.
1979
Oncorhynchus
mykiss
2.6
g
F,
M,
T,
D
194
285.42
Chakoumakos
et
al.
1979
Oncorhynchus
mykiss
4.3
g
F,
M,
T,
D
194
133.33
Chakoumakos
et
al.
1979
Oncorhynchus
mykiss
9.2
cm,
9.4
g
F,
M,
T,
D
194
230.21
Chakoumakos
et
al.
1979
Oncorhynchus
mykiss
9.9
cm,
11.5
g
F,
M,
T,
D
194
171.88
Chakoumakos
et
al.
1979
Oncorhynchus
mykiss
11.8
cm,
18.7
g
F,
M,
T,
D
194
205.21
Chakoumakos
et
al.
1979
Oncorhynchus
mykiss
13.5
cm,
24.9
g
F,
M,
T,
D
194
535.42
Chakoumakos
et
al.
1979
Oncorhynchus
mykiss
13.4
cm,
25.6
g
F,
M,
T,
D
194
253.13
Chakoumakos
et
al.
1979
Oncorhynchus
mykiss
6.7
cm,
2.65
g
F,
M,
T
9.2
2.80
Cusimano
et
al.
1986
Oncorhynchus
mykiss
134
g
F,
M,
T
120
80.00
Seim
et
al.
1984
Oncorhynchus
mykiss
parr
F,
M,
T,
D,
I
31
90.00
Mudge
et
al.
1993
Oncorhynchus
mykiss
swim­
up,
0.29
g
F,
M,
T,
D
36.1
19.60
Cacela
et
al.
1996
G­
4
Appendix
G.
Hardness
Slopes
Species
Lifestage
Method
Hardness
(
mg/
L
as
CaCO3)
LC50
or
EC50
Total
(
ug/
L)
Reference
Oncorhynchus
mykiss
swim­
up,
0.25
g
F,
M,
T,
D
36.2
12.90
Cacela
et
al.
1996
Oncorhynchus
mykiss
swim­
up,
0.23
g
F,
M,
T,
D
20.4
5.90
Cacela
et
al.
1996
Oncorhynchus
mykiss
swimup,
0.23
g
F,
M,
T,
D
45.2
37.80
Cacela
et
al.
1996
Oncorhynchus
mykiss
swim­
up,
0.26
g
F,
M,
T,
D
45.4
25.10
Cacela
et
al.
1996
Oncorhynchus
mykiss
swim­
up,
0.23
g
F,
M,
T,
D
41.9
17.20
Cacela
et
al.
1996
Oncorhynchus
tshawytscha
alevin,
0.05
g
F,
M,
T
23
26.00
Chapman
1975,
1978
Oncorhynchus
tshawytscha
swim­
up,
0.23
g
F,
M,
T
23
19.00
Chapman
1975,
1978
Oncorhynchus
tshawytscha
parr,
9.6
cm,
11.58
g
F,
M,
T
23
38.00
Chapman
1975,
1978
Oncorhynchus
tshawytscha
smolt,
14.4
cm,
32.46
g
F,
M,
T
23
26.00
Chapman
1975,
1978
Oncorhynchus
tshawytscha
3
mo,
1.35
g
F,
M,
T,
I
13
10.20
Chapman
&
McCrady
1977
Oncorhynchus
tshawytscha
3
mo,
1.35
g
F,
M,
T,
I
46
24.10
Chapman
&
McCrady
1977
Oncorhynchus
tshawytscha
3
mo,
1.35
g
F,
M,
T,
I
182
82.50
Chapman
&
McCrady
1977
Oncorhynchus
tshawytscha
3
mo,
1.35
g
F,
M,
T,
I
359
128.40
Chapman
&
McCrady
1977
Oncorhynchus
tshawytscha
swim­
up,
0.36­
0.45
g
F,
M,
T,
D
36.6
7.71
Welsh
et
al.
2000
Oncorhynchus
tshawytscha
swim­
up,
0.36­
0.45
g
F,
M,
T,
D
34.6
13.02
Welsh
et
al.
2000
Oncorhynchus
tshawytscha
swim­
up,
0.36­
0.45
g
F,
M,
T,
D
38.3
14.90
Welsh
et
al.
2000
Oncorhynchus
tshawytscha
swim­
up,
0.36­
0.45
g
F,
M,
T,
D
35.7
19.06
Welsh
et
al.
2000
Pimephales
promelas
adult,
40
mm
S,
M,
T
103
310.00
Birge
et
al.
1983
Pimephales
promelas
adult,
40
mm
S,
M,
T
103
120.00
Birge
et
al.
1983
Pimephales
promelas
adult,
40
mm
S,
M,
T
262
390.00
Birge
et
al.
1983;
Benson
&
Birge
1985
Pimephales
promelas
­­­
S,
M,
T
52
55.00
Carlson
et
al.
1986
Pimephales
promelas
­­­
S,
M,
T
52
85.00
Carlson
et
al.
1986
Pimephales
promelas
­­­
S,
M,
T
36
180.00
Carlson
et
al.
1986
Pimephales
promelas
­­­
S,
M,
T
36
95.00
Carlson
et
al.
1986
Pimephales
promelas
<
24
h
S,
M,
T
290
15.00
Schubauer­
Berigan
et
al.
1993
Pimephales
promelas
<
24
h
S,
M,
T
290
44.00
Schubauer­
Berigan
et
al.
1993
Pimephales
promelas
<
24
h
S,
M,
T
290
200.00
Schubauer­
Berigan
et
al.
1993
Pimephales
promelas
<
24
h,
0.68
mg
S,
M,
T
19
4.82
Welsh
et
al.
1993
Pimephales
promelas
<
24
h,
0.68
mg
S,
M,
T
19.5
8.20
Welsh
et
al.
1993
Pimephales
promelas
<
24
h,
0.68
mg
S,
M,
T
16.5
31.57
Welsh
et
al.
1993
Pimephales
promelas
<
24
h,
0.68
mg
S,
M,
T
17
21.06
Welsh
et
al.
1993
Pimephales
promelas
<
24
h,
0.68
mg
S,
M,
T
19
35.97
Welsh
et
al.
1993
Pimephales
promelas
<
24
h,
0.68
mg
S,
M,
T
17
59.83
Welsh
et
al.
1993
Pimephales
promelas
<
24
h,
0.68
mg
S,
M,
T
17
4.83
Welsh
et
al.
1993
Pimephales
promelas
<
24
h,
0.68
mg
S,
M,
T
17.5
70.28
Welsh
et
al.
1993
Pimephales
promelas
<
24
h,
0.68
mg
S,
M,
T
18.5
83.59
Welsh
et
al.
1993
Pimephales
promelas
<
24
h,
0.68
mg
S,
M,
T
18.5
182.00
Welsh
et
al.
1993
Pimephales
promelas
larval,
0.32
g
S,
M,
T
173
290.00
Dwyer
et
al.
1995
Pimephales
promelas
larval,
0.56
g
S,
M,
T
173
630.00
Dwyer
et
al.
1995
Pimephales
promelas
larval,
0.45
g
S,
M,
T
173
400.00
Dwyer
et
al.
1995
Pimephales
promelas
larval,
0.39
g
S,
M,
T
173
390.00
Dwyer
et
al.
1995
Pimephales
promelas
3.2­
5.5
cm,
0.42­
3.23
g
S,
M,
T
165
450.00
Richards
&
Beitinger
1995
Pimephales
promelas
2.8­
5.1
cm,
0.30­
2.38
g
S,
M,
T
159
297.00
Richards
&
Beitinger
1995
Pimephales
promelas
1.9­
4.6
cm,
0.13­
1.55
g
S,
M,
T
168
311.00
Richards
&
Beitinger
1995
Pimephales
promelas
3.0­
4.8
cm,
0.23­
1.36
g
S,
M,
T
167
513.00
Richards
&
Beitinger
1995
Pimephales
promelas
<
24
h
S,
M,
T,
D
45.540586
62.23
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
45.540586
190.50
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
44.539694
68.58
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
44.539694
168.91
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
44.539694
94.62
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
45.540586
143.51
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
45.04014
120.65
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
45.04014
196.85
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
138.123096
133.35
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
151.134692
184.15
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
138.123096
304.80
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
139.123988
292.10
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
47.041924
133.35
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
37.033004
92.71
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
60.05352
152.40
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
76.067792
177.80
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
103.091876
203.20
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
103.091876
190.50
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
107.095444
196.85
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
134.119528
234.95
Erickson
et
al.
1996a,
b
G­
5
Appendix
G.
Hardness
Slopes
Species
Lifestage
Method
Hardness
(
mg/
L
as
CaCO3)
LC50
or
EC50
Total
(
ug/
L)
Reference
Pimephales
promelas
<
24
h
S,
M,
T,
D
45.04014
146.05
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
46.041032
171.45
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
45.04014
152.40
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
45.04014
184.15
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
44.039248
203.20
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
45.04014
203.20
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
46.041032
203.20
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
189.168588
222.25
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
46.041032
146.05
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
75.0669
139.70
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
46.041032
139.70
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
74.066008
152.40
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
133.118636
203.20
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
76.067792
196.85
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
134.119528
266.70
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
52.046384
99.06
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
51.045492
111.13
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
50.0446
78.74
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
51.045492
92.71
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
51.045492
85.09
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
53.047276
123.19
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
53.047276
165.10
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
52.046384
190.50
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
47.041924
165.10
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
47.041924
127.00
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
47.041924
92.08
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
47.041924
66.68
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
140.12488
393.70
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
88.078496
317.50
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
59.052628
107.95
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
41.036572
67.95
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
27.024084
45.72
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
43.038356
177.80
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
25.0223
13.97
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
107.095444
304.80
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
87.077604
71.12
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
85.07582
83.82
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
88.078496
104.78
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
87.077604
139.70
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
87.077604
152.40
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
87.077604
260.35
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
87.077604
488.95
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
87.077604
203.20
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
251.223892
704.85
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
252.224784
952.50
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
252.224784
1244.60
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
251.223892
1485.90
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
200.1784
781.05
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
140.12488
476.25
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
90.08028
273.05
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
19.016948
22.23
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
34.030328
24.13
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
51.045492
36.83
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
29.025868
27.94
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
30.02676
26.67
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
27.024084
20.32
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
27.024084
26.67
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
90.08028
190.50
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
60.05352
109.86
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
120.10704
203.20
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
180.16056
209.55
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
91.081172
146.05
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
90.08028
165.10
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
93.082956
254.00
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
92.082064
311.15
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
91.081172
165.10
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
144.128448
920.75
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
292.260464
1073.15
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
440.39248
1003.30
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
217.193564
933.45
Erickson
et
al.
1996a,
b
G­
6
Appendix
G.
Hardness
Slopes
Species
Lifestage
Method
Hardness
(
mg/
L
as
CaCO3)
LC50
or
EC50
Total
(
ug/
L)
Reference
Pimephales
promelas
<
24
h
S,
M,
T,
D
218.194456
742.95
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
212.189104
1879.60
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
S,
M,
T,
D
92.082064
266.70
Erickson
et
al.
1996a,
b
Pimephales
promelas
adult
F,
M,
T
198
470.00
Mount
1968
Pimephales
promelas
­­­
F,
M,
T
31
75.00
Mount
&
Stephan
1969
Pimephales
promelas
5.6
cm,
1.6
g
F,
M,
T
200
440.00
Geckler
et
al.
1976
Pimephales
promelas
4.7
cm
F,
M,
T
200
490.00
Geckler
et
al.
1976
Pimephales
promelas
fry,
6
wk,
2.2
cm
F,
M,
T
202
490.00
Pickering
et
al.
1977
Pimephales
promelas
subadult,
6
mo,
5.5
cm
F,
M,
T
202
460.00
Pickering
et
al.
1977
Pimephales
promelas
­­­
F,
M,
T
48
114.00
Lind
et
al.
Manuscript
(
1978)
Pimephales
promelas
­­­
F,
M,
T
45
121.00
Lind
et
al.
Manuscript
(
1978)
Pimephales
promelas
­­­
F,
M,
T
46
88.50
Lind
et
al.
Manuscript
(
1978)
Pimephales
promelas
­­­
F,
M,
T
30
436.00
Lind
et
al.
Manuscript
(
1978)
Pimephales
promelas
­­­
F,
M,
T
37
516.00
Lind
et
al.
Manuscript
(
1978)
Pimephales
promelas
­­­
F,
M,
T
87
1586.00
Lind
et
al.
Manuscript
(
1978)
Pimephales
promelas
­­­
F,
M,
T
73
1129.00
Lind
et
al.
Manuscript
(
1978)
Pimephales
promelas
­­­
F,
M,
T
84
550.00
Lind
et
al.
Manuscript
(
1978)
Pimephales
promelas
­­­
F,
M,
T
66
1001.00
Lind
et
al.
Manuscript
(
1978)
Pimephales
promelas
30
d,
0.15
g
F,
M,
T,
D
43.9
96.00
Spehar
&
Fiandt
1986
Pimephales
promelas
60­
90
d,
3.3
cm,
0.7
g
S,
M,
T
101
252.00
Bennett
et
al.
1995
Pimephales
promelas
<
24
h
F,
M,
T,
D
47.041924
31.75
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
F,
M,
T,
D
243.216756
117.48
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
F,
M,
T,
D
255.727906
48.26
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
F,
M,
T,
D
47.041924
73.03
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
F,
M,
T,
D
45.04014
59.06
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
F,
M,
T,
D
45.04014
78.74
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
F,
M,
T,
D
45.540586
22.23
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
F,
M,
T,
D
49.043708
6.99
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
F,
M,
T,
D
45.04014
22.23
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
F,
M,
T,
D
43.038356
107.32
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
F,
M,
T,
D
45.540586
292.10
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
F,
M,
T,
D
45.04014
81.28
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
F,
M,
T,
D
45.04014
298.45
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
F,
M,
T,
D
45.540586
241.30
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
F,
M,
T,
D
45.04014
133.35
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
F,
M,
T,
D
44.039248
93.98
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
F,
M,
T,
D
44.039248
67.95
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
F,
M,
T,
D
22.52007
4.76
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
F,
M,
T,
D
24.021408
13.97
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
F,
M,
T,
D
23.020516
29.85
Erickson
et
al.
1996a,
b
Pimephales
promelas
<
24
h
F,
M,
T,
D
21.519178
59.69
Erickson
et
al.
1996a,
b
Lepomis
macrochirus
3.58
cm,
0.63
g
R,
M,
D
85
2291.67
Blaylock
et
al.
1985
Lepomis
macrochirus
12
cm,
35
g
F,
M,
T
45
1100.00
Benoit
1975
Lepomis
macrochirus
10.3
cm,
18.6
g
F,
M,
T
200
8300.00
Geckler
et
al.
1976
Lepomis
macrochirus
10.1
cm,
19.2
g
F,
M,
T
200
10000.00
Geckler
et
al.
1976
Lepomis
macrochirus
2.8­
6.8
cm
F,
M,
T
25.9
1000.00
Cairns
et
al.
1981
Lepomis
macrochirus
3.58
cm,
0.63
g
F,
M,
D
85
1354.17
Blaylock
et
al.
1985
G­
7
Appendix
G.
Hardness
Slopes
SUMMARY
OUTPUT
Overall
Slope
Regression
Statistics
Multiple
R
0.714033268
R
Square
0.509843507
Adjusted
R
Square
0.508287455
St&
ard
Error
0.744214128
Observations
317
ANOVA
df
SS
MS
F
Significance
F
Regression
1
181.4715328
181.4715328
327.651897
1.05959E­
50
Residual
315
174.4642206
0.553854669
Total
316
355.9357534
Coefficients
St&
ard
Error
t
Stat
P­
value
Lower
95%
Upper
95%
Intercept
­
1.34057E­
15
0.04179923
­
3.20717E­
14
1
­
0.082240968
0.082240968
X
Variable
1
0.958366107
0.052945018
18.10115734
1.05959E­
50
0.854195537
1.062536676
G­
8
