PB82-225038
Cadmium an.1 Endrin Toxicity to Fish in
Waters Containing Mineral Fibers
(U.S.) Environmental
Duluth, MN
Research Lab
Hay 82
U.S. Department of Commerce
National Technical Information Service
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EPA-bOO/3-b2-u53
May S.98'2
CADMIUM AND ENDRIN TOXICITY TO FISH
IN WATERS CONTAINING MINERAL FIBERS.
Anthony R. Carlson, Jiines A. Tucker. Vincent R. Matcson. Gary L.
Philip M. Cook, Gayle F. Olson, and Frank A. Puglisi
Environment a 1 Research Laboratory-Duluth
Duluth, Minnesota 55804
U. S. ENVIRONMENTAL PROTECTION AGENCY
ENVIRONMENTAL RESEARCH LARORATORY-DULUTH
OFFICE OF RESEARCH AND DEVELOPMENT
DULUTH, MINNESOTA 55804
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TECHNICAL REPORT DATA
c read Insinjciitins vn Ihi' fi'i v/ir PI'/'""' t'
1. REPORT NO,
EPA-600/3-82-053
ORD Report
3. RECIPIENT'S ACCESSION NO.
-± -o-'j o'*>
*. TITLE AND SUBTITLE
S. REPORT OATE
May 1982
CADMIUM AND ENORIN TOXICITY TO FISH IN WATERS
co:iTAi'!i:!R nriEPAL n^cns.
F PERFORMING ORGANIZATION CODE
7. AUTHOH(S)
9. PERFORMING ORGANIZATION REPORT NO.
Carlson, A.R. , J.A. Tucker, V.R. Mattson, G-L. Phippe,
P.M. Cook. G.F. Olson, and '-.A. Puglisi
9, PERFORMING ORGANIZATION NAME AND ADDRESS
U-S. Environmental Protection Agency
Environmental Research Laboratory-Duluth
6201 Congdon Boulevard
Duluth, Minnesota 55804
19 PROGRAM ELEMENT NO.
11. CONTRACT/GRANT NO.
12. SPONSORING AGENCY NAME AND ADDRESS
Same as above
13. TYPE OF REPORT ANDPEHIOD COVERED
14. SPONSORING AGENCY CODE
15. SUPP-EMENTARY NOTES
1G ABSTRACT
Taconite tailings and their component asbestiform minerals in Lnke Superior water had
no demonstrable effect on the chronic toxlcity of cadmiun to the flatfish, Jordanella
flo^'idae. Maximum acceptable toxicant concentrations determined in life cycle tests,
where effects on survival, growth, reproduction, and bioconcentration were u<=ed is
endpoints, were between 3-3-7-4, 3-0-fa.b, and 3.A-/.3 ug cadmiun/liter at 0.004, 0-08,
and 0.95 nig/liter taconite tailings concentrations, respectively. Similarly, in two
tests (A and ]i) exposing recently hatched fathead minnows, Pimcphales promelas, for 45
dnys to several concentrations of cndrin, taconite tailings had no observable~effect on
survival, growth and bioconcentration. No observable effect concentrations (NOEC(s))
for endrin at 0.2 mg/liter taconite tailings concentration were between 0.30 and 0.60
pg/liter lor group _A fish and 0.075 and 0.15 pR/liter for group JJ fish. At 0.05
mg/liter taconite tailings concentration, the NOEC(s) were between 0.30 and 0.60
pg/liter for group /i and 0-15 and 0.30 ug/liter for group JJ fish. At 0-95 mg/liter
taconite tailings the NOEC(s) were between 0.15 and 0.30 ug/Hter for group A fish and
0-30 and 60 ug/liter for group 15 fish. ~
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18. DISTRIBUTION STATEMENT
RELEASE TO PUBLIC
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2J SECURITY CLASS I This pair/
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DISCLAIMER
This report has been reviewed by the Environmental Research Laboratory-
Duluth, U.S. Environmental Protection Agency, and approved for publication.
Mention of trade names or commercial products does not constitute endorsement
or recommendation for use.
11
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ABSTRACT
Taconite tailings and their component asbe?t ifortn minerals in Lake
Superiijr water had PC demonstrable effect on the chronic toxicity of cadmium to
the flagfish, Jordanclla flondae. Maximum acceptable toxicant concentrations
determined in life cycle tests, whert effects on ',-urvival, growth, reproduc-
tion, and bioconcentration were used as cndpoints, were between 3.3-7.4,
3.0-6.5, and 3.4-7.3 ug cadmium/liter nt 0.004, 0.08, and 0.95 rag/liter
taconite tailings concentrations, respcc11 ve ly. ^ Similarly, \n two tests (A and
B) exposing recently hatched fathead minnows, Pimepha 1es pr omelas, for 43 days >
Xj
to several concentrations of endrip, taconite tailings had no observable effect
on survival, »rowh and b loccncentrat ion. No observable effect concentrations
(NOEC(s)) for endrin at 0.2 mg/liter taconite tailings concentration were
between 0.30 and 0.60 i;g/liter for group _A fish and 0.075 and 0.15 ug/liter for
group B_ fish. At 0.05 mg/liter taconite tailings concentration, the NOEC(s)
were between 0.30 and 0.60 |Jg/liter for group_A fish and 0.15 and 0.30 ug/liter
Cor group ^ fish. At 0.95 mg/Liter taconite tailings the NOEC(s) were between
0.15 and 0.30 iig/liter for srciup A fish and 0.30 and 60 UR/liter for group B
fish.
O
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INTRODUCTION'
Lake Superior is the source of the experimental water supply at the
Environmental Research Laboratory (ERL) of the U.S. Environmental Protection
Agency located at Buluth, Minnesota. The suspended solids content of this
water varies with cllmatological conditions. It contains diatom fragments,
organic debris, quartz particles, some cliy minerals, and araphibole particles
ranging from blocky cleavage frajrwents to asbestiforn fibers. The source of
these amphibole particles has been traced to the. Reserve Mining Company
effluent to the lake at Silver Bay, Minnesota (Cook, et al. 1974). These
materials are known as taconite tailings.
The amphiboles are hydrated silicates which include the commercially
important asbestos minerals: amosite, crocidolite, anthophyllite, treraolite,
and nctinolitc (Cock, et al. 1974). The predominant amphibole present in the
tailings fraction of the suspended solids is cummingtonlte-grunerite [(MR,
Fe)y Si,^22 (QH^l with smaller amounts of tremolite-actinolite and
hornblende present. Commercial amosite is an asbestiform
curnraingtonite-j>runerite amphlbole ami is nondistingulshable by present methods
of identification from many of the asbestiforn mineral fibers found in the
tailings fraction of suspended solids in the lake water.
In general, little Is known about the effects of suspended fine particles,
or, more specifically, the asbestiform amphibole minerals in the aquatic
environment. It is not known how fine particles, such as those present in the
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tailings, Influence the toxicity of a substance to fish, yet such Information
is important in establishing meaningful water quality criteria.
Daily analysis of amphibole (taronite tailings) and suspended solids
concentrations in Duluth's water supply were begun in 1973 at the ERL-Duluth.
Duluth's water intake is located approximately 4 km up-current frora the intake
of the ERL-Diiluth experimental water supply and is similar in design and
placement in the lake. Intermittent comparative measurements have indicated
that the taconite tailings content of these water supplies were similar, even
though at tines suspended solid concentrations varied considerably. The
taconite tailings concentration of the municipal w.iter supply, treasured dally
from March 1973 to January 1974, was used •'s a guide in planning tills study.
The average tailings concentration during this period was 0.19 rng/liter and
ranged from 0-04 to 0.8 mg/.llrer; the total suspended solid concentrations
averaged 0.83 mg/liter.
Thp purpose of this st-.urfy w.-i.= to determine if the presence of the taconite
tailings in Lake Superior water significantly alters the toxicity of an
Inorganic (cad.niuni) and organic (endrln) conpound to fish-
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MATERIALS AND METHODS
EXPOSURE SYSTEM
The exposure apparatus for both the cadmium and endrin studies consisted
of three modified proportional dilutcrs (Mount and Brungs, 1967) with
independent cycling and toxicant delivery systems. Synchronization of the
three diluter delivery cycles was achieved by wiring the diluters in series
through the cycling micro-switches. Flows were split 4 ways with each chamber
receiving approximately 500 ml per diluter cycl£. Each diluter delivered five,
p
toxicant concentrations and a control to duplicate 30 x 60 x 30 cm. glass
spawning and offspring growlh chambers -:ach containing 43 liters of test wa«ar
at a depth of 23 cm.
Prior to entering the spawning chambers, the water passed through a 28 x
30 x 9 cm glass chamber containing 6 liters of water which was used for embryo
incubation. The intermittent flow rate to each chamber was 11 liters ocr hour
and water temperatures were maintained near 25 0. Temperatures in the test
chambers uere usually within 1 C of each other; minimum and maximum daily
temperatures over the test period were 21.0 and 26.6 C.
A combination of Sylvania Gro-Lux and Dura-Test Vj.ta Lite flourestent
lamps was the main source of illumination. The lamps were used in conjunction
with incandescent bulbs controlled by a timer that regulated the photopcriod,
16 hours of light and 8 hours of dark, with simulated sunrise and sunser
(Drummond and Dawson, 1970).
O c>
3
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TEST WATER PHYSICAL-CHEMICAL CHARACTERISTICS
Suspended Solids and Taconite Tailings; Cadmium Study--The preheated test
water in the cadmium experiments was unfiltered lake water which is routinely
used in Laboratory toxicity tes'.s, and filtered lake water with nearly complete
removal of suspc'.nded solids. The unfi leered lake water (ambient) was
com inually monitored and maintained at or below 3 Jackson turbidity units for
the first 62 days of the cadmium exper iment by us ini» a 151,AGO liter reservoir
during periods of high turbidity. Thereafter, because of the danger of the
reservoir freezing, its use was discontinued and instead lake, water was passed
through a 5 inn prefil'ter and was used in the ambient or unfiltcred test water
anil unfiltercd plus tailings test water svstctns. Filtration of the lake water
to remove suspended solids was accomplished by passing it through a sand filter
(granite)., a Filterite 5 urn bleached white cotton prefilter, with final
filtration through a Nucleporc 0.2 urn "GO" membrane filter.
Taconite tailings were obtained from the Reserve Mining Company plant at
Silver Kay, Minnesota and transported to the laboratory in 19 liter polyethy-
lene IURS and stored at -10 C. Prior to use, the. contents of the iugs were.
thoroughly mixed and then fractionated by sedimentation for 2h hours to remove
particles with a settling size larger thar. 2 urn (Cook 1975). The resulting
suspensions were combined and stored in 380 liter Nalgene holding tr.nks
equipped with rec i rculit i ng pumps to kt-ep the particles thoroughly mixed.
Tailings from the holding tanks were transferred by hand in 19 liter
polvi-thvIene jugs as needed to maintain a tailings supply in a 200 liter
rec i re j lat i ng f inert-lass reservoir (10 C) from which a peristaltic pump metered
the tailings into head boxes feeding the appropriate diluter systems.
Three taconire tailings Lest concentrat ions were maintained. These test
concentration!; were selected to ran.'.'.e from l?ss than the minimum and greater
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than the maximum concentrations previously measured In the Duluth municipal
water supply.
For the cndmlum study, 0-2 V'm filtered water was delivered to one of the
three dlluters, while a second dlluter received the unfiltered lake water for
the first 62 days and thereafter water filtered through the 5 um Filterlte
filters- The third diluter systea received water of the same quality as the
second diluter system plus approximately 1 rag/liter taconlte tailings.
Total suspended solids and taconlte tailings concentrations of samples
from a control in each diluter system were ueasured by gravimetric analysis
and x-ray diffraction at least once a week using methods described by Cook
(1975). The menu + standard deviations and ranges of the measurements are
presented in Table 1.
Suspended Solids and Taconlte Tailings: Endrin Study—The test water used
for the endrin study was essentially the same ns that used for the cadmium
experiment with the exception thnt the unf1 1.t^red (ambient) water was not
treated by passing it through a 5 um prefilter as was necessary in the cadmium
exposure. Filtration of the lake water was accomplished as described for the
cadmium experiment except a Nucleporc 0.2 um "QR" membrane filter was used in
place of the "GO" filter for the final filtration. Test water conditions were
modified for the endrin study so that 0.2 micron filtered water was delivered
to one of the three diluter systems, with the second diluter system receiving
filtered water plus approximately 1 mg/liter tailings. The third diluter
system received uuflltered lake water.
Measurement for total suspended solids and tailings concentrations of five
day composite samples from each diluter system were made as described for the
cadmlun test and are presented In Table 2. Daily turbidity measurements were
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Table 1. Taconite Tailings and Total Suspended Solids Concentrations (nvj/i) in Test Wator in Cadnium Stuuy
Test Water
Character! st ic
Tai l
TC
TSS
TC
TSS
Period of Teot
1-62 day1"
1-62 day
62-90 dayc
62-90 day
Total-90 day
Total-90 da/
Fi Itered (0.2
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Table 2. Tacc.•' •" Tailings, Total Suspended Solids Concentrations (yij/L) and Turbidity In Tec.1
Water In Endrln Study
Wjlor
Filtered (0.2 un)
Unfiltercd
FIMarod plus tailings
(1 mn,/L)
ractar 1st Ic
^,
*_SDa'b Rnng
Mejn
Rango
Rango
Tai lings
Concentration (mg/l)
To1-3l Suspended
Sol Ms USS)
Turb!dltyd
0.
0.
0.
02-"). G.
30+0. ,25
2 HO. 07
0.01- 0.
0.06-0.
O.H-0.
04
38
35
0.05+0.02 0.03-0.10 0.93K). 19
0.72+0.48 0.33-1.59 1.43*0.52
0.61»0.?6 0.36-1.50 2.47+0.36
0.62-1.26
0.83-2.32
1.60-3.40
3SO - standard deviation
bN = 10
= 8
"Nophoootric turbidity unils
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also made in 2 chambers at each taronite tailings concentration in order to
monitor suspended solid concentrations; results are presented in Table 3.
Problems were encountered with the filtration system during the entire
testing period resulting in tailings concentrations in the filtered water
approximately 5 times the mean level for the filtered water for the cadmium
test (see Table 1).
Incoming lake water was passed through an Aquafine Model MP-2-SI (1200GPH)
ultraviolet Water Sterlizer to minimize bacterial contamination of the filters
and test chambers. . . • .
Water Chemistry: Cadmium Test--Water chemistry measurements were made at
approximate 2 week intervals in at least one control and one high, toxicant
»-
..•oncentrat ion in esrh diluter system. The results did not vary appreciably
M.
between systems or between experiments. Dissolved oxygen concentrations for
the cadmum test ranged *ro"> 7,'-9.2 m»/l'rer. Thf ran»es for total hardness,
total alkalinity and acididty were 44-51, 37-49, and 0.2-2.0 mg/liter as
CaC02, r< ;pectively. The pH measurements ranged from 7.4 to 8.2.
Water Chemistry: Endrin Study— Analvsis for chemical characteristics of
the test water were made weekly using methods recommended by the American
Public Health Association et al. (197J). The dissolved oxygen concentrations
ranged from 7.3-3.5 mg/liter. Ranges for total hardness, total alkalinity and
acidity were 40-44, 38-42, and 1.65-4.2 me/liter as CaCC^, respectively. The
pH measurements ranged from 7.12 to 7.94.
Cadmium Dosing ^nd Measurement—The cadmium stock solution consisted of
106 mg CdCl2 (reagent grade) and 0.1 ml concentrated HC1 oer liter of
filtered (0.2 urn) lake water. This stock was contained in a Mariotte bottle
which provided a constant ^ydrostati^ head delivering the toxicant to all three
diluter systems where the dilution ratio was adjusted to provide a control and
8
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five cadmium concentrations with a 0.5 diluter factor. Each dilutee was
equipped with a toxicant metering device (McAllister et al., 1972), providing :j
mean.1; of independent adjustment of toxicant flow to the diluters.
Measurements for cadmium in test water samples were made weekly on an
alternating basis between duplicate tanks (Table 3)- Analysis for cadmium were
conducted on ,3 Perkin-Elmer Model 403 atomic absorption spectrophotoneter using
'he flaraeless method for detection. The mean percent recovery and standard
deviation (SO) for 45 spiked cadmium water samples was 93-39 + 11.08. Analysis
of cadmium reference standards with known concentrations of 1.8 and 16 ng/llter
cadmium were run concurrently with the unknowns. Mean recovery and SD for the
standard were 1.84 + 0.08 with 17.8 + 1.23 (n = 21) respectively.
Whole fish, that survived from test initiation to test end (90 days), in
the controls and the nominal cadmium concentrations of 3.7 and 7.5 ug/liter
were analyzed for cnd-.nium using a wet digestion, atomic absorption spectro-
uliuiomei ei' metlioj described by Leonard (1971), Cadniu^t tissue conccntra t {or. of
duplicate conposite samples of offsprings which were exposed through embryo
,development and for 30 days post hatch were also determined.
Endrin Dosing and Measurement—A saturation apparatus (Veith and Constock
1975) was utilized to maintain a stock solution of endrin which was delivered
to each diluter by n chemical metering device (Mount and Warner 1965). Water
containing five concentrations of endrin with a 0.5 dilution factor and a
control were delivered to duplicate test tanks. Endrin concentrations in test
water samples were analyzed using «os ehromatojxraphic procedures for
chlorinated pesticides (Thompson, 1974). Measurement of toxicant concentration
in one of each of the duplicate tanks was made each week on an alternatInp,
basic. The mean + standard deviation (SD) and ranges for endrin in test water
are shown in Table t. The mean + SD recovery of endrin from spiked control
water samples was 99.81% + 3.02 (n = 19).
9
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Tabl» 3. Mes^ur i»d Cadmium Concentrations (ug/L) In Tsst Water ,-jt Thr»» Taconlts
Tailings Concentrations
N'can Taccyiito Tailings (mg/lltor)
Uom I n.T 1
concentration
(ug/L)
30,0
15,0
7,5
3.7
1.3
0,0
(control )
0.004
Moan
+ SO3 Range
n = 24
31.2+_3.9 25,7-41.9
16.9_H.O 14.7-19.5
7,4+_1,5 5.4-11.2
3,V0.3 2.7-5,8
2.0J_0,3 1.5-2.7
0.0 C-0-trace
0,03
jf_ S0a Fjngo
n = 24
29.9+2.5 26.5-33.9
15.V|_1,4 14,0-18,0
6,5_*_1.2 4.3-9.0
3.0_10..' 2.5-3.5
1,C_»p.2 1.2-2,1
0,0 0,0-trac»
0.95
J_ SDS Rango
n = 24
32.5+_4.6 23.7-45.4
16.9_^2.2 12.8-21.6
7.3+J.3 5.1-9.5
7.4*0.3 7.1-4.0
1.8+0.2 1.4-2,5
0.0 O.O-traca
3SO = standard d9vlati»n
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Table 4. Measured Endrin Concentrations (ug/L) In Tes1 Water at Thrto Taeonit* Tailings Concentrations
o
Norn 1 n a 1
concenti avion
(ug/L)
0.60
0.30
0.15
0-075
0.037
0.00 (control)
Mean T3coni te Tal I ini;s (mg/ liter)
Number of
Analyst s
14
12-14
5-7
6-7
8-0
14
Mrt=vi and
+_ SDa
0.57+0.39
0.25jK),012
0.13J_0.01
0.078+0,05
0,04^+0.04
0,0
0.02
Range
0.52-0.66
0.23-0.28
0.12-0- 15
0.065-0.00
0.037-0,^1
Mean and
0.60_*0.04
0.28+0.02
p. 15+_0.03
J.032+_0.6V04
0.042+0,003
0.0
0,05
Mean and
Range + SO3
0.5!>-0.70 0.57+0,05
0.25-0.33 0.23_+_0.03
0.1>0.21 0. 14+_0.02
0.077-0.087 0.074+0.008
l
0.037-0.045 0.04^+0.002
0.0
0.93
Range
0.52-0,66
0.20-0.30
0.11-0.16
0.063-0.088
0.042-0.051
3SD = standard deviation
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Endrin tissue concentrations were determined by grinding the fish in a
blender with sufficient anhydrous sodium sulfate to produce a dry powdery
mixture. This mixture was extracted in a Soxhlet extractor for six hours with
a 50-50 acetone-r,-hexane solvent mixture. The extract was evaporated to near
dryness then diluted to 20 ml with n-hexane. Eight ml (40%) of the extract was
placed on a 20 g Flonsil column and eluded with 15% Ethyl-ether in n-hexane.
After discarding the first 50 ml of eluant the next 200 ml was collected in a
volumetric ?lask. This sample was analyzed, by gas chromatography, "as is" or
diluted to bring the endrin concentration into the linear range of the
analytical standard (concentrating samples was not required). The mean +
standard deviation recovery of endrin 'was 97.6% + 7.14 for ondrin spiked tissue
samples. A de. ;rmmation of the percent fat (lipids) was carried out on 2 ml
(10%) of the extract. Fat was defined as the residue remaining after 15 mm in
a 130 C drying oven.
BIOLOGICAL METHODS
Cadmium Study--Flapfish Jordanella floridae brood stock was obtained from
the University of Michigan and held in 0.2 um filtered water and used as a
source of ycung test fish for the following experiment.
The experiment wa.s initiated by placing 25 flagfisa larvae up to 2 days
old (group 1) into the spawning chambers. The next day 20 larvae (group 2)
from the same lot were placed into the growth chambers (replicates of the
spawning chambers) of each of the three exposure systems. In the system
receiving the addit i->rial taconite tailings, group 1 and 2 control fish were
combined after the second week of exposure and the rest of group I discarded.
To obtain reproductive information, the number of fish was reduced on the 30th
and 45th day of exposure so that each chamber contained no more than 15 or 2
12
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males and 5 females, respectively. Additional larvae (20 per chamber) were
placed in the vacated chambers and exposed for 14 days. For details of the
test procedure see Spehar (1976).
The fish we.'e fed live San Francisco brine shrimp, nauplii Atcmia salina,
for the first 30 days and trozen brine shrimp supplemented with a commercial
trout food thereafter. The fish were not fed for 24 hours prior to sacrifice.
Measurement of the effects of cadmium on survival, growth, reproduction,
and bioconcentration was used to determine a maximum acceptable foxicant
concentration (MATC) (Mount and Stephan, 1967) at the three levels of taconite
tailings.
Endrin Study—Fathead minnow Pimephalcs promclas embryos used were
producec by a brood sroel- maintained at the University of Minnesota, St. Paul,
and were hatched in the filtered Lake Superior water. Twenty-five larvae, 2-3
days post-hatch (Group A), were placed into tach cii.nnbi: f,
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DATA ANALYSIS
Survival, growth, and reproduction data were statistically analyzed using
a one-way analysis of variance classification. Dunnett's test for comparing
all means with a control was used with 5 or aore treatments - Duncan's multiple
range test was used when treatments compared were four or less (Steel and
Torrie, 1960). A significance level of P=0.05 was established for all
statistical tests. Nominal toxicant and mean taconite tailings concentrations
will be used hereafter for discussion purposes.
14
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RESULTS AND DISCUSSION
Exposure of flagfish to 30.0 ug/liter cadmium at the three taconite
tailings concentrations caused high mortality within the first 30 days of
exposure and all were dead by test end (Table 5). At the 15 ug/lltcr cadmium
concentrations, effects on survival were evident after 30 days of exposure and
only a few survived t'o test end. Exposure at this cadmium concentration caused
fish behavioral aberrations which consisted of apparent muscle spasms followed
by a period of uneonslousness, then recovery. This aberrant behavior appeared
to be initiated by disturbances such as chamber cleaning and was first observed
17 days after testing began. Only one spawn occurred at this cadmium
concentration (Table 6) and was in the 0.00^ mg/liter taconite tailings
concentvat ion. At 7.5 ug/liter cadmium, survival wns reduced during the
spawning period (day 45-90) at the high taconire tailings concentration (0.93
mg/liter). Mean egg production per female (Table 6) was reduced by exposure to
7.5 ug/liter cadmium at the two lower taconite tailing concentrations (0.004
and 0.08 mg/liter) and the percentage hatch was lessened by exposure to 7.5
ug/liLer cadmium at the hi^h taconite tailings concentration (0.95 mg/liter).
Survival and growth of offspring in the control at the 0-95 mg/liter taconite
tailings concentration was less than at the 1.8-7,5 ug/liter cadmium
concentrations (Table 7).
The growth measurements for parental fish were highly variable between
many duplicates and treatment levels (Table 8) and only one difference from
15
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Table 5. Survival of Fish (Group 2) Exposed to Severs! Concentrations of Cadmium at Throo Concentrations of
Taconite TaiI Ings
Percentage survival
to 30 days3
Percentage survival
fron 30th to 45th (joy"
Percentage survival
fror. 45th to 90th day
Mean taconlto tailings (mg/L)
Ncxnlnal
cadm 1 um
concentration
(uq/L)
30
15
7.5
3.7
1.8
0.0
(control )
Dupl icate
chambers
A
a
A
B
A
B
A
0
A
B
A
B
0.004
5C
Oc
50C
60C
90
95
100
95
95
85
95
100
0.08
5C
Oc
55
45
95
yO
95
70
65
90
90
75
0.95
0
5
30
30
100C
85C
85
7?
05
50
15"
45b
0.004
0(1)
40.0(10)c
33.?(12)c
100.0(i5)
93.1(15)
100.0(15)
100.0(15)
100.0(15)
100.0(15)
100.0(15)
100.0(15)
0.08
0(1)
27.3C(1I)
44.4C(9)
100.0(15)
100.0(15)
100.0(15;
93.3(15)
100.0(15)
100.0(15)
100.0(15)
100.0(15)
0.95
OC)
33.JC(6)
50.0C(6)
93.3(15)
86.7(15)
100.0(15)
100.0(15)
100.0(15)
100.0(15)
100.0(15)
100.0(15)
0,004
_
50.0C(4)
50.0C(4)
100.0(7)
100.0(7)
100.0(7)
85.7(7)
85.7(7)
85.7(7)
85.7(7)
100.0(7)
0.08
_
Oc(3)
25.0C(4)
100.0(7)
71.4(7)
83.3(6)
88.7(7)
100.0(7)
100.0(7)
100.0(7)
85,7(7)
0.95
.
Oc(2)
Oc(4)
57, 1C(7)
71.4C(7)
100.0(7)
100.0(7)
100.0(7)
100.0(6)
100.0(7)
100.0(6)
'initially 20 fish »ere placed In oach chamber.
"N Is In paronlheses.
cV,«an of duplicates signflcantly different from control nea.n (Bennetts' test, P = 0.05).
"Data doduced. No mortality occurred after combining group 1 and 2 fish on the Nth day of testing.
-------
P«L>Io ». Surviving
^LTxtt-lrvjdlJirtt }otmt UJJ (»t-r 4W v I V IIV)
t- n J (»j I • /1 «ud I i>) p* oJii L I I *Ji IU9WII *
» uicur.trJl Ion Dufl It «»1«
Jl'il/' ) ^t^ZJSf* ^-00* gjjd p.9» O.W4 O.Ott O.V
15 A - 0/^u »/«> o/y yy o 0 4^ - - 39,^(M-
u i/1 a/1 0/9 0 fl o , »" - - - ..
/-? * y/5 ^/) I/J 7,V'0 l,M.'o' 4,/Vil 3V/" )4K- VV5 ^V, 11V) A7,tf(^> 40. M1 U"
U 7/5 2/^ !/•* . . l,Ji)< <,^1« >,*t-f «bO" »>5" ?7; »J,>H5) A9.01 11) M.ft(«»*
3,7 A y/j 7/J y/5 4«,06) *,WIO «,4l2> •» I,0u0 l,t« *9.7f M> 59.7(IO) 4O,5t l)t
L> -/* 3/4 2/> ^,/»' 9,9'1 9,9«4l >UJ 1,777 I,VW1 66,91! I > 70.0( )•) ^5,^^IC)
1,tf A 1/5 2/^ 7/:> 5,6t>3 11,^10 f ,MJ -*fcG 2,2*> 1,901 60.JU2) 7<,7( l») }3.7(15I
y 2/4 2/5 2/^ *,5.'fr »,*^9 t,7M 66* l,^4» |,«»| >b,>(l5> 77,2()» :»,7
Control A 1/5 7/5 7/> 5,IOb »,M7 7,651 l,l»» l,fc7U 5X) ft^.3()4) 73-3(19) ft1.5(tf>
0 7/9 7/4 2/' f>,3t>7 l, IL6 p,»«i> 1,071 1,311 l,7lM 5).4( 17) 5o,9( H) 6(j,6l ItO
-J
I ln> woi Lil I CtV l«l<>0 f r ud j Jj ui 1 **J prvxJuC I lofi rfi li fi «dl v*>t *l nod try »uto If *C 1 I 'KJ 1'i" «»jL( x*3* P**^ 'unwil " '<" ud> Ivl * I pToJut I Itxi
^4 I b l>> por oo^ f*i*^ ob r
J*n> *«lo survive,) ID jpj^o Uil Jlwd
MKuo » ly;i I Mc«"1 I 1 dl
'
-------
Tablo 7. Gro-th and Survival Datj of Flagfish Offspring After 30 Days of Exposure (Cadmium Study)
00
Percent survival3
Nominal
cad™ nn
concert t.-j t i on
(•jg/O
15
7.5
3,7
1.8
Control
Mean total length
and weight
(nvVgm)
Moon Taconite Tailings (mg/ liter)
Dup 1 i cat o
charters
A
B
A
B
A
B
A
B
A
B
0.004
82b
-
85
95
90
95
100
85
90
75
O.OB
„
-
55
50
75
95
85
70
80
95
0.95
_
~
95C
95C
8^c
80C
90C
100C
55
55
O.Q04
20. 2/1 7. 5b
-
24. 6/36, 4d
22.9/24.6
22.3/22.8
2^.5/22.6
24.5/26.7
22.4/24.0
20,1/14.8
22.2/20,1
0.03
. _
19.9/14.9
22.0/21.3
20.1/13.6
22,1/18.1
22.8/24.5
20.4/16.0
20.8/16.9
18.9/11.9
0.95
_
-
23,7c/23.6
24.6c/26.3
23.3c/24.2
23- 1c/27.3
23.0c/22,8
23.3c/26.3
21.3/17.8
21.7/21.1
3Twenty larvae wero placed in each chamber shortly afctcr hatch, except whore noted below.
^Elevan larvae wore placed in this ctiomber,
cMed'i of duplicate!- is signl f icanl ty di'ferent thar, controi mean (Ducan's t»,st( P = 0.05).
Oa'3 based on 10 fish. Ten others were removed by misto»e before 30 days of exposure.
-------
Table 8. Mean Total Length (mm) and Live '/eight (gm) of Spanning Adult
Flagfish at Test End (Cadnium Study)
Mean total length
and weight of
females (nm/g^)
Maan total length
and w»ljht of
males (ren/gm)
Mean Taconite Tailings (mg/llter)
Ncmi nal
cad^i urn
concentrat [ on
(ug/l)
15
7,5
3,7
1.8
Control
Djpl teat*
chambers
A
3
A
B
A
B
A
B
A
B
0,004
43-5/1.8*
29.0/0.8*
43,0/1.7
«2.*/l,5
41,4/1.6
43.7/1.7
43.0/1,7
42.7/1,7
41.8/1.6
44,0/1.8
0,08
_
39.2/1.4
33.0/1.2
42.4/1,1
39,0/1.3
44,8/1.9
41.2/1.4
42.0/1.6
40.0/1.4
0.95
_.
37.0/1.0
39,7/1,3
42.4/1.6
42.2/1,5
41.2/1.5
41.7/1.9
39.2/1.2
41.0/1.6
0.004
45.0/1,7*
58.0/3.6
58,5/4,3
56,3/3.5
55.0/3,8
57.0/3.8
60.5/4.6
58,0/4.4
53.6/3.0
0.08
_
47.0/1,9''
51.5/2,7b
57,0/3,8
60.5/4.5
57.5/4.1
56.0/2.4
56,5/3.4
59,0/3.7
0.95
_
54.0/3,0
59,0/4,3
54,0/3.4
52.5/2.8
55.5/3.5
55.5/3.4
56.0/3.7
59.0/4,0
3Thi_'S9 values were not included in the statistical
bM»»n of duplicates significantly different from control (Duncan's test, P = 0.05),
-------
the control could bo demonstrated. This was reduced weight of males exposed to
7.5 ug/llter cadmium at 0.08 mg/liter taconite tailings.
The uptake of cadmium by the parental fish and offspring at 3.7 and 7-5
ug/liter was not demonstrably affected by taconite tailings concentration
(Table 9).
The maximum acceptable toxicant concentrations (MATC) for cadmium and
flagfish, based on the above survival, growth, reproductive, and bloconccnfra-
tion data for Lake Superior water with 0.004, 0-08, and 0.05 mg/liter
concentrations of taconite taili'-'gs, are between 3.3-7.4, 3.0-6.5, and 3-4-7.3
'ig Cd/liter, respectively. These MATC values arc similar to those determined
by Spehar (1976). H^ found the MATC for rlagfish-to lie between 4-1 and 8.1 ug
Cd/liter under ambient conditions (unmeasured) of suspended solids in Lake
8~
Superior water. Mean concentrations of cadmium measured in the tissues of the.
Si*
parental fish exposed for 90 days at cadmium concentrations near 3.7 and 7.5
ur;/]iter in the present study ranged from 5.6 to 8.2 and 9.2 to 12.1 ng/g of
fish tissue, respectively. Spehar (1976) found similar cadmium tissue
concentrations (moans near 6 and 10 njj/g) after 100 days of exposure at 4.1 and
8.1 mg/llter cadnium. Offspring tissue concentrations were also similar
between studies .
The percentage survival of fl^?,fish after two weeks of testing was
markedly less in control charabery receiving the additional taconite tailings
(Table 10) but was not affected thereafter (Table 6). In two of these
chambers, where accurate cumulative counts of dead larvae were made, most
deaths occurred within the first tt'iree days of testing (2-5 days post-hatch).
Several moribund fish were removed from these tanks and were examined under a
microscope and found to be infected with fungus. Generally fish exposed to
cadmium at concentrations up to 15 ug/llter survived in greater numbers than in
O o
the disease affected controls. This indicates that the addition of taconite
20
-------
Table 9. Mean and St.ind.ird Deviation (!i in Parentheses) of Cadmium <
Whole Body Tissue Samples (ug/g Wet Weight) for Flngfish Exposed to
Two Concentrations of Cadmium at Three Taconite Tailings
Concentrations. Parents uvrc Exposed for 90 Days,
and Offspring for 30 Days After Hatch-
Mean Taconitc Tailings (rag/lite-)
0.004
0.08
0.95
Nominal
cadnium
conrintrntion
in water (ug/1)
7.5
3.7
Control
Cadnium tls>~'je
conccntra tion
(Mg/g)
Cadrjlura tissue
concentratIon
Parc?.t£
10.1 + 1.4 (3) 9.4 + 0.9 (8)
5.9 + 1.6 (8) 5-6 + 1.7 (8)
0.2 + 0.05 (4) 0.3 + 0.09 (4)
Cadmium tissue
concentration
C'Jg/8)
12.1 + 2.7 (5)
8.2 + 2.9 (8)
0.8 t- 0-8 (4)
7.5
3.7
Control
Offspring
12.7 + 1.8 (4) 13.0 + 3.6 (4)
10.0 -^ 1.0 (4) 13.7 + 0.5 (4)
0.1 + 0.03 (4) 0-2 + 0.04 (4)
18.3 + 6.1 (4)
11 .8 + 3.4 (4)
0.1 + 0.03 (4)
21
-------
Table 10. Surv'.al of risn 1o 13 or !4 Days cf t/;.-;urfe to $e/«ral Cad.-.;:,m Concentrations at 3 Coocentral icns of
l.'.x-.ir.nl
C^i'ii S u*n
concen1r»1 Ion
(UG/L)
30
15 0
7.5
3.7
1.8
0.0
('.'/Mr ',!)
'Vuj.-i ol S'j.M Icalci is
9io 1 leva
A
B
A
3
A
B
A
B
A
B
A
il jn H ic^n! 1 >
Or 0.,
FiSTi-m
T.iconi 1C
0.004
4!
99
52
96
100
76
92
92
84
88
dlffcrcM
n 1 < Ish
a.:o t-Tvi
1 a i i i no s
0.08
16
24
88
88
72
88
100
88
88
60
63
i,',
"?XS^ T hi)
val
/>
"eirfc>
.-»,:> r (
n?j;e •,
,. h
•jrvival
C
Perr
tri::/!.) Tjco.nile tji lin^s (nq/L) T^coni
0.95
0
8
84
84
84
84
76
16
96
88
&
:c>n'ryl n
0.004
Oa
90
30
)0
100
95
35
1 //
t.i > (!?«n
0.08
20*
90
80
95
90
95
70
65
90
90
'( /''
n.tlj-
0.95
0
5
60
45
100
i 85
85
75
85
50
$
'i
Tcsl , P » 0
0.004
35
20
100
90
75
100
100
85
75
90
80
.05)
r.xjf, 3 Hsr>
t'i1»:o survi
1 o 1 a i I i nqs
0.08
303
25a
95
100
85
100
100
90
85
55
"
IVjl
< nq/L )
0.95
35
20
90
190
100
95
95
90
85
}?
N* fish from 1h9 controls of group 1 and 2 ware combined «i1h respective duplicates and us»d to producs
growth and reproductive data.
cTxolvo doad larvae wore removed on the third doy of testing.
"Thlrtoon do.id larvao woro romovod on tho third day of to-itinq.
-------
tailings to the water increased the incidence of infection and cadmium exposure
reduced the incidence of infection. A causal disease organism was not
identified but may have been fungus which can be a primary or secondary source
of infection. The disease is probably not. due to the direct effect of taconite
tailings because later testing spanning the 2-5 day post-hatch life stage at
approximately 4 times the taconite tailings concentrations (Table 11) did not
elicit similar disease symptoms or affect survival-
ENDRtN STUDY .
No fish survived in groups A or B fish at 0.60 'jg/liter endrin (Tables 12
and 13). Survival was not affectcd .bv endrin at 0.30 :ig/liter or lower
concentrations.
Fish growth in group A fish (Table 12) was reduced at 0.30 ug/Hter endrin
only in water containing the highest concentrations of t.iconite tailings (0.93
mg/liter). Contrasting results were evident in group B fish (Table 13).
Growth was reduced at 0.30 yg/liter .=ndrin concentrations in water containing
0.02 and 0.05 mg/liter taconite tailings, respectively, but was not reduced in
water containing 0.93 mg/liter taconite tailings. Growth also was reduced at
the 0.15 ug/liter endrin concentration in water containing 0.02 mg/liter
taconite tailings.
The uptake of endrin in the fathead minnow tissues, whole body (Table 14)
and fat (Figure 2), was directly proportional to the endrin concentration in
the test water ?nd was not demonstrably affected by taconite tailings
concentrst ion.
The no observable effect toxicant concentration (NOEC) for endrin-exposev'-
minnows at the 0.02 mg/liter taconite tailings concentration was between 0.30
and 0.60 yg/liter for group A fish and between 0.075 and 0.15 ug/liter for
23
-------
Tabla 11. Survival of FlagfIsh Larva* After Six Days of Exposur* to Several
Concentrations of "(aconite Tailings
Nominal
taconlta tal 1 ! ng
concentration
(mg/L)
4.0
1,0
I.U
0.5
0.25
0.02
0.004
Mean measured Turbidity Percentage
cone. (mg/L) (N.T.U. Units) survival3
Taconita Suspended
tailings solids Mean Range Moan
n = 2*> n = 14
3.78 3,l<2 5.6 (5,1-6.3) 92.5
<1i i( ( 'lU-iii") ur>. ,'
l.y (t. ()-:.:> 95.0
1.1 (0.9-1.4) 93.7
0.7 (0.6-0.8) 90
0.012 0.44 0.4 (0.3-0.5) 83.7
0.010 0.18 0.2 (0.1-0.2) 90.0
Range
90-95
Ull . |l)|)
90-100
80-100
85-90
65-95
65-100
8T"cnty larvaa varo plscod In cich of 4 ropllcjto chanbcrs gt eoch concontratIon,
"rjumbor of onalysis.
-------
Table 12. Survival and Growth Data for Fathead Minnows Exposed t» Several Concentrations of Endrlnt at Threo Taconltc Tailings
Concsntrat Ions fcr 45 Days (Group A).
fo
Percantag* survival3
Norn IjDa 1
cndrin
concsntrat I on
(ug/l)
0.60
0,30
0,15
0.075
0.038
Control
Mean
Mojn T»ccnlt» Tailing
Dupl lest*
chambers
A
B
A
B
A
. U
A
B
A
B
A
B
0.02
or-
Oc
92
88
100
100
96
100
100
96
95
100
0.05
Oc
Oc
100
76
92
84
83
84
92
100
88
0,93
Oc
Oc
92
96
HO
60
69
34
84
88
72
28
0.02
^
35.5+2.3
36.435.8
37,1+_3.4
36.4+3.1
36.5+_2.8
36.6+3.2
36.93>.0
37.5+3.7
36.9_*3, 1
total |9ngth_+SDb
(mm)
D (mj/litor)
0.05
_
36. '+4. 2
37,033.9
38,2+4.1
39-4+2,8
38.3+_3.7
38.8+2.7
38.13>,4
37.3+3.7
39.0+_1.8
0.93
-
36.7+2,7c
37. 13>.7C
37.9+-1.3
38.2T4.7
38.9+3.6
37.1JM.6
38.5+3.0
38.43>.7
39.2+4.1
39.1_*_3.8
Moan «»lght _+_ SDb (gm)
0.02
-
.43+. 09
,47+_. 14
.51+. 14
.44T. 15
.47+. 14
.47+. 12
.48+. 12
.50T. 14
.50+, 14
.473;, 12
0.05
-
.51+. 22
.52+. 20
.60+. 19
.50~T. 12
.63+, 12
.58_>_. 19
.60+. 12
.563;. 16
.51+. 16
.58+. 17
0.93
_
.49+.09c
.513;. 15C
. 58T. 23
,61+. 17
.52+;. 20
.58+, 15
.57T. 17
.64+. 23
,60_*_,09
9Twenty-flv» larvae placed In each test chamber.
bSD = standard dsvlation.
cM»an of duplicates is significantly different frcxn control moan (Cunn»1ts" tost, P = 0.05).
-------
Table 13- Survival and Growth Data for Fathead Minr»«s txposed to Soveral Concentrations at Endrin at Three Taeonlte Tailings
Concentrations for 45 Days (Group B).
Percentage survival
Nominal
endr 1 n
concern r at i on
(ug/l)
0.60
0.30
0.15
0.075
0,038
Control
-rotal length + SDb
(mm)
Mean wolght _+_ SDb (gn)
Mean raconltu Tailings (mg/litar)
Dupl Icate
chambers
A
B
A
B
A
B
A
B
A
B
A
B
0,02
Oc
Oc
75
90
CO
100
100
85
95
95
100
103
0.05
Oc
Oc
65
75
'JO
95
?5
95
90
95
80
100
0.93
Oc
Oc
90
100
90
80
100
70
75
80
85
60
O.u2
-
36.3+3.8c
36,>_>3.2C
36. 1+'..6C
36.3jM.2c
37.6+3.2
36, 7T2. 9
36.6+3.4
37.2j>.2
37.7+5.7
33.0j|_2.7
0,05
• - .
J4.8+5.1C
3>.5_t_3.3c
37.6+4.2
36.2^3.5
33.4+3.9
37.4+_3.3
39.6+3.2
39.133.8
38.2+4.0
39,8+2.6
0.93
35.3+2.7
34.9_*3.5
37.3+3.7
38.7+_2.8
37.6+3.2
38.4j^2,7
37.5+2,9
35.6+3,3
36.6+2.6
38.0j»_3. 1
0.02
-
,44+,13c
. 46T, 1 3C
,45+,17c
.48^.20"
.51 + . 14
. 48_+, 1 2
.50+. 17
.51+. 10
.52+. 20
,52^.10
0,05
-
.39+.18c
.43T.12C
.53+. 19
.47jf_, 13
.60+. 19
,51j+_. 15
.62+. 17
-60+_. 19
.58+. 18
.553;. 13
0,93
-
.43+- 12
.43^,13
,54+. 15
.57T.11
,54+. 13
. 57V. 1 2
.51+.]*
. 45"+". 1 3
.48+. 09
.61+. 15
^wanty larvae placed In each tost chamber,
bSO = standard deviation.
cMean of duplicates is significantly different from control r.ean (Ounnetts1 tost, P = 0.05).
-------
Table 14, Concentrations of Endrln In Duplicate (A and 8) Ccmposlte Whole Body Tissue Samples (ug/g Wet rfolght) and Percent Fat
(In Parentheses) for Fathead Minnows Exposed to Several Concentrations of Endrln »t Three Taconlte Tailings Concentrations
Mean Taconite Tailings (rag/liter)
0.02 0.05
Nominal
endrln Endrln tissue Endrin tissue
concentration concentration concentration
In water (ug/l) (uj/g)' Moan (jg/g)a Mean
A B n B
0.30 4,120(8.9) 3,760(8,7) 3,940 3,290(6,5) 4,130(8.8) 3,710
0.15 1,950(7.7) 2,240(10.2) 2,095 2,400 <9.6) 2,240(10.1) 2,320
0.075 ',050(11.1) 1,100(9.1) 1,075 1,240(11,1) 1,160(10.1) 1,200
0.037 552(8.0) 578(10,0) 550 573(11.2) 522(11.0) 548
'J 0,00 70(9,1) 55(10.8) 61 36( 0.7) 45(11.3) 40
(control)
0.93
Endrin tissue
concentration
(yg/g)' Mean
A D
3,400(9.4) 3,590(8.2) 3,495
1,950(10,6) 2,110(10,0) 2,030
998(10.8) 1,110(10.6) 1,054
570(10.7) 461(8,2) 515
T8(9,4) 85(10.3) 82
'Porcont fat In parontheses.
-------
SO.OOOr
oo
,UMO,000
en
c.
.£ 30,000
c
o
s
*-
20,000
c
o
O
c
v_
TJ
C
UJ
10,000
Taconite Tailings (mg/liter)
• — 002
* — 005
.—0.93
210
240
270
300
Endrin Concentration in Water (ng/l)
Figure 1. Linear relationship of endrin in fat of'fathead minnows exposed to
several concentratIons of endrin at three taconlte tailings
ccncentra t Ions.
-------
group U fish. At 0.05 rag/Liter '...icon i te tailings concentration, the NOEC was
between 0.30 and 0.60 ug/liter for group A fish ond between 0.15 rind 0.30
Ug/liter for group JJ fish. At the 0.95 rag/liter uaconite tailings
concentration the NOEC for endrin was between 0.15 and 0,30 ug/liter for group
_A fish and between 0,30 and 0.60 ug/liter for group _B fish. Although these
values are quite variable, they are similar to the following results obtained
in previous endrin toxicity tests at this l.iborat ory. The NOEC for fathead
minnows exposed to endrin for 300 days was between 0.14 nnd 0.25 ug/liter
(Jarvinen and Tyo, 1978). A MATC determined for endrin exposed flagfish was
between 0.22 and 0.30 ug/liter (Herm.inutz, 1978).
CONCLUSION
The differences in toxicant responses measured for fish in these
experiments could be caused by inherent biological variability of the fish
and/or experimental error. However, uiiese experiments ifo reflect the presence
of effect threshold concentrations unaffected by the rrinpe of taconite tailings
concentrations maintained in these studies. It is concluded, under the
conditions of these experiments, Lh.it mean taconite tailings concentrations
ranging from 0.004 to 0.95 mg/liter had no demonstrable effect on the chronic
toxicity of cadmium to the flagfish. Similarly, me-n taconite tailings
concentrations ranging from 0,02 to 0.93 mg/lucr had no demonstrable effect on
endrin toxicity to the fathead minnow. There were also no effects of taconite
tailings on the bioconcentrat ion of cadmium or endrin by the fish studied. The
information j.:uied from these toxicity tests in regard to the direct effects of
taconite tailings are inconclusive. Therefore, it is our conclusion that
taconite tailings at the concentrations tested did not demonstrsbly alter the
effect threshold concentrations for cadmium and endrin determined by previous
experience in this laboratory.
29
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