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November 17, 1972
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Mr. Elmer P. Wheeler Manager, Environmental Health Monsanto Company 800 N. Lindbergh Boulevard St. Louis, Missouri 63166
Dear Mr. Wheeler:
Re: IBT No. A8570 - Four-Day Fish Toxicity Studies with Aroclor 1242, Aroclor 1254, and Aroclor 1260 in Bluegills and Channel Catfish ________ _________
We are submitting herewith our laboratory report dated
November 17, 1972, prepared in connection with the above study.
.Very truly yours
JCC/mp
J. C. Calandra President
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REPORT TO MONSANTO COMPANY
FOUR-DAY FISH TOXICITY STUDIES WITH AROCLOR 1242, AROCLOR 1254, AND AROCLOR 1260
IN BLUEGILLS AND CHANNEL CATFISH
NOVEMBER 17, 1972
IBT NO. A8570
I. Introduction
At the request of Monsanto Company, four-day fish toxicity studies using bluegills (Lepomia mac ro'chlrua). and channel catfish
(Ictalurua punctatua) were conducted with the following three materials:
1. Aroclor 1242, Lot No. AK-255
2. Aroclor 1254, Lot No. AK-38
3. Aroclor 1260, Lot No, AK-3
Water samples were collected under the following three bioaasay
vessel conditions:
1. Aroclor and fish exposure
2. Blank exposure {Aroclor concentration under Aroclor and fish
exposure, but without fish). 3. Control exposure (fish but no Aroclor).
013721
The water collection schedule is presented in Table I.
WATER PCB-SD0000036063
TABLE I TEST MATERIALS: Aroclor 1242, Aroclor 1254* and Aroclor 1260
Four-Day Fifth Toxicity Studiea
Bioaftftay Veaael Condition*
Aroclor-Fiah Expoaure (Aroclor plua Fiah)
Water Collection Interval* ________ (Hour*)_____________
'0
1 2 3 6 12
24 48
72
No Survival Concentration Partial Survival Concentration Total Survival Concentration
X X X XX XXX X X X
Blank Expoaure (Aroclor but no Fiah)
No Survival Concentration Partial Survival Concentration Total Survival Concentration
i Control Expoaure (Fiah but no Aroclor)
X X X X XX X X X X X X XX X X X X X X XX X X
X X X X X X X X.
X X X
X
015
96
X X X X X
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II. Summary
The results of the four-day fish toxicity studies are summarized
below.
Test Material
Four -Day TLS0 (ppm)
Bluegills
Channel Catfish
Aroclor 1242
0.24
0. 13
Aroclor 1254 Aroclor 1260
> 100.0 >100.0
s* 0. 20 >100.0
Respectfully submitted, INDUSTRIAL BIO-TEST LABORATORIES. INC.
Report prepared by:
Kenneth Ebbens, B. S. Assistant Toxicologist Acute Toxicity
Report approved by:
1
Carmen Mastri, B. S. Senior Group Leader Acute Toxicity
.
M.' L. Keplinge/T Ph.
Manager, Toxicology
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III, Procedure
14
017
The procedure wai generally* the same for each sample and species
o fish.
Healthy, young fish with an average length of 35 to 75 millimeters
were used as test animals. All fish were kept under observation for a
period of not less than ten days prior to experimental use. During this
period, the fish were observed for general health and suitability as
test animals. The fish were held in large aerated stock tanks and were
fed frozen brine shrimp or Purina Trout Chow 2 until three days
immediately prior to testing. The channel catfish and bluegills were
maintained at a temperature of 18 C. All stock tanks and bioassay
vessels contained reconstituted, deionized water. The following com
pounds were added in the amounts stated per liter of deionized water:
30 mg calcium sulfate 30 mg magnesium sulfate 48 mg sodium bicarbonate
2 mg potassium chloride
Preliminary screening was conducted in order to find the general
levels of toxicity of Aroclor 1242, Aroclor 1254 and Aroclor 1260.
Once these levels had been determined, three to ten concentrations of *
each test material were selected for the respective bioassays. Bio
assay vessels were filled with 12. 5 liters of well aerated reconstituted
water. Ten fish were tested at each concentration.
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After a 24-hour acclimatization period, calculated amounts of each Aroclor were dispersed into the respective vessels. The test dilutions were prepared within one hour of the bioassay.
Following the introduction of the test material, the fish were ob served for 96 hours, during which time all mortalities and/or untoward behavioral reactions were recorded. The concentration of dissolved oxygen was determined for all test solutions in which mortalities occurred to determine that adequate, oxygen was available to the test specimens. * Concentrations of dissolved oxygen of 4 mg/1 (4 ppm) for bluegills and channel catfish were considered to be minimal for valid testing. The pH of the teat solution was also determined when mortalities were recorded. All fish were collected, rinsed with deionized water, and preserved by freezing for future residue analysis.
At the end of the observation period, the four-day median tolerance limits (TL50'i) for each Aroclor were calculated employing the technique of Litchfield and Wilcoxon**.
In addition, water samples were collected at selected time intervals as listed in Table I.
013725
* Kach OX-4-P Dissolved Oxygen Test Kit, Hach Chemical Company,
Ames, Iowa.
** J. T. Litchfield, Jr. and F. Wilcoxon, "A Simplified Method of
Evaluating Dose-Effect Experiments," J. Pharm. k Exp. Thcr.
96,99 (1949).
,
WATER PCB-SD0000036067
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IV, Results
^ Ql9
A, Survival, DO, and pH Data
The survival, dissolved oxygen content, and pH data for
bluegills and channel catfish are presented in Tables 11 through VII.
013726
WATER PCB-SD0000036068
Concentration* (ppra)...
0.01 0.10 0.18 0.32 0.56 1.0
TABLE 11 TEST MATERIAL? Aroclor 1242
O^Q
Four-Day Fish T oxicity Study - Bluegills
Survival* DO* and pH Data
Number of Survivors (Hours)
1-6 24 48 72 96
Percent 1-6 Hrs. Survival DO PH
Dissolved Oxygen (ppm) and pH 24 Hrs. 48 Hrs. 72 Hrs.
DO pH DO pH DO pH
10 10 10 10 10 100
10
10 10 10
7
70 --
.-
--
10
10 10 9
7
70 -
- 57
10
10 10 8
6
60 - -
--
-
57
10 9 5 3 1 10 - - 5 7 4 7 4 7
10 4 0
- 0 - 5 7 5 7- .
96 Hre. DO pH
57 57 57 .4 7
--
Four-Day TL^q = 0.24 ppm Note: The test material was dispensed into the bioassay vessels in the form of a 0.1 percent (w/v) dilution in aceton<
- = Not determined.
;
Concentrations are expressed in terms of Aroclor 1242.
1
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Concentration* -iBBfHl.. ,
0.01 0.18 0.32 0.56 1.0 1.8 3.2 5.6 10.0 100.0
TABLE III TEST MATERIAL; Aroclor 1254
21
Four-Day Fish Toxicity Study - BluegiUs
Survival, DO, mad pH Data
Number ol Survivor* (Hours)
1-6 24 48 72 96
Percent Survival
1-6 Hrs. DO pH
Dissolved Oxygen (ppm) and pH
24 Hr*. 48 Hrs.
72 Hra.
DO pH DO pH DO pH
10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 . 10 10
100 100 100 100 100 100 100 100 100 100
--- - -- - - - ---- -
-- - ---- - -- -- -- -- - ---- ------- - -- ---- -
96 Hre DO pi
_ --
------
Four-Day TLjq > 100. 0 ppm
Note#; The test material was dispensed into the bioassay vessels in the form of 0. 1, 1.0, and 10.0 percent (w/v) dilutions in acetone for levels 0.01-0.56 ppm, 1.0-10.0 ppm, and 100.0 ppm, respectively.
- = Not determined.
* Concentrations are expressed in terms of Aroclor 1254.
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WATER PCB-SD0000036070
Concentration* (PPTM) .
10.0 100.0
TABLE IV
TEST MATERIAL: Aroclor 1260
Four-Day Fish Toxicity Study - Bluegills
Survival, DO, and pH Data
Number of Survivors . (Hours) 1-6 24 48 72 96
Percent Survival
. 1-6 Hrs. DO pH
Dissolved Oxygen (ppm) and pH 24 Hr. 48 Hrs. 72 Hrs.
DO pH DO pH DO pH
10 10 10 10 10 10 10 10 10 10
100 100
96 Hrs. DO pH
Four-Day T L^q > 100. 0 ppm Note: The teat material wai dispensed into the bioaaaay vellels in the form of a 10. 0 percent (w/v) dilution in aceton
Not determined. Concentrations are expressed in terms of Aroclor 1260.
I
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WATER_PCB-SD0000036071
Concentration*
____________
' 0.10 0.18 0.32 0.56 1.0 1.8 3.2 5.6
10.0
TABLE 1
TEST MATERIAL: Aroclor 1254
* 023
Four-Day Fish Toxicity Study - Channel Catfish Survival, DO, and pH Data
'
Number of Survivors
(Hours) 1-6 24 48 72 96
Percent Survival
1-6 Hr*. DO pH
Dissolved Oxygen (ppm) and pH
24 Hr*. 48 Hr.
72 Hr*.
DO pH DO pH DO pH
96 Hr DO pH
10 10 10 10 10
10 10 10 9 6 10 10 10 9 3 10 9 9 6 1 10 9 8 2 1 10 10 10 2 2 10 10 8 0 10 10 7 2 2 10 10 5 1 1
100 60 30 10
10 20
0* 20 10
6 7 5- - - - - -
7
7 7 6- - - - - -
7
- - 87777777
- - 8 77777 77
6 7 '- - - - - -
--
6 7 6 7- - - -
--
6 7 6 7- - - -
--
--
--
6 `1
7
6
7
Four-Day TLjQ-arO. 20 ppm Note: The test material was dispensed into the bioassay vessels in the form of 0. 1 and 1.0 percent (w/v) dilutions i
acetone for levels 0. 1 ppm through 0. 56 ppm and 1.0 ppm through 10. 0 ppm, respectively.
- = Npt determined. * Concentrations are expressed in terms of Aroclor 1254.
I
t
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Concentration*
____________
0.032 0.056 0. 10 0. 18 0.32 0.56 1.0
'
TABLE V
TEST MATERIAL? Aroclor 1242 Four-Day Fish Toxicity Study - Channel Catfish
024
Survival, DO, and pH Data
Number of Survivors (Hours)
1-6 24 48 72 96
Percent Survival
10 10 10 10 10
10 10 10 10
8
10 10 8 7 6
10 10 2 1 0
10 7 2 1 1
10 2 0 - -
10 0 -
100 80 60 0 10 0 0
1-6 Hrs, DO pH
Dissolved Oxygen (ppm) and pH 24 Hrs. 48 Hrs. 72 Hrs.
DO pH DO pH DO pH
- - ----- -
- - - - 6767
- - - - 6767
- -776767
- -7767- -
7 7
*" ** -*
96 Hrs. DO pH
57 57 57
--*- ~
Four-Day TL<jq = 0. 13 ppm
Note: The test material was dispensed into the bioassay vessels in the form of a 0.1 percent (w/vj dilution in aceton
- = Not determined. if
Concentrations are expressed in terms of Aroclor 1242.
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Concentration* (ppm)1-6
10.0 100.0
TABLE VII TEST MATERIAL: Aroclor 1260
02 &
Four-Day Fish Toxicity Study - Channel Catfish
Survival* DO, and pH Data
Number of Survivors (Hours) 24 48 72 96
Percent 1-6 Hrs. Survival DO pH
Dissolved Oxygen (ppm) and pH
24 Hrs.
48 Hrs. 72 Hrs.
DO pH DO pH DO pH
96 Hrs. DO pH
10 10 10 10 10
100
10 10 10 10 10 . 100
Four-Day T> 100.0 ppm Note: The test material was dispensed into the bioassay vessels in the form of a 10. 0% (w/v) dilution in acetone. - = Not determined. * Concentrations are expressed in terms of Aroclor 1260.
i
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1 13
B. Reactions The reactions noted following introduction of Aroclor 1242,
Aroclor 1254, and Aroclor 1260 are presented for bluegills and channel catfish in Tables VIII through XIII, respectively.
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TABLE VIII
TEST MATERIAL: Aroclor 1242 Four-Day Fish Toxicity Study-Bluegills
27
Summary of Reactions
Concentration . <PPm>_________
0. 01
Reaction None
Time of Onset Following Dose Administration
(hours)
-
Duration of
Reaction (hours)
-
Time of Death Following Dose Administration
(hours) '
-
0. 10 0. 18 0. 32 0. 56
Hypoactivity
,
Hypoactivity
Hypoactivity
Hypoactivity . Loss of equilibrium
96
72 n 12
1 -6 12
No recovery noted
No recovery noted
No recovery noted
No recovery noted 36
96 72 - 96 72 - 96 24 - 96
_
1. 0 if
Hypoactivity Loss of equilibrium Prostration
1-6 1 -6 12
Until death
24 - 48
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Concentration
(PPm)
..
0.01 0.18 0. 32 0. 56 1.0 1.6 3.2 5.6 10.0 100.0
TABLE IX
TEST MATERIAL: Aroclor 1254 Four-Day Fish Toxicity Study-Bluegills
Summary of Reactions
Reaction
Time of Onset
Duration
. Following Dose
of
Administration
Reaction
(hours)
(hours)
None None None None None None
None None None None
-#
'
-
-
-
-
-
-
-
#28
Time of Death Following Dose Administration
(hours)
-
-
i
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Concentration (PPm)....... 10.0
100.0
TABLE X TEST MATERIAL: Aroclor 1260
029
Four-Day Fish Toxicity Study-Bluegills Summary of Reactions
Reaction
Time of Onset Following Dose Adminis tration
(hours)
Duration of
Reaction (hours)
Time of Death Following Dose Administration
(hours)
Hypoactivity
72
No' recovery noted
-
Hypoactivity
72
No recovery noted
-
------------------- 1------------- -
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Concentration ____ feBm)-
0. 032 0. 056 0. 10 0.18 0.32
0. 56
1. 0
TABLE XI
%Sdi)
TEST MATERIALS- Aroclor 1242
Four-Day Fish Toxicity Study-Channel Catfish
Summary of Reactions
Reaction None
Time of Onset Following Dose. Admini s tration
(hours)
-.
Duration of
Reaction (hours)
-
Time of Death Following Dose Adminis tration
(hours)
Hypoactivity Lon of equilibrium
96 96
No recovery noted
96
Hypoactivity Loss of equilibrium
48 48
No recovery noted 24
48 - 96
Hypoactivity Prostration
48 72
Until death
48 - 96
Hypoactivity Prostration Surfacing Vertical swimming
.
Hypoactivity Prostration Vertical swimming
Hypoactivity Prostration Vertical swimming
1-6 24 48 48
No recovery noted 24 24 24
1-6 24
24 013737
I-6 1 -6 I -6
Until death
Until death
24 - 72 24 * 48 24
WATER PCB-SD0000036079
Concentration . (PPm)
0. 10 ' 0. 18 0. 32 0.56 1.0
1.8
3. 2
5.6
10.0
TABLE XII
TEST MATERIAL:' Aroclor 1254
Four-Day Fish Toxicity Study-Channel Catfish
Summary of Reactions
Reaction None
Time of Onset Following Dose Administration .......................... (hours)
Duration of
Reaction (hours)
-
. Time of Death Following Dose Administration (hours)
-
Hypoactivity
72
No recovery noted
72 - 96
Hypoactivity Hypoactivity
. .
72 24 '
No recovery noted No recovery noted
72 - 96 24 - 96
Hypoactivity Prostration
24
No recovery noted
24 - 96
24 48
Hypoactivity Prostration
24
No recovery noted
72
24 48
Hypoactivity Prostration
24
Until death
48 - 72
24 48
Hypoactivity 013738
Prostration
24
No recovery noted 48
48 - 72
Hypoactivity Prostration
. 24 24
No recovery noted 48
48 - 72 -
_________ ______1___
WATER PCB-SD0000036080
Concentration . (PPm)
10.0
100.0
TABLE XIII
TEST MATERIAL: Arodor 1260
Four*Day Fish Toxicity Study-Channel Catfish Summary of Reactions
Reaction
Time of Onset Following Dose Administration
(hours)
Duration of
Reaction (hours)
/ Time of Death Following Dose Administration (hours)
Hypoactivity Hypoactivity
72
No recovery noted
-
72
No recovery noted
-
o
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