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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 Hrrt*r IBT NO. A8570
I. Introd uc tion At the request of Monsanto Company, four-day fish toxicity
studies using bluegilis (Lepomis macrochirus),and channel catfish (Ictalurus punctatus) were conducted with the following three materials:
1. Aroclor 1242, Lot No. AK-255 r * 2. Aroclor 1254, Lot No. AK-38 3. Aroclor 1260, Lot No. AK-3 S'" Water samples were collected under the following three bioassay 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). The water collection schedule is presented in Table 1.
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TABLE I
TEST MATERIALS} Aroclor 1242, Aroclor 1254, and Aroclor 1260
Four-Pay Fish Toxicity Studies
Bioassay Vessel Conditions
Water Collection Intervals ______(Hour a}
0 1236
12
24 48
72
Aroclor-Fish Exposure (Aroclor plus Fish]
No Survival Concentration Partial Survival Concentration Total Survival Concentration
X X X XX XXX X X X
Blank Exposure (Aroclor but no Fish)
No Survival Concentration Partial Survival Concentration Total Survival Concentration
X X X X XX X X X X X X XX X X X X X X XX X X
X X X
Control Exposure (Fish but no Aroclor)
X X X X XX X X
X
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 Aroclor 1242. Aroclor 1254 Aroclor 1260
Four-Day TLjo (ppm)
Bluesills
Channel Catfish
0.24
0.13
>100.0
^0.20 -
>100.0
> 100.0
Respectfully submitted, INDUSTRIAL BIO-TEST LABORATORIES, INC.
Report prepared by:
Kenneth Ebbens, B.S. Assistant Toxicologist Acute Toxicity
Report approved by:
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Carmen Mastri, B.S. Senior Group Leader Acute Toxicity
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M. L, Keplin Manager, Toxicology
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III. Procedure The procedure was generally the same for each sample and species
of 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 HZ 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 Z mg potassium chloride
Preliminary screening was conducted in order to find the general levels of toricity 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 test 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 (TLgo's) 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 1.
,
* Hach 0X-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,M J. Pharm. k Exp, Thor. 96,99 {1949).
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IV, Results
-
A, Survival, DO, and pH Data
The survival, dissolved oxygen content, and pH data for
bluegills and channel catfish are presented in Tables II through VII,
J>/
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Concentration* (opm)
0. 01 0. 10 0.18 0.32 0.56 1.0
TABLE II
TEST MATERIAL! Aroclor 1242
Four-Day Fish Toxicity Study - BluegUl*
1 Survival, DO, and pH Data
Number of Survivors (Hours)
1-6 24 48 72 96
Percent Survival
10 10 10 10 10 100
10
10 10 10
7
70
10
10 10 9
7
70
10
10 10 8
6
60
10 9 5 3 1
10
10 4 0 *
0
1-6 Hra. DO pH
Dissolved Oxygen (ppm) and pH 24Hra. 48 Hra. 72 Hra, DO oH DO pH DO oH
.
.. .
..
-
.
. ...
. 57
. . . .. . 5 7
- 5 74 74 7
- - 5757
96 Hrs. DO pH
57 57 57 47 *
Four-Day TL50 * 0> 24 ppm
Note: The tost material was dispensed into the bioassay vessels in the form of a 0.1 percent (w/v) dilution in acetone.
* Not determined.
.
' Concentrations are expressed In terms of Aroclor 1242.
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TABLE III
TEST MATERIALS Aroelor 1254
Four-Day Fish Toxicity Study - Bluegills
' Survival, DO, and pH Data
..............Number^/Survivors
Concentration* (ppm)
(Hours) 1-6 24 48 72 96
Percent Survival
1-6 Hre. DO pH
Dissolved Oxygen (ppm) and pH
24 Hrs. 48Hrs. DO pH DO pH
72 Hrt. DO pH
96 Hrs. DO pH
0,01 0, 18 0.32 0.56 1.0 1.8 ' 3.2 5.6 10.0 100.0
10 10 10 10 10 100 10 10 10 10 10 100 10 10 10 10 10 100 10 10 10 10 10 100 10 10 10 10 10 100 )0 10 10 10 10 100 10 10 10 10 10 100 10 10 10 10 10 100 10 10 10 10 10 100 10 10 10 10 10 100 .
Four-Day TLgQ > 100, 0 ppm
Note*. The teat material was dispensed into the bioassay vessels in the form o 0.1, 1.0, and 10,0 percent (w/v) dilutions in acetone for levels 0. 01-0. 56 ppm, 1.0-10,0ppm, and 100,0 ppm, respectively.
Not determined.
Concentrations are expressed in terms of Aroelor 1254.
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Concentration* (ppm)
10.0 100.0
TABLE IV
TEST MATERIAL: Aroclorl26D
Four-Day Fish Toxicity Study - Bluegllls
> Survival, DO, and pH Data
Number of Survivors (Hours)
1-6 24 48 72 96
Percent Survival
10 10 10 10 10
100
10 10 10 10 10 . 100
Dissolved Oxygen (ppm) and pH 1-6 Hrs. 24Hrs. 48 Hra. 72 Hrs. DO pH DO dH DO pH DO pH
96Hrs. DO pH
Four-Day TLjq > 100. 0 ppm Note: Tho test material was dispensed into the bioassay vellcle In the form of a 10. 0 percent (w/v| dilution In acetone. - = Not determined. * Concentrations are expressed lit terms of Aroclor 1260.
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AS 3 A * 130
Concentration* (DDm)
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
> 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 ]
10 2 0 -
10 0
'
100 80 60 0 10 0 0
1 -6 Hrs. DO i>H
.._ .. ."*
Dissolved Oxygen (ppm) and pH 24 Hrs. 48 Hrs. 72 Hrs.
DO pH DO pH DO pH
._676 -_ 6 767 7 7 6767 7 7 67 77 -
96 Hrs. DO pH
57 57 57 .. --
*
Four-Day TLgg * 0.13 ppm Note: The test material was dispensed Into the bioassay vessels in tho form of a 0. 1 percent (w/v) dilution in acetone. * Not determined,
Concentrations are expressed in terms of Aroclor 1242.
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Concentration0 (pom)
0.10 0. 18 0. 32 0.56 1.0 1.8 3.2 5. 6 >0. 0
TEST MATERIAL: Aroclor 1254
Four.Day Fiah Toxicity Study - Channel Catfish 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 Hrs. 48 Hrs.
72 Hrs.
DO pH DO pH DO pH
96 Hrs. DO pH
to 10 10 10 10
100
10 10 10 9 6
60 - - - . - - 6 7 5 7
10 10 10 9 3
30 - - - - - - 7 7 6 7
10 9 9 6 1
10 - - 8 7 7 7 7 7 7 7
to 9 8 Z 1
10 - - 8 7 7 7 7 7 7 7
10 10 10 2 2
20 - - - - - - 6 7
10 10 8 0 .
0 - -- -6767-
10 10 7 2 2
20 . . . - 6 7 6 7 -
10 10
5
1
1
10 ` ` ` 6 7 6 7
O m in
t
Four-Day TL^q-vO. 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 in acetone for levels 0. 1 ppm through 0. 56 ppm and 1.0 ppm through 10. 0 ppm, respectively.
- ~ Not determined.
* Concentrations arc expressed in terms of Aroclor 1254.
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Concentration* (ppm)
10.0 100.0
TABLE VJI
TEST MATERIAL: Aroclor 1260
Four-Day Fish Toxicity Study Channel Catfish
' Survival. DO, and pH Data
Number of Survivors (Hours)
1-6 24 48 72 96
Percent Survival
Dissolved Oxygen (ppm) and pH
1-6 Hrs. 24 Hrs,
48Hrs. 72 Hrs.
DO oH DO oH DO nH DO pH
10 10 10 10 10
100
10 10 10 10 10 . 100
96 Hrs. DO pH
Four-Day TLjq > 100. 0 ppm Note: The test material was dispensed into the bloassay vessels In the form of a 10.0% (w/v) dilution in acetone. - * Not determined. Concentrations are expressed in terms of Aroclor 1260.
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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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Concentration (ppm) 0.01 0. 10 0. 18 0. 32 0. 56
1.0
TAB LE VIII
TEST MATERIAL* Aroclor 1242
Four-Day Fish Toxicity Study-Bluegills
Summary ol Reactions
Reaction
Tima of Onset Following Dose Administration
(hours)
Duration of
Reaction (hours)
Time of Death Following Dose
Administration (hours)
None
- --
Hypoactivity
,
96
No recovery noted 96
Hypoactivity
72 No recovery noted 72 - 96
Hypoactivity
12
No recovery noted 72 - 96
Hypoactivity , Lost oi equilibrium
1-6 : 12
No recovery noted 36
24 - 96
Hypoactivity Lost oi equilibrium Prostration
1 -6 1-6 12
Until death
24 - 48
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Concentration (ppm)
0. 01 o. la 0. 32 0. 56 1. 0 i.e 3. 2 5.6 10,0 100.0
TABLE IX
TEST MATERIAL! Aroclor 1254
Four Day Fish Toxicity Study-Bluegillfl
Summary of Reactions
Reaction
None None None None None None None None None None
' Time of Onset Following Dose Administration (hours)
. -. -. -
'-
'
Duration of
Reaction (hours)
.
-
'
Time of Death Following Dose Administration
(hours)
`
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MO-
Concentration fpnml 10. 0
100.0
TABLE X
TEST MATERIAL: Aroclor 1260
Four-Day Fish Toxicity Study-Bluegllls , Summary of Reactions
Reaction
` Time of Onset Following Dose
Administration (hours)
Duration ' of
Reaction (hours)
Time of Death Following Dose Administration
(hours)
Hypoactlvity
72
No recovery noted
Hypoactlvity
72
No recovery noted
.
;*t ui
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Concentration 0. 032 0.056 0. 10 0. 18 0. 32
0. 56
1. 0
TABLE XI
,
TEST MATERIAL:' Aroclor 1242
Four-Day Fish Toxicity Study-Channel Catfish
. Summary of Reactions
Reaction
None
Hypoactivity Loss of equilibrium
Hypoactivity Loss of equilibrium
Hypoactivity Prostration
Hypoactivity Prostration Surfacing Vertical swimming
Hypoactivity Prostration Vertical swimming
Hypoactivity
Vertical swimming
Time of Onset Following Dose, Administration
(hours)
Duration of
Reaction (hours)
Time of Death Following Dose Administration
(hours)
---
<56
No recovery
96
96 noted
48
No recovery noted
48 - 96
48 24
48
Until
48 - 96
72 death
1 -6 24 48 48
No recovery noted 24 24 24
24 - 72
1 -6 24 24
1 -6 1-6 I -6
Until death
Until death
24 - 48
24 063661
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HttclmlniaL B I O - T E S T Ja b ta fo A icI, Hue,
mcentr&tion (ppm) 0. 10 0. 18 0. 32 0. 56 (.0
1.8
3.2
5.6
10.0
TAB LE XII
TEST MATERIAL? ' Aroclor U54 Four-Day Fish Toxicity Study-Channel Catfish
.
Reaction None Hypoactivity Hypoactivity
Summary of Reactions
' Time of Onset Following Dose Administration (hours)
Duration of
Reaction (hours)
Time of Death Following Dose Administration
(hours)
---
72
No recovery noted
72 - 96
72
No recovery noted
72 - 96
Hypoactivity
Hypoactivity Prostration
Hypoactivity Prostration
Hypoactivity Prostration
Hypoactivity Prostration
Hypoactivity Prostration
24
No recovery noted
24 - 96
24
No recovery noted
24 - 96
24 48
24
No recovery noted
72
24 48
24
Until death
48 - 72
24 48
24
No recovery noted
48 - 72
24 48
24
No recovery noted
48 72
24 48
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9nJujliiaL B IO -T E S T
Concentration (ppm)
10. 0
100.0
TABLE XIII
TEST MATERIAL:' Aroclor 1260
Pour-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
-
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