EPA-600/3-76-061b
July 1976
Ecological Research Series
STANDARD TEST FISH DEVELOPMENT
Parti
Environmental Research Laboratory
Office of Research and Development
U.S. Environmental Protection Agency
Duluth, Minnesota 55804
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RESEARCH REPORTING SERIES
Research reports of the Office of Research and Development, U.S. Environmental
Protection Agency, have been grouped into five series. These five broad
categories were established to facilitate further development and application of
environmental technology. Elimination of traditional grouping was consciously
planned to foster technology transfer and a maximum interface in related fields.
The five series are:
1. Environmental Health Effects Research
2. Environmental Protection Technology
3. Ecological Research
4. Environmental Monitoring
5. Socioeconomic Environmental Studies
This report has been assigned to the ECOLOGICAL RESEARCH series. This series
describes research on the effects of pollution on humans, plant and animal
species, and materials. Problems are assessed for their long- and short-term
influences. Investigations include formation, transport, and pathway studies to
determine the fate of pollutants and their effects. This work provides the technical
basis for setting standards to minimize undesirable changes in living organisms
in the aquatic, terrestrial, and atmospheric environments.
This document is available to the public through the National Technical Informa-
tion Service. Springfield, Virginia 22161.
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EPA-600/3-76-06lb
July 1976
STANDARD TEST FISH DEVELOPMENT
PART II
Chronic Toxicity of Guthion to the Fathead Minnow
(Pimephales promelas Refinesque)
by
Ira R. Adelman
and
Lloyd L. Smith, Jr.
Department of Entomology, Fisheries, and Wildlife
University of Minnesota
St. Paul, Minnesota 55108
Grant No. R800940
Project Officer
Robert W. Andrew
Environmental Research Laboratory
Duluth, Minnesota 55804
U.S. ENVIRONMENTAL PROTECTION AGENCY
OFFICE OF RESEARCH AND DEVELOPMENT
ENVIRONMENTAL RESEARCH LABORATORY
DULUTH, MINNESOTA 55804
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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.
Approval does not signify that the contents necessarily reflect the views
and policies of the U.S. Environmental Protection Agency, rior does mention
of trade names or commercial products constitute endorsement or recommen-
dation for use.
ii
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ABSTRACT
(£>
Three chronic bioassays of Guthioir^were conducted with fathead minnows.
All tests were begun with eggs and the longest lasted 20 days after
termination of spawning, a total of 250 days. Parameters measured were
survival, growth, fecundity, and growth and survival of second genera-
tion fry and eggs. The most sensitive criteria for effect of the
toxicant was fecundity. The maximum acceptable toxicant concentration
(£>
was between 0.33 and 0.51 pg/liter Guthion, and the application factor
was between .0017 and .0027 based on a soft water acute and a hard
water chronic bioassay.
iii
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CONTENTS
Page
Abstract iii
List of Tables vi
Acknowledgements V11
Sections
I Conclusions 1
II Recommendations 2
III Introduction 3
IV Materials and Methods 4
Number of Experiments 4
Apparatus 4
Fish 5
Chemical Analysis 9
V Results 11
(
Egg and Fry Survival 11
Growth 13
Reproduction 13
Second Generation 16
VI Discussion 17
VII References 18
VIII Publications 20
IX Glossary 21
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TABLES
No. Page
1 Analysis of Well Water 6
2 Means and Standard Deviations (in Parentheses) of
Conditions in All Test Chambers 7
3 Percentage Survival of Fathead Minnow Eggs and Young
Exposed to Various Concentrations of Guthion^ 12
4 Mean Weight of Fathead Minnows at Various Time
Intervals After Spawning (Number of Fish Weighed in
Parentheses) 14
5 Fecundity of Fathead Minnows Exposed to Various
Concentrations of Guthion^ 15
6 Effect of Guthiori^on Survival and Growth of Second
Generation Eggs and Fry 16
vi
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ACKNOWLEDGEMENTS
The authors wish to thank Richard W. Frenzel and particularly
Gary D. Siesennop for assistance in conducting the experiments.
vii
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SECTION I
CONCLUSIONS
The results described in the following report permit certain conclusions
concerning the toxicity of Guthiorr to fathead minnows.
1. The maximum acceptable toxicant concentration for fathead minnows is
between 0.33 and 0.51 yg/liter Guthion.
2. The application factor comparing an acute bioassay in soft water and
a chronic bioassay in hard water is between .00017 and .00027.
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SECTION II
RECOMMENDATIONS
(iv)
Guthion^ concentrations in the environment should not exceed
0.33 yg/liter.
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SECTION III
INTRODUCTION
GuthiorP (azinphosmethyl) is an important and widely used insecticide
on deciduous fruits and cotton as well as many other fruits and vege-
tables. Although acute toxicity data have been reported by many workers
for a variety of fish species (Katz, 1961; Pickering, Henderson, and
Lemke, 1962; Meyer, 1965; and Macek and McAllister, 1970), no informa-
tion has been reported on the effect of exposures over a complete life
cycle. Since data on chronic toxicity are important for establishment
of water quality standards, fathead minnows were exposed to various
®
concentrations of Guthion over a complete life cycle, from egg through
fry in the second generation, in order to determine a maximum acceptable
toxicant concentration (MATC) (Mount and Stephan, 1967).
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SECTION IV
MATERIALS AND METHODS
NUMBER OF EXPERIMENTS
Three experiments were conducted, subsequently referred to as tests I,
II, and III. Tests I and II were preliminary and were terminated after
26 and 68 days, respectively, when it became apparent that the lowest
concentration was causing significantly greater mortality than the
control. Furthermore chemical analysis of small Guthion concentrations
had not been perfected, and residues on glassware and water sampling
equipment made analyzed concentrations inaccurate. This problem was
particularly true for test II where desired concentrations were low
enough to be influenced. Therefore, information from the first two
tests is only roughly quantitative but does indicate an area of possible
effect. For test III all glassware was soaked in a strong sodium hy-
droxide solution (pH = 12"^ for at least two days prior to use and no
residues were found in blank analyses.
APPARATUS
A proportional diluter (Mount and Brungs, 1967) that dispensed seven
toxicant concentrations with a 60% dilution factor and one control was
used for administration of the toxicant in a flow-through system.
Technical grade Guthioir^ (93%) was added directly to a single mixing
chamber by the injector described by DeFoe (1975). An air dispersion
stone and baffles thoroughly mixed the Guthion^with incoming water
before it was siphoned to the toxicant cells in the diluter. The test
chambers for first generation young and adult fish were constructed of
-------
glass and G.E. Silicone Seal, measured 50 x 25 x 20 cm, and contained
20 liters. With a flow rate of 250 ml/min, 90% replacement of water
occurred in approximately 3-1/4 hr. Temperature was maintained at a
constant 25 C by a hot water exchange system in the head tank where
aeration also occurred. All test chambers in test III contained an air
dispersion stone in order to maintain oxygen concentrations. A dense
bacterial growth occurred in the two highest concentrations apparently
using the Guthion^ as a nutrient and the aeration was necessary to
replace oxygen consumed by the bacterial respiration. Chambers were
siphoned daily except when fry were present to keep this bacterial
growth to a minimum. A complete analysis of hard water from a deep
well is reported in Table 1. Those water quality characteristics which
varied in individual test chambers are listed in Table 2.
Fry in the second generation were reared in glass chambers measuring
20 x 20 x 21 cm and containing 6.4 liters. For each concentration
water flowed from the outlet of the adult test chamber into the first
fry chamber, then from the outlet of that to a second fry chamber so
that two groups of offspring from the same adults could be reared simul-
taneously. Eggs were hatched in a small jar with a nylon screen bottom.
One jar in each adult test chamber was oscillated so that water flowed
past the eggs without raising them from the screen.
All adult chambers were exposed to the direct illumination of a 40-watt
cool white fluorescent tube, suspended 50 cm above the water surface.
Second generation fry chambers received only indirect illumination from
laboratory light. Tests were begun with a photoperiod equivalent to
January 1 at Evansville, Indiana and adjustments were made every 15 or
16 days.
FISH
Fathead minnows originally obtained from the Environmental Research
Laboratory at Duluth, Minnesota were reared in a laboratory culture
system. All tests were started with eggs from five spawning pairs which
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Table 1. ANALYSIS OF WELL WATER
(milligrams/liter)
Item
Concentration
Total hardness as CaCO,
Calcium as CaCO_
Iron
Chloride
Sulfate
Sulfide
Fluoride
Total phosphates
Sodium
Potassium
Copper
Manganese
Zinc
Cobalt* nickel
Cadmium, mercury
Ammonia, nitrogen
Organic nitrogen
220
140
0.02
<1
<5
0.0
0.22
0.03
6
2
0.0004
0.0287
0.0044
<0.0005
<0.0001
0.20
0.20
taken from well head before aeration and heating;
pH 7.5.
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Table 2. MEANS AND STANDARD DEVIATIONS (IN PARENTHESES) OF CONDITIONS IN ALL TEST CHAMBERS
Item
Test chamber
12345678
Guthiori^Xug/l) 0 7.6(1.3)
Temp. (C) 24.8(.6) 25.0(.6)
pH (reading) 7.73(.13) 7.53(.11)
DO (mg/1) 6.60(.66) 5.95(.24)
Guthion®(yg/l) 0 1.8(.9)
Temp. (C) 24.6(1.7) 24.6(1.8)
pH (reading) 7.75(.09) 7.58(.11)
DO (mg/1) 6.28(.77) 5.54(.45)
Guthion®(yg/l) .04(.OS)-'' .10(.04)
Temp. (C)
pH (reading)
DO (mg/1)
24.1(1.1) 24.4(1.2)
6.96(.34) 6.52(.57)
Test I
11.7(1.9) 20.5(1.9) 34.5(3.2)
24.9(.6) 25.0(.6) 25.0(.6)
7.52(.10) 7.49(.09) 7.51(.ll)
5.55(.56) 5.03(.05) 5.20(.74)
Test II
1.9(1.0) 2.8(1.1) 3.4(1.2)
24.6(1.9) 24.4(1.9) 24.6(1.9)
7.57(.10) 7.60(.09) 7.60(.09)
5.55(.45) 5.58(.31) 5.36(.38)
Test III
Adult chambers
.16(.05) .24(.06) .33(.08)
24.4(1.2) 24.4(1.2) 24.6(1.4)
8.07(.ll) 8.06(.ll) 8.05(.ll)
6.49(.34) 6.27(.64) 5.79(.59)
61.4(2.4) 96.7(6.6) 165.9(13.1)
24.9(.6) 25.1(.6) 25.1(.6)
7.52(.16) 7.50(.09) 7.56(.17)
4.48(1.28) 4.46(.61) 5.13(.51)
4.8(.8) 6.5(.8) 15.0(2.2)
25.4(.6) 25.3(.6) 25.4(.6)
7.58(.08) 7.60(.09) 7.63(.10)
5.22(.57) 5.17(.61) 4.90(1.07)
Ml)
24.5(1.4)
8.05(.ll)
5.77(.90)
.72(.15)
24.4(1.2)
8.06(.12)
:.86)
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Table 2 (continued). MEANS AND STANDARD DEVIATIONS (IN PARENTHESES) OF CONDITIONS
IN ALL TEST CHAMBERS
00
Test chamber
Item
1
2
3
4
5
6
7
8
Test III
Fry chamber 1
Guthion^yg/l)-^ . 04 ( . 03)^ . 10 ( . 04)
Temp . (C)
pH (reading)
DO (mg/1)
Temp. (C)
pH (reading)
DO (mg/1)
23.6(.8)
8.22(.08)
7.30(.41)
23.0(.6)
8.23(.08)
7.46(.32)
23.8(.5)
8.17(.04)
6.86(.22)
23.0(.6)
8.16(.05y
6.96(.32)
.16(.05)
23.8(.4)
8.15(.05)
6.81(.30)
Fry
22.9(.4)
8.16(.05)
7.09(.29)
.24(.06)
23. 8(.
8.16(.
6.65(.
chamber
23. 0(.
8.15(.
6.99(.
5)
04)
33)
2
6)
05)
34)
.33(
23. 9(
8.12(
6.17(
23. 1(
8.13(
6.45(
.08)
.4)
.02)
• 41)
.6)
.07)
.51)
.51(.
23. 6(.
8.15(.
6.00(.
23.2(1
8.18(.
6.49(.
11)
6)
10)
76)
.0)
08)
84)
.72(.
23. 6(.
8.14(.
5.87(.
22.4(1
8.15(.
6.06(.
15)
4)
08)
60)
.4)
08)
83)
a/
j-,Concentration may be high by as much as 50% for the four^lowest concentrations.
—/Unknown source of contamination resulted in some Guthion^ in control.
— Concentrations measured in adult chambers.
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were distributed in a random stratified manner to egg chambers in each
toxicant concentration and control. Initial numbers of eggs were 90,
70, and 100 in tests I, II, and III, respectively. Eggs were in the
middle to late blastula stage when first exposed to the toxicant and
more than 99% were fertile. Mortality of eggs and fry was recorded at
24-hr intervals in tests I and II. In test III eggs were checked at
24-hr intervals but mortality of young was recorded only after 22 days
to avoid increased stress from handling. At that time two runts or
malformed fish were removed from each chamber and fish were randomly
thinned to 15 fish.
Ten-cm sections of transite pipe were split lengthwise to form a
spawning substrate. After 90 days six of these sections were placed in
each aquarium, three on each end with an open area in the center. As
secondary sexual characteristics developed, males were removed at bi-
weekly intervals to keep no more than five per chamber. Spawning sub-
strates were checked at 1000 hours each day and if spawning occurred,
eggs were counted and a random sample of 50 was placed in the egg basket
for determination of survival through hatching unless the basket was
already occupied. Two groups of fry from each concentration were reared
for 50 days, otherwise all hatched fry were discarded.
Fry were fed ground Glencoe trout pellets and live brine shrimp twice a
day for the first 30 days. Older fish were fed Oregon Moist frozen
pellets twice a day and live brine shrimp once a day.
CHEMICAL ANALYSIS
Concentrations of Guthion6^ in each adult test chamber were usually
analyzed twice per week for the first 5 months of test III and usually
once per week thereafter. Samples from either or both fry chambers were
analyzed periodically. Since the percentage difference between fry
chambers and adult chambers analyzed at the same time ranged from -24%
to +21% with a mean percentage difference of iO%, the analyses from
adult chambers were used.
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A 1-liter sample of water from each test chamber was extracted with 10
ml benzene (pesticide grade) and the benzene was then evaporated to 0,5
to 2 ml, depending on the concentration. With each series of samples
(§)
1 liter of deionized water was spiked with 1 ml Guthion^ in acetone and
this sample was treated the same as all others to determine a recovery
efficiency which averaged 99.2 - 6.5%. Samples were analyzed on a Beck-
man GC 72-5 gas chromatograph with a 122-cm glass column packed with 50%
DC 200 on 80/100 mesh Gas Chrom-Q and a nonradioactive electron capture
detector. Standards were prepared from 93% technical grade Guthion^ and
were injected alternately with unknowns. Final Guthion*^concentrations
were corrected for the recovery efficiency and the technical grade pro-
duct.
The pH was measured with a meter twice per week in adult chambers and
once per week in fry chambers when they were occupied. Temperature was
measured with a thermister three times per week in all chambers and dis-
solved oxygen was analyzed weekly by the azide modification of the
Winkler analysis.
10
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SECTION V
RESULTS
EGG AND FRY SURVIVAL
Survival of eggs was probably not directly affected by any concentration
/g\
of Guthion** tested. In Test I percentage survival through hatching
®
decreased from 90 to 29 to 17% as Guthion concentration increased
from 34.5 to 61.4 to 96.9 yg/liter but then survival increased to 76%
®
as Guthion concentration increased to 165.9 yg/liter (Table 3). This
decreased survival was caused by eggs in these concentrations being
/&,
enmeshed in a bacterial network that apparently used Guthioir* as an
energy source and grew in proportion to the concentration. Dead eggs
in chambers with reduced survival were partly hatched as the fry seemed
to become enmeshed in the bacteria and could not break free. Since the
chorion of these eggs softened, fry may also have been weakened. Sur-
vival in the highest concentration was apparently greater because most
eggs hatched 1 day sooner, when the bacteria were less dense and before
the chorion had softened.
At 22 days there was decreased survival of fry in comparison to controls
at 7.6 yg/liter in Test I, and at approximately 6.5 yg/liter in Test II
(Table 3). By 57 days survival decreased at approximately 1.8 yg/liter
(R)
Guthion^ in Test II but did not decrease at 0.51 yg/liter in Test III.
In Test III Guthion caused no decrease in survival at any life history
stage with the possible exception of the highest concentration where
fry were lost wheh the chamber overflowed. The MATC in terms of sur-
vival was between 0.51 yg/liter in Test III and a maximum of 1.8 yg/liter
11
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Table 3. PERCENTAGE SURVIVAL OF FATHEAD MINNOW EGGS AND YOUNG EXPOSED
TO VARIOUS CONCENTRATIONS OF GUTHIOt
Test chamber
Item
Guthion^Cug/l)
% hatched
% survival
22 days
Guthion (yg/1)
% hatched
% survival
22 days
57 days
98 days
/**>
Guthion^Xyg/l)
% hatched
% survival
22 days
from 22-
60 days
1
0
99
54
0
91
57
54
52
2
7.6
84
43
1.8
92
48
42
35
.O^.IO
86
61
100
84
80
100
3
Test
11.7
97
29
Test
1.9
90
57
34
21
Test
.16
82
74
93
4
I
20.5
95
16
a/
II-
2.8
90
43
5
0
III
.24
85
82
100
5
34.5
90
1
3.4
87
48
3
0
.33
85
76
100
6
61.4
29
0 -
4.8
93
48
0
0
.51
86
67
93
7 8
96.9 165.9
17 76
0 0
6.5 15.0
63 84
37 35
0 0
0 0
.72
83
c/
v/ _
88
a/
— Concentration may be high by as much as 50% for the four lowest con-
centrations .
— Unknown source of contamination resulted in some Guthion in the
control.
c7
— Tank overflowed and fry were lost.
12
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where increased mortality occurred in Test II.
GROWTH
In test II fish were ewighed twice en masse during the experiment and
at termination. Each time the control fish weighed more than those in
the lowest concentration, 1.8 yg/liter (Table 4). In test III the first
two weighings were of individual fish that were randomly thinned from
the experiment. The third weighing was of fish that had remained in the
experiment. Individual t-test comparisons indicated no significant dif-
ference between the control and any of the treatments (p>.05) at 22 days
(Table 4). At 60 days the 0.10 and 0.24 yg/liter treatment levels
weighed significantly more than the controls (p<.02) but since the
0.16 yg/liter treatment level was not different from the control,
Guthion was probably not the cause of the above differences. After
90 days there was no significant difference (p>.05) between the control
and any treatment. Although dissolved oxygen concentrations were lower
with increasing Guthioir^ concentrations, these oxygen levels did not
seem to affect growth.
By 120 days males in some treatments started to defend territories and
fish were not weighed to avoid any influence of handling on spawning
behavior. At termination of the experiment after 250 days there was no
consistent pattern of growth related to Guthioir concentration. There-
fore, Guthion5^ affected growth of fathead minnows through 90 to 100 days
at essentially the same concentrations that caused mortality, or between
0.72 and 1.8 yg/liter.
REPRODUCTION
Interpretation of fecundity data was complicated by the variation in sex
ratios in test chambers. Although an attempt was made to maintain five
males and seven females in each chamber, in some chambers that many
males were not produced, in some immature males could not be identified,
and in some individuals were lost to accidental death or escapement.
13
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Table 4. MEAN WEIGHT OF FATHEAD MINNOWS AT VARIOUS TIME INTERVALS AFTER SPAWNING
(NUMBER OF FISH WEIGHED IN PARENTHESES)
Item
Test chamber
12 3 4 5 6 7 8
Guthion® (ug/1)
Weight (mg)
57 days
85 days
98 days
Guthion® (yg/1)
Weight (mg)
1.8
Test II
1.9 2.8
3.4
4.8
162(33) 111(29) 55(21) 72(3) 20(2)
320(33) 250(24) 230(14)
380(33) 350(24) 300(13)
0.4
0.10
Test III
0.16 0.24
0.33
0.51
6.5
0.72
a/
j-.Fish randomly thinned from experiment.
—.Fish remaining in experiment.
— Termination.
15.0
22
60
90
250
days^
days-
days—
days— - male
female
24(8)
179(15)
580(15)
2834(3)
1434(9)
24(27)
242(15)
580(15)
3047(5)
1594(7)
28(25)
216(13)
640(15)
3455(5)
1458(5)
27(19)
231(15)
660(14)
3051(5)
1353(5)
23(21)
260(15)
600(15)
2628(5)
1558(5)
21(11)
165(13)
600(13)
2965(6)
1360(6)
_
_
720(15)
3136(5)
912(1)
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Spawning occurred after 121 days when the day length reached 12 hr and
45 min. Spawning ceased after 230 days when the day length was reduced
to 12 hr. Intensive spawning occurred first at 0.33 yg/liter GuthionY
followed 12 to 14 days later by spawning at 0.24, 0.16 and 0.10 yg/liter.
Intensive spawning by the control fish occurred 29 days later and 56
days later in 0.51 and 0.72 yg/liter Guthiort-' although at 0.51 yg/liter
spawning occurred infrequently during the test. The number of spawnings
and eggs were greatest in the control chamber (Table 5). This result
may have been caused by the highest female to male sex ratio throughout
the test. The more significant measurements in terms of GuthiorHeffect
were the number of eggs per spawning and per female since these nor-
malize the dissimilar sex ratios. On this basis Guthioff-'affected
fecundity between 0.33 yg/liter which was similar to controls and 0.51
yg/liter where there was about a 50% reduction in eggs per spawning and
per female (Table 5). The decreased oxygen concentrations with increas-
ing Guthion-'was apparently not responsible for the effect on fecundity
since the oxygen concentration at 0.33 and 0.51 yg/liter GuthioA-^was
similar.
Table 5. FECUNDITY OF FATHEAD MINNOWS EXPOSED TO VARIOUS CONCENTRATIONS
OF GUTHION5^
GuthiorP
yg/l
0.04
0.10
0.16
0.24
0.33
0.51
0.72
Number of
spawnings
88
54
57
49
52
21
27
Number
of eggs
16401
t
8541
9567
7091
8590
1791
2408
Number of
eggs per
spawning
186
158
168
145
165
85
89
Number of
eggs per
female^/
1691
1220
1611
1239
1718
256
782
a/
— Computed by intervals between thinnings.
15
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SECOND GENERATION
Random samples of 50 eggs were taken from 12 spawnings from each test
chamber except the second highest where samples from seven spawnings
were taken. These eggs were hatched in the oscillating egg baskets. As
with the first generation eggs, there was no effect of Guthiott^ at the
tested concentrations (Table 6).
Table 6. EFFECT OF
ON SURVIVAL AND GROWTH OF SECOND GENERATION
EGGS AND FRY
GuthionS/ concentration, yg/1
Item
% survival - eggs
% survival - fry-
Is t group
2nd group
Mean weight (mg)-
1st group
2nd group
0.04
86.2
79
69
94
143
0.10
84.2
97
89
130
102
0.16
90.9
100
88
94
135
0.24
86.2
97
89
126
85
0.33
82.3
89
100
115
144
0.51 0.72
86.4 90.2
- -1 78
100
78 128
222 80
a/
— After 50 days from hatching.
— Some fry lost due to overflow.
Survival or growth of fry was not consistently affected by the tested
Guthion*^ concentrations (Table 6). Survival was poorest among controls.
Considerable differences in growth occurred between test chambers but
these differences were not related to Guthiori-' concentrations (Table 6).
16
-------
SECTION VI
DISCUSSION
The lowest concentration of Guthion^ that had an adverse effect on fat-
head minnows was 0.51 yg/liter where fecundity was much reduced (Table
5). An estimation of the maximum acceptable toxicant concentration
based on this parameter is between 0.33 and 0.51 yg/liter Guthionr
Effects on survival and growth during early life history stages occurred
at slightly higher concentrations (Tables 3 and 4). Although no effect
occurred in test III at the highest concentration (0.72 yg/liter) growth
and survival were reduced at the lowest concentration (1.8 yg/liter) in
test II.
The 96-hr LC50 for 11-week-old fathead minnows tested in a relatively
soft water was 1900 yg/liter (Part I of this report). An application
factor based on this acutely toxic concentration and the MATC in the
present study is between .00017 and .00027. Although this application
factor is extremely small compared to factors found for many other
toxicants (U.S. Environmental Protection Agency, 1973), it would prob-
ably be even smaller if the acute tests were conducted in hard water.
Henderson, Pickering and Tarzwell (1960) found that the 96-hr LC50 of
Guthioir-^to fathead minnows was approximately 25 to 40% less in soft
water than in hard (20 ing/liter vs. 400 mg/liter total hardness). This
differential in hardness was more extreme than the acute vs. chronic
test in the present study, but if the application factor was based on an
acute test conducted in hard water, the implication is that the factor
would be even smaller.
17
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SECTION VII
REFERENCES
DeFoe, D.L. 1975. Multichannel Toxicant Injection System for Flow-
through Bioassays. J. Fish. Res. Board Can. 32;544-546.
Henderson, C., Q.H. Pickering, and C.M. Tarzwell. 1960. Toxicity of
Organic Phosphorus and Chlorinated Hydrocarbon Insecticides to Fish.
In: Biological Problems in Water Pollution. Trans. 1959 Seminar.
Cincinnati, R.A. Taft Sanit. Eng. Center, p. 76-92.
Katz, M. 1961. Acute Toxicity of Some Organic Insecticides to Three
Species of Salmonids and to the Three-spine Stickleback. Trans. Amer.
Fish. Soc. 90(3):264-268.
Macek, K.J., and W.A. McAllister. 1970. Insecticide Susceptibility of
Some Common Fish Family Representatives. Trans. Amer. Fish. Soc. 99:
20-27.
Meyer, F.P. 1965. The Experimental Use of Guthion as a Selective Fish
Eradicator. Trans. Amer. Fish. Soc. 94(3);203-209.
Mount, D.I., and W.A. Brungs. 1967. A Simplified Dosing Apparatus for
Fish Toxicology Studies. Water Res. 1(1):21-29.
Mount, D.I., and C.E. Stephan. 1967. A Method for Establishing Accept-
able Toxicant Limits for Fish - Malathion and the Butoxyethanol Ester of
2,4-D. Trans. Amer. Fish. Soc. 96:185-193.
18
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Pickering, Q.H., C. Henderson, and A.F. Lemke. 1962. The Toxicity of
Organic Phosphorus Insecticides to Different Species of Warm Water
Fishes. Trans. Amer. Fish. Soc. 91(2);175-184.
U.S. Environmental Protection Agency. 1973. Water Quality Criteria
1972. Ecol. Res. Ser. EPA-R3-73-033. 594 p.
19
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SECTION VIII
PUBLICATIONS
Adelman, I.R., L.L. Smith, Jr., and G.D. Siesennop. Chronic Toxicity
of Guthion to the Fathead Minnow (Pimephales promelas Rafinesque).
Bulletin of Environmental Contamination and Toxicology (submitted).
20
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SECTION IX
GLOSSARY
Adult - A fish older than 4 months.
Application Factor - The ratio of the MATC to the 96-hr LC50.
Bioassay - A toxicity test; the estimation of the strength of a poison
by its effect on a living organism.
Bioassay, acute - A toxicity test of short duration, usually less than
3 weeks.
Bioassay, chronic - A toxicity test of long duration, usually 1 month
or longer.
Bioassay, flow-through - A toxicity test where the toxicant and diluent
water are continuously replaced by fresh material.
Biastula - Early embryonic stage where cellular division has resulted
in cells arranged as a hollow spherical body.
Chorion - The outer membraneous covering of a fish egg.
Fry - A fish younger than about 50 days.
LC50 - The concentration of poison that will kill 50% of the test
organisms at a specified time.
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Maximum Acceptable Toxicant Concentration (MATC) - A concentration of
toxicant determined from a chronic bioassay and considered an estimation
of the level at which no harm will occur in the natural environment.
It is bracketed by the highest level of no effect and the lowest level
of effect in the bioassay.
22
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TECHNICAL REPORT DATA
(Please read Instructions on the reverse before completing)
1. REPORT NO.
EPA-600/3-76-061b
3. RECIPIENT'S ACCESSION>NO.
4. TITLE AND SUBTITLE
STANDARD TEST FISH DEVELOPMENT, PART II
5. REPORT DATE
July 1976 (Issuing Date)
6. PERFORMING ORGANIZATION CODE
7. AUTHOR(S)
Ira R. Adelman and Lloyd L. Smith, Jr.
8. PERFORMING ORGANIZATION REPORT NO
9. PERFORMING ORGANIZATION NAME AND ADDRESS
Department of Entomology, Fisheries, and Wildlife
University of Minnesota
St. Paul, Minnesota 55108
10. PROGRAM ELEMENT NO.
1BA608
11. CONTRACT/GRANT NO.
Grant R800940
12. SPONSORING AGENCY NAME AND ADDRESS
U.S. Environmental Protection Agency
Office of Research and Development
Environmental Research Laboratory
Duluth, Minnesota 55804
13. TYPE OF REPORT AND PERIOD COVERED
T7TNAL
14. SPONSORING AGENCY CODE
EPA-ORD
15. SUPPLEMENTARY NOTES
See Part I, EPA-600/3-76-061a
16. ABSTRACT
Three chronic bioassays of GuthiorTwere conducted with fathead minnows.
All tests were begun with eggs, and the longest lasted 20 days after termination
of spawning, a total of 250 days. Parameters measured were survival, growth,
fecundity, and growth and survival of second generation fry and eggs. The most
sensitive criteria for effect of the toxicant was fecundity. The maximum ^
acceptable toxicant concentration was between 0.33 and 0.51 yg/liter Guthion^
and the application factor between .00017 and .00027.
17.
KEY WORDS AND DOCUMENT ANALYSIS
DESCRIPTORS
b.lDENTIFIERS/OPEN ENDED TERMS C. COS AT I Field/Group
Bioassay
Criteria
Fecundity
Growth
Minnows
Water quality
Survival
Guthion
Chronic toxicity
Bioassay survival
Application factor
Maximum acceptable
toxicant concentration
06C
06F
06S
18. DISTRIBUTION STATEMENT
RELEASE TO PUBLIC
19. SECURITY CLASS (ThisReport)
UNCLASSIFIED
21. NO. OF PAGES
31
20. SECURITY CLASS (Thispage)
UNCLASSIFIED
22. PRICE
EPA Form 2220-1 (9-73)
23
•OUSGPO: 1976 — 657-695/5461 Region 5-11
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