&EFA
Protection
DuJuth WIN
EPA 600 3 80 006
January 1980
Research and Development
Sublethal Effects of
Toxaphene on
Daphnids, Scuds and
Midges
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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 nine series. These nine broad cate-
gories were established to facilitate further development and application of en-
vironmental technology Elimination of traditional grouping was consciously
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The nine series are:
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5. Socioeconomic Environmental Studies
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This report has been assigned to the ECOLOGICAL RESEARCH series. This series
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This document is available to the public through the National Technical Informa-
tion Service, Springfield, Virginia 22161.
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EPA-600/3-80-006
January 1980
SUBLETHAL EFFECTS OF TOXAPHENE ON
DAPHNIDS, SCUDS, AND MIDGES
by
Herman 0. Sanders
Fish-Pesticide Research Laboratory
Fish and Wildlife Service
United States Department of the Interior
Columbia, Missouri 65201
Contract No. EPA-IAG-141(D)
Project Officer
Leonard Mueller
Environmental Research Laboratory
Duluth, Minnesota 55804
ENVIRONMENTAL RESEARCH LABORATORY
OFFICE OF RESEARCH AND DEVELOPMENT
U.S. ENVIRONMENTAL PROTECTION AGENCY
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, nor does mention of
trade names or commercial products constitute endorsement or recommendation
for use.
ii
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FOREWORD
This report describes the toxicity of a pesticide, toxaphene, to three
species of aquatic invertebrates. Toxaphene has diverse uses; as a piscicide
for controlling fish populations; an insecticide for controlling insect
pests on livestock; most extensive use is on agricultural lands as an
insecticide on crops.
Toxaphene is persistent and highly toxic to aquatic organisms but little
is known about the chronic effects on growth or reproduction. Research
reported here helps determine chemical pollution affects to aquatic life and
bioaccumulation in aquatic organisms.
This report studies the effects of the insecticide toxaphene on repro-
duction in daphnids (Daphnia magna); on growth in length of scuds (Gammarus
pseudolimnaeus); and on emergence of midges (Chironomus plumosus); when
continuously exposed to five different concentrations.
J. David Yount, Ph.D.
Deputy Director
Environmental Research Laboratory-Duluth
til
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ABSTRACT
Daphnids (Daphnla magna), scuds (Gammarus pseudolimnaeus), and midge
larvae (Chlronomus plumosus) were continuously exposed to toxaphene in a flow-
through system. Exposure of daphnids for a complete life cycle (21 days) to
0.12, 0.28, 0.54, and 1.0 ug/1 of toxaphene significantly (P<0.05) reduced
production of young; the no-effect concentration was 0.07 ug/1. Toxaphene
concentrations of 0.25 ug/1 and greater significantly (P<0.05) reduced growth
of scuds and concentrations of 3.2 ug/1 and greater significantly (P<0.05) re-
duced emergence of midges. The no-effect concentrations were 0.13 ug/1 for
growth of scuds and 1.0 ug/1 for emergence of midges. Daphnids continuously
exposed to toxaphene accumulated residues after 7 days that were 4,000 times
(based on organism wet weight) the water concentration of 0.06 ug/1. Whole
body residues in midge larvae were below the minimum detection limit of 0.1
ug/g. Maximum acceptable toxicant concentrations (MATC) of toxaphene for the
three species of aquatic invertebra'tes were estimated using reproduction of
daphnids, growth of scuds, and emergence of midges as indicators of toxic
effects. The MATC was estimated to be between 0.07 and 0.12 ug/1 for daphnids,
between 0.13 and 0.25 ug/1 for scuds, and between 1.0 and 3.2 ug/1 for midges.
This report was submitted in partial fulfillment of Contract Number EPA-IAG-
141 (D) by the Fish-Pesticide Research Laboratory, Fish and Wildlife Service
U.S.D.I. under the sponsorship of the Environmental Protection Agency.
Work was completed March, 1976.
iv
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CONTENTS
Foreword ill
Abstract iv
Tables vi
Acknowledgments vii
1. Introduction • 1
2. Conclusions 2
3. Recommendation 3
4. Materials and Methods 4
Water Characteristics 4
Toxicant Information 4
Acute Toxicity Procedures...» 4
Chronic Toxicity Procedures 6
Reproduction o f daphnids 6
Growth of s cuds 6
Emergence of midges 6
Residue accumulation 7
Calculation of Application Factors 7
5. Results 8
Acute Toxicity. 8
Reproduction of Daphnids 8
Growth of Scuds 8
Emergence of Midges 10
Residue Accumulation 10
6. Discussion 12
References 14
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TABLES
Number Page
1 Chemical characteristics of well water at the Columbia National
Fisheries Research Laboratory 5
2 Effects of Different Concentrations of Toxaphene on Reproduction of
Daphnia magna After Continuous Exposures of 14 and 21 Days at
18 ± 1 C 9
3 Cumulative Percentages of Midges (Chironomui^ plumosus) that Emerged
After Continuous Exposure of the Larvae to Different Concen-
trations of Toxaphene at 22 ± 1 C 11
vi
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ACKNOWLEDGMENTS
I thank David Hayes for his assistance in developing a midge rearing
technique and construction of a flow-through system; Foster Mayer for his
assistance in developing a technique for determining growth of scuds; Philip
Lovely and James Wencker for their aid in conducting the chronic studies and
maintenance of invertebrate cultures; and James Johnson for conducting the
toxaphene residue analyses.
vii
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SECTION 1
INTRODUCTION
Toxaphene (technical chlorinated camphene containing 67-69% chlorine) is
a synthetic organochlorine insecticide that contains numerous isomers. It has
been used as a piscicide for controlling fish populations in lakes*-, , in-
cluding sea lamprey populations in the upper Great Lakes3 and for controlling
insect pests on livestock^. However, its most extensive use has been on
agricultural lands for controlling a variety of insect pests on cotton, grains,
fruits, and forage. Of the more than 30 million pounds of this insecticide
used annually in the United States, over half is applied for controlling in-
sect pests on cotton^.
Toxaphene is a complex compound and a highly persistent chemical in the aquatic
environment , 6. it has been reported to persist in hydrosoil at concentrations
of 0.2 to 1 mg/kg for over 6 years7. Since many aquatic invertebrates live
in or on the surface of the hydrosoil, the presence of low toxaphene concen-
trations may reduce growth and inhibit reproduction, which in turn may affect
fish and other animals that feed on these organisms.
Only limited data have been published concerning the effects of toxaphene on
aquatic invertebrates and most of the information has resulted from observa-
tions following field application of this chemical in fish control projects
and fish population studies""^". Laboratory studies with toxaphene and
aquatic invertebrates have been concerned primarily with acute toxicities11"13,
and few data are available on the chronic effects of the compound on repro-
duction, metamorphosis, or growth. Since the biological significance of
toxaphene residues in aquatic invertebrates is largely unknown, the present
study was undertaken to evaluate the effects of toxaphene on reproduction in
daphnids (Daphnia magna); on growth in length of scuds (Gammarus pseudolimnaeus);
and on emergence of midges (Chironomus plumosus). These organisms make up
a significant portion of the macroinvertebrate fauna in many freshwater
habitats, and they are Important food of many species of fish and waterfowl.
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SECTION 2
CONCLUSIONS
Daphnid reproduction was significantly (P<0.05) reduced at toxaphene
concentrations of 0.12 ug/1 or higher, but not at 0.07 ug/1.
Growth of scuds was significantly (P<0.05) decreased at toxaphene concen-
trations of 0.25 ug/1 or higher; the no-effect concentration was 0.13 ug/1.
Continuous exposure of midge larvae through a complete life cycle to
toxaphene concentrations of 3.2 ug/1 and higher significantly (P<0.05) re-
duced pupation and emergence; the no-effect concentration was 1.0 ug/1.
Toxaphene residues accumulated by daphnids during a 7-day exposure were
4000 times (based on organism wet weight) the water concentration of 0.06 pg/1.
In contrast midge larvae accumulated little or no toxaphene; whole body
residues were below the minimum detection limit of 0.1 ug/g.
Reproduction of daphnids was the most sensitive indication of in-
vertebrate species susceptibility to chronic exposure to toxaphene.
Based on chronic tests evaluating reproduction of daphnids, emergence of
midges, and growth of scuds, the maximum acceptable toxicant concentration was
estimated to be between 0.07 and 3.2 ug/1.
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SECTION 3
RECOMMENDATIONS
Inasmuch as reproductive Impairment in daphnids was found to be a
sensitive criterion for evaluating the chronic toxicity of toxaphene to an
aquatic invertebrate, Daphnia magna should be used in future chronic toxicity
studies until methods for culturing and testing other species of daphnids
can be developed.
Although length measurement, used in this study for determining growth,
was found to be a sensitive method for evaluating the chronic toxicity of
toxaphene to the scud Gammarus pseudolimnaeus, further research on laboratory
rearing of this organism is needed. Techniques for maintaining a healthy
reproducing population of scuds through a complete life cycle must be im-
proved .
The midge Chironomus plumosus should be considered as a test organism in
chronic toxicity studies because it adapts well to laboratory conditions, is
widely distributed in a variety of aquatic habitats, has easily recognizable
development stages, and is an important food of young and adult fish.
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SECTION 4
MATERIALS AND METHODS
Test organisms included two crustaceans and an aquatic insect; a daphnid
(Daphnia magna); a scud (Ganmiarus_ pseudolimnaeus) ; and the larvae of a midge
(Chironomus plumosus). The rearing method described by Sanders and Cope^-l
was used in maintaining a continuous supply of daphnids. Scuds were from
cultures maintained in the laboratory-"- . Rearing techniques described by
Biever and Ivlera were used to maintain a continuously reproducing
population of midges. Daphnids and scuds were tested at 18 ± 1 C and midge
larvae at 22 ± 1 C. A combination of Duro-test and wide-spectrum Growlux
bulbs provided light over the cultures and tests. The photoperiod was
automatically controlled for 16 hours light and 8 hours dark.
WATER CHARACTERISTICS
The water used for cultures and all toxicity tests was from a deep well.
Chemical characteristics of this water are summarized in Table 1. During the
chronic exposure, the dissolved oxygen concentration and water temperature
were measured dally in test containers. The pH and hardness were measured at
the beginning and termination of each test.
TOXICANT INFORMATION
An experimental-use sample of toxaphene (X-16189-49) was furnished by
Hercules Inc. Stock solutions of toxaphene were prepared in ethanol and
further diluted with water in a flow-through system modeled after Mount and
Brungs^ , and were delivered by the apparatus designed by Chandler et al.^8.
All concentrations, as well as the controls, contained 0.1 ml/1 of ethanol.
Prior to initiating the tests, the flow-through systems were operated
for 24 h to allow for concentration equilibrium and to establish that
toxaphene water concentrations were constant. Toxaphene concentrations in
water were measured in the high, medium, low, and control containers before
the introduction of test organisms and at termination of the tests. Methods
used for water residue analysis were described by Stalling and Huckins .
ACUTE TOXICITY PROCEDURES
Acute toxicity tests were conducted under static conditions; methods used
were those recommended for standardized laboratory toxicity tests by the
Committee on Methods for Toxicity Tests with Aquatic Organisms^. Daphnids
were first instar; scuds in an early instar; and midges In the early fourth
instar. The measure of acute toxicity for daphnids and midge larvae was
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TABLE 1. CHEMICAL CHARACTERISTICS OF WELL WATER AT THE COLUMBIA NATIONAL
FISHERIES RESEARCH LABORATORY
Parameter
Ca
Mg
K
804
NO 3
N02
NH4/N
Phenol
ci2
Cl
F
CN
Fe
Cu
Zn
Cd
Cr
Pb
Alkalinity
Hardness (EDTA)
PH
Specified sensitivity
limits, mg/liter
0.1
0.1
0.5
0.01
0.05
0.05
0.01
0.001
0.001
0.01
0.01
0.005
0.01
0.0001
0.001
0.001
0.01
0.001
1.0
1.0
0.1
Concentration ,
mg/liter
70
27
3.9
4.4
<0.05
<0.036
0.066
<0.001
<0.001
29
0.34
0.006
0.014
0.0045
<0.001
<0.0005
<0.01
0.0015
237
272
7.4
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the 48 h median effective concentration (48-h EC50) based on immobilization.
The toxicity of toxaphene to scuds was expressed in terms of LC50, the
calculated concentration of chemical in water which produces a 50% mortality
of test organisms during a specific time'.
CHRONIC TOXICITY PROCEDURES
Reproduction of Daphnids
The daphnid reproduction studies were begun by introducing 10 first-in-
star daphnids up to 24 h old into duplicate exposure vessels containing 1
liter of water. Thus, 20 daphnids per treatment were exposed continuously
through a complete life cycle (21 days) to toxaphene concentrations of 0,
0.07, 0.12, 0.28, 0.56, or 1.0 ug/1. Organisms were fed a suspension of yeast
in sufficient amounts to support a stable population. Reproductive success
was determined by counting the offspring produced in each concentration after
the parent daphnids had been exposed for 21 days. The mean number of young
produced per adult was determined by averaging the number of young produced
in two replicate tests. Data were analyzed by analysis of variance, and
significant differences among treatments were determined by a multiple means
comparison test (least significant difference* ).
Growth of Scuds
The study of chronic toxicity of toxaphene to scuds was started with
young collected firom gravid females. Ten young scuds were placed in each
container and fed maple leaves that had been previously soaked in water for
several weeks. The scuds (5-10 days old) were exposed to toxaphene for 30
days in a flow-through system1'. Flow-splitting chambers designed by Benoit
and Puglisi" were used to mix and divide each toxaphene concentration into
four exposure chambers. The average measured concentrations were 0, 0.06,
0.13, 0.25, 0.50, and 1.0 ug/1.
A method proposed by McKim and Benoit^ for measuring lengths of juvenile
fish from photographs was used to measure growth of the scuds. Scuds were
photographed at 0 and 30 days of exposure. Measurements were made on the
photo-enlarged image of the scuds, and lengths of animals were determined
from interpretation of the photographs.
Emergence of Midges
The study of chronic toxicity of toxaphene to midge was started with first-
instar larvae (1.5 mm long and up to 24 h old). One hundred larvae, counted
with the aid of a 10X lens, were placed in duplicate exposure contaminers that
had been previously prepared by adding 13 g of sand and 0.3 g of a commercial
dog candylS to 1 liter of water. The larvae were exposed to toxaphene
concentrations of 0, 1.0, 3.2, 5.6, 10, and 32 ug/1. During the test larvae
were fed 0.3 g of the candy every 5 days until they transformed into the pupal
stage. The test was ended after 30 days, when about 80-95 percent of the
control larvae had completed metamorphosis into the adult form. Cast pupal
skins at the water surface in test containers were counted and removed daily
to determine adult emergence. The effects of toxaphene on midge emergence
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were determined by conducting an analysis of variance on the arcsin transfor-
mation for portions (angle « arcsin percentage) followed by a least
significant difference test^l.
Residue Accumulation
Accumulation studies with daphnids and midge larvae were conducted in a
flow-through system: the toxaphene was delivered with the apparatus described
by Chandler et al. . Daphnids were exposed to measured toxaphene concen-
trations of 0.06 and 0.12 ug/1 and midge larvae to concentrations of 0.25,
1.8, and 3.2 ug/1. These concentrations, selected on the basis of the results
of the chronic-toxicity studies, represent the lowest toxaphene concentration
that produced an effect on the organisms and a concentration in which no ef-
fect would be expected. Daphnids and midge larvae (500 mg of each) were
sampled from each concentration after 1, 7, and 14 days of exposure. Residue
analyses were performed according to the method of Stalling and Huckins .
Samples were prepared for analysis by homogenizing 100-500 mg of organisms
with 8 g of anhydrous sodium sulfate. The samples were extracted by per-
colation in 1 cm i.d. glass columns with 50 ml portions of 5% diethyl ether
in petroleum ether. Sample cleanup was accomplished by adding the sample
extract to 2 g of heated Florisil in a 1 cm i.d. column and eluting toxaphene
with 45 ml of 5% diethyl ether in petroleum ether. The samples were con-
centrated to 0.5 ml and toxaphene residues were quantified by gas liquid
chromatography (GLC) with 6^Ni-electron capture detection. A 160 cm long
x 2.0 mm i.d. glass column packed with 3% (w/w) OV-7 on Chromosorb W-H.P.
was used, with a 30 ml/min flow of nitrogen carrier gas. The column was
operated at 200 C. The minimum detection limit of a toxaphene standard was
0.5 ng. For sample quantities of 300-500 mg the detection limit was 0.1 ug/g.
CALCULATION OF APPLICATION FACTORS
A method proposed by Mount and Stephen2^ for establishing acceptable
toxicant limits for aquatic organisms under continuous exposure conditions was
used to calculate an application factor for determining a toxicant concentra-
tion that has no adverse effect on reproduction of daphnids, emergence of
midges, or growth of scuds. The application factor consists of the laboratory
determined maximum acceptable toxicant concentration (MATC) that has no ef-
fect on the test organisms during chronic exposure, divided by the 48-h EC50
for daphnids and midges and the 96-h LC50 for scuds.
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SECTION 5
RESULTS
ACUTE TOXICITY
In static tests the toxiclty of toxaphene varied widely among the in-
vertebrates tested, with the Crustacea being the more susceptible. The 48-h
ECSO's ranged from 10 ug/1 for daphnids to 180 ug/1 for midges. The 96-h
LC50 for scuds was 24 ug/1.
REPRODUCTION OF DAPHNIDS
Continuous exposure of daphnids through a complete life cycle (21 days)
to toxaphene concentrations of 0.12, 0.28, 0.54, and 1.0 ug/1 significantly
reduced (P<0.05) production of young (Table 2). Production of young in the
0.7 ug/1 concentration was similar to that in controls after 21 days of ex-
posure. Survival of adult daphnids at the end of the tests was 90 - 95% in
all toxaphene concentrations and controls, except at 1.0 ug/1, in which adult
survival was 70%.
The 48-h EC50 of 10 ug/1 for first-instar daphnids was approximately 80
times greater than the lowest toxaphene concentration (0.12 ug/1) that af-
fected daphnid reproduction. Based on the chronic exposure of daphnids to
toxaphene, the MATC was estimated to be between 0.07 and 0.12 ug/1 and the
application factor was between 0.007 and 0.01.
GROWTH OF SCUDS
Growth of scuds as measured by increases in length, was significantly
decreased (P<0.05) in toxaphene concentrations of 0.25, 0.50 and 1.0 ug/1
after 30 days of exposure. Growth of scuds at 30 days was similar in controls
and toxaphene concentrations of 0.06 and 0.13 ug/1. Survival after the 30
day exposure was 90-100% in all concentrations and controls. However, data
on survival of scuds beyond 30 days were too variable for sound statistical
analyses.
The 96-h LC50 of 24 ug/1 for scuds was about 180 times greater than the
lowest toxaphene concentration (0.13 ug/1) that affected scud growth. The
MATC for scuds was estimated to be between 0.13 and 0.25 ug/1 and the ap-
plication factor was between 0.001 and 0.01.
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TABLE 2. EFFECTS OF DIFFERENT CONCENTRATIONS OF TOXAPHENE ON REPRODUCTION OF
DAPHNIA MA.GNA AFTER CONTINUOUS EXPOSURES OF 14 AND 21 DAYS AT 18
+ 1 C.
Toxaphene
concentration
Oig/D
0
0.07
0.12
0.28
0.54
1.0
Number of offspring after:
14 days
308
310
186a
165a
100a
76a
21 days
596
542
289a
212a
154a
110a
Significantly different from controls (P<0.05), n - 2.
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EMERGENCE OF MIDGES
Emergence of midges was significantly reduced (P<0.05) after 30 days
exposure to toxaphene concentrations of 3.2, 5.6, 10, and 32 ug/1 (Table 3).
Emergence was also significantly delayed (P<0.05) in the 1.0 ug/1 concentration
at 20 days; however, at 30 days the emergence time was similar to that in the
controls. Pupation was progressively reduced and the length of development
was increased in concentrations of 5.6 ug/1 through 32 ug/1. Larvae held at
these concentrations behaved abnormally and appeared unable to build well
defined larval tubes, within which pupation occurs. At the end of the study,
some of these larvae had not pupated, and appeared to be stunted. This re-
duction in pupation was directly related to the reduction in emergence of
adults at the higher concentrations.
The 48-h EC50 of 180 ug/1 was about 60 times greater than the lowest
toxaphene concentration (3.2 ug/1) that significantly reduced emergence. The
MATC for midges was estimated to be between 1.0 and 3.2 ug/1 and the ap-
plication factor was between 0.005 and 0.01.
RESIDUE ACCUMULATION
The magnitude of toxaphene accumulation from water by daphnids during
continuous exposure was proportional to the toxaphene concentration in water.
After a 7 day exposure to a measured toxaphene concentration of 0.12 ug/1,
daphnids concentrated toxaphene 4,000 times (0.5 ug/g; wet weight) the level
in water. When daphnids were exposed to 0.06 ug/1 of toxaphene, they accumu-
lated total body concentrations 4,000 times (0.25 ug/g) that of water. Up-
take at both toxaphene concentrations reached a peak after 7 days of exposure.
However, residues in daphnids did not stabil.ize at the peak concentration
with additional exposure (14 days); rather, the residues were eliminated at
rates exceeding the accumulation rate. Residues in daphnids exposed at 0.12
ug/1 declined by 50 percent between 7 and 14 days, but residues in those ex-
posed at 0.06 ug/1 declined by only 10 percent.
When midge larvae were exposed continuously to toxaphene concentrations
in water of 1.8, 3.2, and 5.6 ug/1 for 1, 7, and 14 days, whole body residues
in all samples remained below the minimum detection limit of 0.1 ug/g for
tissues. These results indicate that the sample size (500 mg sample of larvae,
representing 250 larvae) was not adequate for residue analyses. It was not
feasible to expose a larger population of larvae because of problems in
rearing large numbers of organisms. Nevertheless, these negative data
suggest that midge larvae accumulate far less toxaphene than daphnids.
10
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TABLE 3. CUMULATIVE PERCENTAGES OF MIDGES (CHIRONOMUS PLUMOSUS) THAT EMERGED AFTER CONTINUOUS
EXPOSURE OF
THE LARVAE
TO DIFFERENT
CONCENTRATIONS OF TOXAPHENE
AT 22 + 1 C.
Days
of
exposure
15
20
25
30
0
9
66
86
88
1.0
0
41a
81
82
Toxaphene
3.2
0
343
54a
56a
Oug/1)
5.6
0
20a
42a
43a
10
0
26a
42a
44a
32
0
12a
20a
20a
aSignificantly different from controls (P<0.01), n = 2.
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SECTION 6
DISCUSSION
Aquatic Invertebrates are often exposed to toxaphene applied directly
to their habitat , or as a result of runoff from treated agricultural lands.
Observations of various aquatic invertebrates after field application of
toxaphene confirm the response of these organisms to exposure to this chemical,
Hilsenoff^ reported that the application of 100 ug/1 of toxaphene for control
of rough fish in a lake resulted in total elimination of the midge population
and that midge larvae did not reappear until 9 months after treatment.
Results from the present study suggest that reductions of a midge population
might occur at much lower concentrations, since a toxaphene concentration
of 3.2 ug/1 would cause a significant reduction in pupation and emergence of
midges.
Laboratory studies have shown that toxaphene is acutely toxic to fish
(96-h LCSO's = 2.0 to 14 ug/125' 2^), whereas daphnids are slightly more
resistant. Sanders and Cope^^reported that the 48-h EC50 immobilization
values for three species of daphnids ranged from 10 to 19 ug/1^. These
results tend to support the observations made by Tanner and Hayes^, who re-
ported that a lake treated with this chemical for rough fish control sup-
ported large populations of daphnids several weeks after treatment but, re-
mained extremely toxic to fish. This greater resistance of daphnids to
toxaphene could enable them to concentrate significant residues and pass these
residues through the food chain to higher trophic levels. This conclusion
also agrees with the findings of Schoettger and Olive27, who reported that
static exposure of daphnids to toxaphene concentrations of 10-20 ug/1 for
312 h did not produce detrimental effects. However, sufficient residues
were accumulated by the daphnids during this exposure to produce complete
mortality in test fish fed these exposed organisms. The results of our up-
take studies with daphnids Indicated that maximum accumulation occurred
during the first 7 days of exposure. Residues in daphnids exposed at a con-
centration of 0.12 ug/1 of toxaphene did not attain a stable equilibrium and
residues declined by half between 7 and 14 days of continuous exposure; how-
ever, residues in daphnids exposed at a concentration of 0.06 ug/1 of tox-
aphene declined only slightly. This relation suggests that the factors that
contribute to metabolism and excretion may have been stimulated at the 0.12
ug/1 concentration and caused the elimination rate to exceed the accumulation
rate. Although the residues accumulated by daphnids were not identified, it
is assumed the loss was caused by excretion and degradation of the parent
compound. Mayer et al. ° reported that toxaphene was degraded in fish and
that the more chlorinated toxaphene isomers are preferentially stored by brook
trout (Salvelinus fontinalis) while the less chlorinated ones are more rapidly
eliminated. Further studies are needed to determine residue data from
12
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various components of simllated or natural food chains.
Chronic toxicity determined for the three species of aquatic invertebrates
indicated that toxaphene is biologically active at concentration's well below
those that are acutely toxic. All organimsm were susceptible to chronic ex-
posure to toxaphene and the no-effect concentrations ranged from 0.07 to 1.0
ug/1. On the basis of chronic tests evaluating reproduction of daphnids*.
emergence of midges, and growth of scuds, the maximum acceptable toxicant
concentration (MATC) of toxaphene was estimated to be between 0.07 and 3.2
ug/1. Dividing the approximate MATC by the appropriate EC50 or LC50 gives an
application factor ranging from 0.001 to 0.01. This range in values ap-
proximates the application factor of 0.01 which has been used for -establish-
ing water quality criteria for organochlorine insecticides^ .
Fishes appear to be more susceptible to toxaphene than the aquatic in-
vertebrates tested; the maximum toxaphene concentration in water acceptable
for brook trout fry, determined by Mayer et al. ° to be below 0.039 ug/1,
should be relatively safe for these freshwater organisms.
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REFERENCES
1. Henegar, D.L. Minimum Lethal Levels of Toxaphene as a Piscicide in North
Dakota Lakes. U.S. Fish Wildlife Service, Washington, D.C. Resour. Publ.
7. 1966. 16 pp.
2. Hooper F.F., and A.R. Grzenda. The Use of Toxaphene as a Fish Poison.
Trans. Am. Fish. Soc. 85; 180-190. 1957.
3. Gaylord, W.E. and B.R. Smith. Treatment of East Bay, Alger County,
Michigan with Toxaphene for Control of Sea Lampreys. U.S. Fish Wildlife
Service, Washington, D.C. Resour. Publ. 11. 1966. 7 pp.
4. Hercules Incorporated. Toxaphene: Use Patterns and Environmental Aspects.
Hercules Inc. Wilmington, Del. November 1970. 172 pp.
5. Terriere, L.C., U. Kiigemagi, A.R. Gerlach, and R.L. Borovicka. The
Persistence of Toxaphene in Lake Water and Its Uptake by Aquatic Plants
and Animals. J. Agr. Food Chem. K/l) :66-69, 1966.
6. Kallman, B.J., O.B. Cope, and R.J. Navarre. Distribution and Detoxification
of Toxaphene in Clayton Lake, New Mexico. Trans. Am. Fish Soc. 91(1) :
14-22, 1962.
7. Johnson, W.D., G.F. Lee, and D. Spyridakis. Persistence of Toxaphene in
Treated Lakes. Int. J. Air Water Pollut. 1.0:555-60, 1966.
8. Hilsenhoff , W.L. The Biology of Chironomus plumosua (Diptera: Chironomidae)
in Lake Winnebago, Wisconsin. Ann. Entomol. Soc. Am. 59(3) :465-473, 1966.
9. Tanner, H.A. , and M.L. Hayes. Evaluation of Toxaphene as a Fish Poison.
Colorado Coop. Fish. Rea. Unit, Quart. Rep. >4: 31-39, 1955.
10. Needham, R.G. Effects of Toxaphene on Plankton and Aquatic Invertebrates
in North Lakes. U.S. Fish Wildlife Service, Washington, D.C. Resource
Publ. 8. 1966. 16 pp.
11. Sanders, H.O., and O.B. Cope. Toxicities of Several Pesticides to Two
Species of Cladocerans. Trans. Am. Fish. Soc. 9[5/2) : 165-169, 1966.
12. Sanders, H.O., and O.B. Cope. The Relative Toxicities of Several
Pesticides to Naiads of Three Species of Stoneflies. Limnol. Oceanogr.
13(1): 112-117, 1968.
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13. Sanders, H.O. Toxiclty of Some Insecticides to Four Species of
Malacostracan Crustaceans. U.S. Fish Wildlife Service, Washington, D.C.
Technical Paper 66. 1972. 19 pp.
14. Sanders, H.O. Toxicity of Pesticides to the Crustacean Gammarus
lacustris. U.S. Fish Wildlife Service, Washington, D.C. Technical
Paper 25. 1969. 18 pp.
15. Biever, K.D. A Rearing Technique for the Colonization of Chironomid
Midges. Ann. Entomol. Soc. Am. 5_8(2): 135-136, 1965.
16. Ivlera, I.V. Mass Cultivation of Invertebrates (Biology and Methods).
U.S. Department of Commerce, National Technical Information Service,
Springfield, Va. 1973. 148 pp.
17. Mount, D.I., and W.A. Brungs. A Simplified Dosing Apparatus for Fish
Toxicology Studies. Water Research 1:21, 1967.
18. Chandler, J.H. Jr., H.O. Sanders, and D.F. Walsh. An Improved Chemical
Delivery Apparatus for Use in Intermittent-flow Bioassays. Bull. Environ.
Contam. Toxicol. 12_(1) :123-128, 1974.
19. Stalling, D.L., and J.N. Huckins. Analysis and 6C-MS. Characterization
of Toxaphene in Fish and Water. U.S. Environmental Protection Agency,
Duluth, Minnesota. Ecological Research Series EPA-600/3-76-076. 1976.
20. Committee on Methods for Toxicity Tests with Aquatic Organisms. Methods
for acute toxicity tests with fish, macroinvertebrates and amphibians.
U.S. Environmental Protection Agency, Corvallis, Oregon. Ecol. Res.
Series No. EPA-660/3-75-009. 1975. 67 pp.
21. Snedecor, G.W. Statistical Methods. Ames, Iowa, Iowa State Univ. Press,
1965. 534 pp.
22. Benoit, D.A., and F.A. Puglisi. A Simplified Flow-splitting Chamber and
Siphon for Proportional Diluters. Water Res. ]_\ 1915-1916, 1973.
23. McKim, J.M., and D.A. Benoit. Effect of Long-term Exposures to Copper
on Survival, Reproduction, and Growth of Brook Trout Salvelinus
fontinalis. J. Fish. Res. Bd. Can. 28:655-662, 1971.
24. Mount, D.I., and C.E. Stephan. A Method for Establishing Acceptable
Toxicant Limits for Fish-Malathion and the Butoxyethanol Ester of 2,4-D.
Trans. Amer. Fish. Soc. 21:185-193, 1967.
25. Macek, K.J., C. Hutchinson, and O.B. Cope. The Effects of Temperature
on the Susceptibility of Bluegills and Rainbow Trout to Selected
Pesticides. Bull. Environ. Contam. Toxicol. 4(3):174-183, 1969.
26. Macek, K.J., and W.A. McAllister. Insecticide Susceptibility of Some
Common Fish Family Representative. Trana. Am. Fish. Soc. 99(1):20-27,
1970,
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27. Schoettger, R.A., and J.R. Olive. Accumulation of Toxaphene by Fish
Food Organisms. Limnol. Oceanogr. 6^(2):216-219, 1961.
28. Mayer, F.L., P.M. Mehrle, and W.P. Dwyer. Toxaphene Effects on
Reproduction, Growth, and Mortality of Brook Trout. U.S. Environmental
Protection Agency, Duluth, Minnesota. Ecological Research Series EPA-
600/3-75-013. 1975. 51 pp.
29. U.S. Environmental Protection Agency. Proposed Criteria for Water Quality,
Vol. I.U.S. Environmental Protection Agency. Washington, D.C. October
1973. 425 pp.
16
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TECHNICAL REPORT DATA
(Please read Instructions on the reverse before completing)
1. REPORT NO.
EPA-600/3-80-006
3. RECIPIENT'S ACCESSION NO.
4. TITLE AND SUBTITLE
Sublethal Effects of Toxaphene on Daphnids, Scuds,
and Midges
5. REPORT DATE
January 1980 issuing date
6. PERFORMING ORGANIZATION CODE
7. AUTHOR(S)
Herman 0. Sanders
8. PERFORMING ORGANIZATION REPORT NO.
9. PERFORMING ORGANIZATION NAME AND ADDRESS
Fish-Pesticide Research Laboratory
Fish and Wildlife Service
U.S. Department of the Interior
Columbia Missouri 65201
10. PROGRAM ELEMENT NO.
1BA021
11. CONTRACT/GRANT NO.
EPA-IAG-14KD)
12. SPONSORING AGENCY NAME AND ADDRESS
Environmental Research Laboratory - Duluth, MN
Office of Research and Development
U.S. Environmental Protection Agency
Duluth, Minnesota 55804
13. TYPE OF REPORT AND PERIOD COVERED
14. SPONSORING AGENCY CODE
EPA/600/03
16. SUPPLEMENTARY NOTES
16. ABSTRACT
Daphnids (Daphnia magna), scuds (Gammarus pseudolimnaeus), and midge larvae
(Chironomus plumosus) were continuously exposed to toxaphene in a flow-through system.
Exposure of daphnids for a complete life cycle (21 days) to 0.12, 0.28, 0.54, and 1.0
ug/1 of toxaphene significantly (P<0.05) reduced production of young; the no-effect
concentration was 0.07 ug/1. Toxaphene concentrations of 0.25 ug/1 and greater
significantly (P<0.05) reduced growth of scuds and concentrations of 3.2 ug/1 and
greater significantly (P<0.05) reduced emergence of midges. The no-effect concen-
trations were 0.13 ug/1 for growth of scuds and 1.0 ug/1 for emergence of midges.
Daphnids continuously exposed to toxaphene accumulated residues after 7 days that
were 4,000 times (based on organism wet weight) and water concentration of 0.06 ug/1.
Whole body residues in midge larvae were below the minimum detection limit of 0.1 ug/g.
Maximum acceptable toxicant concentrations (MATC) of toxaphene for the three species of
aquatic invertebrates were estimated using reproduction of daphnids, growth of scuds,
and emergence of midges as indicators of toxic effects. The MATC was estimated to be
between 0.07 and 0.12 ug/1 for daphnids, between 0.13 and 0.25 ug/1 for scuds, and
between 1.0 and 3.2 ug/1 for midges.
17.
KEY WORDS AND DOCUMENT ANALYSIS
DESCRIPTORS
b.lDENTIFIERS/OPEN ENDED TERMS C. COSATI Field/Group
Growth
Reproduction (biology)
Mortality
Pesticides
Invertebrates
Emergence
Daphnids (Daphnia magna)
Scuds (Gammarus
pseudolimnaeus)
Midge (Chironomus
plumosus)
life cycle, toxaphene,
continuous exposure,
chronic effects
06 F,T
18. DISTRIBUTION STATEMENT
Release to Public
19. SECURITY CLASS (THI* Report)
Unclassified
21. NO. OF PAGES
25
20. SECURITY CLASS (This page)
Unclassified
22. PRICE
EPA Form 2220-1 (R«v. 4-77) previous EDITION is OBSOLETE
*U.I.MVEIMMBITnMTIM OfflCC MO .tS7-
17
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