&EPA
United States
Environmental Protection
Agency
Office of Water
Regulations and Standards
Criteria and Standards Division
Washington DC 20460
EPA 440/5-80-046
October 1980
Ambient
Water Quality
Criteria for
Endosulfan
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AMBIENT WATER QUALITY CRITERIA FOR
ENDOSULFAN
Prepared By
U.S. ENVIRONMENTAL PROTECTION AGENCY
Office of Water Regulations and Standards
Criteria and Standards Division
Washington, D.C.
Office of Research and Development
Environmental Criteria and Assessment Office
Cincinnati, Ohio
Carcinogen Assessment Group
Washington, D.C.
Environmental Research Laboratories
Corvalis, Oregon
Duluth, Minnesota
Gulf Breeze, Florida
Narragansett, Rhode Island
Agency
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DISCLAIMER
This report has been reviewed by the Environmental Criteria and
Assessment Office, U.S. Environmental Protection Agency, and approved
for publication. Mention of trade names or commercial products does not
constitute endorsement or recommendation for use.
AVAILABILITY NOTICE
This document is available to the public through the National
Technical Information Service, (NTIS), Springfield, Virginia 22161.
11
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FOREWORD
Section 304 (a)(l) of the Clean Water Act of 1977 (P.L. 95-217),
requires the Administrator of the Environmental Protection Agency to
publish criteria for water quality accurately reflecting the latest
scientific knowledge on the kind and extent of all identifiable effects
on health and welfare which may be expected from the presence of
pollutants in any body of water, including ground water. Proposed water
quality criteria for the 65 toxic pollutants listed under section 307
(a)(l) of the Clean Water Act were developed and a notice of their
availability was published for public comment on March 15, 1979 (44 FR
15926), July 25, 1979 (44 FR 43660), and October 1, 1979 (44 FR 56628).
This document is a revision of those proposed criteria based upon a
consideration of comments received from other Federal Agencies, State
agencies, special interest groups, and individual scientists. The
criteria contained in this document replace any previously published EPA
criteria for the 65 pollutants. This criterion document is also
published in satisifaction of paragraph 11 of the Settlement Agreement
in Natural Resources Defense Council, et. al. vs. Train, 8 ERC 2120
(D.D.C. 1576), modified, 12 EMf 1833 (D.D.C. 1979).
The term "water quality criteria" is used in two sections of the
Clean Water Act, section 304 (a)(l) and section 303 (c)(2). The term has
a different program impact in each section. In section 304, the term
represents a non-regulatory, scientific assessment of ecological ef-
fects. The criteria presented in this publication are such scientific
assessments. Such water quality criteria associated with specific
stream uses when adopted as State water quality standards under section
303 become enforceable maximum acceptable levels of a pollutant in
ambient waters. The water quality criteria adopted in the State water
quality standards could have the same numerical limits as the criteria
developed under section 304. However, in many situations States may want
to adjust water quality criteria developed under section 304 to reflect
local environmental conditions and human exposure patterns before
incorporation into water quality standards. It is not until their
adoption as part of the State water quality standards that the criteria
become regulatory.
Guidelines to assist the States in the modification of criteria
presented in this document, in the development of water quality
standards, and in other water-related programs of this Agency, are being
developed by EPA.
STEVEN SCHATZOW
Deputy Assistant Administrator
Office of Water Regulations and Standards
111
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ACKNOWLEDGEMENTS
Aquatic Life Toxicology:
William A. Brungs, ERL-Narragansett
U.S. Environmental Protection Agency
David J. Hansen, ERL-Gulf Breeze
U.S. Environmental Protection Agency
Mammalian Toxicology and Human Health Effects:
Thomas L. Ferguson (author)
Midwest Research Institute
Steven D. Lutkenhoff (doc. mgr.)
ECAO-Cin
U.S. Environmental Protection Agency
Bonnie Smith (doc. mgr)) ECAO-Cin
U.S. Environmental Protection Agency
C. Stuart Baxter
University of Cincinnati
Patrick Durkin
Syracuse Research Corporation
Rolf Hartung
University of Michigan
Betty Herndon
Midwest Research Institute
John Laseter
University of New Orleans
Fumio Matsamura
Michigan State University
Robert E. Menzer
University of Maryland
R. F. Robinson
Purdue University
Carl C. Smith
University of Cincinnati
Woodhall Stopford
Duke University Medical Center
Jonathan Ward
University of Texas Medical Branch
Technical Support Services Staff: D.J. Reisman, M.A. Garlough, B L Zwayer
P.A. Daunt, K.S. Edwards, T.A. Scandura, A.T. Pressley, C.A. Cooper
M.M. Denessen.
Clerical Staff: C.A. Haynes, S.J. Faehr, L.A. Wade, D. Jones, B.J. Bordicks
B.J. Quesnell, P. Gray, R. Swantack
IV
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TABLE OF CONTENTS
Page
Criteria Summary
Introduction A-l
Aquatic Life Toxicology B-l
Introduction B-l
Effects B-2
Acute Toxicity B-2
Chronic Toxicity B-6
Plant Effects B-8
Residues B-8
Miscellaneous B-10
Summary B-10
Criteria B-ll
References B-31
Mammalian Toxicology and Human Health Effects C-l
Exposure C-l
Ingestion from Water C-l
Ingestion from Food C-10
Inhalation C-32
Dermal C-36
Pharmacokinetics C-36
Absorption C-36
Distribution C-37
Metabolism and Excretion C-38
Effects C-45
Acute, Subacute, and Chronic Toxicity C-45
Synergism and/or Antagonism C-58
Teratogenicity C-60
Mutagenicity C-62
Carcinogenic! ty C-63
Criterion Formulation C-76
Existing Guidelines and Standards C-76
Current Levels of Exposure C-77
Special Groups at Risk C-80
Basis and Derivation of Criterion C-80
References C-89
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CRITERIA DOCUMENT
ENDOSULFAN
CRITERIA
Aquatic Life
For endosulfan the criterion to protect freshwater aquatic
life as derived using the Guidelines is 0.056 ug/1 as a 24-hour
average and the concentration should not exceed 0.22 ug/1 at any
time.
For endosulfan the criterion to protect saltwater aquatic
life as derived using the Guidelines is 0.0087 ug/1 as a 24-hour
average and the concentration should not exceed 0.034 ug/1 at any
time.
Human Health
For the protection of human health from the toxic properties
of endosulfan ingested through water and contaminated aquatic or-
ganisms, the ambient water criterion is determined to be 74 ug/1.
For the protection of human health from the toxic properties
of endosulfan ingested through contaminated aquatic organisms
alone, the ambient water criterion is determined to be 159 ug/1.
VI
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INTRODUCTION
Endosulfan is a broad-spectrum insecticide of the group of
polycyclic chlorinated hydrocarbons called cyclodiene insecti-
cides. It was discovered and developed in 1954 by Farbwerke
Hoechst AG. in Germany and introduced under the registered trade-
mark Thiodan®. The trade names of endosulfan include Beosit®,
Chlorthiepin®, Cyclodan®, Insectophene®, Kop-Thiodan®, Malix®, Thi-
for®, Thimul®, Thioden®, Thionex® (Berg, 1976).
Annual production of endosulfan in the United States was esti-
mated in 1974 at three million pounds. It is presently on the U.S.
EPA'S restricted list which limits its usage. However, significant
commercial use of endosulfan for insect control on vegetables,
fruits, and tobacco continues.
Endosulfan is a light to dark brown crystalline solid with a
terpene-like odor, having the molecular formula C9C16H603S,
a molecular weight of 406.95, and a vapor pressure of 9 x 10~3
mm Hg at 80°C (Brooks, 1974; Whetstone, 1972). It exhibits a solu-
bility in water of 60 to 150 ug/1 and is readily soluble in organic
solvents (Braun and Frank, 1973). The chemical name for endosulfan
is 6,7,8,9,10,10-hexachloro-l,5,5a,6,9,9a-hexahydro-6,9-methano-
2,4,3-benzodioxathiepin-3-oxide. It is prepared through the Diels-
Adler addition of hexachlorocyclopentadiene with cis-butene-1,4-
diol to form the bicyclic dialcohol, followed by esterification and
cyclization with SOC12 (Windholz, 1976).
Technical grade endosulfan has a purity of 95 percent and is
composed of a mixture of two steroisomers referred to as alpha and
A-l
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beta or I and II. It has a melting point range of 70 to 100°C and
a density of 1.745 at 20°C (Burchfield and Johnson, 1965). The
endosulfan isomers are present in the ratio 70 percent isomer I to
30 percent isomer II. Impurities present in technical grade endo-
sulfan consist mainly of the degradation products and may not ex-
ceed 2 percent endosulfandiol and 1 percent endosulfan ether.
Endosulfan is commercially available in the form of wettable pow-
ders, emulsified concentrates, granules, and dusts of various con-
centrations (Berg, 1976). It is a powerful contact and stomach
insecticide used to control a wide spectrum of insects.
Endosulfan is stable to sunlight, but is susceptible to oxi-
dation and the formation of endosulfan sulfate in the presence of
growing vegetation (Cassil and Drummond, 1965). Technical grade
endosulfan is sensitive to moisture, bases, and acids and decom-
poses slowly by hydrolysis to SO2 and endosulfan alcohol.
In the environment, endosulfan is metabolically converted by
microorganisms, plants, and animals to endosulfan sulfate, endo-
sulfandiol, endosulfan ether, endosulfan hydroxyether, and endo-
sulfan lactone (Martens, 1976; Chopra and Mahfouz, 1977; Gorbach,
et al. 1968).
A-2
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REFERENCES
Berg, H. 1976. Farm Chemicals Handbook. Meister Publishing Co.,
Willoughby, Ohio.
Braun, H.E. and R. Frank. 1973. Unpublished data. In; Endosul-
fan: Its effects on environmental quality. Natl. Res. Counc.
Can., Ottawa.
Brooks, G.T. 1974. Chlorinated Insecticides. CRC Press, Cleve-
land, Ohio.
Burchfield, H.P. and D.E. Johnson. 1965. Guide to the analysis
of pesticide residues. U.S. Government Print. Off., Washington,
D.C.
Cassil, C.C. and P.E. Drummond. 1965. A plant surface oxidation
product of endosulfan. Jour. Econ. Entomol. 58: 356.
Chopra, N. and A. Mahfouz. 1977. Metabolism of endosulfan I, en-
dosulfan II, and endosulfan sulfate in tobacco leaf. Jour. Agric.
Food Chem. 25: 32.
Gorbach, S.G., et al. 1968. Metabolism of endosulfan in milk
sheep. Jour. Agric. Food Chem. 16: 95.
A-3
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Martens, R. 1976. Degradation of (8,9,-C-14) endosulfan by soil
microorganisms. Appl. Environ. Microbiol. 31: 853.
Whetstone, R.R. 1972. Kirk-Othmer Encyclopedia of Chemical Tech-
nology. John Wiley and Sons, Inc., New York.
Windholz, M. (ed.) 1976. The Merck Index. Merck and Co., Inc.
Rahway, N.J.
A-4
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Aquatic Life Toxicology*
INTRODUCTION
Endosulfan is a broad spectrum chlorinated cyclodiene insecticide. Al-
though restrictions on the use of endosulfan in the United States have been
proposed, significant commercial use continues for insect control on vegeta-
bles, fruits, alfalfa, and tobacco. Technical endosulfan is a 94 to 96 per-
cent mixture of two stereoisomers, endosulfan I and II, in the approximate
ratio of 70:30. Both isomers are readily metabolized to endosulfan sulfate
by a wide variety of organisms (Maier-Bode, 1968). Toxicity of the isomers
may be different, but insufficient data are available to determine which
isomer is more toxic. In addition, the relative toxicity of the two isomers
may vary with the species tested.
Technical grade endosulfan or formulations containing technical endosul-
fan have been used for most toxicity testing. Data reported herein were
largely based on tests using technical grade endosulfan or one of the two
isomers of endosulfan. Tests using formulations such as emulsifiable con-
centrates were not used for criteria derivation because of possible effects
of other components of the formulation.
The acute toxicity of endosulfan to freshwater and saltwater organisms
has been well studied, particularly in the 1960's and 1970's, although most
acute studies were carried out under static conditions with unmeasured con-
centrations. Freshwater chronic tests have been conducted on one inverte-
brate and one fish species. No measured steady-state freshwater bioconcen-
*The reader is referred to the Guidelines for Deriving Water Quality Crite-
ria for the Protection of Aquatic Life and Its Uses in order to better un-
derstand the following discussion and recommendation. The following tables
contain the appropriate data that were found in the literature, and at the
bottom of each table are calculations for deriving various measures of tox-
icity as described in the Guidelines.
B-l
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tration test data are available, and only one value for a freshwater plant
effect is available. Saltwater chronic tests have been conducted on one in-
vertebrate and one fish species. Data on the bioconcentration of endosulfan
by saltwater organisms are available for two fish species.
Two freshwater studies have been conducted on the effect of temperature
on endosulfan toxicity, and one study was conducted on the effect of water
hardness. In general, based on the limited data, toxicity seemed to in-
crease with increasing temperature, but hardness had no effect.
Data from three reports (Lemke, 1980; Nebeker, et al. 1980; Schimmel,
1980) comprise a substantial amount of the toxicity information available
for endosulfan and freshwater and saltwater organisms. These three reports
summarize results from interlaboratory comparison studies (round robins) of
static and flow-through tests on fathead minnows and rainbow trout (Lemke,
1980) and on a copepod (Acartia tonsa), mysid shrimp and sheepshead minnow
(Schimmel, 1980); chronic tests with Daphnia magna comprised the third study
(Nebeker, et al. 1980).
EFFECTS
Acute Toxicity
All acute tests on endosulfan with freshwater organisms were static,
except for the study by Macek, et al. (1976) (Table 6) and the interlabora-
tory comparison study of Lemke (1980) (Table 1). The concentration of endo-
sulfan in freshwater tests was measured only in the static test of Herzel
and Ludemann (1971) (Table 6) and the interlaboratory comparison tests
(Lemke, 1980). Values for the freshwater tests with five invertebrate and
five fish species are given in Table 1.
B-2
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Acute values for freshwater invertebrate species range from 2.3 yg/1 for
a stonefly, Pteronarcys californica, to 740 yg/1 for a cladoceran, Daphnia
magna (Table 1).
Several of the authors cited in Tables 1 and 6 reported freshwater acute
test values of other pesticides in addition to endosulfan. With inverte-
brate species, endosulfan had intermediate toxicity among the chlorinated
hydrocarbon insecticides. Sanders (1969, 1972) found endosulfan to be less
toxic than DOT and endrin, but more toxic than lindane, toxaphene, chlor-
dane, heptachlor, and dieldrin for two species of scud, Gammarus fasciatus
and Gammarus lacustris. Sanders and Cope (1968) found somewhat different
results for the stonefly, Pteronarcys californica. Endosulfan was less
toxic than endrin, dieldrin, and heptachlor, but more toxic than lindane,
DOT, and chlordane; toxaphene had toxicity similar to endosulfan. Ludemann
and Neumann (1962) found endosulfan less toxic than DDT and chlordane but
more toxic than heptachlor for a midge, Chironomus plumosus, with lindane
being about as toxic as endosulfan.
Freshwater fish species are, in general, more sensitive to endosulfan
than are invertebrate species. Acute values for fish species range from
0.17 yg/1 for rainbow trout to 4.4 ug/1 for bluegill (Table 1). With fish
species endosulfan was second in toxicity only to endrin in acute studies
with both organophosphate and organochlorine insecticides (Ludemann and Neu-
mann, 1960; Macek, et al. 1969). With fishes, endosulfan was consistently
one of the most toxic pesticides tested.
Pickering and Henderson (1966) studied the effect of water hardness on
toxicity of endosulfan and observed no significant effect. Ninety-six-hour
LCejg values for the bluegill exposed to technical-grade endosulfan in soft
and hard water were 3.3 and 4.4 yg/1, respectively.
B-3
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In contrast to the effect of hardness, toxicity of endosulfan generally
increased with increasing temperature, although the observed differences may
be at least partially attributable to experimental variability. Macek, et
al. (1969) found an almost twofold increase in toxicity to rainbow trout
when tested at 7.2 and 12.7'C as compared to 1.68C. Schoettger (1970b)
found that endosulfan toxicity increased threefold with temperature for
rainbow trout tested at 10°C as compared to 1.5°C. He also found that endo-
sulfan toxicity increased 16 percent with temperature for white sucker and
twofold with temperature for Daphnia magna, when tested at 19°C compared to
10°C. The only exception was the damselfly, Ischnura sp., which showed a
twofold decrease in toxicity when tested at 19*C as compared to 8°C. Al-
though not shown in the tables, the differences in toxicity with temperature
were usually greater at 24 hours than at 96 hours.
The absence of flow-through tests with measured concentrations in most
of the freshwater studies is primarily a function of the technology and
state-of-the-art of aquatic toxicology at the time when much of the testing
was done. Herzel and Ludemann (1971) studied the effect of aeration on the
results of static tests (Table 6). They found a greater than sixfold de-
crease in the measured concentrations of endosulfan at the end of a 96-hour
static unaerated exposure, and greater than a 40-fold decrease in an aerated
test, compared to the initial concentration at the start of the test. These
results indicate the potential problems of determining the effective expo-
sure concentration in static tests and of interpreting and comparing the
results of static tests.
In the freshwater interlaboratory study (Lemke, 1980) of acute toxicity,
with side-by-side comparison of static and flow-through tests, endosulfan
was three times more toxic to rainbow trout and two times more toxic to fat-
B-4
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head minnows in flow-through tests than in static tests, based on measured
toxicant concentrations.
Species mean acute values are listed in Table 3. The values for the
rainbow trout and fathead minnow were calculated from the measured values
for flow-through tests of the interlaboratory study (Lemke, 1980) and were
the two lowest freshwater species mean acute values. The Freshwater Final
Acute Value for endosulfan, derived from the species mean acute values
listed in Table 3 using the procedure described in the Guidelines, is 0.22
wg/1.
Twenty-three acute values have been reported for endosulfan and seven
saltwater invertebrate species (Table 1). Nineteen of the values were pro-
vided by the saltwater interlaboratory comparison study (Schimmel, 1980).
Additional results of shorter acute toxicity tests with three invertebrate
species are shown in Table 6. The acute values range from 0.032 yg/1 for
the copepod, Acartia tonsa, (Schimmel, 1980) to 730 yg/1 for the annelid
worm, Neanthes arenaceodentata (U.W. EPA, 1980). Four other arthropod spe-
cies were tested and only one Palaemon macrodactylus, had an LCgQ value
higher than 2 wg/1. Eastern oysters were less sensitive than arthropods,
with EC,.- values (based on decreased shell deposition) of 65 (Butler,
1963) and 380 ug/1 (Butler, 1964). The increased toxicity shown in the 1963
study may be related to a higher test temperature of 28"C versus 19°C in the
1964 study.
Sixteen acute toxicity tests have been conducted with five species of
saltwater fishes from five fish families (Table 1). Of the five species
tested, the LCg_ values range from 0.09 ug/1 for spot (Schimmel, et al.
1977) to 3.45 yg/1 for the sheepshead minnow (Schimmel, 1980). Table 6 pro-
vides 48-hour LC5Q data for the spot and the white mullet.
B-5
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The Saltwater Final Acute Value for endosulfan, derived from the species
mean acute values listed in Table 3 using the procedure described in the
Guidelines, is 0.034 pg/1.
Chronic Toxicity
Freshwater invertebrate chronic data are available with Daphnia magna
from the study of Macek, et al. (1976) and from the inter!aboratory compari-
son study of Nebeker, et al. (1980) (Table 2). Based on the effects of en-
dosulfan on survival of Daphnia magna through the first two generations, the
chronic limits for endosulfan in the study by Macek, et al. (1976) were 2.7
to 7.0 ug/1 in a three-generation flow-through test. Interlaboratory com-
parison data from replicated renewal chronic Daphnia magna tests from two
different laboratories gave chronic limits of 35.3, 72.8, 75.2, and 154.4
yg/1 at one laboratory and 20.0, 32.0, 32.0, and 48.0 ug/1 at the other lab-
oratory for effects on reproduction (Nebeker, et al. 1980). It should be
noted that the chronic limit values from the study by Macek,, et al. (1976)
are 10-fold less than those from the study by Nebeker, et al. (1980). This
difference in chronic values may be due to the use of different procedures
by Macek as compared to Nebeker and to the variability between laboratories
indicated by the extremes of data from the round-robin study of Nebeker, et
al. (1980). Acute-chronic ratios derived for Daphnia magna range from 4.4
to 39. Acute values used for these calculations were those obtained in
tests at the same laboratories at which the chronic studies were conducted.
Because two acute tests and two chronic tests were conducted at each
laboratory, arithmetic means of the two acute and of the two chronic tests
at each laboratory were used to calculate the ratio (Table 2).
The only available chronic data for freshwater fish species are those of
Macek, et al. (1976) with the fathead minnow (Table 2). The life-cycle test
lasted 40 weeks, and survival, growth, and reproduction were monitored.
B-6
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Based on no statistically significant adverse effects on parental fish or
offspring at 0.20 ug/1 and observed poor hatchability of control eggs
hatched in 0.40 ug/1, the chronic limits for fathead minnows were 0.20 and
0.40 ug/1, which result in a chronic value of 0.28 ug/1 (Table 2). Although
there is no 96-hour LC5Q value for fathead minnow from a flow-through test
by the same investigator using measured concentrations in the same water, an
acute-chronic ratio of 3.0 was calculated for the fathead minnow using the
fathead minnow species mean acute value of 0.83 ug/1 (Table 3).
An endosulfan 28-day life-cycle study was conducted with a saltwater
mysid shrimp (Mysidopsis bahia). In that study (U.S. EPA, 1980), the
chronic limits were 0.33 and 0.71 ug/1; the geometric mean of these gives a
chronic value of 0.48 ug/1. Both survival and reproduction (number of young
per female) were affected at 0.71 ug/l but not at 0.33 ug/1 (U.S. EPA,
1980). The acute-chronic ratio for the species is 2.8, based on an acute
value obtained from the same laboratory that conducted the chronic test
(Table 2).
Sheepshead minnows were continuously exposed to endosulfan for 28 days
starting with newly-fertilized eggs to the juvenile stage (Table 2). Survi-
val was significantly less than that of the controls in juveniles exposed to
concentrations >1.3 ug/1. Embryos were apparently unaffected by any concen-
tration tested. Average standard lengths of fish exposed to concentrations
j>0.6 ug/1 were significantly less than that of controls. Statistical
analyses failed to demonstrate adverse effects at concentrations £0.27
ug/1. Based on the results of this test, specifically the effects on growth
of juvenile fish, the chronic limits are 0.27 and 0.6 ug/1, giving a chronic
value of 0.40 ug/1. The acute-chronic ratio for the species is 2.4, using
an acute value obtained from a test conducted at the same laboratory in
which the early life stage test conducted (Table 2).
B-7
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The acute-chronic ratios for endosulfan range from 11 for Daphnia magna
(the geometric mean of three values) to 2.4 for the sheepshead minnow (Table
2). The resulting Final Acute-Chronic Ratio is 3.9. The Freshwater and
Saltwater Final Chronic Values, obtained by dividing the repsective Final
Acute Values by the Final Acute-Chronic Ratio, are 0.056 and 0.0087 ug/l,
respectively (Table 3).
Plant Effects
The only freshwater plant effect data were obtained from studies by
Gohrbach and Knauf (1971) and Knauf and Schulze (1973) (Tables 4 and 6). In
the metabolism study by Gohrbach and Knauf (1971) green alga, Chlorella vul-
garis, in 10,000 yg/1 solutions of 14C endosulfan took up the endosulfan
rapidly and began excreting endosulfan-alcohol to the water with no observed
effect on growth of the alga. In the study by Knauf and Schulze (1973)
Chlorella exposed to endosulfan as a 35 percent emulsifiable concentrate
showed growth inhibition 2,000 pg/1. Although the above data do not provide
a Final Plant Value, they indicate a lack of sensitivity of a green alga,
Chlorella vulgaris, to endosulfan toxicity.
The only saltwater plant datum available (Table 6) is that of Butler
(1963), who reported an 86.6 percent decrease in productivity of natural
phytoplankton communities (as measured by 14C uptake during a four-hour
exposure) when exposed to 1,000 ug/1, which is more than 1,000 times higher
than those that produced deleterious effects on fish or invertebrate species
in acute studies.
Residues
No appropriate bioconcentration studies with endosulfan were conducted
with any freshwater fish species.
Roberts (1972, 1975) investigated the rates of uptake, depuration, and
translation and the bioconcentration factor (BCF) of endosulfan, using the
B-8
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saltwater bivalve Mytilus edulis (Table 6). In both studies he reported very
low BCF values (12 after 112 days and 29 after 14 days); however, no mention
was made of the analysis for endosulfan sulfate, the metabolite of technical
endosulfan. Analyses for the metabolite are important because Knauf and
Schulze (1973) have shown that metabolites of endosulfan that contain the
sulfur atom exhibit toxicities to aquatic vertebrate and invertebrate spe-
cies that are similar to those of the technical material.
Several studies were conducted to determine the bioconcentration of en-
dosulfan by saltwater organisms (Table 5). Schimmel, et al. (1977) studied
the uptake, depuration and metabolism of endosulfan by the striped mullet.
When concentrations of endosulfan I and II and endosulfan sulfate were com-
bined to determine the BCF value, Schimmel, et al. (1977) reported an aver-
age BCF of 2,429 for the edible portion and an average whole body BCF of
2,755; nearly all of the endosulfan measured was in the form of the sul-
fate. Although the uptake portion of the study was conducted for 28 days,
the authors questioned whether a steady-state condition was reached since
the highest residue was reported on the 28th day of exposure. After two
days in an endosulfan-free environment, no endosulfan or sulfate was detect-
able in the exposed mullet. Sheepshead minnow juveniles, exposed from the
embryonic stage for 28 days, were analyzed for endosulfan residues (U.S.
EPA, 1980). The average whole body concentration factor for these fish was
328. Nearly all of the detectable endosulfan was that of the two isomers.
This result contrasts sharply with those of Schimmel, et al. (1977) in which
the BCF was nearly five times higher and endosulfan sulfate was the predomi-
nate residue. Reasons for the disparity are unclear but appear to due to a
capacity of mullet to metabolize the pesticide; sheepshead minnows apparent-
ly are unable to do so.
B-9
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Because no maximum permissible tissue concentration is available for en-
dosulfan, no Final Residue Value can be generated.
Miscellaneous
Other data for freshwater and saltwater effects of endosulfan are listed
in Table 6. None of the data in these studies indicate that the final acute
and chronic values calculated for endosulfan are inappropriate.
Summary
Data on acute toxicity of endosulfan are available for 10 freshwater
fish and invertebrate species that are involved in diverse community func-
tions. Acute toxicity values ranged from 0.17 ug/1 for rainbow trout to 740
ug/1 for Daphnia magna, with invertebrate species generally being less sen-
sitive than fish species. Except for recent data from an interlaboratory
comparison study, most of the data are from static tests with unmeasured
concentrations.
Five chronic tests with Daphnia magna gave chronic values ranging from
4.3 to 108 ug/1, and a single fathead minnow chronic test gave a value of
0.28 ug/1. Acute-chronic ratios ranged from 39 for one of the Daphnia tests
to 3.0 for the fathead minnow test.
The Freshwater Final Acute Value for endosulfan, based on 10 species, is
0.22 ug/l» and the Freshwater Final Chronic Value is 0.056 yg/1. No residue
data are available for endosulfan and any freshwater fish or invertebrate
species.
Plant data are available for only one species of freshwater alga, Chlo-
re 11 a vulgaris, and indicated that the alga was more resistant to endosul-
fan than were the other freshwater organisms tested.
Data on acute toxicity of endosulfan are available for 12 saltwater fish
and invertebrate species. Acute toxicity values ranged from 0.032 wg/l for
a copepod to 730 ug/1 for an annelid worm. Chronic data are available from
B-10
-------
a life-cycle study on the mysid shrimp and an early life stage study with
the sheepshead minnow. The Saltwater Final Acute Value for endosulfan,
based on 12 species, is 0.034 yg/1, and the Saltwater Final Chronic Value is
0.0087 yg/1.
Limited information on effects of endosulfan on saltwater plants indi-
cates that, as was true for freshwater, phytoplankton were much more resis-
tant than the saltwater fish and invertebrate species tested.
Bioconcentration factors for endosulfan are available for two saltwater
fish species and range from 328 to 2,755. No maximum permissible tissue
concentration or wildlife chronic feeding study is available to calculate a
Final Residue Value.
CRITERIA
For endosulfan the criterion to protect freshwater aquatic life as de-
rived using the Guidelines is 0.056 yg/1 as a 24-hour average, and the con-
centration should not exceed 0.22 ug/1 at any time.
For endosulfan the criterion to protect saltwater aquatic life as de-
rived using the Guidelines is 0.0087 yg/1 as a 24-hour average, and the con-
centration should not exceed 0.034 yg/1 at any time.
B-ll
-------
Table 1. Acute values for endosulfan*
Species
Cladoceran,
Daphnla tnagna
Cladoceran,
Daphnla magna
Cladoceran,
Daphnla magna
Cladoceran,
Daphnia magna
Cladoceran,
Daphnla magna
Cladoceran,
Daphnla magna
W
' Cladoceran,
ro Daphnla magna
Cladoceran,
Daphnla magna
Cladoceran,
Daphnla magna
Cladoceran,
Daphnla magna
Cladoceran,
Daphnja magna
Cladoceran,
Daphnla magna
Cladoceran,
Daphnla magna
Cladoceran,
Daphnia magna
Method**
LC50/EC50
(ug/l)
Species Mean
Acute Value
(ug/l) Reference
FRESHWATER SPECIES
s,
s.
s,
s.
s.
s,
s.
s,
s,
s.
s,
s,
s,
s.
M
M
M
M
M
M
M
M
M
M
M
M
U
U
218
282
250
630
740
378
266
158
372
328
343
271
166
132
Lemke,
Lemke,
- Lemke,
Lemke,
Lemke,
Lemke,
Lemke,
Lemke,
Lemke,
Lemke,
Lemke,
Lemke,
Macek,
1980
1980
1980
1980
1980
1980
1980
1980
1980
1980
1980
1980
et a
Schoettger,
-------
Table t. (Continued)
ro
t->
10
Species
Cladoceran,
Daphnla magna
Scud,
Gammarus fasclatus
Scud,
Gammarus lacustrls
Stonefly (naiad),
Pteronarcys ca 1 1 f orn 1 ca
Damsel fly (naiad),
Ischnura sp.
Damsel fly (naiad),
Ischnura sp.
Rainbow trout.
Sal mo galrdnerl
Rainbow trout.
Sal mo galrdnerl
Rainbow trout,
Sal mo galrdnerl
Rainbow trout.
Sal mo galrdnerl
Rainbow trout.
Sal mo galrdneri
Rainbow trout.
Sal mo galrdnerl
Rainbow trout.
Sal mo galrdnerl
Rainbow trout.
Sal mo galrdnerl
Method**
S, U
s, u
S, U
s. u
s, u
s, u
FT, M
FT, M
FT, M
FT, M
FT, M
FT, M
FT, M
FT, M
Species Mean
LC50/EC50 Acute Value
(Mg/l> (wa/D
62 261
6.0 6.0
5.8 5.8
2.3 2.3
71.8
107 88
0.86
0.81
0.17
0.29
0.30
0.27
0.26
0.41
Reference
Schoettger, 1970b
Sanders, 1972
Sanders, 1969
Sanders & Cope, 1968
Schoettger, 1970b
Schoettger, 1970b
Lemke, 1980
Lemke, 1980
Lemke, 1980
Lemke, 1980
Lemke, 1980
Lemke, 1980
Lemke, 1980
Lemke, 1980
-------
Table I. (Continued)
I
M
*-
Species
Rainbow trout,
Salmo galrdnerl
Rainbow trout,
Salmo galrdnerl
Rainbow trout,
Salmo galrdnerl
Rainbow trout,
Salmo galrdnerl
Rainbow trout,
Salmo galrdnerl
Rainbow trout,
Salmo galrdnerl
Rainbow trout,
Salmo galrdnerl
Rainbow trout,
Salmo galrdnerl
Rainbow trout,
Salmo galrdnerl
Rainbow trout,
Sal mo galrdnerl
Rainbow trout,
Salmo galrdnerl
Rainbow trout,
Salmo galrdnerl
Rainbow trout,
Salmo galrdnerl
Rainbow trout,
Salmo galrdnerl
Method**
FT,
FT,
FT,
FT,
s,
s.
s.
s,
s.
s.
s.
s.
s,
s.
M
M
M
M
M
M
M
M
M
M
M
M
M
M
LC50/EC50
(yg/l)
0.32
0.42
0.26
0.24
1.21
0.94
0.49
0.80
1.34
2.43
1.30
0.63
1.69
1.63
Species Mean
Acute Value
(ug/l) Reference
Lemke,
Lemke,
Lemke,
Lemke,
- Lemke,
Lemke,
- Lemke,
Lemke,
Lemke,
Lemke,
Lemke,
Lemke,
Lemke,
Lemke,
1980
I960
1980
1980
1980
1980
1980
1980
1980
1980
1980
1980
1980
1980
-------
Table 1. (Continued)
do
M
Ul
Species
Rainbow trout.
Sal mo gairdneri
Rainbow trout.
Sal mo gairdneri
Rainbow trout,
Sal mo gairdneri
Rainbow trout.
Sal mo qalrdneri
Rainbow trout,
Sal mo gairdneri
Rainbow trout.
Sal mo gairdneri
Rainbow trout.
Sal mo gairdneri
Fathead minnow,
Plmephales promelas
Fathead minnow,
Plmephales promelas
Fathead minnow,
Plmephales promelas
Fathead minnow,
Plmephales promelas
Fathead minnow,
Plmephales promelas
Fathead minnow,
Plmephales promelas
Fathead minnow,
Method**
S, M
S, M
S, U
S, U
S, U
S, U
S, U
FT, M
FT, M
FT, M
FT, M
FT, M
FT, M
FT, M
Species Mean
LC50/EC50 Acute Value
(uq/l) (wa/D
0.69
0.79
2.6
1.7
1.5
0.8
0.3 0.34
1.20
1.01
0.29
0.45
0.76
0.73
0.81
Reference
Lemke, 1980
Lemke, 1980
Macek, et al
Macek, et al
Macek, et al
Schoettger,
Schoettger,
Lemke, 1980
Lemke, 1980
Lemke, 1980
Lemke, 1980
Lemke, I960
Lemke, 1980
Lemke, 1980
. 1969
. 1969
. 1969
1970b
1970b
Plmephales promelas
-------
Table 1. (Continued)
w
Species
Fathead minnow,
Plmephales promelas
Fathead minnow,
Plmephales promelas
Fathead minnow,
Plmephales promelas
Fathead minnow,
Plmephales promelas
Fathead minnow,
Plmephales promelas
Fathead minnow,
Plmephales promelas
Fathead minnow,
Plmephales promelas
Fathead minnow,
Plmephales promelas
Fathead minnow,
Plmephales promelas
Fathead minnow,
Plmephales promelas
Fathead minnow,
Pimephales promelas
Fathead minnow,
Plmephales promelas
Fathead minnow,
Plmephales promelas
Fathead minnow.
Method**
FT, M
FT,
FT,
FT,
FT,
s.
s.
s,
s,
s,
s.
s.
s.
s.
M
M
M
M
M
M
M
M
M
M
M
M
M
LC50/EC50
(ug/l)
0.80
1.67
1.57
0.75
1.00
2.35
3.45
2.10
3.20
1.70
1.48
1.90
0,97
1.35
Species Mean
Acute Value
(ug/l) Reference
Lemke,
Lemke,
Lemke,
Lemke,
Lemke,
Lemke,
- Lemke,
Lemke,
Lemke,
Lemke,
Lemke,
Lemke,
LefRke,
- Lemke,
1980
1980
1960
1980
1980
1980
1980
1980
1980
1980
1980
1980
1980
1980
Plmephales promelas
-------
Table I. (Continued)
tt>
Species
Fathead minnow,
Plmephales promelas
Fathead minnow.
Plmephales promelas
Fathead minnow.
Plmephales promelas
White sucker.
Catostomus commerson i
White sucker,
Catostomus commerson 1
Guppy,
Poecilia reticulata
Bluegill,
Lepomis macrochirus
Bluegill,
Lepomis macrochirus
Annel id worm,
Neanthes arenaceodentata
Eastern oyster.
Crassostrea virgin lea
Eastern oyster,
Crassostrea vlrglnica
Copepod ,
Acartla tonsa
Copepod,
Acartia tonsa
Method**
S, M
S, M
S, M
S, U
S, U
S, U
S, U
S, U
S, U
FT, U
FT, U
S, U
S, U
Species Mean
LC50/EC50 Acute Value
(ug/l) (pq/D
1.20
3.20
2.50 0.83
3.5
3.0 3.2
3.7 3.7
3.3
4.4 3.8
SALTWATER SPECIES
730 730
65
380 157
0.12
0.05
Reference
Lemke, 1980
Lemke, 1980
Lemke, 1980
Schoettger, 19705
Schoettger, 1970b
Pickering &
Henderson, 1966
Pickering &
Henderson, 1966
Pickering &
Henderson, 1966
U.S. EPA, 1980
Butler, 1963
Butler, 1964
Schlmmel, 1980
Schlmmel, 1980
-------
Table 1. (Continued)
ro
i
CD
Species
Copepod,
Acartia tonsa
Copepod,
Acartia tonsa
Copepod,
Acartia tonsa
Copepod,
Acartia tonsa
Mysld shrimp,
Mysldopsis bah la
Mysld shrimp,
Mysldopsis bah la
Mysid shrimp,
Mysidopsls bah la
Mysld shrimp,
Mysldopsis bah la
Mysld shrimp,
Mysldopsis bah la
Mysid shrimp,
Mysldposls bah la
Mysid shrimp,
Mysidopsis bahla
Mysid shrimp,
Mysidopsis bahla
Mysid shrimp,
Mysldopsis bahia
Mysid shrimp,
Mysidopsis bahia
Method"
S, U
s, u
S, U
s, u
s, u
s, u
s, u
s, u
s, u
FT, M
FT, M
FT, M
FT, M
FT, M
LC50/EC50
-------
Table 1. (Continued)
(fl
I
Species
Korean shrimp,
Palaemon macrodacty 1 us
Korean shrimp,
Palaemon macrodacty 1 us
Grass shrimp.
Pal aemonetes puglo
Pink shrimp,
Penaeus duorarum
Sheepshead minnow,
Cyprlnodon variegatus
Sheepshead minnow,
Cyprlnodon variegatus
Sheepshead minnow,
Cyprlnodon variegatus
Sheepshead minnow,
Cyprlnodon variegatus
Sheepshead minnow,
Cyprlnodon variegatus
Sheepshead minnow,
Cyprlnodon variegatus
Sheepshead minnow,
Cyprlnodon variegatus
Sheepshead minnow,
Cyprlnodon variegatus
Sheepshead minnow,
Cyprlnodon variegatus
Sheepshead minnow.
Method**
s,
FT,
FT,
FT,
s.
s,
s.
s.
s.
s.
FT,
FT,
FT,
FT,
U
U
M
M
U
U
U
U
U
U
M
M
M
M
Species Mean
LC50/EC50 Acute Value
(ug/l) (ug/l) Reference
17.1
3.4
1.31
0.04
2.7
1.4
1.2
2.87
3.45
2.81
1.10
0.34
0.60
0.88
Schoettger, 1970a
7.6 Schoettger, 1970a
1.31 Schlmmel, et al. 1977
0.04 Schlnmel, et al. 1977
Schlmmel, 1980
Schlmmel,
Schimmel,
Schlmmel,
Schlmmel,
Schlmmel,
Schimmel,
Schlmmel,
Schlmmel,
Schlmmel,
1980
1980
1980
1980
1980
1980
1980
1980
1980
Cyprlnodon variegatus
-------
Table 1. (Continued)
LC50/EC50
Species Method** (yg/l)
Sheepshead minnow, FT, M 1.15
Cyprlnodon varlegatus
Sheepshead minnow, FT, M 0.83
Cyprlnodon varlegatus
Striped bass, FT, U 0.10
Morone saxati 1 Is
Plnflsh, FT, M 0.30
Lagodon rhomboides
Spot, FT, M 0.09
Leiostomus xanthurus
Striped mullet, FT, M 0.38
Mugl 1 cephalus
Species Mean
Acute Value
(ug/D Reference
Schlmmel, 1980
0.76 Schimmel, 1980
0.10 Korn 4 Earnest, 1974
0.30 Schlmmel, et al. 1977
0.09 Schlmmel, et al. 1977
0.38 Schirmiel, et al. 1977
to
o
* Technical grade endosuI fan
**S = static, FT = flow-through, U = unmeasured, M = measured
-------
Table 2. Chronic values for endosulfan*
NJ
Species
Cladoceran,
Daphnla magna
Cladoceran,***
Daphnla magna
Cladoceran,***
Daphnla magna
Cladoceran,****
Daphnla magna
Cladoceran,****
Daphnla magna
Fathead minnow,
Plmephales promelas
Mysld shrimp,
Mys 1 dops 1 s bah 1 a
Sheepshead minnow,
Cyprlnodon varlegatus
Limits Chronic Value
Test«« (ug/D
-------
Table 2. (Continued)
Acute-Chronic Ratios
to
to
Spec 1 es
Cladoceran,
Daphnla magna
Cladoceran,
Daphnla magna
Cladoceran,
Daphnla magna
Fathead minnow,
Plmephales promelas
Mysld shrimp,
Mysldopsis bah la
Sheep shead minnow,
Cyprlnodon varlegatus
Acute
Value
(ug/i)
166
250*
350*
0.83
1.37**
0.95**
Chronic
Value
(ug/l)
4.3
32.2*
79.4*
0.28
0.48
0.40
Ratio
39
7.8
4.4
3.0
2.8
2.4
* Arithmetic mean of replicate tests by same Investigator In the same water for both
acute and chronic tests.
**Acute value from test by same investigator In the same water source as for chronic
value.
Geometric mean of acute-chronic ratios for Daphnla magna = 11
-------
Table 3. Species mean acute values and acute-chronic ratios for endosulfan
ink* Species
FRESHWATER
10 Cladoceran,
Daphnla magna
9 Damsel fly (naiad),
Ischnura sp.
8 Scud,
Gammarus fasciatus
7 Scud,
Gammarus lacustrls
6 Bluegill,
Lepomis macrochirus
5 Guppy,
Poec Ilia ret i cu 1 ata
4 White sucker,
Catostomus commer son i
3 Stonefly (naiad),
Pteronarcys ca 1 1 f orn 1 ca
2 Fathead minnow,
Plmephales promelas
1 Rainbow trout.
Sal mo gairdneri
SALTWATER
12 Annel i d worm,
Neanthes arenacondentata
1 1 Eastern oyster,
Crassostrea virginica
Species Mean
Acute Value
-------
Table 3. (Continued)
w
10
Rank*
9
8
7
6
5
4
3
2
1
Species
Grass shrimp,
Pa 1 aemonetes puglo
Mysld shrimp,
Mysidopsls bah la
Sheep shead minnow,
Cyprlnodon varlegatus
Striped mul let,
Mugl 1 cephalus
Plnflsh,
Lagodon rhomboldes
Copepod,
Acartla tonsa
Striped bass,
Morone saxatl 1 Is
Spot,
Lelostomus xanthurus
Pink shrimp,
Penaeus duorarum
Species Mean
Acute Value
(U9/D
1.31
0.83
0.76
0.38
0.30
0.14
0.10
0.09
0.04
Species Mean
Acute-Chronic
Ratio
2.8
2.4
* Ranked from least sensitive to most sensitive based on species mean
acute value.
Final Acute-Chronic Ratio = 3.9
Freshwater Final Acute Value = 0.22 ug/l
Freshwater Final Chronic Value = 0.22 ug/l t- 3.9 = 0.056 ug/l
-------
Cd
to
tn
Table 3. (Continued)
Saltwater Final Acute Value = 0.034 ug/l
Saltwater Final Chronic Value = 0.034 ug/l t- 3.9 = 0.0087 ug/l
-------
Species
FRESHWATER SPECIES
a
i
K)
ffl
Table 4. Plant values for endosulfan
Result
Chemical Effect (ug/D Reference
Green alga,
Chi ore! la vulgarly
Fndosu 1 fan
>4C- labeled
None observed
on growth
10,000 Gohrbach
1971
& Knauf,
-------
Species
Table 5. Residues for endosuI fan*
Tissue
Llpld Bloconcentration
(?) Factor
Duration
(days) Reference
SALTWATER SPECIES
Sheep shead minnow, Whole body 3.6** 328
Cyprlnodon varlegatus
Striped mul let. Edible tissue - 2,429***
Mug) 1 cephalus
Striped mullet. Whole body 1.0 2,755***
Mug II cephalus
28 U.S. EPA, 1980
28 Schlmnel, et al. 1977
28 Schlmnel, et al. 1977
td
* Technical grade endosuI fan
** Percent lip Id data from Hansen, 1980
*** Bloconcentration factor includes bloconcentratlon of the metabolite, endosulfan sulfate.
-------
Table 6. Other data for endosulfan
Chemical*
Duration
Effect
Result
(tig/1) Reference
FRESHWATER SPECIES
Green alga,
Ch 1 ore 1 1 a vu 1 gar 1 s
Midge (larva),
Chlronomus plumosus
Tubl field worm.
Tub If ex tub if ex
Rainbow trout (fry),
Sal mo galrdnerl
Northern pike
(f Ingerl ing),
Esox lucius
Carp,
Cyprlnus carplo
w
w Cal"p«
CD Cyprlnus carpio
Carp (f Ingerl Ing),
Cyprlnus carp_|o_
Fathead minnow,
Plmephales promelas
Mosqultoflsh,
Gambusla af finis
Mosqultoflsh,
Gambusla af finis
Guppy,
Poecl lia retlculata
Guppy,
Poeci 1 la retlculata
Guppy,
Poec Ilia ret i cu 1 ata
Endosul fan
35 EC*»
Techn 1 ca 1
grade
Techn 1 ca 1
grade
Techn 1 ca I
grade
Techn 1 ca 1
grade
2* EC
2* EC
Technical
grade
Techn 1 ca 1
grade
Thlodan *l
2* EC
Thlodan «l 1
2*. EC
Techn 1 ca 1
grade
Techn 1 ca 1
grade
Techn I ca 1
grade
120 hrs
24 hrs
96 hrs
24 hrs
24 hrs
24 hrs
24 hrs
48 hrs
7 days
24 hrs
24 hrs
5 hrs
96 hrs
96 hrs
Inhibited growth
LC50
100* mortality
100* mortality
100* mortality
70* mortal Ity
60* morta 1 1 ty
LC50
Incipient LC50
6* morta 1 1 ty
24* mortal ity
100* mortality
85* mortality
55* mortality
>2,000
53
10,000
10
5
10
25
It.O
0.86
0.1
1 bs/acre
0.1
1 bs/acre
50
4.2
4.2
Knauf & Schulze, 19
Ludemann & Neumann,
1962
Ludemann & Neumann,
1962
Ludemann & Neumann,
1961
Ludemann & Neumann,
1961
Mulla, et al. 1967
Mulla, et al. 1967
Ludemann & Neumann,
1960
Macek, et al. 1976
Mulla, 1963
Mulla, 1963
Jones, 1975
Herzel & Ludemann,
1971
Herzel & Ludemann,
1971
-------
Table 6. (Continued)
w
Species
Blueglll,
Lepomls macroch 1 rus
Bullfrog (tadpole),
Rana catesbelana
Bullfrog (tadpole),
Rana catesbelana
Mai lard (young).
Anas platyrhynchus
Natural phytoplankton
communities
Common mussel,
Mytl lus edul Is
Common mussel,
Mytl lus edul Is
Blue crab,
Calllnectes sapldus
Brown shrimp,
Penaeus aztecus
Brown shrimp,
Crangon crangon
Grass shrimp,
Palaemonetes puglo
Pinflsh,
Lagodon rhomboides
Spot,
Lelostomus xanthurus
Chemical
Techn 1 ca 1
grade
Thlodan 1
2% EC
Thlodan II
2% EC
Techn 1 ca 1
grade
Technical
grade
Technical
grade
Technical
grade
Techn 1 ca 1
grade
Technical
grade
Techn I ca 1
grade
Techn 1 ca 1
grade
Techn 1 ca 1
grade
Techn 1 ca 1
grade
Duration
Unspecified
24 hrs
96 hrs
5 days
SALTWATER
4 hrs
112 days
14 days
2 days
2 days
2 days
4 days
4 days
4 days
Effect
50* Inhibition
of brain Mg-ATPase
60% morta 1 1 ty
lOJf mortality
50% mortality
SPECIES
86.6$ decrease
in productivity
1,000 14C
B ioconcentrat Ion
factor = 12
B Ioconcentrat Ion
factor = 29
EC50***
EC50***
LC50
Bioconcentration
factor = 175****
Bioconcentration
factor = 1,173****
Bioconcentration
factor = 779****
Result
(ug/t)
6,050
0.1
Ibs/acre
0.1
Ibs/acre
1,050
mgAg
1,000
35
0.4
10
Reference
Yap, et al. 1975
Mulla, 1963
Mulla, 1963
Hill, et al. 1975
Butler, 1963
Roberts, 1972
Roberts, 1975
Butler, 1963
Butler, 1963
Portman & W i 1 son ,
1971
Schimnel, et al. 1977
Schlmmel, et al. 1977
Schlmnel, et al. 1977
-------
Table 6. (Continued)
tfl
i
CJ
o
Species
Spot,
Leiostomus xanthurus
White mul let.
Mug it curema
Striped mul let,
Mugl 1 cephalus
Chemical
Technical
grade
Techn i ca 1
grade
Technical
grade
Duration
2 days
2 days
4 days
Effect
LC50
LC50
Blocortcentrat Ion
factor = 1,115****
Result
(ug/l)
0.6
0.6
Reference
Butler, 1964
Butler, 1963
Schimmel, et at. 1977
* Thlodan = formulation trademark; EC = emulsiflable concentrate.
** Formulation 35* emulsiflable concentrate, unmeasured.
*** Loss of equilibrium.
****Bloconcentration factor includes bioconcentration of the metabolite, endosulfan sulfate.
-------
REFERENCES
Butler, P.A. 1963. Commercial fisheries investigations, pesticide-wildlife
studies, A review of Fish and Wildlife Service investigations during 1961
and 1962. U.S. Dept. Int. Fish Wildl. Circ. 167: 11.
Butler, P.A. 1964. Pesticide-Wildlife Studies, 1963: A review of Fish and
Wildlife Service investigations during the calendar year. U.S. Dept. Int.
Fish Wildl. Circ. 199: 5.
Gohrbach, S. and W. Knauf. 1971. Endosulfan und Umwelt: Oas Rueskstands-
verhalten von Endosulfan und seine Wirkung auf organsimen, die im Wasser le-
ben. Schriftenreihe des Vereins fiir Wasser-, Boden- und Lufthygiene.
34: 85.
Hansen, 0. 1980. Memo to C.E. Stephan. June 1980.
Herzel, F. and 0. Ludemann. 1971. Verhalten und Toxizitat von Endosulfan
in Wasser unter verschiedenen Versuchsbedingungen. Z. Angew. Zool. 58: 57.
Hill, E.F., et al. 1975. Lethal dietary toxicities of environmental pollu-
tants to birds. U.S. Fish Wildl. Serv. Spec. Sci. Rep. Wildl. 191.
Jones, W.E. 1975. Detection of pollutants by fish tests. Water Treat.
Examin. 24: 132.
B-31
-------
Knauf, W. and E.F. Schulze. 1973. New findings on the toxicity of endosul-
fan and its metabolites to aquatic organisms. Meded. Fac. Lanbouwwet,
Rijksuniv. Gent. 38: 717.
Korn, S and R. Earnest. 1974. Acute toxicity of 20 insecticides to striped
bass Morone saxatilis. Calif. Fish Game. 60: 128.
Lemke, A.E. 1980. Comprehensive report. Interlaboratory comparison acute
testing set. U.S. Environ. Prot. Agency, Environ. Res. Lab., Duluth,
Minnesota.
LiJdemann, D. and H. Neumann. 1960. Versuche uber die akute toxische Wir-
kung neuzeitlicher Kontaktinsektizide auf einsommerige Karfern (Cyprinum
carpio L.) A. Angew. Zool. 47: 11.
Ludemann, D. and H. Neumann. 1961. Versuche uber die akute toxische Wir-
kung neuzeitlicher Kontaktinsektizide auf Susswassertiere. 1. Angew. Zool.
48: 87.
Ludemann, D. And H. Neumann. 1962. Uber die Wirkung der neuzeitlichen Kon-
taktinsektizide auf die Tiere des Susswassers. Anz. Shadlingsunde. 35: 5.
Macek, K.J., et al. 1969. The effects of temperature on the susceptibility
of bluegills and rainbow trout to selected pesticides. Bull. Environ. Con-
tarn. Toxicol. 4: 174.
Macek, K.J., et al. 1976. Toxicity of four pesticides to water fleas and
fathead minnows. U.S. Environ. Prot. Agency, EPA 600/3-76-099.
B-32
-------
Maier-Bode, H. 1968. Properties, effect, residues and analytics of the
insecticide, endosulfan. Residue Rev. 22: 1.
Mulla, M.S. 1963. Toxicity of organochlorine insecticides to the mosquito
fish Gambusia affinis and the bullfrog Rana catesbeiana. Mosq. News.
23: 299.
Mulla, M.S., et al. 1967. Evaluation of organic pesticides for possible use
as fish toxicants. Prog. Fish-Cult. 29: 36.
Nebeker, A.V., et al. 1980. Comparative sensitivity of rainbow trout, fat-
head minnow, and Daphnia magna to silver and endosulfan. Manuscript.
Pickering, Q.H. and C. Henderson. 1966. The acute toxicity of some pesti-
cides to fish. Ohio Jour. Sci. 66: 508.
Portman, J. E. and K.W. Wilson. 1971. The toxicity of 140 substances to
the brown shrimp and other marine animals. Ministry of Agriculture,
Fisheries and Food. Shellfish Information Leaflet. No. 22.
Roberts, D. 1972. The assimilation and chronic effects of sublethal con-
centrations of endosulfan on condition and spawning in the common mussel
Myti.lus edulis. Mar. Biol. 16: 119.
Roberts, D. 1975. Differential uptake of endosulfan by the tissues of
Mytilus edulis. Bull. Environ. Contam. Toxicol. 13: 170.
B-33
-------
Sanders, H.O. 1969. Toxicity of pesticides to the crustacean Gammarus
lacustris. U.S. Bur. Sport Fish. Wild!. Tech. Pap. 25.
Sanders, H.O. 1972. Toxicity of some insecticides to four species of mala-
costracan crustaceans. U.S. Bur. Sport Fish. Wildl. Tech. Pap., 66.
Sanders, H.O. and 0. B. Cope. 1968. The relative toxicities of several
pesticides to naiads of three species of stoneflies. Limnol. Oceanogr.
13: 112.
Schimmel, S.C. 1980. Final report on results of the acute toxicity round
robin using estuarine animals. U.S. Environ. Prot. Agency, Environ. Res.
Laboratory, Gulf Breeze, Florida.
Schimmel, S.C., et al. 1977. Acute toxicity to and bioconcentration of en-
dosulfan by estuarine animals. Aquatic Toxicology and Hazard Evaluation,
ASTM STP 634, Am. Soc. Test. Mater.
Schoettger, R.A. 1970a. Fish-Pesticide Research Laboratory: Progress in
sport fishery research. U.S. Dept. Int. Bur. Sport Fish Wildl. Resour.
Publ. 106.
Schoettger, R.A. 1970b. Toxicology of thiodan in several fish and aquatic
invertebrates. Investigations in fish control. U.S. Bur. Sport Fish.
Wildl. 35.
B-34
-------
U.S. EPA. 1980. Unpublished laboratory data. Environ. Res. Laboratory,
Gulf Breeze, Florida.
Yap, H.H., et al. 1975. In vitro inhibition of fish brain ATPase activity
by cyclodiene insecticides and related compounds. Bull. Environ. Contam.
Toxicol. 14: 163.
B-35
-------
Mammalian Toxicology and Human Health Effects
EXPOSURE
Ingestion from Water
Schulze, et al. (1973) presented data from the U.S. Geologi-
cal Survey program for monitoring pesticides in the streams of the
western United States for the period October 1968 to September
1971. At 20 sampling stations, water samples were collected at
monthly intervals and analyzed for residues of endosulfan and
other pesticides by gas chromatography. No attempt was made to
separate suspended sediment from the water for separate analysis.
The low detection limit for endosulfan was TO.005 ug/1. In a
total of 546 water samples analyzed, one sample (from the Gila
River at Gillespie Dam, Arizona) contained an endosulfan residue
of 0.02 ug/lf along with residues of five other organochlorine
insecticides.
FMC (1971) reported endosulfan levels in runoff water in
North American agricultural areas. Water samples from a pond lo-
cated in a field treated with endosulfan contained no detectable
residues «10 ug/l). Mud samples from the bottom of the pond,
however, contained a maximum of 0.05 mg/kg ^endosulfan and 0.07
mg/kg endosulfan sulfate. These samples were taken approximately
280 days after the last endosulfan application.
In a subsequent study, irrigation runoff was monitored from a
field in California treated at a rate of 1.12 kg/ha (FMC, 1972).
Water residues (^- and X-endosulfan) were approximately 15 Wg/l
following the first irrigation but dissipated to below the detec-
tion limit (0.005 ug/1) after 15 days.
C-l
-------
Miles and Harris (1971) measured insecticide residues in the
water of a creek flowing into Lake Erie and in ditches draining an
agricultural area near Lake Erie. In these water systems, water
was sampled weekly and bottom mud was sampled monthly from mid-
April to mid-October 1970. No endosulfan residues were found in
the creek. In the drainage ditch, endosulfan residues in the
water ranged from <2 to 32 ng/1 at the pumphouse, where the water
was lifted into Lake Erie when necessitated by the water level,
and from <2 to 187 ng/1 one mile upstream from the pumphouse.
Endosulfan residues in bottom mud were <1 to 1 ug/kg (dry weight
basis) at the pumphouse and ranged from 4 to 62 ug/kg upstream.
In 1971, in order to compare residue contributions from areas
of differing insecticide use, Miles and Harris (1973) determined
insecticide residues in water systems draining agricultural,
urban-agricultural, and resort areas in Ontario, Canada. Water,
bottom mud, and fish samples from these water systems were col-
lected between mid-April and mid-October and analyzed for endosul-
fan residues by gas-liquid chromatography. Endosulfan residues in
individual water samples ranged from <1 to 11 ng/1 in Big Creek
and from <1 to 3 ng/1 in the Thames River (average level for all
samples was <1 ng/1); no residues were found in the Muskoka River
(limit of detection 1 ng/1). No endosulfan residues were detected
in 18 samples of bottom mud or in a total of 57 fish collected
from the three water systems.
The National Research Council of Canada (NRCC) (1975) reports
unpublished data (Frank, 1973) on endosulfan residues in water
samples collected four times per year between 1968 and 1973 in six
C-2
-------
southern Ontario rivers and municipal water supplies. Over this
period, endosulfan was detected only in one sample, at a level of
0.012 ug/1. In 1973, five water and three sediment sampling sites
were monitored at 2-week intervals from late March to mid-Septem-
ber, and monthly thereafter. Endosulfan residues were detected
only in water during one sampling period at levels of 0.047 to
0.083 ug/1.
Frank, et al. (1977) subsequently published the results of
pesticide analysis of 50 sediment samples collected on a grid from
Lake St. Clair in 1970 and 1974. In 1970, endosulfan residues
were present in the sediments at a mean residue of 0.2 ug/kg
(ranging from nondetectable levels (<0.2 ug/kg) to 2.2 ug/kg).
Only 20 percent of the samples, however, contained endosulfan and
these residues (c^- and /^-endosulfan with traces of endosulfan
sulfate) were confined to sediments from the lower reaches of the
ship channel between Lake St. Clair and the Detroit River and off-
shore from the mouth of the Thames River. Endosulfan was not de-
tected in any of the 1974 samples.
Endosulfan residues in Lakes Erie and Ontario have been re-
ported by the Environmental Quality Coordination Unit (1973) of
the Canada Centre for Inland Waters. Of 40 samples of surface and
bottom water from Lake Erie, 5 contained endosulfan concentrations
ranging from 0.005 to 0.014 ug/1. Of 40 Lake Ontario samples, 6
contained endosulfan at concentrations of 0.005 to 0.051 ug/1.
Residues in the sediment samples and in the other water samples
were below the detection limits, 0.005 ug/1 of water and 5 to 10
ug/kg of sediment.
C-3
-------
Wong and Donnelly (1968) measured pesticide concentrations in
the St. Lawrence River and in the Bay of Quinte which empties into
the northern shore of Lake Ontario. Endosulfan was generally non-
detectable in the St. Lawrence River, but a few samples contained
endosulfan residues between 0.020 and 0.060 ug/1.
Several laboratories studied the occurrence of endosulfan
residues in the Rhine River in West Germany and in the Netherlands
following a massive endosulfan-caused fish kill in the Rhine in
June 1969 due to an accidental point source contamination. This
episode was the result of accidental discharge of approximately
220 Ib of endosulfan into the river system rather than from run-
off (NRCC, 1975}.
Seivers, et al. (1972) monitored the concentrations of endo-
sulfan in the Rhine and Main Rivers in West Germany from June to
December 1969. The endosulfan concentrations found in these sam-
ples were within the following ranges:
Endosulfan Concentration
Range (ng/1)
<100
100-500
500-1,000
1,000-10,000
>10,000
Total
Number of Samples from
Rhine
55 (100%)
Main
21 (38%)
27 (49%)
4 ( 7%)
3 ( 6%)
0 ( 0%)
3 (14%)
1 ( 4%)
4 (18%)
9 (41%)
5 (23%)
22 (100%)
C-4
-------
Many communities along the Rhine draw their water supplies
from the river. Endosulfan residues in 35 samples of Rhine shore
filtrates collected between June 1969 and February 1970 contained
endosulfan concentrations ranging from <10 to 35 ng/1.
Greve and Wit (1971) determined endosulfan concentrations in
about 320 samples of surface water and 35 samples of drinking
water collected between June 24 and August 31, 1969, from the
Dutch section of the Rhine and its tributaries following a massive
fish kill in June. Endosulfan was identified by gas-liquid chro-
matography. The maximum concentration of endosulfan (c^ +x£ ) found
in river water in the Netherlands was 0.70 ug/1 on the first day
of sampling. From this maximum value, a steady decrease was
observed; one month after initiation of sampling, the endosulfan
concentration had fallen below the limit of detectability, 0.01
ug/1.
Greve and Wit (1971) found that river silt readily adsorbs
endosulfan. Of the endosulfan present in raw river water samples,
82 to 85 percent could be removed by filtration or centrifugation.
Ferric hydroxide gel and activated carbon (used in the treatment
of drinking water) were still better adsorbents for endosulfan.
Ferric hydroxide gel not only adsorbed endosulfan, but also cat-
alyzed its hydrolysis.
In a more extensive monitoring study, Greve (1972) measured
endosulfan residues in the Dutch section of the Rhine River from
September 1969 to March 1972. During this period, water samples
were collected three times a week in the Waal River, the main
branch of the Rhine River in the Netherlands. Endosulfan
C-5
-------
residues were found in 75 percent of the samples, ranging from
<0.01 to 0.88 ug/1/ the average and median endosulfan concentra-
tions were 0.10 and <0.01 ug/1, respectively, and the upper and
lower deciles were <0.01 to 0.29 ug/1.
Wegman and Greve (1978) monitored the Dutch aquatic environ-
ment from September 1969 to December 1975 for organochlorine pes-
ticides. Some 1,492 samples were analyzed, including surface
water, rainwater, groundwater, and drinking water. The results of
these analyses were as follows:
No. of Sample Sets Analyzed
Endosulfan* Total Maximum Endosulfan*
Year Containing No. Residue (ug/D
H69" 17 32 0.81
1970 36 45 0.40
1971 9 22 0.25
1972 7 35 0.09
1973 9 22 0.10
1974 1 3 0.02
1975 1 1 0-02
*c^- and/^-endosulfan; practical detection limit is 0.01 ug/1.
Herzel (1972) monitored organochlorine insecticides in sur-
face waters in the Federal Republic of Germany. Samples of unfil-
tered water and suspended solids were analyzed from about 25 sites
sampled in May 1971, and unfiltered water was analyzed from seven
sites sampled monthly between April 1970 and June 1971. All sam-
ples were analyzed by gas chromatography , and the detection
C-6
-------
limits for c^- and /<7-endosulfan were 10 to 30 ng in 30 ml of hex-
ane extract. Of 120 samples of unfiltered surface waters ana-
lyzed, eight contained residues of <^ -endosulfan ranging from 10
to 100 ng/1, and three contained residues of ^-endosulfan ranging
from 20 to 95 ng/1. These endosulfan concentrations were found in
samples from the Rhine, the lower Main, and the Regnitz and,
according to the investigator, originated from industrial efflu-
ents.
Of 20 samples of suspended solids, two contained c^ -endosul-
fan at concentrations of 22 and 24 ng, and one contained ^-endo-
sulfan at a concentration of 9.6 ng. These values are expressed
in terms of the quantities of each endosulfan isomer (in nano-
grams) found in the solids suspended in one liter of water.
Tarrant and Tatton (1968) studied the presence of organo-
chlorine pesticides in rainwater in the British Isles. The total
precipitation collected in each 3-month period at seven sampling
stations was analyzed by thin-layer and gas-liquid chromatography.
The detection limit for endosulfan was about 1 ng/1. NO endosul-
fan residues were detected in any of the 28 composite samples of
rainwaters analyzed.
Gorbach, et al. (1971a) investigated the presence and persis-
tence of endosulfan residues in East Java in a river system
(Brantas River) and in ponds and seawater following large-scale
use of endosulfan on rice in the delta region of the Brantas
C-7
-------
River. The concentration of endosulfan residues in the water sam-
pled as determined by gas chromatography were as follows:
Endosulfan Residues (ug/D
£7S ,0 Sulfate
Canals
Fish Ponds
River system
Madura Sea
Average
Range
Average
Range
Average
Range
Average
Range
<0
<0
<0
<0
<0
<0
<0
<0
.13
.01-5.8
.03
.01-0.25
.01
.01-5.0
.02
.01-0.09
<0
<0
<0
<0
<0
<0
<0
<0
.12
.01-2.4
.02
.01-0.08
.11
.01-2.0
.02
.01-0.07
<0
<0
<0
<0
<0
<0
<0
<0
.18
.01-0
.06
.01-0
.19
.01-0
.08
.01-0
.55
.44
.45
.28
The highest residue levels (5.8 and 2.4 ug/1 of - and -
endosulfan, respectively) were detected in a canal that drained
treated fields shortly after an endosulfan application. Within
two days, these high levels decreased to about 0.2 ug/1 by degra-
dation and/or dilution with uncontaminated water. Total endosul-
fan residues (^ +
-------
After treatment, the initial water concentration of total
endosulfan residues in one field was 68 ug/lf declining to the
pretreatment value of 0.5 to 0.8 ug/1 within five days. In the
mud of both submerged test fields, maximum total endosulfan resi-
dues were 0.053 and 0.008 mg/kg, respectively, directly after
treatment, declining to about 0.01 to 0.02 mg/kg by the fifth day
post-treatment. In an adjacent dry rice field, a maximum endosul-
fan residue of 1.9 mg/kg was found. The sulfate equivalent in the
total endosulfan residues increased with time, pointing to conver-
sion of the parent compound in the presence of water.
Several fish kills attributable to endosulfan have been re-
ported from other countries. One major, widely publicized and
investigated episode occurred in the Rhine River in West Germany
in 1969.
Osmond (1969) reported on an endosulfan-related fish kill
which took place in Ontario, Canada, in August of 1969. Analysis
of water samples collected where fish had been killed from the
Thames River and a tributary where the contamination occurred
showed endosulfan concentrations of 0.096 and 0.260 ug/1, respec-
tively. Two other samples taken from upstream on the Thames River
and from further up the tributary had endosulfan levels of 0.022
and 0.026 ug/1.
A second fish kill occurred in a pond near Simcoe, Ontario,
in 1972 (Frank, 1972). Endosulfan could not be detected in the
pond water (limits of detection 0.001, 0.002, and 0.01 ug/1 for
c/) -endosulfan, x^-endosulfan, and endosulfan sulfate, respec-
tively). However, bottom sediment from one end of the pond
C-9
-------
contained 0.9, 1.0, and 1.1 ug/kg (dry weight) of C"5-endosulfan,
^''-endosulfan, and the sulfate, respectively. Sediment from the
other end of the pond contained 1.2 ug/kg (dry weight) endosulfan
sulfate.
Ingestion from Food
Endosulfan is a broad-spectrum insecticide and acaricide that
is registered in the United States for use in the control of over
100 different insect pests occurring in over 60 food and nonfood
crops.
Official U.S. tolerances for pesticide residues in raw agri-
cultural commodities are published in the Code of Federal Regula-
tions, Title No. 40, and in the Federal Register. Appropriate
food additive tolerances for processed commodities are published
in Title No. 21 of the Code of Federal Regulations. U.S. toler-
ances for endosulfan and its metabolite are listed in Table 1.
Endosulfan tolerances that have been set by other countries
are summarized in Table 2.
The acceptable daily intake (ADI) is defined by Lu (1973) as
the daily intake of a pesticide which during an entire lifetime
appears to be without appreciable risk on the basis of all known
facts at the time of evaluation. It is expressed in milligrams of
the chemical per kilogram of body weight (mg/kg).
The ADI for pesticides is established jointly by the Food and
Agricultural Organization (FAO) Working Party on Pesticide Resi-
dues and the World Health Organization (WHO) Expert Committee on
C-10
-------
TABLE 1
U.S. Tolerances for Endosulfan*9
mg/kg
Crop
mg/kg
Crop
mg/kg
Crop
n
i
0.3
1
0.2b
1
2
2
2
O.lb
0.2b
2
O.lb
2
2
2
0.2
0.2
2
2
2
2
0.2
1
2
Alfalfa (fresh) 2
Alfalfa hay 0.2b
Almonds 0.2
Almond hulls
Apples 2
Apricots 0.2
Artichokes
Barley grain 0.2
Barley straw
Beans 2
Blueberries 2
Broccoli 0.2b
Brussels sprouts 2
Cabbage 0.5C
Carrots 2
Cattle (meat, fat, meat 0.2b
by-products) 2
Cauliflower O.lb
Celery 0.2b
Cherries 2
Collards 2
Corn, sweet (kernels plus 2
cobs with husks removed) 0.2b
Cottonseed 2
Cucumbers 2
Eggplants
Filberts
Goats, (meat, fat, meat
by-products)
Grapes
Hogs (meat, fat, meat
by-products)
Horses (meat, fat, meat
by-products)
Kale
Lettuce
Macadamia nuts
Melons
Milk fat
Mustard greens
Mustard seed
Nectarines
Oats, grain
Oats, straw
Peaches
Pears
Peas (succulent type)
Pecans
Peppers
Pineapples
2
0.21
2
2
0.2
b
O.lb
0.2b
0.2b
0.2
2
2
O.lb
0.5
2
2
0.2
24d
2
2
0.2b
2
O.lb
0.2b
Plums
Potatoes
Prunes
Pumpkins
Rapeseed
Rye grain
Rye straw
Safflower seed .
Sheep (meat, fat,
meat by-products)
Spinach
Strawberries
Sugar beets (with-
out tops)
Sugarcane
Summer squash
Sunflower seeds
Sweet potatoes
Tea, dried
Tomatoes
Turnip greens
Walnuts
Watercress
Wheat grain
Wheat straw
Winter squash
*Source: 40 CFR 180.182, 1977; 21 CFR 193.170, 1977
alncludes its metabolite, endosulfan sulfate
Negligible residue
cNegligible residue in milk
additive tolerance
-------
TABLE 2
Tolerances Reported by Other Countries*
Country
Commodity
Tolerances (mg/kg)
o
i
i—
to
Australia3
Canadab
New Zealand0
Netherlands0
South Africab
Fat of meat of cattle and sheep
Milk and milk products
Fruits, grain, vegetables, cottonseed
Peas
Artichokes, beans, cauliflower, celery, cucum-
cumber, eggplant, grapes, melons, peppers,
pumpkins, squash, strawberries, tomatoes,
watercress
Apples, apricots, broccoli, Brussel sprouts,
cabbage, cherries, lettuce, nectarines,
peaches, pears, plums, prunes, spinach
Fruits and vegetables
Berries, mushrooms
Fruit (except berries) and vegetables
Potatoes
Cabbage, green beans, boysenberries, young-
berries, tomatoes, cucurbits, peas, citrus
Peaches, apples, pears
*Source: WHO, 1975
alncludes c^ - and /£ -endosulfan and endosulfan sulfate
^Includes ct\ - and x^-endosulfan
cResidues not specified
0.2
0.5 (fat basis)
1.0
0.5
1.0
2.0
2.0
1.0
0.5
0.1
2.0
0.5
-------
Pesticide Residues, and thus is not an officially recognized stan-
dard in the United States. The ADI for endosulfan is 0.0075 mg/kg
(FAO, 1975).
Corneliussen (1970, 1972) reported the residue levels of sev-
eral chlorinated insecticides in various foods before and after
processing by a dietician. The effect of processing on residues
of endosulfan (includes the two isomers and sulfate) are reported
for only one food class, leafy vegetables. Corneliussen (1970)
reported the residues as 0.011 mg/kg before processing and 0.006
mg/kg after processing. Corneliussen (1972) reported the residues
as 0.007 and 0.002 mg/kg, respectively.
McCaskey and Liska (1967) studied the effect of processing on
the residues of endosulfan and endosulfan sulfate in milk. One
group of milk samples came from cows fed 500 to 2,000 mg/day endo-
sulfan for 7 to 11 days; the other group of milk samples contained
endosulfan which had been added in solution in ethyl alcohol. The
investigators were not able to detect endosulfan in the milk from
the cows fed endosulfan, but the milk did contain endosulfan sul-
fate. The residues were reported on a milk fat basis since mois-
ture was being removed from the milk during processing. The re-
sults are presented here.
C-13
-------
Residue (mg/kg, fat basis)
Endosulfan
Product Sulfate* Endosulfan**
Raw milk 15.2 15.9
Forewarmed milk 12.7 12.7
Condensed milk 12.6 11.4
Spray-dried milk 8.8 10.1
Evaporated milk 8.8 9.9
Drum-dried milk 4.5 8.0
* Detected in milk from cows administered endosulfan
**Added to milk in alcohol solution
Li, et al. (1970) reported a study in which two dairy cows
were given 1 mg/kg/day endosulfan for two weeks. Analyses of the
dairy products (pasteurized milk, cream, butter, cheese, dried and
condensed whole milk, etc.) indicated only a very small (not quan-
tified) concentration of <^-endosulfan. Endosulfan sulfate, how-
ever, was not detected.
Johnson, et al. (1975) studied the effects of freeze-drying
on the residues of endosulfan in tobacco. The treated samples
were analyzed for both endosulfan isomers and endosulfan sulfate.
The results are presented in Table 3.
The reduction in endosulfan residues amounted to 34 to 43
percent on a weight basis compared to the control samples. The
two types of freeze-drying had about the same effect; the percent
reduction in residue was about the same for both high and low in-
itial residue levels. The percent reduction was greater for
C& -endosulfan than for /^-endosulfan or endosulfan sulfate.
Beck, et al. (1966) conducted four cattle feeding tests for
the purpose of determining residues of endosulfan. In the four
tests endosulfan in the tissue, milk, alfalfa, grass, and silage
C-14
-------
TABLE 3
The Effect of Freeze-drying on Endosulfan Residues on Tobacco*
n
i
i — '
01
Initial
Pesticide Freeze-drying
Level Treatment
Low Control
Standard
Freeze-drying
Extraction +
f reeze-drying
High Control
Standard
Freeze-drying
Extraction +
f reeze-drying
-Endosulfan
0.12
0.05
0.05
0.25
0.11
0.10
Residues
-Endosulfan
0.98
0.56
0.59
2.35
1.27
1.29
(mg/kg)a
Endosulfan
Sulfate
2.43
1.59
1.68
7.65
4.45
4.94
Total
Endosulfan
3.53
2.20
2.32
10.24
5.83
6.33
*Source: Adapted from Johnson, et al. 1975
aThe analytical method was electron-capture gas chromatography
-------
was determined by colorimetric analysis. In one test, the inves-
tigators fed alfalfa treated with endosulfan to Hereford steers
and analyzed for residues of endosulfan in the omental fat. Two
steers were used in each experiment at treatment levels of 0.15,
1.10, 2.50, and 5.00 mg endosulfan/kg body weight/day. Two steers
(treatment levels 5 mg/kg/day and 2.5 mg/kg/day) developed muscle
convulsions (after 2 and 13 days, respectively); the experiments
at these levels were discontinued.
After 60 days, one of the steers receiving the 0.15 rag/kg
treatment showed no residues of endosulfan in the fat tissue, but
one of the steers receiving the 1.10 mg/kg treatment showed 1.0
mg/kg endosulfan in the fat tissue. Two other steers were also
fed 1.10 mg/kg of endosulfan in acetone solution twice daily, and
after seven days, urine and fecal samples were taken. The two
steers excreted endosulfan at the rate of 6.7 and 5.0 ing/day in
the feces and 18.5 and 15.9 mg/day in the urine. This rate of ex-
cretion accounted for only 7.4 and 4.9 percent of the daily dose
administered. Since most of the endosulfan was not excreted or
accumulated in the body fat, the investigators concluded that it
must have been metabolized.
Beck et al. (1966) grazed Hereford steers on Coastal Bermuda
grass that had been treated with endosulfan. No residues were
found in the fat of any of the steers which had been, grazing from
31 to 36 days on the treated grass. The levels of endosulfan in
the grass varied from 102 mg/kg (dry weight basis) on the first
day after treatment (when one test group began grazing) to 1.53
mg/kg on the day the last test group had completed grazing.
C-16
-------
Beck, et al. (1966) also fed groups of cows silage made from
Coastal Bermuda grass treated with endosulfan. The maximum resi-
due of endosulfan in the silage was 6.43 mg/kg (dry basis), which
appeared in the grass treated at 1 Ib Al/acre. There were no de-
tectable residues of endosulfan in the milk between the groups of
cows which received treated silage and a control group of cows
which received untreated silage.
McCaskey and Liska (1967) examined the effect of processing
on the residues of endosulfan in milk. The investigators were not
able to detect any endosulfan in the milk of cows fed up to 2,000
mg/day for 11 days. However, they did detect 0.6 ppm endosulfan
sulfate in a raw milk sample, but the investigators did not state
the treatment rate for the cow which produced that sample.
The Food and Drug Administration (FDA), Department of Health,
Education and Welfare, monitors pesticide residues in the nation's
food supply through two programs. One program, commonly known as
the "total diet" or "market basket" program involves the examina-
tion of food ready to be eaten. This investigation measures the
amount of pesticide chemicals found in a high consumption varied
diet. The samples are collected in retail markets and prepared
for consumption before analysis. The other program involves the
examination of large numbers of samples obtained when lots are
shipped in interstate commerce to determine compliance with toler-
ances. These analyses are complemented by observations and in-
vestigations in the growing areas to determine the actual prac-
tices being followed in the use of pesticide chemicals (Duggan, et
al. 1971).
C-17
-------
A majority of the samples collected in these programs are
categorized as "objective" samples. Objective samples are those
collected about which there is no suspicion of excessive residues
or misuse of the pesticide chemicals. All samples of imported
food and fish are categorized as objective samples even though
there could be reason to believe excessive residues may be found
on successive lots of these food categories.
Market basket samples for the total diet studies are pur-
chased bimonthly from retail stores in five regions of the United
States. A shopping guide totaling 117 foods for all regions is
used, but not all foods are represented in all regions because of
differences in regional dietary patterns. The food items are
separated into 12 classes of similar foods (e.g., dairy products;
meat, fish, and poultry; legume vegetables; and garden fruits) for
more reliable analysis and to minimize the dilution factor. Each
class in each sample is a "composite." The food items and the
proportion of each used in the study were developed in cooperation
with the USDA and represent the high consumption level of a 16- to
19-year-old male. Each sample represents a 2-week supply of food
(Duggan, et al. 1971).
Surveillance samples are generally collected at major har-
vesting and distribution centers throughout the United States and
are examined in 16 FDA district laboratories. Some samples may be
collected in the fields immediately prior to harvest. Surveil-
lance samples are not obtained in retail markets. Samples of im-
ported foods are collected as they enter the United States (Dug-
gan, et al. 1971).
C-18
-------
The results of these FDA testing programs are intermittently
published in Pesticides Monitoring Journal. Pesticide residues
are analyzed by multi-residue methods. The residues of endosulfan
(total of. c^ - and /^-isomers and sulfate) reported in the total
diet program are listed in Table 4. The average endosulfan resi-
dues in raw agricultural products are listed in Table 5. The
average incidence and daily intake of endosulfan based on these
data for a 6-year period are listed as follows (Duggan and Cor-
neliussen, 1972).
No. of Positive Daily
Year* Composites Examined
1965
1966
1967
1968
1969
1970
216
312
360
360
360
360
Composites (%) Intake (mg)
-
1.6
0.3
0.8
4.2
5.3
-
<0.001
<0.001
<0.001
0.001
0.001
*Annual test period is from June of previous year to April
of year listed.
A number of studies have been reported concerning the pres-
ence of endosulfan residues in tobacco and tobacco products. The
following paragraphs briefly summarize results from these
studies.
C-19
-------
TABLE 4
Endosulfan Residues in Total Diet Sample*
Class of Food
Number of Composites
Containing Endosulfan
Amount (mg/kg)
Time Period of Study
and Source
n
i
to
o
Leafy vegetables
Garden fruits
Fruits
Leafy vegetables
Leafy vegetables
Garden fruits
Oils, fats, and shortening
Leafy vegetables
Potatoes
Garden fruits
Fruits
Leafy vegetables
Garden fruits
Fruits
Oils, fats, and shortening
Leafy vegetables
Potatoes
Garden fruits
Fruits
Fruits
Potatoes
Leafy vegetables
Garden fruits
1
3
1
1
1
1
8
2**
4
7
5
3
1
15
2
2
5
6
1
7
6
0.016
<0.001, 0.002, 0.006
0.014
0.003
0.014
0.008
0.0134
<0.001-0.042
0.004-0.011
<0.001, 0.001, 0.002,
0.007
0.002, 0.010
<0.001-0.040
0.001-0.005
0.008-0.010
0.185
<0.001-0.063
<0s001f- 0,007
<0.001, 0.061
<0.001-0.045
<0.001-0.006
<0.001
<0.001-0.008
<0.001
June 1965-April 1966
Duggan, et al. (1967)
June 1966-April 1967
Martin & Duggan (1968)
June 1967-April 1968
Corneliussen (1969)
June 1968-April 1969
Corneliussen (1970)
June 1969-April 1970
Corneliussen (1972)
June 1970-April 1971
Manske & Corneliussen (1974)
June 1971-July 1972
Manske & Johnson (1975)
-------
TABLE 4 (Continued)
Class of Food
Number of Composites
Containing Endosulfan
Amount (mg/kg)
Time Period of Study
and Source
n
i
NJ
Potatoes
Leafy vegetables
Garden fruits
Fruits
Potatoes
Leafy vegetables
Garden fruits
Leafy vegetables
Garden fruits
4
17
4
4
6
3
3
5
2
<0.001-0.015
<0.001-0.439
<0.001-0.002
<0.001-0.007
<0.001-0.016
<0.001-0.012
<0.001
<0.001-0.022
<0.001-0.006
August 1972-July 1973
Johnson & Manske (1976)
August 1973-July 1974
Manske & Johnson (1977)
August 1974-July 1975
Johnson & Manske (1977)
**
Total endosulfan
Endosulfan sulfate only
- , and sulfate)
-------
TABLE 5
Average Endosulfan3 Residues in Raw Agricultural Products
During 5-year Study (1964-1969)*
0
1
to
M
Domestic
Average
No. of Incidence Residue No. of
Class of Food Samples % (mg/kg) Samples
Large fruits 6,763 0.8 <0.001 2,495
Small fruits 2,695 2.0 <0.001 496
Leafy and stem 13,864 4.9 0.01 153
vegetables
Vine and ear 8,072 1.4 <0.001 1,791
vegetables
Imported
Incidence
%
0.4
2.4
4.0
6.7
Average
Residue
(mg/kg)
<0.001
<0.001
0.03
<0.001
Source: Duggan, et al. 1971
aTotal includes
-------
Borough and Gibson (1972) reported the residue levels of
- and //-endosulfan and endosulfan sulfate in three brands of
cigarettes purchased in the years 1970 to 1972. The residues were
determined by gas chromatography; this method has a detection
limit of 0.01 mg/kg. In 1970 and 1971 the residues were all below
the detection limit. The results for 1972 were as follows:
Endosulfan Residue (mg/kg)
Brand
Regular A
Regular B
Filter B
Filter C
Menthol C
Average
C*.
0.01
0.01
0.01
0.01
0.01
0.01
X
0.12
0.14
0.10
0.09
0.10
0.11
Sulfate
0.27
0.30
0.21
0.25
0.30
0.26
Total
0.40
0.45
0.32
0.35
0.41
0.38
Domanski and Sheets (1973) reported the levels of endosulfan
residues (total for ^ - and /7-endosulfan plus endosulfan sulfate)
in several varieties of 1970 U.S. auction market tobacco. The
results are presented in Table 6.
Endosulfan residues on various U.S. tobacco products were
reported by Domanski, et al. (1973) for 1971 products and by Do-
manski, et al. (1974) for 1973 products. Much of the tobacco for
the 1971 cigarette samples had been in storage for two or more
years. The results are presented in Table 7.
C-23
-------
TABLE 6
Endosulfan Residues, U.S. Auction Market Tobacco (1970)*
o
i
Type
Flue-cured
Burley
Dark air-cured
Light air-cured
Dark fire-cured
Location
Tobacco Belt
Georgia- Florid a
North-South
Carolina border
Eastern
Middle
Old
States
North Carolina
Tennessee
Kentucky
Tennessee
Kentucky
Maryland
Tennessee
Kentucky
Virginia
Total Endosulfan
Range
<0. 2-11.1
0.2-21.9
<0. 2-5.0
<0.2-4.5
<0.2-2.7
<0.2-3
<0.2
1.4-14.3
0.3-12.5
5.8-13.6
<0.2-3.3
1.4-4.6
2.8-11.9
0.4-6.5
Residue average
(mg/kg)a
3.6
3.9
1.5
1.0
0.7
0.1
<0.2
8.6
5.7
8.5
1.5
3.2
6.0
3.3
'Source: Adapted from Domanski and Sheets, 1973
aTotal of c^ ~ and ^-endosulfan and endosulfan sulfate; the analytical method was
electron-capture gas chromatography
-------
TABLE 7
Endosulfan Residues on U.S. Tobacco Products (1971 and 1973)
n
I
NJ
tn
Product
Cigarettes
Cigars
Little cigars
Smoking or pipe tobacco
Chewing tobacco
Snuff
Total Endosulfan Residues (mg/kg)*
Range (average) __
1971**
<0.2-0.4 (0.2)
<0.2-1.1 (0.4)
0.3-0.5 (0.4)
<0.2-0.2 (<0.2
<0.2-0.5 (0.2)
<0.2 «0.2)
1973***
0.36-1.27 (0.83)
0.03-1.03 (0.37)
0.15-0.26 (0.22)
0.08-0.61 (0.37)
0.06-0.86 (0.36)
0.06-0.17 (0.12)
* The analytical method was electron-capture gas chromatography
** Source: Domanski, et al. 1973
***Source: Domanski, et al. 1974
-------
Domanski and Guthrie (1974) reported endosulfan residue
levels (total fortA- and/^-endosulfan plus endosulfan sulfate and
several other insecticides) in six brands of cigars purchased in
1972. The residues were determined by gas chromatography. The
results for endosulfan were as follows:
Brand Total Endosulfan Residues (mg/kg)
1 0.64
2 0.26
3 0.63
4 0.36
5 0.49
6 <0.20
Average 0.41
Gibson, et al. (1974) reported residues of endosulfan in Ken-
tucky Burley tobacco for the years 1963 to 1972. The residues for
endosulfan included the two isomers and the sulfate. Endosulfan
residues were not detected until 1968. The residues in tobacco
from auction warehouses in Kentucky and residues in the Kentucky
Burley tobacco pool were as follows:
From Auction Warehouse in In Kentucky Burley
Kentucky ^__ Tobacco Pool
Year Residue (mg/kg )" Residue (mg/kg)~
1968 0.23 Not reported
1969 0.30 0.86
1970 4.19 2.68
1971 4.60 Not reported
1972 4.10 Not reported
Thorstenson and Dorough (1976) reported residue levels of
C/ - and /^-endosulfan and endosulfan sulfate in "reference" and
"alkaloid" cigarettes prepared by the Tobacco and Health Research
Institute for the years 1969 and 1974. The "reference" cigarette
C-26
-------
is a composite which reflects a blend of an "average" domestic un-
filtered cigarette; the "alkaloid" cigarettes were composites
which contained blends of low-nicotine Burley and flue-cured
tobaccos. There were not detectable residues of endosulfan in the
1969 samples. The range and average endosulfan residues in the
1974 samples, which consisted of one alkaloid and three reference
cigarettes, were as follows:
Residue (mg/kg)
Compound Range Average
^ -Endosulfan 0 0
^-Endosulfan 0.4-0.7 0.5
Endosulfan sulfate 0.4-1.1 0.7
Total endosulfan 0.8-1.5 1.2
Schimmel, et al. (1977) reported on both short- and long-term
exposures of marine species to endosulfan. Pink shrimp, Penaeus
duorarum, did not show any uptake at all when exposed to 0.089
ug/1 endosulfan for 96 hours, while grass shrimp, Palaemonetes
vulgaris, had 96-hour bioconcentration factors ranging from 164 at
0.40 ug/1 (the highest concentration with 0 percent mortality) to
245 at 1.75 ug/1 (65 percent mortality). Maximum bioconcentration
factors after 96 hours for marine fish were 1,299 for pinfish,
Lagodon rhomboides, 895 for spot, Leiostomus xanthurus, and 1,344
for striped mullet, Mugil cephalus) . The mullet was also used in
a long-term exposure test for 28 days followed by 28 days in clean
water. After exposure to 0.035 ug/1 endosulfan for 28 days, the
bioconcentration factors were 2,429 for edible tissue and 2,755
for whole body. After two days in clean water, endosulfan was not
detected (limits of detection = 0.01 ug/g in tissues).
C-27
-------
The investigators noted that in all exposure tests endosulfan sul-
fate was the predominant and often sole form of endosulfan found
in the tissues.
Roberts (1972) studied the accumulation of endosulfan in com-
mon mussels, Mytilus edulis, when exposed to levels of 0.1, 0.5,
and 1.0 mg/1 endosulfan in seawater. The concentration factors
determined from these tests were as follows:
Endosulfan Exposure Period (days)
Concentrations \A_ 2_7 4_2 56_ 70_ 8j> 100 112
(mg/1)
Bioconcentration Factors
0.1 13.0 17.0 13.5 19.3 22.5 16.1 17.0 17.0
0.5 4.7 5.8 4.9 8.3 6.9 7.0 7.8 11.0
1.0 2.8 3.3 3.7 3.9 6.5 7.4 7.1 8.1
Roberts (1972) also reported a rapid fall in tissue residue levels
(1 to 2 mg/kg for all three exposure levels) within 58 days of re-
moval from endosulfan-containing waters.
In further studies, Roberts (1975) investigated the differen-
tial uptake of endosulfan by tissue of M. edulis. Eighty mussels
approximately 60 mm (2.4 in.) in length were exposed to 0.1 mg en-
dosulfan/1 in slowly flowing seawater for 36 days, then trans-
ferred to clean seawater for a further period of 23 days. Weekly
samples of six mussels were taken for determination of endosulfan
residues in the digestive gland, the mantle plus gonad, the gills,
and the remaining tissue, consisting of pedal retractor muscles,
foot, and anterior and posterior adductor muscles.
Results showed that the major site of concentration of endo-
sulfan is the digestive gland. The approximate maximum endosulfan
C-28
-------
residues found and the times at which they occurred (in number of
days after initial exposure) were as follows, expressed as ug
endosulfan (both isomers) per g wet weight:
Digestive gland 6.1 after 7 days
Mantle and gonad 1.8 after 36 days
Gills 2.1 after 15 days
Remaining tissue 2.3 after 36 days
Mean residue level 2.5 after 36 days
Upon removal of the mussels to clean seawater, the endosulfan
residue levels initially declined fairly rapidly in all tissues;
the greatest decline occurred in the digestive gland during the
first 14 days of elution. During the final six days of elution,
the rate of residue loss was similar for all tissues.
Ernst (1977) also evaluated the bioconcentration of endosul-
fan in M. edulis in static tests. The inital concentration of
endosulfan was 2.05 ug/1, and it reached a steady-state concentra-
tion of 0.14 ug/1. The concentration factor for tf -endosulfan
calculated from the tissue levels of the steady-state concentra-
tion in the water was 600. The half-life for elimination of the
residue was calculated to be 33.8 hours.
Roberts (1975) also conducted endosulfan uptake studies with
the scallop, Chlamys opercularis. In this species, endosulfan
concentrations in the digestive gland and in the foot, and ante-
rior and posterior adductor muscles were similar to those seen in
M. edulis. However, the endosulfan level in the gills of M.
edulis was almost five times that in gills of £. opercularis,
while the reverse was true in the gonad and mantle tissues. The
mean tissue residue levels for both species, estimated from the
summated values for the separate tissues, were very similar
despite the difference in distribution between tissues.
C-29
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Gorbach (1972) referred to an experiment in which goldfish,
Carassius auratus, were exposed for five days to 1 ug/1 14C-
labeled endosulfan in water. The fish attained endosulfan
concentrations of 0.4 ug/g or a bioconcentration factor of 400.
It was also stated that the parent compound as well as all
metabolites were excreted within 14 days when the fish were
transferred to fresh water.
Oeser, et al. (1971) held goldfish for five days in a test
solution containing mean residues of 350 ug/1 14C-labeled en-
dosulfan. The average ratio of body residues to skin residues of
205:1 indicated that most of the endosulfan was in the fish body,
not the skin. After 14 days in fresh water, the test fish had ex-
creted 96 percent of the radioactivity that had been absorbed in
the test solution.
Investigations by Schoettger (1970) with 14C-labeled en-
dosulfan indicated the compound is taken up and deposited in var-
ious tissues of fish at varying rates. The liver and gut (with
feces) contained the most pesticide whereas the heart, blood,
gill, kidney, and brain showed slower uptake rates; less was found
in gut (empty), skin, and muscle. The investigator noted that in
general those tissues containing relatively large amounts of blood
contained the higher concentrations of residues, with the excep-
tion of the gut and feces. Radiotracer and chemical analysis
techniques showed a water-soluble metabolite of endosulfan in the
bile of western white suckers (Catostomus commersoni), northern
creek chubs (Semotilus atromaculatus) , and goldfish., Schoettger
C-30
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(1970) suggests that endosulfan degrades to its alcohol, which is
then conjugated with glucuronic acid and excreted with the feces.
A bioconcentration factor (BCF) relates the concentration of
a chemical in aquatic animals to the concentration in the water in
which they live. The steady-state BCF for .a lipid-soluble com-
pound in the tissues of various aquatic animals seem to be propor-
tional to the percent lipid in the tissue. Thus, the per capita
ingestion of a lipid-soluble chemical can be estimated from the
per capita consumption of fish and shellfish, the weighted average
percent lipids of consumed fish and shellfish, and a steady-state
BCF for the chemical.
Data from a recent survey on fish and shellfish consumption
in the United States were analayzed by SRI International (U.S.
EPA, 1980). These data were used to estimate that the per capita
consumption of freshwater and estuarine fish and shellfish in the
United States is 6.5 g/day (Stephan, 1980). In addition, these
data were used with data on the fat content of the edible portion
of the same species to estimate that the weighted average percent
lipids for consumed freshwater and estuarine fish and shellfish is
3.0 percent.
One laboratory study, in which percent lipids and a steady-
state BCF were measured, has been conducted on endosulfan. The
BCF value, after normalization to 1 percent lipids, is 91.1 (see
Table 5 in Aquatic Life Toxicology, Section B) . An adjustment
factor of 3 can be used to adjust the normalized BCF to the 3.0
percent lipids that is the weighted average for consumed fish and
shellfish. Thus, the weighted average bioconcentration factor for
C-31
-------
endosulfan and the edible portion of all freshwater and estuarine
aquatic organisms consumed by Americans is calculated to be 270.
Inhalation
According to the American Conference of Governmental Indus-
trial Hygienists (ACGIH, 1977), the Threshold Limit Value-Time
Weighted Average (TLV-TWA) for endosulfan is 0.1 mg/m3. The
tentative value for the Threshold Limit Value-Short Term Exposure
Limit (TLV-STEL) is 0.3 mg/m3. The TLV-TWA is based on a normal
8-hour workday or 40-hour workweek, day-after-day exposure. The
TLV-STEL is the maximum concentration to which a worker may be
exposed continuously for as long as 15 minutes without irritation,
chronic or irreversible tissue changes, or narcosis sufficient to
increase the inclination to accident or to affect self-rescue or
work efficiency. Up to four such exposures may occur per day pro-
vided at least 60 minutes elapse between the exposures and pro-
vided the TLV-TWA is not exceeded in the time lapses.
Apparently neither Occupational Safety and Health Administra-
tion (OSHA) exposure limits nor National Institute for Occupa-
tional Safety and Health (NIOSH) recommended exposure limits have
been established for endosulfan (NIOSH, 1978). Further, a recent
international comparison of hygienic standards for chemicals in
the work environment did not list standards for endosulfan
(Winell, 1975).
Lee (1976) summarized the results of intensive ambient air
sampling at selected locations over the nation in which samples
C-32
-------
were analyzed for pesticide residues. Samples were collected dur-
ing 1970, 1971, and 1972. The results of these tests for endosul-
fan-containing samples are given in Table 8.
Wolfe, et al. (1972) evaluated potential respiratory exposure
for a number of pesticides. Tests were conducted by sampling
spraymen operating tractor-drawn power air-blast equipment as they
applied pesticides to fruit orchards. Endosulfan was applied as a
0.08 percent spray. The estimated respiratory exposure was 0.01
to 0.05 mg/hour (average 0.02 mg/hour).
Exposure to endosulfan was found by respirator pad analysis
to be greater during mixing operations than in spraying opera-
tions. With a 5-minute exposure time, 182,800 ng of endosulfan
were detected on the respirator pad during a mixing operation;
only 4,664 ng were detected during a 30-minute spraying operation
(Oudbier, et al. 1974).
Tessari and Spencer (1971) analyzed air samples from human
environments for pesticide residues. Nylon screens were placed
inside and outside the homes of 12 men occupationally exposed to
pesticides, including endosulfan, for a period of one year, five
days each month. Endosulfan residues were found in 13 of 52 in-
door air samples from the formulators1 households. In the posi-
tive samples, endosulfan residues ranged from 0.22 to 4.52 ug/m2
of filter; the mean of the positive samples was 1.77 ug/m2. The
positive samples came from only two households, and the house-
holders in both cases were formulators who had handled endosulfan.
C-33
-------
TABLE 8
from 16 States*
O
I
Co
Rnmm?rv «f Endosulfan Residues in Air adm^es I.LUIH o.« <>..<,..» _
1970
Concentration (hg/m-*)
No. of
State Residue Name Samples
Arkansas
Illinois
Kansas
Kentucky
Montana
North Carolina
All 16 states
-Endosulfan
-Endosulfan
-Endosulf an
-Endosulfan
-Endosulfan
-Endosulfan
-Endosulfan
-Endosulfan
-Endosulfan
72
53
64
68
48
54
-
Positive
Samples Arithmetic
(«) Mean
6.94
11.11
7.55
12.50
32.35
16.67
9.26
6.61
1.02
1.1
2.4
2.2
5.5
159.4
13,9
0.7
13.0
0.2
1972
Mean of positive
Positive Maximum No. of Samples
Samples Value Samples (%)
15.5
22.0
28.8
43.8
492.8 2
83.5
7.2
111.9 2
22.0
27.1 60 ND
54.5 ND
39.5 36 ND
70.7 49 ND
,256.5 43 ND
211.7 36 ND
10.9 41 ND
,256.5 - ND
54.5 ND
1971
Concentration (ng/m3)
Mean of
Arithmetic Positive Maximum
Mean Samples Value
ND ND ND
ND ND ND
ND ND ND
ND ND ND
ND ND ND
ND ND ND
ND ND ND
ND ND ND
ND ND ND
Concentration (ng/m-1)
Arkansas
Illinois
Kansas
Kentucky
Montana
North Carolina
All 16 states
-Endosulfan
-Endosulfan
-Endosulfan
-Endosulfan
-Endosulfan
-Endosulfan
-Endosulfan
-Endosulfan
-Endosulfan
64
59
65
66
69
64
-
ND
ND
ND
ND
ND
ND
ND
ND
NU
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
*Source: Lee, 1976
-------
No endosulfan residues were found in the outdoor air near any of
the formulators1 households, or in the indoor or outdoor air at
the farmers' households.
The National Research Council of Canada (NRCC, 1975) reported
unpublished data (Boelens and Frank, 1973) on endosulfan spray
drift from an aerial spray application of endosulfan to a tobacco
field in Norfold County, Ontario, Canada (endosulfan formulation
and rate of application not given in secondary source.). Residues
in various parts of the field were determined based on levels de-
tected in pans filled with water. Endosulfan levels detected in
the water within the field were 55.0 mg/1 between tobacco rows;
8.5 mg/1 under plants in the rows; 20.0 mg/1 in an opening in the
field; and 0.01 mg/1 at the edges of the field. No detectable
(detection limit not reported) endosulfan residues were found at
the edge of the field at the soil surface. Water in an adjacent
stream contained endosulfan residues ranging from traces to 0.22
mg/1. Sediment from the stream contained 2.7 mg/kg of endosulfan
in a sample collected opposite the field's drainage and 0.23 and
0.37 mg/kg in two other samples collected nearby. Aquatic mono-
cotyledonous plants contained 0.01 mg/kg.
Keil, et al. (1972) observed endosulfan spray drift in a
field test on tobacco in South Carolina. Three treatments of en-
dosulfan (formulation and AI content not given) at the rate of 0.5
Ib Al/acre per treatment were applied by ground equipment to
single-row (12-ft) plots separated by guard rows. Each treatment
or control plot was replicated four times in a completely ran-
domized design. Samples of tobacco foliage were collected for
C-35
-------
residue analysis at 11 post-treatment intervals ranging from one
day after the first application to 18 days after the third.
Even though the experimental design included guard rows, en-
dosulfan residues ranging from 0.037 to 0.679 mg/kg resulting from
spray drift were found on plots treated with another insecticide,
and on untreated control plots. However, in most instances (18 to
22 samples from plots not treated with endosulfan), the residue
from drift was less than the least significant difference at the
95 percent probability level, 0.363 mg/kg endosulfan.
Dermal
The 1977 listing of TLV values showed a "skin" designation
for endosulfan (ACGIH, 1977). This designation refers to the po-
tential contribution to the overall exposure by the cutaneous
route including mucous membranes and eyes, either by airborne en-
dosulfan or by direct contact with it.
Wolfe, et al. (1972) also evaluated potential dermal exposure
of spraymen applying a 0.08 percent endosulfan spray. The esti-
mated dermal exposure was 0.6 to 95.3 mg/hour (average 24.7
mg/hour).
Possible intoxication due to the dermal exposure was sug-
gested by Kazen, et al. (1974) who analyzed hexane hand rinsings
and found that endosulfan persisted on exposed workers' hands for
1 to 112 days after exposure.
PKARMACOKINETICS
Absorption
Undiluted endosulfan is slowly and incompletely absorbed in
the mammalian gastrointestinal tract (Maier-Bode, 1963). However,
C-36
-------
when endosulfan is dissolved in a carrier vehicle such as cotton-
seed oil, the oil and the insecticide are readily, though not com-
pletely, absorbed by rats (Boyd and Dobos, 1969) and other mammals
(Maier-Bode, 1968). The /^-isomer is more readily absorbed than
the (T^-isomer (Demeter, et al. 1977).
Alcohols, oils, and emulsifiers also accelerate the absorp-
tion of endosulfan by the skin (Maier-Bode, 1968).
Inhalation is not considered to be an important route of up-
take of endosulfan because of its low vapor pressure (9 x 10""^
mm Hg) (Maier-Bode, 1968).
When endosulfan is dissolved in chloroform and painted on the
shaven skin of rabbits, it is readily absorbed (Gupta and Chandra,
1975).
A 1:1,000 dilution of endosulfan instilled on the conjunctiva
of rabbits' eyes caused neither pain nor subsequent inflammation,
which was apparently because of rapid removal by the lacrimal
fluid (Hoechst, 1967a) .
Distribution
After ingestion, endosulfan is first distributed to the liver
and then to the following organs: brain, heart, kidneys, lungs,
spleen, testes, thymus gland, suprarenal glands, mammary glands,
skeletal muscles, and the remainder of the gastrointestinal tract
(Boyd and Dobos, 1969; Maier-Bode, 1968).
Two reports of individuals committing suicide by ingesting
endosulfan present some data regarding the distribution of endo-
sulfan in man. Demeter, et al. (1977) report on one victim who
ingested a preparation containing 12.4 percent - and 8.1 percent
C-37
-------
^-endosulfan. The order of distribution was as follows: stomach
contents > small intestine contents > liver > kidneys > urine >
blood.
Table 9 summarizes the data reported by Coutselinis, et al.
(1978) from three suicide cases.
Metabolism and Excretion
The metabolism of endosulfan in mammalian species has been
widely investigated. The generalized metabolic pathway for endo-
sulfan in animals is given in Figure 1.
Demeter and Heyndrickx (1978) have detected endosulfan sul-
fate as a metabolite in humans by analysis of two human post-
mortem cases. Both were male, and both had taken a 20 percent en-
dosulfan product by mouth within hours of death, in one case under
three hours. Endosulfan sulfate was not detectable in blood or
urine but was present in liver (3.4 mg/kg average), brain (0.5
mg/kg), and kidney tissue (0.4 mg/kg).
No other information was found regarding the metabolism of
endosulfan (or endosulfan sulfate) in humans.
In a review by Matsumura (1975) a pathway for metabolism in
rats, mice, and insects was presented which differed somewhat from
that given by Knowles shown in Figure 1. Matsumura did not show
the transformation of the ether to the hydroxyether but indicated
the hydroxyether was formed directly from either the diol or the
lactone (Matsumura, 1975).
The results of a study using 14C-labeled endosulfan indi-
cated the sulfate to be the metabolite most commonly present in
organs, tissues, and feces of rats whether dosed with the
-------
TABLE 9
Concentration Levels of Endosulfan in Biological Material*
Case
1
2
3
Blood
(mg/100 ml)
0.4
0.8
0.7
Liver
(mg/100 g)
0.08
0.1
0.14
Kidney
(mg/100 g)
0.24
0.32
0.28
Brain
(mg/100
0.025
0.03
0.028
g)
C-39
-------
CH2—
CH2—O'
SO,
Cl
Endosulfan ether
Endosulfan lactone
FIGURE 1
Metabolism of Endosulfan in Animals
Source: Knowles, 1974; Menzie, 1974
C-40
-------
/^-isomer (Whitacre, 1970). The feces .also contained large
amounts of unchanged endosulfan. Endosulfan diol, <^\ -hydroxy-
ether, and lactone were recovered from both urine and feces of
rats fed either endosulfan isomer.
When the rats were administered the diol and the -hydroxy-
ether, both were partially transformed to the lactone and excreted
in urine. The diol was also transformed to hydroxyether in the
small intestine and in feces.
Feces usually had the highest radioactivity and must be con-
sidered to be the principal route of elimination in the rat.
The metabolism of 14C-labeled endosulfan was also studied
in BALB/C strain mice by Deema, et al. (1966). The 14c-
labeled endosulfan used was labeled in the hexachlorocyclodiene
ring at carbons 5 and 6. The compound was composed of 58.3 per-
cent ^-endosulfan and 35.6 percent ^endosulfan, 6.0 percent of
the ether, and 1.0 percent of the alcohol.
Two male mice and four female mice were studied with groups
of two each being fed 0.30, 0.25, or 0.20 mg labeled compound in
300 mg food. After 24 hours the amount of the labeled compound
in 1 g of organ or excreta was greatest in feces (98,452 cpm) ,
followed by visceral fat (7,053 cpm), urine (3,746 cpm), liver
(2,883 cpm), kidney (1,390 cpm), brain (424, cpm), respired C02
(302 cpm), and blood (92 cpm). Total recovery of the labeled
endosulfan was approximately 65 percent.
Purified unlabeled endosulfan was also fed at 0.3 mg/mouse in
a 300 mg food pellet. After 24 hours large amounts of endosulfan
sulfate were found in the liver, small intestine, and visceral fat
C-41
-------
with a trace in skeletal muscle and kidney. The endosulfan was
found only in the stomach, small intestine, and feces. By chroma-
tography, a metabolite that appeared to be identical with endo-
sulfan alcohol was detected in urine.
When mice were fed only the -isomer of endosulfan, the
material was detected in the stomach, small intestine, and feces
although endosulfan sulfate was detected in the liver, small in-
testine, visceral fat, and feces. Endosulfan alcohol was found in
urine. Neither the parent compound nor any of its metabolites
were detected in the brain.
When the ^-isomer was fed, endosulfan sulfate was found in
the liver, kidney, small intestine, muscle, and visceral fat. The
alcohol was detected in the urine, but neither the x^-isomer nor
any metabolites were detected in the brain.
When 10 mice were fed diets containing 10 mg/kg purified en-
dosulfan for 28 days, endosulfan sulfate was detected in the liver
and visceral fat of all animals although lower amounts were de-
tected than in organs of other test mice 24 hours after they had
been fed a single 0.3 mg dose. Endosulfan isomers or metabolic
products were not detected in the brain, but a product having the
same retention time as endosulfan alcohol was detected in the
urine. When the feces were analyzed, both isomers, endosulfan
sulfate, endosulfan alcohol, and endosulfan ether were detected.
Endosulfan alcohol was detected in the urine of animals fed
either endosulfan sulfate, endosulfan ether or endosulfan diol.
C-42
-------
The principal metabolic products produced in the mouse under
the conditions of this study were endosulfan sulfate and endosul-
fan alcohol (Deema, et al. 1966).
Dogs (unspecified breed or number of each sex) were admin-
istered^- and ^-endosulfan for 28 days at 0.35 and 1.75 mg/kg/
day (FMC, 1963). Upon analysis only traces of cA - and ^-endosul-
fan were detected in the urine (0.02 to 0.1 ppm), but large
amounts (13 to 25 percent of the endosulfan fed) were detected in
the feces.
In a study with two East Frisian milk sheep, radiolabeled
endosulfan (65 percent ch and 35 percent/^) that was administered
as a single oral dose of 0.3 mg/kg was almost entirely eliminated
in 22 days (Gorbach, et al. 1968). About 50 percent of the radio-
label was excreted in the feces, 41 percent in the urine, and 1
percent in the milk. On the 22nd day the level in the milk was 2
ug/l.
The highest blood concentrations of radiolabel were reached
after 24 hours (4.3 to 4.5 x 10~4 percent of administered
activity) .
The maximum elimination in feces was observed on the second
day (20.8 and 18.6 percent of administered dose). The unchanged
isomers were detected in the feces. The lactone, diol, and hy-
droxyether of endosulfan were not detected in the feces.
Radioactivity peaked in urine in the first 24 hours (18.5
percent of the dose) and then decreased. Two metabolites were
detected in urine, one characteristic of endosulfan alcohol and
the other characteristic of the hydroxyether. Of the activity, 70
C-43
-------
percent was present in the alcohol and 30 percent in the hydroxy-
ether.
After 40 days the organ with the highest concentration of
radiolabel (0.03 ug/g) was the liver.
The investigators noted that no fat-soluble metabolite other
than endosulfan sulfate was detected in the milk of the test ani-
mals and that no major metabolite was retained in fat or in the
organs for long periods of time (Gorbach, et al. 1968) .
In another study, between 0.1 and 0.2 mg/1 endosulfan sulfate
was detected in the milk of cows that had been given 2.5 mg/kg
ch -endosulfan, 2.5 mg/kg x^-endosulfan, and 5 mg/kg endosulfan
sulfate in the feed for 30 days (FMC, 1965). Less than 0.005
mg/1 endosulfan sulfate was detected in the milk 20 days after
administration of the insecticide was stopped.
The half-life of endosulfan in the milk of cows that survived
poisoning was reported to be 3.9 days (Braun and Lobb, 1976).
These residues were present primarily as endosulfan sulfate.
The endosulfan isomers were detectable in milk for six days in one
animal and 13 days in another, with a detection limit of 0.001
mg/1. Endosulfan sulfate residues were detected for 35 days in
both animals. Blood contained detectable amounts of the sulfate
metabolite (0.025 mg/1) for one day after exposure. Parent
isomers were not found in blood.
In sheep given single oral doses of 14c-labeled endosul-
fan at 14 mg/kg, the half-life of radiolabeled endosulfan in the
feces and urine of sheep was reported to be about two days (Kloss,
et al. 1966) .
C-44
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Dorough, et al. (1978) studied the fate of endosulfan in fe-
male rats given the insecticide by esophageal intubation. Five
days after a single radiolabeled dose, 88 percent of the ^-isomer
and 87 percent of the /^isomer were recovered in the urine (13
percent) and the feces (75 percent). Two days after a single dose
was given to rats with cannulated bile ducts, 47 percent of the
^5-isomer and 29 percent of the ^- isomer were secreted in bile.
After another group of these rats had eaten diets containing
endosulfan for 14 days, the half-life of the residues was deter-
mined to be approximately seven days.
The last group of rats was fed 5 mg/kg endosulfan metabolites
(the sulfate, diol, c^-hydroxyether, lactone, and ether deriva-
tives) for 14 days. The organs containing the greatest amounts of
endosulfan derivatives were the kidneys (1 ug/g) and the liver (3
ug/g).
EFFECTS
Acute, Subacute, and Chronic Toxicity
Values for the LD50 of technical endosulfan (an 2:1
mixture of
-------
TABLE 10
Acute Toxicity of Endosulfan
Tesl animal
(sex) (strain)
Rat (-) (S(irague-Dawley)
Rat, H (Sliennjn)
Rat, F (Sherman)
Rat (-) (-)
Rat, M (Wlstar)
Rat (-) (-)
Rat (-) (-)
K.lt, M (-)
Rat, M (-)
O Rat, F (-)
1 Rat. F (-)
£ Rat (-) (-)
Rat (-) (-)
Rat, M (Sherman)
Rat, F (Sherman)
Endosul fan or
formulation
Purified
Technical
Technical
Technical
Technical
a-endosul fan
p-endosulfan
Technical
Technical
Technical
Teclinlcal
HOE 2671 (201 AI)
IIOE 2671 (sol.
powder)
Technical
Teclinlcal
Solvent (carrier)
Corn oil
Peanut ol 1
Peanut oil
—
"
~
"
Alcohol
101 alcohol In peanut oil
Alcohol
1O1 alcohol In peanut oil
Alcohol
Alcohol
Xylenc
Xylene
Number of anlmala
per test group
60 total
70 total
16/treatment group
16/ treatment group
16/treatment group
16/treatment group
6/ treatment group
4/treatment group
60 total
70 total
Route of
administration
Ora
Ora
Ora
Ora
Ora
Ora
Intraperltnneal
Intraperltoneal
Intraperitoiieal
Intraperltoneal
Intraperltoneal
Intraperltoneal
Dermal
Dermal
U>50 (mg Al/kg)*/
40-50
43 (41-40)
18 (15-21)
35
121 (+ 16)
76 mg/kg
240 mg/kg
46.7 (36.4-51.8)
89.4 (73-107.4)
22.1 (18.6-26.9)
48.6 (36.4-51.8)
8 (6.1-10.1)
13.5 (9.5-19.3)
130 (104-163)
74 (58-94)
Source
Llndi|ulst and Dahm (1957)
Galnes (1969)
Gal lies (1969)
Junes et at. (1968)
Boyd and Dobos (1969)
Hoechsl (I967b)
llo.-clisi (I9b>b)
Gupta (1976)
Gupta (1976)
Gupta (1976)
Gupta (1976)
Leiidle (Iui6)
Lendle (1956)
Guinea (1969)
GaJnes (1969)
Rat, H (-)
Mouse, H (-)
Mouse, M (-)
House, f (-)
Mouse, f (-)
Rabbit, F (Albino)
Rabbit, F (Albino)
ThloJan®
Teclinlcal
Teclinlcal
Technical
Technical
Technical (90%)
Technical (> 911)
Inhalation, 4 hours 350 mg/m3
Alcohol
101 alcohol In peanut oil
Alcohol
IOX alcohol in peanut oil
Chloroform
Chloroform
16/treatment group
16/trrutment group
16/treatment group
16/treatmeut group
',/treatment group
4/trQatmcnt group
Intraperttoneal
Intraperltoneal
Intraperltoneal
Intraiierttoneal
Dermal
Dennal
6.9 (5.4-8.9)
12.6 (9.4-16.8)
7.5 (5.3-10.1)
13.5 (10.6-16.8)
182 (+ 36)
167 (T 21)
Ely et al. (1967)
Gupta (1976)
Gupla (1976)
Gupta (1976)
Gupta (1976)
Gupta and Gh.-milra (19/5)
Gupta fiii'l Chandra (19/5)
o/ AI - Active ingredient.
-------
Some difference in toxicity occurs whenever different vehi-
cles are used as the carrier. Lendle (1956) quoted an LD50 of
only 8 mg/kg when endosulfan was dissolved in ethyl or isopropyl
alcohol and given intraperitoneally to rats, but similar animals
treated with endosulfan in cottonseed oil have an LD50 as high
as 48.6 mg/kg.
In another study (Gupta, 1976), male rats given endosulfan in
alcohol exhibited an LD50 at 46.7 mg/kg, but similar males
given the material in 10 percent alcohol in peanut oil exhibited
an LD50 at 89.4 mg/kg. While the amount of endosulfan neces-
sary to yield an LD50 was less for female rats, the twofold
difference between administration in the two different vehicles
remained the same for both sexes.
Boyd and Dobos (1969) estimated the largest nonlethal dose
(LD0) to be 60 mg/kg and the smallest totally lethal dose of
endosulfan (LD50) to be 180 mg/kg in Wistar rats.
Truhaut, et al. (1974) demonstrated that there were differ-
ences in the toxicities of endosulfan to different rodents: the
LD50 of 96 percent pure endosulfan administered orally to rats
and hamsters was 64+4 mg/kg in the rat and 118 + 16 mg/kg in the
hamster. The maximum dose without fatality was 40 mg/kg for the
rat and 70 mg/kg for the hamster. Biochemical measurements, or
effects of endosulfan dosing on enzyme levels, showed that in the
hamster, endosulfan inhibited cholinesterase significantly, where-
as there was no effect on rat cholinesterase. On the other hand,
C-47
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the activities of enzymes GPT and LDH were significantly elevated
by endosulfan dosage in the rat, but in the hamster they were un-
affected.
The difficulty in extrapolating LD5Q data from one animal
to another was demonstrated in a study by Li, et al. (1970), who
estimated (based on rat data) that 12.5 mg/kg would be an accept-
able dose for (Brown-Swiss and Holstein) dairy cattle. Within 10
hours of dosing, however, the two treated cows were in an extreme
state of excitation, and six days after dosing one of the animals
(Brown-Swiss) died.
The effects of accidental dermal exposures of cattle to endo-
sulfan were reported by Thompson (1966). Two hundred and fifty
cattle (breed, age, and sex not reported) were acciclently sprayed
with a 5 percent endosulfan miscible oil concentrate diluted
1:300, giving a wash concentration of approximately 0.12 percent
endosulfan. The cattle were sprayed early in the morning. Signs
of toxicity were noted in 50 of the 250 animals by about noon.
Four cattle were dead by 4 p.m. and six more died by the next
morning. The symptoms of exposure were those of hyperexcitability
(Thompson, 1966).
An accidental poisoning of three cows with endosulfan was re-
ported. The poisoning occurred when the animals ate grass which
had been sprayed 10 months earlier with an endosulfan emulsion
spray (reported as 35 percent endosulfan). Analysis of the organs
of one of the animals with gas chromatography showed the presence
C-48
-------
of £/ -endosulfan at 7 to 9 ug/kg, /^-endosulfan at 3.5 to 4.5
ug/kg, and metabolites as high as 9 mg/kg (Schmidlin- Meszaros and
Romann, 1971) .
Four of five crossbred male and female calves, weighing 60 to
170 kg, died within 24 hours after being dusted with a 4 percent
dust formulation of endosulfan. Symptoms of toxicity included
frenzied activity, violent convulsions, blepharospasm, and overall
extreme hyperexcitability. One of the animals was necropsied, and
no gross lesions were seen. Laboratory analysis revealed 0.73,
3.78, and 0.10 mg/kg endosulfan in the brain, liver, and rumen
contents, respectively (Nicholson and Cooper, 1977). This report
indicates excellent skin absorption in cattle, and probably a
toxic dosage much lower than that reported for rats, for which 110
mg/kg is an experimental fatal dose (Dreisbach, 1974). Milk and
tissue were also analyzed from another dairy herd which was ex-
posed to endosulfan; 9 of 18 animals exposed died (Braun and Lobb,
1976). Liver, kidney, and muscle tissue contained endosulfan sul-
fate at a level of 4.2, 1.1, and 0.6 mg/kg, respectively. Milk
from one of the survivors contained 1 ug/kg endosulfan sulfate at
the end of five weeks, at the time a blood sample contained 0.025
mg/kg endosulfan. The symptoms of exposure were like those
described in the first report.
The signs of toxicity observed in rabbits were similar to
those in rats and mice, the onset occurring three to six hours
after exposure. Hyperexcitability, dyspnea, decreased respira-
tion, discharge from the eyes, and tremors were followed by con-
vulsions. The convulsions appeared at intermittent or regular
C-49
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intervals. The animals preferred to rest on the sternum with the
forelimbs extended. Eventually the animals lost response to pain-
ful stimuli. This loss first occurred in the hindlimbs and then
spread to the forelimbs, followed by loss of motility, loss of
corneal reflex, a deep coma, and death (Gupta and Chandra, 1975).
In cattle dermally exposed to endosulfan the signs of toxi-
city consisted of listlessness, blind staggers, restlessness, hy-
perexcitability, muscular spasms, goose-stepping, and violent
"fits" (Thompson, 1966).
Three other reports of accidental animal poisoning (species
not specified) describe the toxic effects of endosulfan exposure
(Panetsos and Kilikidis, 1973; Utklev and Westbye, 1971; Schmid-
lin-Meszaros and Ronann, 1971; all cited by Demeter and Heyn-
drickx, 1978). The effects reflected an induced neurotoxicity and
were roughly analogous to those in endosulfan-poisoned humans.
A survey by California veterinarians reported on the occur-
rence of domestic animal poisoning by organochlorines, including
the death of calves following contamination of feed bunks with
endosulfan. No specific instances or dose levels were reported,
but signs of poisoning and treatment were tabulated (Maddy and
Riddle, 1977). Signs of poisoning included apprehension, hyper-
sensitivity and spasms of the eyelids and front quarters progress-
ing to the hind quarters; these spasms may be continuous or inter-
mittent. Clonic-tonic seizures, loss of coordination, circling
frontward or backward, and abnormal posturing is seen. The animal
may become comatose. The veterinary treatment emphasizes agents
C-50
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to control particularly violent neuromuscular activity in severe
poisonings (Maddy and Riddle, 1977).
Ely, et al. (1967) report that the inhalation 4-hour LC50
of endosulfan was 0.35 mg/1 for male rats. Under similar test
conditions the 4-hour LC50 for female rats was 0.08 mg/1.
Whether these values are from work done by Ely, et al. or are
quoted from some other report, however, is not clear. Details on
procedures, numbers of animals, etc., were not given.
Gupta and Chandra (1975) studied the eye irritation proper-
ties of endosulfan. When aqueous suspensions of 5, 10, and 20
percent endosulfan were instilled into one eye each of six rabbits
(two per group), no irritation or congestion was observed in any
of the animals.
A 1:1,000 endosulfan dilution instilled in rabbit eyes caused
neither pain nor subsequent inflammation (Hoechst, 1967a).
Skin irritation and skin sensitization studies have apparent-
ly not been made with endosulfan, although one report notes that
the skin of rabbits treated dermally with endosulfan at 100 mg/kg
did not exhibit any cutaneous abnormalities (Gupta and Chandra,
1975).
Signs of poisoning in dogs dosed orally with 200 and 500
mg/kg body weight were increased saliva formation, vomiting, and
tonic and clonic cramps (Hazleton Laboratories, 1967). Signs of
toxicity in endosulfan-exposed cattle have been described earlier
in this section.
Gross necropsy of rats fed endosulfan at near the LD50
range (see Table 10) revealed congestion of the brain and an acute
C-51
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gastroenteritis. Dark reddish areas were often seen in the kid-
neys, liver, spleen, and thymus. The skin was of normal appear-
ance. Edema of the interstitial tissue of the testes was noted.
A loss of organ weight was observed in most animals, but sig-
nificantly so in cardiac muscle, stomach, kidneys, liver, skin,
spleen, testes, and thymus (Boyd and Dobos, 1969).
Gupta and Chandra (1975) report that following an acute der-
mal exposure of rabbits to endosulfan at 100 mg/kg of body weight,
necropsy revealed congestion in the kidneys, peritoneum, and the
muscles underlying the skin. No other gross pathological condi-
tions were observed. Microscopic examination of the liver re-
vealed marked congestion and dilation of sinusoids., In oome of
the lobules hepatocytes were observed undergoing degenerative
changes around central veins. Sections of the kidneys from
treated animals showed groups of glomeruli with shrunken tufts and
thickened Bowman's capsules. Occasionally the epithelium of the
proximal convoluted tubules were necrotic and desquamated. The
adrenals of treated animals exhibited cell disruption, foamy cyto-
plasm, and eccentric nuclei in the zona reticularis.
Necropsy of cattle that died following an accidental (dermal)
exposure to endosulfan did not reveal any great pathological
changes, although congestion and edema of the lungs along with
froth in the trachea were observed (Thompson, 1966).
The liver was the principal target, with increased weight and
an apparent increase in drug metabolizing enzym.es (Gupta and
Gupta, 1977a,b). Rats that were dosed on either 7 or 15 consecu-
tive days with 2.5 or 5.0 mg/kg technical endosulfan showed liver
C-52
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effects. Neither testes nor adrenals of the endosulfan-treated
animals differed in weight from the controls.
The kidney, stomach, and intestine of fish were adversely
affected by exposure to a 35 percent emulsifiable concentrate for-
mulation of endosulfan at levels of 0.4 and 0.8 ug/1, and the same
dose also severely damaged the liver (Amminikutty and Rege, 1977;
1978). Acute treatment involved observation of histological
change that occurred in fish 96 hours after the formulation was
added to the fish water. The 96-hour LC50 was 1.6 ug/1, and
renal tubular cells were affected. Both stomach and intestinal
mucosa were severely damaged. Fish, chronically exposed to levels
of 0.4 and 0.53 ug/1 for 16 weeks showed hyperplasia of the kidney
and necrosis of intestinal mucosa cells (Amminikutty and Rege,
1978). The same dose levels and time produced vacuolated and
ruptured hepatic cells, as well as frequent total destruction of
pancreatic islet cells (Amminikutty and Rege, 1977).
A toxic effect unreported in other studies, testicular atro-
phy in male Osborne-Mendel rats, was seen in the recent carcino-
genicity bioassay [National Cancer Institute (NCI), 1978]. Testi-
cular pathology occurred in 18/47 (38 percent) of the group re-
ceiving 445 mg/kg endosulfan of 98.8 percent purity in the diet
and in 24/47 (51 percent) of the group receiving 952 mg/kg. The
pathology was characterized by degeneration and necrosis of the
germinal cells lining the seminiferous tubules. Three of 19, or
16 percent, of the control rats had testicular atrophy in this
study. Male mice of the B6C3F1 strain, receiving 6.9 and 3.5
mg/kg in the diet, in the same study, showed a slight indication
C-53
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of testicular atrophy with pathology in 3 of 50 high dose and 2 of
50 low dose animals. Control mice had neither testicular inflam-
mation nor atrophy.
Protein-deficient male Wistar strain rats were reported to be
four times as susceptible to the toxic effect of technical grade
endosulfan as rats having adequate protein nutrition. The toxi-
city of the pesticide was determined after the rats had been fed
for 28 days on a purified diet low in protein. Test animals were
compared to rats on the purified diet with normal protein and to
rats on standard laboratory chow.
With the purified diet containing no additional protein the
LD50 in rats was 5.1 + 1.4 mg/kg. At dietary protein levels
of 3.5, 9.0, 26.0, and 81 percent (28 days' feeding), endosulfan
LD50s in rats were 24 + 10, 57 + 4.0, 102 + 16, and 98 + 7
mg/kg, respectively. The LD50 value for endosulfan when given
in standard laboratory chow was 121 + 16 mg/kg (Boyd and Dobos,
1969; Boyd, et al. 1970).
Toxicity of endosulfan sulfate to mammals is about the same
as the parent compound. However, the endosulfan alcohol, hydroxy-
ether, and lactone have LD50s ranging from 150 to 1,500 mg/kg
in the rat (Gorbach, 1972).
Dorough, et al. (1978) determined the acute oral toxicities
of endosulfan and its apolar metabolites to female albino mice.
The approximation method used resulted in values that correlated
very closely with LDso values. The most toxic compounds were
endosulfan sulfate (8 mg/kg), (A-endosulfan (11 mg/kg), and ^-en-
dosulfan (36 mg/kg). With these compounds, no symptoms of poison-
C-54
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ing were seen until the lethal dose was almost reached, and the
lethal doses caused convulsions and death within one hour. Four
other metabolites were tested: endosulfan c* -hydroxyether, endo-
sulfan lactone, endosulfan ether, and endosulfandiol, with acute
lethal doses of 120, 120, 270, and over 2,000 mg/kg, respectively
(Table 11).
Rats were reported to tolerate endosulfan at oral doses of up
to 3.2 mg/kg/day for three months without observed injury (Czech,
1958).
The no-effect level for dogs was considered to be 30 mg/kg
feed (^0.75 mg/kg/day) (FMC, 1967).
A no-effect level for endosulfan in rats was studied with
respect to induction of microsomal liver enzymes (Den Tonkelaar
and Van Esch, 1974). The activities of aniline hydroxylase,
aminopyrine demethylase, and hexobarbital oxidase in experimental
groups each consisting of six Wistar male rats were compared with
those of six control animals. Results from aniline hydroxylase
induction studies indicated that when endosulfan was fed in the
diet at 200 mg/kg for two weeks the activity of the enzyme was 123
percent of the control (statistically greater); at 50 mg/kg the
activity of the enzyme in treated animals was nearly the same as
the control (slightly less). Treatment with endosulfan at a
dietary level of 200 mg/kg also statistically increased the activ-
ity of aminopyrine demethylase, but not the activity of hexobar-
bital oxidase. The no-effect dietary level for endosulfan for
rats was considered to be 50 mg/kg.
A 6-week toxicity study, dosing 98.8 percent pure endosulfan
in the diet, was performed at five dose levels on B6C3F1 mice,
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TABLE 11
Approximate Lethal Dose of Endosulfan
and Apolar Analogs to Mice*
Compound Dose (mg/kg)
2,000
*Source: Adapted from Dorough, et al. 1978
C-56
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five males and five females per dose, and a similar number of
Osborne-Mendel rats (NCI, 1978). Concentrations of endosulfan in
the rat group were 178, 316, 562, 1,000 and 1,780 mg/kg, and in
the mouse groups 3.2, 5.6, 10, 18, and 32 mg/kg. Animals were
dosed six weeks, then observed two more weeks while on regular
diet. A control group for each species received the vehicle and
normal lab chow.
In male rats, a 9 percent depression in mean body weight
occurred at 562 mg/kg, and a 20 percent depression at 1,000 mg/kg.
No depression in body weight as a function of dose occurred in
female rats. In both sexes of mice, depression in mean body
weight was observed at concentrations of 5.6 mg/kg and above.
Deaths and the endosulfan dose levels:
Rats: 3/5 males, 1,780 mg/kg
1/5 females, 316 mg/kg
4/5 females, 562 mg/kg
Mice: 1/5 males, 10 mg/kg
1/5 females, 5.6 mg/kg
Weight gain of young female rats fed either 5 or 50 mg/kg
endosulfan in the diet for 15 days was used as an indicator of the
compound's effect on animals exposed to the insecticide subacute-
ly. Both groups gained weight at the control rate, and there was
no difference in the weight of livers or kidneys of endosulfan-
exposed rats when compared to control (Dorough, et al. 1978).
The compounds used in this test were purified ^ - and ^ -
endosulfan added as an acetone solution to ground animal feed.
Feed was checked by extraction and chromatography when freshly
C-57
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prepared and after remaining in the feeding cup 24 hours. The
four feeding groups were:
13 rats, ^-endosulfan isomer-5 mg/kg
13 rats, ^-endosulfan isomer-5 mg/kg
4 rats,d-endosulfan isomer-25 mg/kg
4 rats, 7:3 mixture of <^ : /-endosulfan-25 mg/kg
Dogs were reported to "tolerate" endosulfan at doses up to
0.75 mg/kg diet for one year (Hazleton Laboratories, 1959a) .
Oral doses of about 10 and 100 mg/kg endosulfan in the diet
were administered to rats for two years (Hazleton Laboratories,
1959b). Low survival of females and reduced testis weight in
males were seen at the low dose. Consistent histopathological
findings were apparent at the high dose level, which produced
renal tubular damage and some hydropic change of the liver.
Synergism and/or Antagonism
The two human fatalities reported by Demeter and Heyndrickx
(1978) both involved endosulfan ingested with alcohol (although
dimethoate was also in one formulation). The authors suggest that
synergism between alcohol and endosulfan is likely, and they
reference the statements of Lendle (1956), who demonstrated an
increased gastrointestinal absorption of endosulfan in the pres-
ence of alcohols.
The acute toxicity of a diethylphosphorothioate (bromophos-
ethyl) was examined when dosed with endosulfan for synergistic
effects. A group of 10 rats was orally dosed with one-half of the
LD50 of endosulfan, or 24 mg/kg, at the same time they re-
ceived one-half of the LD50 of bromophos-ethyl. The mortality
C-58
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expected was 5/10, or 50 percent; 6/10 died within the one week
observation period, which indicates no synergistic activity oc-
curred (Muacevic, 1973).
Endosulfan was reported by Gupta and Gupta (1977a) to de-
crease the pentobarbital-induced sleeping time in endosulfan-
treated rats. Animals receiving the two higher doses of endosul-
fan showed significant increases in time to sleep induction and
shortening of the sleeping time. Although the blood and brain
concentrations of pentobarbital were significantly reduced at 30
minutes (reflecting the reduced response observed), there were no
differences in concentrations of pentobarbital in blood and brain
in control and treated animals when the rats awoke. This indi-
cated to the authors that the inhibitory effect on pentobarbital
by endosulfan is not due to a change in the sensitivity of the
brain, but could be due to enhanced metabolism of pentobarbital.
The influence of endosulfan on rat hepatic drug metabolizing
enzymes and lipid peroxidation was also measured to define how
endosulfan modifies the action and metabolism of other compounds
by affecting the mixed-function oxidase system (Agarwal, et al.
1978) . A marked increase in the activity of aminopyrine-N-de-
methylase, aniline hydroxylase, and tyrosine amino-transferase was
found, as well as an increase in spontaneous lipid peroxidation.
The increases were all dose dependent at the levels of 2.5 and 5.0
mg/kg (Agarwal, et al. 1978). The increase of the demethylase as
well as the hydroxylase enzyme suggests that endosulfan is a non-
specific inducer of drug metabolism.
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Teratogenicity
Technical grade endosulfan was tested for teratogenic and em-
bryotoxic effects in rats by Gupta, et al. (1978). The insecti-
cide was suspended in corn oil and given orally from day 6 through
day 14 of gestation in doses of 0.0, 5.0, and 10.0 mg/kg. On day
21 of gestation, both dams and fetuses were examined for pathol-
ogy. There was a significant increase in fetal mortality and
resorption sites in endosulfan-treated rats: control rats had 5.5
percent resorption without any dead fetuses, whereas endosulfan-
treated rats had 20 to 22.8 percent resorptions. No malformations
of any significance were noted in 463 fetuses from 59 dams. The
authors conclude that the study demonstrated no teratogenic
effect, but that the administration of endosulfan to pregnant rats
produced a dose-related increase in maternal toxicity, which they
attributed to a possible effect on the female sex hormones (Gupta,
et al. 1978).
Pure endosulfan was tested for embryotoxicity in the fertile
eggs of White Leghorn chickens at levels of 10 to 500 mg/kg. In-
jections were made to the center of the yolk using corn oil or
acetone as the carrier. At 100 mg/kg, endosulfan in acetone re-
duced hatchability by 54 percent compared to controls; 100 mg/kg
endosulfan in corn oil reduced hatchability by 24 percent compared
to controls (Dunachie and Fletcher, 1969). Endosulfan at 500
mg/kg in acetone showed 53 percent hatchability compared to con-
trols .
In similar studies, Smith, et al. (1970) evaluated the em-
bryotoxic effects of endosulfan on chickens. When 72 eggs per
C-60
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treatment and six treatment levels were studied (0.07 to 1.5
mg/egg yolk injection) hatchability was reduced from the zero con-
trol level of 80.0 percent to 77.3 percent at 1.5 mg/egg. At the
lowest concentration tested, percent hatchability was not affected
(Smith, et al. 1970).
In other tests 5 mg endosulfan per egg reduced hatchability
to 60 percent (Dunachie and Fletcher, 1966).
Lutz and Lutz-Ostertag (1972) conducted a study in which eggs
from hens of mixed breeding (Rhode Island Red-Wyandotte White and
Rhode Island Red-Wyandotte White—Light Sussex crosses) were
dipped into or sprayed with endosulfan in alcohol or acetone solu-
tions at concentrations from 0.5 to 5 percent. Following treat-
ment the eggs were incubated normally. Gonads from male and female
chick embryos at days 8 and 9 of incubation were explanted on agar
medium to which three drops of a 0.5 to 1.0 g/1 solution of endo-
sulfan were added.
These investigators reported that the spray and dip treat-
ments of the eggs resulted in alterations in the gonads of the
embryos in both males and females. The cultured gonad underwent
hypertrophy and became vacuolized; thus, there was a tendency to
sterility of the gonads.
Lutz-Ostertag and Kantelip (1970; 1971) performed similar ex-
periments on quail eggs, Coturnix coturnix japonica. They con-
cluded that endosulfan had no teratogenic effect on the quail at
the doses employed, but the male and female embryos were steri-
lized, and, according to the authors, this was due to the anti-
mitotic toxicity exhibited by endosulfan.
C-61
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Mutagenicity
Endosulfan, of unreported concentration, purity, and other
detail, was positive as a base-pair substitution mutagen in direct
Salmonella tests (without microsomal activation). The microbio-
logical tests employed the Salmonella typhimurium histidine auxo-
trophs TA1535, TA1536, TA1537, and TA1538 (Adams, 1978).
Neither the isomers of endosulfan nor the metabolites endo-
sulfan ether and endosulfan sulfate were active in the Salmonella
mutagenicity test with or without the S-9 liver homogenate.
Metabolites endosulfan diol, ^-hydroxyether, and the lactone
severely inhibited bacterial growth even at 10 ug per plate, so
the Ames test on these compounds produced inconclusive results
(Dorough, et al. 1978). All compounds were screened using the
four Salmonella typhimurium strains TA98, TA100, TA1535, and
TA1978 following dose response tests at 10, 100, 500, and 1,000 ug
per plate and were compared to a positive control, 2-acetylamino-
fluorene.
Endosulfan gave negative results when tested for mutagenicity
in Saccharomyces cerevisiae (mitotic gene conversion at the ade
2 and trp 5 loci), Escherichia coli (forward mutation to strepto-
mycin resistance at the str A gene locus), and Serratia marcescens
strains a 21 and a 742 (back mutation to prototrophy). Test dose
levels were not given (Fahrig, 1974).
The most relevant tests for predicting risk to humans are
positive results from in vivo mammalian tests which assess the
chemical's tendency to produce germ cell mutations. The heritable
translocation test in rodents is probably the best test to show
C-62
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chromosomal rearrangements, although the difficult and expensive
specific locus test in inbred mice is also satisfactory.
For assessing risk to man on the mutagenicity of endosulfan,
data that are necessary also include the demonstration that the
proposed mutagenic metabolite actually can reach the germ cells of
mammals when the compound is dosed. Further, knowledge of the
comparative metabolism of endosulfan in the test species versus
that of man is needed.
No tests have been run which define mammalian suppression of
DNA repair, disturbed segregation of chromosomes,, or outright pro-
duction of gene mutations or chromosomal aberrations.
Studies have been conducted that include Ames tests on endo-
sulfan isomers and proposed metabolites using four common Sal-
monella typhimurium strains and liver homogenate S-9 fraction. No
mutagenicity was seen in defined systems, although three of five
metabolites were toxic to the bacteria.
Carcinogenicity
Two bioassay tests by the NCI have been run on endosulfan.
In the first test (Kotin, et al. 1968; Innes, et al. 1969) a 96-
percent pure mixture of the isomers of endosulfan was administered
to mice by two routes: either as an injection in dimethylsulfox-
ide (DMSO) on the 28th day of age (2.15 mg/kg, subcutaneously) or
by stomach tube orally on days 7 to 28 (2.15 mg/kg in gelatin),
following which the compound was mixed with ground feed at levels
of 3 and 6 mg/kg feed.
The mice, C57B1/6 and C3H/AnfFl strains of both sexes, showed
incidences of tumors during the nearly 18 months of feeding as
C-63
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tabulated (Figure 2). Innes, et al. (1969) summarized the statis-
tical analyses and concluded there was no evidence of endosulfan
carcinogenicity.
In the second NCI bioassay on endosulfan (NCI, 1978), tech-
nical grade endosulfan of 98.8 percent purity was dissolved in
corn oil and mixed with the feed for 50 Osborne-Mendel rats of
each sex and 50 B6C3F1 mice of each sex. Chemical administration
was for 78 weeks, after which rats were observed for 33 additional
weeks and mice for 14 additional weeks. The trials on male rats
were terminated early, week 82 for high dose and week 74 for low
dose. Time-weighted average concentrations of endosulfan in the
diets for the entire study are tabulated as follows:
Osborne-Mendel Rats B6C3Fi Mice
MaleFemaleMale Female
(mg/kg) (mg/kg) (mg/kg) (mg/kg)
High dose 952 445 6.9 3.9
Low dose 408 223 3.5 2.0
The doses of endosulfan used in these studies were toxic to
the kidney of rats of both sexes and to male mice. Male rats also
had testicular atrophy, and high early death rates occurred in
both species of male mice. Due to these early deaths, the bio-
assay was not conclusive with regard to males, but enough females
survived to conclude that technical grade endosulfan is not a car-
cinogen to female B6C3F1 mice or to female Osborne-Mendel rats.
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O
I
CTi
U1
E 2
O {"
Gastric
Papilloma
Hepatoma
Pulmonary
a> D Adenoma
O l/>
Other
Tumors
2.9 Cf
2.9Q
2.9
2.9
CT
12
Cf
24.
9
•59
8CT
19 O
11. lQ
14.3 U
036
Concentration Endosulfan in Feed, mg/Kg Feed
FIGURE 2
Tabulation of Mouse Tumor Data from NCI Bioassay on Endosulfan
Source: Kotin, et al. 1968
-------
The official NCI summary recommended against retest of
endosulfan based on the early male animal mortality, since in the
female test animals, the chemical was noncarcinogenic.
Interesting relationships that were not discussed in the
official summaries appear when the data are examined closely.
Table 12, which presents tumors by site and ignores the early
deaths, shows that there were more liver and lung tumors in the
male mice than in matched controls; but this increased occurrence
of tumors is not dose-dependent: there were 6/49 liver tumors in
the low dose males but only 2/50 in the high dose and 1/20 in the
matched controls. Again, in the occurrence of alveolar/bron-
chiolar carcinoma, the matched controls had 0/20, but both high
and low dose male mice had 2/50 and 2/49, respectively.
Early mortality occurred in the males of both rats and mice,
but was a particular problem in the rats. A generalized toxic
nephropathy probably contributed most significantly to the early
deaths, but signs commonly associated with aging in group-housed
laboratory rats were reported in equal numbers in both dosed and
control animals during the last six months. Table 13 summarizes
the early mortality. In necropsies of the early deaths, several
lesions were found, but no actual dose-response pattern was evi-
dent, so no cause was ascribed by the authors to the early animal
deaths. The most prevalent lesions include: nephropathy, para-
thyroid hyperplasia, and testicular atrophy in male rats. Canni-
balism was the most common cause of early death in male mice.
The 95 percent confidence intervals on the relative risk of
developing a tumor furnish additional insight into the statistical
C-66
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TABLE 12
Target Organs for Endosulfan-Induced Tumors*
(incidence/population)
O
I
Osborne-Hendel High dosea
rats Male Low dose*3
Controls
High dosec
Female Low dose^
Controls
High dosee
Male Low dose^
Controls
B6C3F1 mice High dose9
Female Low dosen
Controls
Lung
0/47
0/50
1/20
1/50
1/50
0/20
2/50
2/49
0/20
0/50
6/50
2/20
Lymphomas/
Leukemias
1/47
2/50
4/20
1/50
3/50
1/20
0/50
0/49
0/20
6/50
10/50
6/20
Kidney
2/47
3/50
2/20
3/50
2/5p
1/20
0/50
0/49
0/20
0/50
0/50
0/20
Liver
0/47
0/50
0/20
1/50
1/50
0/20
2/50
6/49
1/20
1/50
0/50
0/20
Endocrine
0/47
1/50
7/20
11/50
19/50
13/20
0/50
0/45
0/20
1/50
0/50
0/20
All
Other Sites
0/47
4/50
0/20
15/50
27/50
14/20
6/50
1/49
3/20
3/50
4/50
1/20
*Source: Summarized from NCI Bioassay Data, 1978
a952 mg/kg feed e6.9 mg/kg feed
b408 mg/kg feed f3.5 mg/kg feed
93.9 mg/kg feed
h2.0 mg/kg feed
C445 mg/kg feed
d223 mg/kg feed
-------
TABLE 13
Animal Survival Times: Tumor Bioassay Studies*
Species
Rat
Mice
Dose Level
Sex or Control
Male High
Low
Control
Female High
Low
Control
Male High
Low
Control
Female High
Low
Control
% Living at Study End
(110 wk=rats 90 wk=mice)
Oa
Ob
25
50
62
70
10
39
15
96
94
85
*Source: Summarized from NCI, 1978
a!5% alive @ wk 74; ended trial 36 wk early
b20% alive @ wk 82; ended trial 28 wk early
C-68
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implications of these data. Many of the confidence intervals, due
to the early mortality, have an upper limit greater than one, in-
dicating the theoretical possibility that the test did not conclu-
sively address the possibility of tumor induction by endosulfan.
In all cases except one, however, the relative risk is unrelated
to the dose of endosulfan received. The occurrence of fibrosar-
coma of subcutaneous tissue in male mice showed a relative risk
greater than one when compared to both pooled controls and with
matched controls, and the risk was dose-related (Table 12) (NCI,
1978). The high incidence of fibrosarcoma of subcutaneous tissue
in all control male mice suggests this difference is unimportant
to the overall carcinogenicity of endosulfan.
Figure 3 illustrates time to tumor data for rats and mice for
this study.
The significance of the negative carcinogenicity data in the
latest NCI bioassay is increased by several factors involved in
the choice of model, which was a stringent test for carcinogeni-
city.
The C3H strain of mouse has one of the highest known inci-
dences of mammary tumors in females and liver tumors in males and
was a parent strain in both the carcinogenesis bioassay of 1968
and that of 1978. Differences in species responses to chemical
carcinogens can often be attributed to differing metabolic path-
ways and metabolites and to an inability of some species to effec-
tively convert the test chemical to an active carcinogen. The
work of Gupta (1978) and Gupta and Chandra (1975) has indicated,
however, that rats, mice, and rabbits all metabolize endosulfan.
C-69
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n
i
MICE
RATS
Hi Dose
Lo Dose
Controls
Hi Dose
Lo Dose
Controls
9
9
cf
cf
cf
1 1
Cf
9
cf
9
I
c?
9
9
i
0
25 50 75
Time to 1st Tumor, Weeks
100
FIGURE 3
Scattergram of Time to Tumor, Rat/Mouse Endosulfan Bioassay
Source: Tabulated from NCI, 1978
-------
The mouse strain used by the NCI in the 1978 carcinogenesis bio-
assay of endosulfan is so prone to the development of liver tumors
with minimal stimulation that two working conferences on the use
of such mice to assess carcinogenicity have been held: in 1969
[International Agency for Research on Cancer (IARC), 1971] and
1975 (Butler and Newberne, 1975). Neither conference has been
able to state conclusively which mouse data should be applied to
risk assessment. When very high levels of test compound (such as
those used for both bioassay trials of endosulfan) are used, tis-
sue injury and repair may be important in the development of
lesions. Other factors such as sex, hormones, and diet have been
suggested as possible modifiers of the carcinogenic activities of
primary carcinogens. The distinct differences in toxicity to males
versus females seen in many endosulfan tests makes it quite likely
that hormonal differences influenced the final test result in the
carcinogenicity bioassays.
The Osborne-Mendel rat, also used in the NCI bioassay of
endosulfan (NCI, 1978), is known to be a strain very resistant to
toxicity, so that high dose levels for extended periods can be
administered. This increases the likelihood for survival and the
appearance of any tumors that would be missed in trials with early
deaths (Tomatis, et al. 1973). The fact that toxicity and early
deaths occurred in the male Osborne-Mendel rats in the 1978 endo-
sulfan bioassays is another indication that doses were more than
adequate to produce an effect if the chance existed to produce
one.
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These animal models give an additionally severe test of car-
cinogenicity in that the parent mice of these inbred strains
carry tumor viruses. In the 1968 NCI bioassay, for example, the
AKR strain has a high rate of leukemia by 6 to 8 months of age.
Use of the C3H strain, with the murine mammary virus,, and the AKR
strains of mice means these bioassays are also testing for promo-
tion mechanisms of the test chemical. There are no known human
tumors that occur by promotion of a human tumor virus, so the use
of these strains to test for carcinogenicity is a severe trial.
In addition, Henschler, et al. (1977) has suggested that mice in
general have a particularly low activity of epoxide hydrase, i.e.,
mice have a decreased ability compared to other animals to detox-
ify reactive epoxides, which are the reactive and toxic intermedi-
ates formed in vivo as metabolites in many industrial chemicals.
Route of exposure, used in the NCI trials (high dietary
levels) , is less relevant to human exposure (dermal and inhalation
of particulates) than it is to domestic animals. While these bio-
assay trials did not measure gastrointestinal absorption, it is
likely that a high concentration of endosulfan reached the liver
by the portal circulation with each meal taken by the test ro-
dents. Endosulfan that reaches the liver complexing and detoxifi-
cation mechanisms by dermal or inhalation routes do so after pass-
age through tissue, the blood stream, and contact with many cellu-
lar mechanisms. The oral route is a particularly severe test for
i
liver effects, and the lack of such effects in these trials is
further indication of a clean bill of health for the carcinogeni-
city of endosulfan. It is also worthy of note that absorption
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from the gastrointestinal tract is the route that is most likely
to ensure that metabolites of endosulfan, as well as the alpha and
beta isomers, impinge upon not only the liver but also other
organs and tissues of the body.
Ely, et al. (1967) reported that one or more convulsions
occurred in each of nine workers exposed to a 50 percent endosul-
fan wettable powder. Six of the nine cases were known not to have
had a history of previous convulsions, but the previous histories
of the other three were uncertain. A causal relationship between
convulsions and exposure to endosulfan was, however, considered
highly likely.
The potential vulnerability of the central nervous systems of
humans to endosulfan was demonstrated in epileptic convulsions and
altered EEC patterns in three subjects exposed to the pesticide.
In one of the patients, occasional EEC alterations were observed
one year after the exposure (Tiberin, et al. 1970) .
Five human deaths due to endosulfan were reported by Terziev,
et al. (1974), two of which were accidental poisonings and three
of which were the result of intentional intake. Details were
lacking, but the most significant post-mortem findings as de-
scribed by Terziev were circulatory disorders, protein dystrophy
in the parenchymal organs, acute lung emphysema, and severe
changes in the neurons.
Two poisoning cases resulting in human fatalities were re-
ported with 20 percent endosulfan products, and both involved
interaction with other chemicals (alcohol in one case and alcohol
with dimethoate in the second). Demeter and Heyndrickx (1978)
C-73
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found
-------
and masks. The symptoms appeared rapidly, within 1 to 2 hours in
the lethal cases, and initially included headache, restlessness,
and increased irritability, followed by vertigo, stupor, disorien-
tation, and epileptiform convulsive seizures. In the workers who
died, there were also loss of consciousness, cyanosis, dyspnea,
foaming at the mouth, and noisy breathing (Israeli, et al. 1969).
It was noted in a later report (Tiberin, et al. 1970) that there
were pathological changes on the electroencephalograms. Hyper-
ventilation improved the EEC picture.
C-75
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CRITERION FORMULATION
Existing Guidelines and Standards
The National Technical Advisory Committee on Water Quality
Criteria (Federal Water Pollution Control Administration, 1968)
did not establish a permissible limit of endosulfan in raw surface
waters for public water supply purposes. The Committee stated,
however, that the 48-hour median tolerance level (TLm) of endo-
sulfan to shrimp is 0.2 ug/1 and, therefore, classified endosulfan
as acutely toxic to shrimp at concentrations of 5 ug/1 or less.
On the assumption that 1/100 of this level represents a reasonable
application factor, the Committee recommended that environmental
levels of endosulfan should not be permitted to rise above 0.05
ug/1. This level is so low that endosulfan cannot be applied
directly in or near the marine habitat without danger of causing
damage.
In the 1972 report of the Committee on Water Quality Criteria
[National Academy of Sciences (NAS), 1972], a maximum concentra-
tion of 0.003 ug/1 of endosulfan is recommended for whole (unfil-
tered) fresh water sampled at any time and at any place. This
concentration was determined by multiplying the acute toxicity
value of endosulfan for the most sensitive native aquatic species
(rainbow trout, Salmo gairdneri) (Schoettger, 1970), by an appli-
cation factor of 0.01. The marine criterion of 0.001 ug/1 was
similarly determined using the LC50 value of the most sensi-
tive marine species (striped bass, Morone saxatilis) (Korn and
Earnest, 1974).
C-76
-------
Revision of the above recommended standards may be indicated
by more recent data. For example, the 96-hour LC50 value of
0.04 ug/1 on pink shrimp, Penaeus duorarum, would, if incorpo-
rated, reduce the saltwater criterion from 0.001 ug/1 to 0.0004
ug/1, using a theoretical application factor of 0.01 (Schimmel, et
al. 1977). This theoretical ratio is used in the absence of an
empirically derived factor. Macek, et al. (1976) have empirically
derived application factors from their work on two fresh water
species, fathead minnows, Pimephales promelas, and water fleas,
Daphnia magna. The 7-day incipient LC50 of 0.86 ug/1 and the
maximum acceptable toxicant concentration (MATC) limits of 0.20 to
0.40 ug/1 for fathead minnows give a derived application factor
(ratio of chronic toxicity to acute or subacute) range of 0.23 to
0.47. MATC limits are the highest concentration for which there
is no effect and the lowest concentration showing an adverse
effect. The 48-hour LC50 of 166 ug/1 and the MATC limits of
2.7 to 7.0 ug/1 for Daphnia magna, however, give a derived factor
range of 0.016 to 0.042.
The recent National Academy of Sciences report on drinking
water did not address water standards for endosulfan (NAS, 1977).
Current Levels of Exposure
Endosulfan has been detected in water samples from the United
States and Canada. Maximum values reported from various studies
include:
0.02 ug/1 in streams of the western United States.(one posi-
tive sample out of 546);
C-77
-------
0.032 ug/1 in drainage ditches from treated agricultural
fields near Lake Erie;
0.011 ug/1 in Canadian water systems;
0.083 ug/1 in Ontario municipal water samples;
0.014 ug/1 in surface and bottom water samples from Lake
Erie;
0.060 ug/1 in the St. Lawrence River;
The detection limit for endosulfan in water, using electron-
capture gas chromatographic methods, is ^0.005 ug/1 (Schulze, et
al. 1973).
Residues in food (^-endosulfan, ^-endosulfan, and endosul-
fan sulfate) result from the use of endosulfan on over 60 food and
nonfood crops.
During the 1965 to 1970 period, daily U.S. intake of endosul-
fan residues was estimated using market basket samples from the
total diet program of the FDA. These samples showed a daily in-
take of endosulfan «* -, j* -, and sulfate) of from <0.001 to 0.001
mg/day.
The acceptable daily intake of endosulfan (i.e., the daily
intake which during an entire lifetime appears to be without
appreciable risk), as established by FAO/WHO, is 0.0075 mg/kg.
This value corresponds to an intake of 0.525 mg/day for a 70-kg
person.
Endosulfan has also been shown to bioconcentrate in the tis-
sue of aquatic species. Bioconcentration data are summarized in
Table 14.
C-78
-------
TABLE 14
Summary of Bloconcentration Data for Endosulfan
Measured
Water Concentration
Test species (rag/liter)
o
1
^j
Common mussel
(Mutilus edulis)
Scallop
(Ghlamys opercularis)
Pink shrimp
(Penacus duorarum)
Grass shrimp
(Palaemonetes vulgaris)
Mullet
(Mugil cephalus)
Spot
(Leiostomus xanthurus)
Pinf ish
(Lagodon rhomboides)
Goldfish
(Garassius auratus)
1,000
100
0.14b
100
0.089
1.75
0.32
0.035
0.076
0.15
1
Exposure
Period
(days)
70
14
14
4
4
4
28
4
4
5
Bioc concentration
Factor8
22.5
28. 5C
600
25. 7C
0
245
1,344
2,755
(2,429)d
895
1,299
400
Source
Roberts
Roberts
Ernst
Roberts
Schimmel
Schimmel
Schimmel
Schimmel
Schimmel
Schimmel
Gorbach
(1972)
(1975)
(1977)
(1975)
, et
, et
, et
, et
, et
, et
al.
al.
al.
al.
al.
al.
(1977)
(1977)
(1977)
(1977)
(1977)
(1977)
(1972)
aHighest bioconcentration factor reported by the respective investigators. Whole body
noted
-Endosulfan steady-state concentration; initial concentration was 2.05 ug/liter
cuased on summated values for separate tissues
dEdible tissue
basis unless otherwise
-------
Endosulfan residues (<^-endosulfan, x/-endosulfan' and endo~
sulfan sulfate) have been detected in most types of U.S. tobacco
products in recent years. The data in Table 15 summarize the
average residue levels (mg residue/kg processed tobacco) detected
in several independent studies.
Air samples from 16 states in 1970 showed an average level of
13.0 ng/m3 c/i-endosulfan and 0.2 ng/m3 /7-endosulfan. None of
the air samples collected in 1971 or 1972, however, contained de-
tectable levels of either isomer.
Special Groups at Risk
Data on the presence of endosulfan residues (^ -endosulfan,
//-endosulfan, and endosulfan sulfate) in food, tobacco, water,
and air have been briefly summarized in the preceding subsection.
These data indicate three human populations that are at risk of
exposure to endosulfan through:
(1) Exposures occuring primarily from: residues in foods as
a result of the use of endosulfan on food crops and feedstuff;
bioconcentration in aquatic species; residues in air adjacent to
sites of manufacture or application; and residues in water.
(2) Residues in processed tobacco products (cigarettes,
cigars, snuff, etc.) resulting from the field use of endosulfan.
(3) Dermal and respiratory exposure occuring during manufac-
ture, formulation/packaging, field application, and harvesting.
Basis and Derivation of Criterion
Establishing a scientific basis for evaluating the hazard of
endosulfan to man is difficult. At very high levels of acute
exposure, humans show central nervous system (CNS) symptoms and
C-80
-------
o
I
00
TABLE 15
Endosulfan Residues in Processed Tobacco
Cigarettes
Cigars
Little cigars
Smoking tobacco
or pipe tobacco
Chewing tobacco
Snuff
Year
1971
1972
1973
1971
1972
1973
1971
1973
1971
1973
1971
1973
1971
1973
Average Residue
(mg/kg)
0.2
0.38
0.83
0.4
0.41
0.37
0.4
0.22
<0.2
0.37
0.2
0.36
<0.2
<0.12
Source
Domanski, et al . (1973)
Dorough and Gibson (1972)
Domanski, et al . (1974)
Domanski, et al. (1973)
Domanski and Guthrie (1974)
Domanski, et al. (1974)
Domanski, et al . (1973)
Domanski, et al . (1974)
Domanski, et al . (1973)
Domanski, et al. (1974)
Domanski, et al . (1973)
Domanski, et al. (1974)
Domanski, et al . (1973)
Domanski, et al . (1974)
-------
may die. Several studies report endosulfan has been ussed for sui-
cides (Terziev, et al. 1974; Couteslinis, et al. 1978). Workers
who failed to use good safety practices (i.e., to cover skin and
use respiratory protection) have died from endosulfan exposure
(Israeli, et al. 1969). In one incident, three persons exposed
showed CNS symptoms; two of them died. It therefore appears that
the most toxic potential effect to man is that of CNS toxicity,
since the available data indicate a lack of carcinogenic or tera-
togenic potential. One study has indicated that endosulfan is a
base-pair substitution mutagen (Adams, 1978). The absence of
reports on toxic effects associated with the proper use of endo-
sulfan (particularly such effects as skin sensitization or other
human symptoms) has been noted (Hoechst, 1966).
There appears to be considerable species variation in toxic
effects. Of the species tested with endosulfan, cattle are the
most sensitive to the neurotoxic effects and would therefore be a
"worst case" model for human toxicity. There are much more con-
trolled toxicity data on rodents, but cattle appear to be closer
in sensitivity and effects to man. Data on CNS toxicity to cattle
are presented in Table 16.
The relevance of these high exposure levels to a water qual-
ity standard presents additional sources of calculation error.
The CNS toxicity in these studies is an acute symptom of high
exposure. All reported human poisonings, however, have resulted
from accident, human error, or suicidal intention. The reported
poisonings of man and the most sensitive other mammal, cattle,
C-82
-------
TABLE 16
CNS Toxicity of Endosulfan in Cattle
n
i
00
U)
Number Time to % Exposed
Animals CNS Toxicity Showing
Dose, Route Exposed (hours) CNS Effects
12.5 mg/kg, oral 2 10 100
0.12% formulation, 250 5 20
dermal
4% dust, dermal 5 2 100
&
Time to Death Exposed
(days) Dying Source
6 50 Li, et al. (1970)
1 4 Thompson (1966)
1 80 Nicholson and
Cooper (1977)
100%
(1976)
-------
have occurred after acute, high level exposure to concentrated
endosulfan. These levels will not occur in drinking water. The
key question then is, are there any data in the toxicology reports
or studies to indicate that CNS effects can occur after chronic,
very low level exposure to endosulfan?
Tiberin, et al. (1970) reported occasional EEC alteration in
one of three men one year after a convulsive seizure following
exposure to endosulfan. Terziev, et al. (1974) report that
autopsy on an endosulfan suicide case showed "changes in the neu-
rons" among lesions in other organs. Rats, although more resis-
tant to toxicity than man or cattle, demonstrate no histopatho-
logical changes in the brain after receiving high doses of endo-
sulfan orally for 78 weeks, or most of a lifetime (NCI, 1978).
Cerebral hemorrhage was reported in seven female rats that
died early in the study (week 21), but the absence of lesions at
even higher and more long-term dosages suggested to the authors
that these deaths were not compound-related. Several lesions were
present in the male rats and mice that died early in these endo-
sulfan feeding studies. The most prevalent lesions included
nephropathy, parathyroid hyperplasia, and testicular atrophy, all
without clear dose response pattern (NCI, 1978).
An important question is, "Do the apolar metabolites of endo-
sulfan remain in the body to produce chronic effects if endosulfan
is ingested in low level quantities over a long term?" No con-
trolled metabolic studies in man have been reported, although
Demeter and Heyndrickx (1978) report that endosulfan sulfate is a
C-84
-------
metabolite in humans. This metabolite is approximately as toxic
to mice as the parent isomers (Dorough, et al. 1978), but no spe-
cific CNS effects were reported (based on toxicity trials on the
pure compound).
The toxicity of endosulfan is somewhat greater in animals
with deficiencies of dietary protein (Boyd and Dobos, 1969; Boyd,
et al. 1970). The differences in even a dose as high as an
LD5Q are not great enough, however, to ascribe any potential
human hazard to this mechanism or to suggest that protein-deprived
humans would be more sensitive to chronic exposure to endosulfan
in drinking water.
It can be concluded that (a) the controlled studies uniformly
report CNS toxicity following acute high level exposure, and (b)
there has been no indication reported of specific lesions in mam-
mals related to mortality following chronic exposure.
A water quality criterion could be based on the lowest no-
observed-effect level (NOEL) reported for endosulfan in test
species. Available data on no-effect levels are summarized in
Table 17.
As no valid experimental results from studies on prolonged
ingestion by man were available, the best available long-term ani-
mal feeding study was used as the basis of criteria formulation.
The selected NOEL is based on a 78-week mouse feeding study at 2.0
mg endosulfan/kg feed concentration. The calculated dose corres-
ponds to 0.4 mg endosulfan/kg body weight/day for a typical 25 g
mouse consuming 5 gs feed/day:
2.0 mg endosulfan 5 g feed mouse „ „
1,000 g feed X mouse-day x 0.025 kg = °'4
C-85
-------
TABLE 17
Summary of Effects of Endosulfan on Different Species and Biochemical Parameters
Species
Rats
Rat
Rat
Rat
Rat
Rat (female
Osborne -Mendel)
Hamsters
Hamsters
' Mice
00 niv-ti
O>
Mice
(Female B6C3F1)
Rabbit
Rabbit
Rabbit
Chickens
uog
Salmonella
typhimurium
Steers
Steers
Target
Organ/Tissue
-
-
Liver
Liver
Embryo
:
Liver
—
_
Eye
Eye
Skin
Egg
Strains TA98,
100, 1534,
and 1978
Effect Observed
Lethality
Lethality
Cholinesterase
inhibition
Microsome enzyme
function
Not teratogenic
Lethality
Lethality
Enzyme inhibition:
GPT , LDH
Height depression
Lethality
Inflammation and
irritation
Inflammation,
irritation
Irritation
Hatchability
Gross and microscopic
lesions
Base-pair substitution
(mutagenicity)
No fat residue
Muscle convulsions
Concentration
or Dose Level
55 mg/kg •= LDn
40 mg/kg = LD0
68 mg/kg minimum
50 ppm diet
10 mg/kg
445 ppm diet
70 mg/kg
134 mg/kg minimum
3.2 ppm diet
2.0 ppm diet
1:1,000 aqueous
20% aqueous solu-
tion
100 mg/kg
0.07 mg/egg
0.75 mg/kg/day
1.0 mg/plate
0.15 mg/kq/day
2.5 mg/kg/day
Route Administered*
Acute Oral
( intragastric)
Acute Oral
Acute Oral
Diet
(2 weeks)
Oral
(gestation day
7-14)
Diet
(78 weeks)
Acute Oral
Acute Oral
Diet
(6 weeks)
Diet
(78 weeks)
Instillation
Instillation
Dermal
Yolk injection
Oral
(52 weeks)
"
Oral (60 days)
Oral (60 days)
Source
Boyd and Dobos
(1969)
Truhaut, et al .
(1974)
Truhaut, et al .
(1974)
Den Tonkelaar and
Van Esch (1974)
Gupta, et al.
(1978)
NCI (1978)
Truhaut, et al .
(1974)
Truhaut, et al .
(1974)
NCI (1978)
NCI (1978)
lloechst (1967a)
Gupta and Chandra
(1975)
Gupta and Chandra
(1975)
Smith, et al. (1970)
FMC (1967)
Dorough, et al .
(1978)
Beck, et al. 1966
Beck, et al. 1966
•Single dose unless otherwise noted
-------
It should be noted that this estimate compares well with a re-
ported 60 day study where steers received endosulfan in their
feed. A 0.15 mg/kg NOEL estimate was observed. A low-observed-
effect level (LOEL) of 1.1 mg/kg was also noted in this study
(Beck, et al. 1966) .
Applying a safety factor of 100 to the derived NCI dosage
gives an upper limit for nonoccupational daily exposure (ADI) of
0.28 mg/kg body weight for a 70 kg person:
Q- 7*
-
kg-day 100 person
= 0.28 mg/day
^' J
For the purpose of establishing a water quality criterion,
human exposure to endosulfan is considered to be based on inges-
tion of 2 liters of water and 6.5 g of fish/day. The amount of
water ingested is approximately 100 times greater than the amount
of fish consumed. The fish bioaccumulation factor for endosulfan,
has been established to be 270 (Stephan, 1980).
The equation for calculating the criterion for endosulfan
content of water is:
(2) (X) + (0.0065) (F) (X) = ADI
where: 2 = amount of drinking water was consumed, I/day
X = endosulfan concentration in water, mg/1
0.0065 = amount of fish consumed, kg/day
F = bioconcentration factor, mg endosulfan/kg fish per
mg endosulfan/1 water
C-87
-------
ADI = limit on daily exposure for a 70 kg person
For F = 270
2X + (0.0065) (270) (X) = 0.28
3.75X = 0.28
X = 0.075 mg/1 or 75 yg/1
Consideration of dietary endosulfan levels (apparently /^>0.01
mg/day or less) and other sources of exposure (ambLent levels,
cigarette smoke, etc.) does not significantly affect this calcula-
tion.
In summary, based on the use of chronic toxicologic test data
for mice and an uncertainty factor of 100, the criterion level for
endosulfan is 75 ug/1. Drinking water contributes 53 percent of
the assumed exposure while eating contaminated fish products ac-
counts for 47 percent. The criterion level can alternatively be
expressed as 159 yg/1 if exposure is assumed to be from the con-
sumption of fish and shellfish products alone.
:-88
-------
REFERENCES
Adams, J.F. 1978. Mutagenicity of some environmental chemicals
in Salmonella test systems without microsomal activation. Mutat.
Res. 53: 142.
Agarwal, O.K., et al. 1978. Effect of endosulfan on drug metabo-
lizing enzymes and lipid peroxidation in rat. Jour. Environ. Sci.
Health. C13: 49.
American Conference of Government Industrial Hygienists. 1971.
Documentation of the threshold limit values for substances in
workroom air. 3rd ed. Cincinnati, Ohio.
American Conference of Government Industrial Hygienists. 1977.
TLVs® threshold limit values for chemical substances and physical
agents in the workroom environment with intended changes for 1977.
1977 TLV Airborne Contaminants Committee. Cincinnati, Ohio.
Amminikutty, C.K. and M.S. Rege. 1977. Effects of acute and
chronic exposure to pesticides, Thiodan 35 E.G. and Agallol "3" on
the liver of widow tetra Gymonocorymbus [sic] ternetzi (Boulen-
ger). Indian Jour. Exp. Biol. 15: 197.
C-89
-------
Amminikutty, C.K. and M.S. Rege. 1978. Acute and chronic effect
of Thiodan 35 E.G. and Agallol "3" on kidney, stomach and intes-
tine of the widow tetra Gymonocorymbus ternetzi Boulenger. Indian
Jour. Exp. Biol. 16: 202.
Beck, E.W., et al. 1966. Residues of endosulfan in meat and milk
of cattle fed treated forages. Jour. Econ. Entomol. 59: 1444.
Boelens, R.G. and R. Frank. 1973. Unpublished data. Water
Resour. Environ. Assessment Plan. Div., Ontario Ministry of the
Environ., Toronto. In; National Research Council of Canada.
Boyd, E.M. and I. Dobos. 1969. Protein deficiency and tolerated
oral doses of endosulfan. Arch. Int. Pharmacodyn. 178: 152.
Boyd, E.M., et al. 1970. Endosulfan toxicity and dietary pro-
tein. Arch. Environ. Health. 21: 15.
Braun, H.E. and B.T. Lobb. 1976. Residues in milk and organs in
a dairy herd following acute endosulfan intoxication. Can. Jour.
Anim. Sci. 56: 373.
Butler, W. and P. Newberne (eds.) 1975. Mouse Hepatic Neoplasia.
Elsevier Scientific Publishing Co., Amsterdam. In; National
Institute of Occupational Safety and Health.
C-90
-------
Corneliussen, P.E. 1969. Residues in food and feed: Pesticide
residues in total diet samples. IV. Pestic. Monit. Jour. 2:140.
Corneliussen, P.E. 1970. Residues in food and feed: Pesticide
residues in total diet samples. V. Pestic. Monit. Jour. 4: 89.
Corneliussen, P.E. 1972. Residues in food and feed: Pesticide
residues in total diet samples. VI. Pestic. Monit. Jour.
5: 313.
Coutselinis, A., et al. 1978. Concentration levels of endosulfan
in biological material (report of three cases). Forensic Sci.
11: 75.
Czech, M. 1958. Medicin u. Chemie. In; ACGIH, 1971. 6: 574.
Deema, P., et al. 1966. Metabolism, storage, and excretion
-^C-endosulfan in the mouse. Jour. Econ. Entomol. 59: 546.
of
Demeter, J. and A. Heyndrickx. 1978. Two lethal endosulfan poi-
sonings in man. Jour. Anal. Toxicol. 2: 68.
Demeter, J., et al. 1977. Toxicological analysis in a case of
endosulfan suicide. Bull. Environ. Contam. Toxicol. 18: 110.
C-91
-------
Den Tonkelaar, E.M. and C.J. Van Esch. 1974. No-effect levels of
organochlorine pesticides based on induction of microsomal liver
enzymes in short-term toxicity experiments. Toxicology. 2: 371.
Domanski, J.J. and F.E. Guthrie. 1974. Pesticide residues in
1972 cigars. Bull. Environ. Contam. Toxicol. 11: 312.
Domanski, J.J. and T.J. Sheets. 1973. Insecticide residues on
1970 U.S. auction market tobacco. Tobacco Sci. 17: 55.
Domanski, J.J., et al. 1973. Insecticide residues on 1971 U.S.
tobacco products. Tobacco Sci. 17: 80.
Domanski, J.J., et al. 1974. Insecticide residues on 1973 U.S.
tobacco products. Tobacco Sci. 18: 111.
Dorough, H.W. and J.R. Gibson. 1972. Chlorinated insecticide
residues in cigarettes purchased in 1970-72. Environ. Entomol.
1: 739.
Dorough, H.W., et al. 1978. Fate of endosulfan in rats and toxi-
cological considerations of apolar metabolites. Pestic. Biochem.
Physiol. 8: 241.
Dreisbach, R.H. 1974. Agricultural poisons: chlorinated hydro-
carbons. In_: Handbook of poisoning, 8th ed. Lange Medical Publi-
cations, Los Altos, California, p. 97.
C-92
-------
Duggan, R.E. and P.E. Corneliussen. 1972. Dietary intake of
pesticide chemicals in the United States (III), June 1968 to April
1970. Pestic. Monit. Jour. 5: 331.
Duggan, R.E., et al. 1967. Residues in food and feed: Pesticide
residues in total diet samples (II). Pestic. Monit. Jour. 1: 2.
Duggan, R.E., et al. 1971. Residues in food and feed: pesticide
residue levels in foods in the United States from July 1, 1963 to
June 30, 1969. Pestic. Monit. Jour. 5: 73.
Dunachie, J.F. and W.W. Fletcher. 1966. Effect of some insecti-
cides on the hatching rate of hens' eggs. Nature. 212: 1062.
Dunachie, J.F. and W.W. Fletcher. 1969. An investigation of the
toxicity of insecticides to birds' eggs using the egg injection
technique. Annu. Appl. Biol. 64: 409.
Ely, T.S., et al. 1967. Convulsions in Thiodan workers: A pre-
liminary report. Jour. Occup. Med. 9: 35.
Environmental Quality Coordination Unit. 1973. Pesticide survey
in Lakes Erie and Ontario. Prepublication manuscript prepared by
Canada Centre for Inland Waters, Burlington, Ontario. In:
National Research Council of Canada.
C-93
-------
Ernst, W. 1977. Determination of the bioconcentration potential
of marine organisms—a steady state approach. I. Bioconcentration
data for seven chlorinated pesticides in mussels (Mytilus edulis)
and their relation to solubility data. Chemosphere. 6: 731.
Fahrig, R. 1974. Comparative mutagenicity studies with pesti-
cides. Int. Agency Res. Cancer Sci. Publ. 10: 161.
Federal Water Pollution Control Administration. 1968. Water
Quality Criteria. Fed. Water Pollut. Control Admin. Natl. Tech.
Adv. Comm. U.S. Dept. Interior. U.S. Govt. Print. Off., Washing-
ton, D. C.
FMC Corp. 1963. Laboratory rep. Niagara Chemical Division, FMC
Corp., Middleport, New York. In; Maier-Bode, 1968. (Unpubl.)
FMC Corp. 1965. Laboratory rep. Niagara Chemical Division, FMC
Corp., Middleport, New York. In; Maier-Bode, 1968. (Unpubl.)
FMC Corp. 1967. Report of Niagara Chemical Division, FMC Corp.,
Middleport, New York. In; Maier-Bode, 1968. (Unpubl.)
FMC Corp. 1971. Project 015: Determination of endosulfan I,
endosulfan II and endosulfan sulfate residues in soil, pond mud
and water. Niagara Chemical Division, FMC Corp., Richmond, Cali-
fornia. In; National Research Council of Canada, 1975. (Unpubl.
rep.)
C-94
-------
FMC Corp. 1972. Project 015: Thiodan residues in soil and irri-
gation runoff water. Niagara Chemical Division, FMC Corp., Rich-
mond, California. In; National Research Council of Canada, 1975.
(Unpubl. rep.)
Food and Agriculture Organization of the United Nations. 1975.
Pesticide residues in food: Report of the 1974 Joint Meeting of
the FAO Working Party of Experts on Pesticide Residues and the WHO
Expert Committee on Pesticide Residues. Agric. Stud. No. 97, Food
Agric. Organ., United Nations, Rome.
Frank, R. 1972. A fish kill near Simcoe, Ontario. Unpublished
report. Ontario Ministry of Agriculture and Food, Guelph, On-
tario. In; National Research Council of Canada, 1975.
Frank, R. 1973. Unpublished data. Ontario Ministry of Agricul-
ture and Food, Guelph, Ontario. In; National Research Council of
Canada, 1975.
Frank, R., et al. 1977. Organochlorine insecticides and PCBs in
sediments on Lake St. Clair (1970 and 1974) and Lake Erie (1971).
Sci. Total Environ. 8: 205.
Gaines, T.B. 1969. Acute toxicity of pesticides. Toxicol.
Appl. Pharmacol. 14: 515.
C-95
-------
Gibson, J.R., et al. 1974. Chlorinated insecticide residues in
Kentucky hurley tobacco: Crop years 1963-72. Pestic. Monit. Jour.
7: 205.
Gorbach, S.G. 1972. Terminal residues of endosulfan. Proc.
2nd Int. Congr. Pestic. Chem. 6: 283.
Gorbach, S.G., et al. 1968. Metabolism of endosulfan in milk
sheep. Jour. Agric. Food Chem. 16: 950.
Gorbach, S.G., et al. 1971a. Residue analyses in the water
system of East Java (River Brantas, ponds, seawater) after con-
tinued large-scale application of Thiodan in rice. Bull. Environ.
Contam. Toxicol. 6: 40.
Gorbach, S.G., et al. 1971b. Residue analyses and biotests in
rice fields of East Java treated with Thiodan. Bull. Environ.
Contam. Toxicol. 6: 193.
Greve, P.A. 1972. Potentially hazardous substances in surface
waters. Part I. Pesticides in the River Rhine. Sci. Total
Environ. 1: 173.
Greve, P.A. and S.L. Wit. 1971. Endosulfan in the Rhine River.
Jour. Water Pollut. Control Fed. 43: 2338.
C-96
-------
Gupta, P.K. 1976. Endosulfan-induced neurotoxicity in rats and
mice. Bull. Environ. Contam. Toxicol. 15: 708.
Gupta, P.K. 1978. Distribution of endosulfan in plasma and brain
after repeated oral administration to rats. Toxicology. 9: 371.
Gupta, P.K. and S.V. Chandra. 1975. The toxicity of endosulfan
in rabbits. Bull. Environ. Contam. Toxicol. 14: 513.
Gupta, P.K. and R.C. Gupta. 1977a. Influence of endosulfan on
pentobarbitone sleeping time and blood and brain concentrations in
male rats. Jour. Pharm. Pharmacol. 29: 245.
Gupta, P.K. and R.C. Gupta. 1977b. Effect of endosulfan pre-
treatment on organ weights and on pentobarbital hypnosis in rats.
Toxicology. 7: 283.
Gupta, P.K., et al. 1978. Teratogenic and embryotoxic effects of
endosulfan in rats. Acta Pharmacol. Toxicol. 42: 150.
Hazleton Laboratories. 1959a. Unpublished report, May 12. Falls
Church, Virginia. In; ACGIH, 1971.
Hazleton Laboratories. 1959b. Unpublished report, May 22. Falls
Church, Virginia. In: ACGIH, 1971.
C-97
-------
Hazleton Laboratories. 1967. Unpublished reports, Mar. 17.
Falls Church, Virginia. In: Maier-Bode, 1968.
Henschler, D., et al. 1977. Carcinogenicity of trichloroethy-
lene: Fact or artifact? Arch. Toxicol. 37: 233.
Herzel, F. 1972. Organochlorine insecticides in surface waters
in Germany - 1970 and 1971. Pestic. Monit. Jour. 6: 179.
Hoechst. 1966. Report of Farbwerke Hoechst A.G., Frankfurt, West
Germany. In; Maier-Bode, 1968. (Unpubl.)
Hoechst. 1967a. Pflanzenschutz Forschung. Laboratory reports.
Farbwerke Hoechst A.G., Frankfurt, West Germany. In_: Maier-Bode,
1968. (Unpubl.)
Hoechst. 1967b. Oral LD50 values for white rats. Report of
Farbwerke Hoechst A.G., Frankfurt, West Germany. In: Demeter and
Heyndrickx, 1978. (Unpubl.)
Innes, J.R.M., et al. 1969. Bioassay of pesticides and indus-
trial chemicals for tumorigenicity in mice: A preliminary note.
Jour. Natl. Cancer Inst. 42: 1101.
International Agency for Research on Cancer. 1971. Liver cancer.
Sci. Publ. No. 1, World Health Organ., Lyon, France.
C-98
-------
Israeli, R., et al. 1969. Endosulfan-Vergiftung, Preliminar-
meldung uber drei Falle. Zentralbl. Arbeitsmed. Arbeitsschutz.
19: 193. (Ger.)
Johnson, R.D. and D.D. Manske. 1976. Residues in food and feed:
Pesticide residues in total diet samples (IX). Pestic. Monit.
Jour. 9: 157.
Johnson, R.D. and D.D. Manske. 1977. Pesticide and other chemi-
cal residues in total diet samples (XI). Pestic. Monit. Jour.
11: 116.
Johnson, W.H., et al. 1975. Effects of freeze-drying on residues
of TDE, DDT, and endosulfan in tobacco. Jour. Agric. Food Chem.
23: 117.
Jones, K.H., et al. 1968. Acute toxicity data for pesticides.
World Rev. Pest. Control. 7: 135.
Kazen, C., et al. 1974. Persistence of pesticides on the hands
of some occupationally exposed people. Arch. Environ. Health
29: 315.
Keil, J.E., et al. 1972. Decay of parathion and endosulfan resi-
dues on field-treated tobacco, South Carolina - 1971. Pestic.
Monit. Jour'. 6: 73.
C-99
-------
Kloss, G.f et al. 1966. Versuche an Schaffen mit C14-mark-
ierten Thiodan. In; Maier-Bode, 1968. (Unpubl.) (Ger.)
Knowles, C.O. 1974. Detoxification of Acaricides by Animals.
In; M.A. Kahn and J.P. Bederka, Jr. (eds.), Survival in Toxic
Environments. Academic Press, New York. p. 155.
Korn, S., and R. Earnest. 1974. Acute toxicity of twenty in-
secticides to striped bass, Morone saxatilis. Calif. Fish Game
60; 128.
Kotin, P., et al. 1968. Evaluation of Carcinogenic, Teratogenic
and Mutagenic Activities of Selected Pesticides and Industrial
Chemicals. Ln; Vol. 1; Carcinogenic Study. Bionetics Res. Lab.
Rep. to Natl. Cancer Inst. PB 223-159. p. 64.
Lee, R.L., Jr. 1976. Air pollution from pesticides and agricul-
tural process. CRC Press, Inc., Cleveland, Ohio.
Lendle, L. 1956. Bericht uber Untersuchungen von Hoe 2671 (Thio-
dan) der Farbwerke Hoechst A.G., July 1956. In; Maier-Bode,
1968. (Ger.)
Li, C.F., et al. 1970. Fate of organochlorine pesticides during
processing of milk into dairy products. Jour. Assoc. Off. Anal.
Chem. 53: 127.
C-100
-------
Lindquist, D.A. and P.A. Dahm. 1957. Some chemical and biologi-
cal experiments with Thiodan. Jour. Econ. Entomol. 50: 483.
Lu, F.C. 1973. Toxicological evaluation of food additives and
pesticide residues and their "acceptable daily intakes" for man:
The role of WHO, in conjunction with FAO. Residue Rev. 45: 81.
Lutz, H. and Y. Lutz-Ostertag. 1972. The action of different
pesticides on the development of bird embryos. Adv. Exp. Med.
Biol. 27: 127.
Lutz-Ostertag, Y. and J.P. Kantelip. 1970. Action de 1'endosul-
fan sur le tractus genital de 1'embryon de poulet et de caille "in
vivo" et "in vitro". Bull. Soc. Zool. Fr. 95: 620. (Fre.)
Lutz-Ostertag, Y. and J.P. Kantelip. 1971. Action sterilisante
de 1'endosulfan (Thiodan) (insecticide organochlore) sur les go-
nades de 1' embryon de poulet et de caille in vivo et in vitro.
C.R. Soc. Biol. 165: 844. (Fre.)
Macek, K.J., et al. 1976. Toxicity of four pesticides to water
fleas and fathead minnows. Ecol. Res. Ser. EPA 600/3-76-099.
U.S. Environ, Prot. Agency, Washington, D.C.
Maddy, K.T. and L.C. Riddle. 1977. Pesticide poisonings in
domestic animals. Mod. Vet. Pract. 58: 913.
0101
-------
Maier-Bode, H. 1968. Properties, effect, residues and analytics
of the insecticide endosulfan (review). Residue Rev. 22: 2.
Manske, D.D. and P.E. Corneliussen. 1974. Residues in food and
feed: Pesticide residues in total diet samples (VII). Pestic.
Monit. Jour. 8: 110.
Manske, D.D. and R.D. Johnson. 1975. Residues in food and feed:
pesticide residues in total diet samples (VIII). Pestic. Monit.
Jour. 9: 94.
Manske, D.D. and R.D. Johnson. 1977. Pesticide and other chemi-
cal residues in total diet samples (X). Pestic. Monit. Jour.
10: 134.
Martin, R.J. and R.E. Duggan. 1968. Pesticide residues in total
diet samples (III). Pestic. Monit. Jour. 1: 11.
Matsumura, F. 1975. In: Toxicology of Insecticides. Plenum
Press, New York. p. 205.
McCaskey, T.A. and B.J. Liska. 1967. Effect of milk processing
methods of endosulfan, endosulfan sulfate, and chlordane residues
in milk. Jour. Dairy Sci. 50: 1991.
C-102
-------
Menzie, C.M. 1974. Metabolism of pesticides: An update. Spec.
sci. rep. Fish Wildl. Serv. Wildl. 184. U.S. Dept. Inter., Wash-
ington, D.C.
Miles, J.R.W. and C.R. Harris. 1971. Insecticide residues in a
stream and a controlled drainage system in agricultural areas of
southwestern Ontario, 1970. Pestic. Monit. Jour. 5: 289.
Miles, J.R.W. and C.R. Harris. 1973. Organochlorine insecticide
residues in streams draining agricultural, urban-agricultural, and
resort areas of Ontario, Canada - 1971. Pestic. Monit. Jour.
6: 363.
Muacevic, G. 1973. Acute toxicity and cholinesterase inhibition
IB vivo of bromophos-ethyl. Toxicol. Appl. Pharmacol. 25: 180.
National Academy of Sciences. 1972. Water Quality Criteria 1972.
Environmental Studies Board, Committee on Water Quality Criteria.
Ecol. Res. Ser. EPA R3.73.033. U.S. Govt. Print. Off., Washing-
ton, D.C.
National Academy of Sciences. 1977. Drinking Water and Health.
U.S. Environ. Prot. Agency, Washington, D.C. PB-269 519. Natl.
Tech. Inf. Serv., Springfield, Virginia.
C-103
-------
National Cancer Institute. 1978. Bioassay of endosulfan for pos-
sible carcinogenicity. Natl. Cancer Inst. Div. Cancer Cause and
Prevention. Bethesda, Maryland. DHEW Pub. No. (NIH) 78-1312.
National Institute for Occupational Safety and Health. 1978.
Criteria and recommended standard—occupational exposure during
the manufacture and formulation of pesticides. U.S. Dept. Hlth.
Edu., Welfare, Washington, D.C.
National Research Council of Canada. 1975. Endosulfan, its
effects on environmental quality. ISSN 0316-0114, NRCC No. 14098.
Ottawa, Canada.
Nicholson, S.S. and G.W. Cooper. 1977. Apparent endosulfan toxi-
cosis in calves. Jour. Am. Vet. Med. Assoc. 170: 319.
Oeser, H., et al. 1971. Endosulfan and the environment. Gior-
nate Fitopathologie (Workshop on Phytopathology) (Sng. transl.).
Udine, Italy, May 1971: 17.
Osmond, D.S. 1969. A fish kill in the Thames River, Ontario.
Ontario Ministry Environ., London, Ontario, Canada. In; National
Research Council of Canada, 1975. (Unpubl. rep.)
Oudbier, A.J., et al. 1974. Respiratory route of pesticide expo-
sure as a potential health hazard. Bull. Environ. Contam. Toxi-
col. 12: 1.
C-104
-------
Panetsos, A. and S. Kilikidis. 1973. Identification of the cause
of animal poisoning in two cases by polarography and gas-liquid
chromatography. Hell. Kteniatr., Iss. April- June: 75. i_n: j.
Demeter and A. Heyndrickx, 1978.
Roberts, D. 1972. The assimilation and chronic effects of sub-
lethal concentrations of endosulfan on condition and spawning in
the common mussel, Mytilus edulis. Mar. Biol. 16: 119.
Roberts, D. 1975. Differential uptake of endosulfan by the tis-
sues of Mytilus edulis. Bull. Environ. Contam. Toxicol.
13: 170.
Schimmel, S.C., et al. 1977. Acute toxicity to and bioconcentra-
tion of endosulfan by estuarine animals. Proc. Am. Soc. Test.
Mater. Symp. Aquat. Toxicol. ASTM Rep. STP 634.
Schmidlin-Meszaros, J. and E. Romann. 1971. Eine accidentelle
Vergiftung von Kuhen mit Endosulfan (Thiodan) [an accidental poi-
soning of cows with endosulfan (Thiodan)]. Mitt. Geb. Lebens-
mittelunters. Hyg. 62: 110. (Ger.)
Schoettger, R.A. 1970. Toxicology of thiodan in several fish and
aquatic invertebrates. Bureau of Sport Fisheries and Wildlife,
U.S. Dept. Inter., Fish Wildl. Serv. Invest. Fish Control.
35: 1.
C-105
-------
Schulze, J.A., et al. 1973. Pesticides in selected western
streams - 1968-1971. Pestic. Monit. Jour. 7: 73.
Sievers, J.F., et al. 1972. Untersuchungen ueber den Endosulfan-
Gehalt in Rhein, Main und in Uferfiltraten (Juni 1969 - February
1970) (Endosulfan content in the Rhine, Main, and in shore fil-
trates, June 1969 - February 1970). Environ. Qual. Saf. 1: 239.
(Ger.)
Smith, S.I., et al. 1970. The effect of injection of chlorinated
hydrocarbon pesticides on hatchability of eggs. Toxicol. Appl.
Pharmacol. 16: 179.
Stephan, C.E. 1980. Memorandum to J. Stara. U.S. EPA. July 3.
Tarrant, K.R. and J.O'G. Tatton. 1968. Organochlorine pesticides
in rainwater in the British Isles. Nature. 219: 725.,
Terziev, G., et al. 1974. Forensic medical and forensic chemical
study of acute lethal poisonings with Thiodan. Folia Med.
(Plovdiv) In; Demeter and Heyndrickx, 1978. 16: 325.
Tessari, J.D. and D.L. Spencer. 1971. Air sampling for pesti-
cides in the human environment. Jour. Assoc. Offic. Anal. Chem.
54: 1376.
C-106
-------
Thompson, G.E. 1966. Poisoning of cattle following accidental
spraying with Thiodan. Jour. S. Afr. Vet. Med. Assoc. 37: 81.
Thorstenson, J.H. and H.W. Dorough. 1976. Chlorinated insecti-
cide residues in the University of Kentucky reference and alkaloid
series cigarettes. Tobacco Sci. 20: 25.
Tiberin, P., et al. 1970. EEC findings in poisoning by endosul-
fan. Electroencephalogr. Clin. Neurophysiol. 28: 642.
Tomatis, L., et al. 1973. The predictive value of mouse liver
tumor induction in carcinogenicity testing. int. Jour. Cancer
12: 1.
Truhaut, R., et al. 1974. Recherches sur les modalites et les
mechanismes d1action toxique des insecticides organochlores.
(Research on the modalities and mechanisms of toxic action of
organochlorine insecticies.) Eur. Jour. Toxicol. Environ. Hyg.
7: 159. (Fre.)
U.S. EPA. 1980. Seafood consumption data analysis. Stanford
Research Institute International, Menlo Park, California. Final
rep., Task II. Contract No. 68-01-3887.
Utklev, H.E. and _C. Westbye. 1971. Poisoning with endosulfan.
Nor. Veterinaertidsskr. in: Demeter and Heyndrickx, 1978.
83: 31.
C-107
-------
Wegman, R.C.C. and P.A. Greve. 1978. Water: organochlorines,
cholinesterase inhibitors, and aromatic amines in Dutch water
samples, September 1969 - December 1975. Pestic. Monit. Jour.
12: 149.
Whitacre, D.M. 1970. Endosulfan metabolism in temperature-
stressed rats. Diss. Abstr. Int. 30: 4435B.
Winell, M. 1975. An international comparison of hygienic stan-
dards for chemicals in the work environment. Ambio. 4: 34.
Wolfe, H.R., et al. 1972. Exposure of spraymen to pesticides.
Arch. Environ. Health. 25: 29.
World Health Organization. 1975. 1974 evaluation of some
pesticide residues in food. WHO Tech. Rep. Ser. 574. Geneva.
Wong, H.F. and J.P. Donnelly. 1968. A preliminary pesticide sur-
vey in the Bay of Quinte and international section of the St.
Lawrence River, August, October 1968. Manuscript Rep. KR-68-4.
Div. Pub. Health Eng., Canada Dept. Natl. Health Welfare, Canada.
In: National Research Council of Canada, 1975.
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